TOPS: a versatile software tool for statistical analysis and visualization of combinatorial gene-gene and gene-drug interaction screens.
Measuring the impact of combinations of genetic or chemical perturbations on cellular fitness, sometimes referred to as synthetic lethal screening, is a powerful method for obtaining novel insights into gene function and drug action. Especially when performed at large scales, gene-gene or gene-drug interaction screens can reveal complex genetic interactions or drug mechanism of action or even identify novel therapeutics for the treatment of diseases.The result of such large-scale screen results can be represented as a matrix with a numeric score indicating the cellular fitness (e.g. viability or doubling time) for each double perturbation. In a typical screen, the majority of combinations do not impact the cellular fitness. Thus, it is critical to first discern true "hits" from noise. Subsequent data exploration and visualization methods can assist to extract meaningful biological information from the data. However, despite the increasing interest in combination perturbation screens, no user friendly open-source program exists that combines statistical analysis, data exploration tools and visualization. We developed TOPS (Tool for Combination Perturbation Screen Analysis), a Java and R-based software tool with a simple graphical user interface that allows the user to import, analyze, filter and plot data from double perturbation screens as well as other compatible data. TOPS was designed in a modular fashion to allow the user to add alternative importers for data formats or custom analysis scripts not covered by the original release.We demonstrate the utility of TOPS on two datasets derived from functional genetic screens using different methods. Dataset 1 is a gene-drug interaction screen and is based on Luminex xMAP technology. Dataset 2 is a gene-gene short hairpin (sh)RNAi screen exploring the interactions between deubiquitinating enzymes and a number of prominent oncogenes using massive parallel sequencing (MPS). TOPS provides the benchtop scientist with a free toolset to analyze, filter and visualize data from functional genomic gene-gene and gene-drug interaction screens with a flexible interface to accommodate different technologies and analysis algorithms in addition to those already provided here. TOPS is freely available for academic and non-academic users and is released as open source.


Postnatal growth following prenatal lead exposure and calcium intake.
The effects on postnatal growth of maternal exposure to low levels of lead during pregnancy have not been well established. In addition, information is limited regarding the protective effect of dietary calcium intake during pregnancy against the effect of lead for fetal and postnatal growth. We investigated the relationship between prenatal exposure to lead and growth at birth and 6, 12, and 24 months postnatal, and evaluated the role of calcium intake against the effect of lead. A total of 1150 pregnant women, and their subsequent offspring, enrolled in a prospective birth cohort study (Mothers and Children's Environmental Health Study), were evaluated. Multivariable regression analysis was conducted to estimate the effects of prenatal maternal blood lead levels on growth at each follow-up. The blood lead levels of participating mothers were <5.0 μg/dL and mean levels were 1.25 μg/dL during the early (before 20 gestational weeks) and late (at delivery) gestational periods. Prenatal exposure to lead, particularly in late pregnancy, was significantly associated with a reduction in infantile growth at 24 months. When pregnant women had dietary calcium intake at mean or upper level, the association was not significant. In contrast, lower than mean level of calcium intake intensified the adverse effect of prenatal lead exposure on growth in children. Prenatal lead exposure <5.0 μg/dL adversely affects postnatal growth and low calcium intake aggravates the effect, indicating more stringent control of lead and sufficient intake of calcium are necessary to help children's health.


Series: The research agenda for general practice/family medicine and primary health care in Europe. Part 6: reaction on commentaries - how to continue with the Research Agenda?
The Research Agenda should be used as a key reference point to which new research should relate its usefulness and added value. Primary care evolves towards more interdisciplinary care, and research should focus more on the core competency of person-centred team care. There is an urgent need to develop clear definitions and appropriate research instruments for this domain. It will be a particular challenge to study comprehensive approaches in primary-care patients with multi-morbidity. The Research Agenda and the commentaries on it show future directions for primary care research. There are challenges related to a changing society, the shared responsibility and guidance of research by professionals and citizens (patients), and the need to fully integrate research as part of primary healthcare provision. There will be a need for a prioritization of spearheads to guide primary care research for the next decade: translational research, research on equity and health differences, on chronic disease and health systems research. This can not be realized without the development and maintenance of a solid research infrastructure: easily maintained and accessed observational databases, helpful information technology, strategies and techniques for patient involvement, advanced research training possibilities, and the development and validation of appropriate research instruments and outcome measures to capture the different challenges. Worldwide, primary care not only is a priority for health care policy, but it needs to become a research priority as well.


Off-service resident education in the emergency department: outline of a national standardized curriculum.
Although many residency programs mandate at least one rotation in emergency medicine (EM), to the best of our knowledge, a standardized curriculum for emergency department (ED) rotations for "off-service" residents has not been developed. As a result, the experiences of these residents in the ED tend to vary during their rotations. To design an off-service EM curriculum, we adopted Kern's six-step approach to curriculum development as a conceptual framework. The resulting program encompasses clinical experience and didactic sessions through which residents are trained in core topics and skills. This knowledge will be applicable in the clinical settings in which residents will continue to train and ultimately practice their specialty. It is flexible enough to be applicable and implementable without being limited by resource availability or faculty strengths.


Dynamics and control of biofilms of the oligotrophic bacterium Caulobacter crescentus.
Caulobacter crescentus is an oligotrophic alpha-proteobacterium with a complex cell cycle involving sessile-stalked and piliated, flagellated swarmer cells. Because the natural lifestyle of C. crescentus intrinsically involves a surface-associated, sessile state, we investigated the dynamics and control of C. crescentus biofilms developing on glass surfaces in a hydrodynamic system. In contrast to biofilms of the well-studied Pseudomonas aeruginosa, Escherichia coli, and Vibrio cholerae, C. crescentus CB15 cells form biphasic biofilms, consisting predominantly of a cell monolayer biofilm and a biofilm containing densely packed, mushroom-shaped structures. Based on comparisons between the C. crescentus strain CB15 wild type and its holdfast (hfsA; DeltaCC0095), pili (DeltapilA-cpaF::Omegaaac3), motility (motA), flagellum (flgH) mutants, and a double mutant lacking holdfast and flagellum (hfsA; flgH), a model for biofilm formation in C. crescentus is proposed. For both biofilm forms, the holdfast structure at the tip of a stalked cell is crucial for mediating the initial attachment. Swimming motility by means of the single polar flagellum enhances initial attachment and enables progeny swarmer cells to escape from the monolayer biofilm. The flagellum structure also contributes to maintaining the mushroom structure. Type IV pili enhance but are not absolutely required for the initial adhesion phase. However, pili are essential for forming and maintaining the well-defined three-dimensional mushroom-shaped biofilm. The involvement of pili in mushroom architecture is a novel function for type IV pili in C. crescentus. These unique biofilm features demonstrate a spatial diversification of the C. crescentus population into a sessile, "stem cell"-like subpopulation (monolayer biofilm), which generates progeny cells capable of exploring the aqueous, oligotrophic environment by swimming motility and a subpopulation accumulating in large mushroom structures.


Clinical and practical considerations in the pharmacologic management of narcolepsy.
Despite published treatment recommendations and the availability of approved and off-label pharmacologic therapies for narcolepsy, the clinical management of this incurable, chronic neurologic disorder remains challenging. While treatment is generally symptomatically driven, decisions regarding which drug(s) to use need to take into account a variety of factors that may affect adherence, efficacy, and tolerability. Type 1 narcolepsy (predominantly excessive daytime sleepiness with cataplexy) or type 2 narcolepsy (excessive daytime sleepiness without cataplexy) may drive treatment decisions, with consideration given either to a single drug that targets multiple symptoms or to multiple drugs that each treat a specific symptom. Other drug-related characteristics that affect drug choice are dosing regimens, tolerability, and potential drug-drug interactions. Additionally, the patient should be an active participant in treatment decisions, and the main symptomatic complaints, treatment goals, psychosocial setting, and use of lifestyle substances (ie, alcohol, nicotine, caffeine, and cannabis) need to be discussed with respect to treatment decisions. Although there is a lack of narcolepsy-specific instruments for monitoring therapeutic effects, clinically relevant subjective and objective measures of daytime sleepiness (eg, Epworth Sleepiness Scale and Maintenance of Wakefulness Test) can be used to provide guidance on whether treatment goals are being met. These considerations are discussed with the objective of providing clinically relevant recommendations for making treatment decisions that can enhance the effective management of patients with narcolepsy.


Msx1 is expressed in retina endothelial cells at artery branching sites.
Msx1 and Msx2 encode homeodomain transcription factors that play a role in several embryonic developmental processes. Previously, we have shown that in the adult mouse, Msx1(lacZ) is expressed in vascular smooth muscle cells (VSMCs) and pericytes, and that Msx2(lacZ) is also expressed in VSMCs as well as in a few endothelial cells (ECs). The mouse retina and choroid are two highly vascularized tissues. Vessel alterations in the retina are associated with several human diseases and the retina has been intensely used for angiogenesis studies, whereas the choroid has been much less investigated. Using the Msx1(lacZ) and Msx2(lacZ) reporter alleles, we observed that Msx2 is not expressed in the eye vascular tree in contrast to Msx1, for which we establish the spatial and temporal expression pattern in these tissues. In the retina, expression of Msx1 takes place from P3, and by P10, it becomes confined to a subpopulation of ECs at branching points of superficial arterioles. These branching sites are characterized by a subpopulation of mural cells that also show specific expression programs. Specific Msx gene inactivation in the endothelium, using Msx1 and Msx2 conditional mutant alleles together with a Tie2-Cre transgene, did not lead to conspicuous structural defects in the retinal vascular network. Expression of Msx1 at branching sites might therefore be linked to vessel physiology. The retinal blood flow is autonomously regulated and perfusion of capillaries has been proposed to depend on arteriolar precapillary structures that might be the sites for Msx1 expression. On the other hand, branching sites are subject to shear stress that might induce Msx1 expression. In the choroid vascular layer Msx1(lacZ) is expressed more broadly and dynamically. At birth Msx1(lacZ) expression takes place in the endothelium but at P21 its expression has shifted towards the mural layer. We discuss the possible functions of Msx1 in the eye vasculature.


Xenobiotics released from fat during fasting produce estrogenic effects in ovariectomized mice.
The pesticide residues 1-(o-chlorophenyl)-1-(p-chlorophenyl)-2,2,2-trichloroethane (o,p'-DDT) and beta-hexachlorocyclohexane (beta-HCH) act as weak estrogens, producing uterotrophic responses in ovariectomized rodents and stimulating human breast cancer cells in culture. Such activity suggests that these compounds may act as tumor promoters in estrogen-responsive tissues. Organochlorine compounds such as o,p'-DDT and beta-HCH are concentrated in body fat. The present report tests whether sufficient compound can be released from fat depots to produce estrogenic effects in uteri of ovariectomized mice. Adult animals were "loaded" with test compound by three daily injections of vehicle (DMSO), 17beta-estradiol (E2), beta-HCH, or o,p'-DDT. Uterotrophic effects were assessed at 24 h after the last loading dose of test compound and at 2 weeks after the loading regimen, with or without a prior 2-day period of fasting. The initial 3-day treatment with either beta-HCH or o,p'-DDT doubled the relative dry weight of the uterus: 102 +/- 8.6 mg/kg body weight (BW) and 104 +/- 4.4 mg/kg BW for beta-HCH and o,p'-DDT, respectively, compared to 49 +/- 1.9 mg/kg BW for vehicle-treated animals. E2-treated animals had uterine dry weights of 228 +/- 11 mg/kg BW. After 2 weeks without further treatment, a 2-day fast produced a decrease in body mass of 4.1 g/animal (fasted, 25.9 +/- 1.89 g versus fed, 30.0 +/- 2.82 g). Animals that had been loaded with beta-HCH and fasted had uterine weights (88 +/- 12 mg/kg BW) significantly greater (P < 0.05) than those of vehicle-loaded, fasted animals (51 +/- 2.9 mg/kg BW) or of beta-HCH-loaded, fed animals (59 +/- 4.6 mg/kg BW). The uterine weights of the fasted and fed o,p'-DDT-loaded or E2-loaded animals were not different from those of control weights. The difference between wet and dry weights showed that fasting of beta-HCH-loaded animals also increased water imbibition in the uterus; there was no effect from fasting in the other groups. Generally, epithelial cell height reflected the same responses as uterine weight with the exception that cell heights of beta-HCH-loaded, fed animals were slightly higher (P < 0.05) than corresponding controls, indicating that there may have been some active compound available to the tissues even without fasting. The effects of fasting show that during periods of lipolysis beta-HCH can be released in quantities sufficient to stimulate estrogen target tissues, suggesting a novel mechanism linking obesity and the progression of estrogen-responsive tumors. The lack of effect from fasting in o,p'-DDT-loaded animals indicates that these compounds are differentially mobilized from fat depots.


Evidence-based measurement in multiple sclerosis: the psychometric properties of the physical and psychological dimensions of three quality of life rating scales.
The selection of measures of quality of life used in clinical trials of multiple sclerosis (MS) should be evidence-based. Head-to-head comparison of measures facilitates the selection of measures. The aim of the study was to compare the psychometric properties of the physical and psychological dimensions in three measures of quality of life to aid choice of the most appropriate scale for use in clinical trials of MS. One hundred and twenty-one people with MS (rehabilitation = 57; steroids = 64) completed a selection of health measures before and after treatment. The psychometric properties of three measures of physical function (MSIS-29 physical, SF-36 physical functioning, FAMS mobility) and three measures of psychological function (MSIS-29 psychological, SF-36 mental health, FAMS emotional well-being) were compared by examining data quality, scaling assumptions, acceptability, reliability, validity and responsiveness. Physical (0.63-0.71) and psychological (0.70-0.75) scales were substantially correlated indicating they measure related constructs. The MSIS-29 physical and psychological scales satisfied all criteria for internal consistency reliability (physical = 0.91; psychological = 0.89) and validity. The SF-36 physical scale had a notable floor effect (20%). The FAMS mobility scale had lower reliability (alpha = 0.78) compared to other measures. The MSIS-29 physical (effect size = 0.91) and psychological (effect size = 0.62) scales were the most responsive. In these three samples, the MSIS-29 had better measurement properties for combined physical and psychological health than the SF-36 and the FAMS.


Comparative BAC-based physical mapping of Oryza sativa ssp. indica var. 93-11 and evaluation of the two rice reference sequence assemblies.
Reference sequences are sequences that are used for public consultation, and therefore must be of high quality. Using the whole-genome shotgun/next-generation sequencing approach, many genome sequences of complex higher plants have been generated in recent years, and are generally considered reference sequences. However, none of these sequences has been experimentally evaluated at the whole-genome sequence assembly level. Rice has a relatively simple plant genome, and the genome sequences for its two sub-species obtained using different sequencing approaches were published approximately 10 years ago. This provides a unique system for a case study to evaluate the qualities and utilities of published plant genome sequences. We constructed a robust BAC physical map embedding a large number of BAC end sequences forrice variety 93-11. Through BAC end sequence alignments and tri-assembly comparisons of the 93-11 physical map and the two reference sequences, we found that the Nipponbare reference sequence generated using the clone-by-clone approach has a high quality but still contains small artifact inversions and missing sequences. In contrast, the 93-11 reference sequence generated using the whole-genome shotgun approach contains many large and varied assembly errors, such as inversions, duplications and translocations, as well as missing sequences. The 93-11 physical map provides an invaluable resource for evaluation and improvements toward completion of both Nipponbare and 93-11 reference sequences.


Launching a multidisciplinary European collaboration towards a cure for HIV: The EU2Cure Consortium.
We felt the urgency to launch the EU2Cure Consortium to support research and find a cure for the human immunodeficiency virus (HIV) infection through intensified collaboration within Europe. This consortium is open to stakeholders on cure in Europe from academia and the community to connect. The aim of this consortium is to intensify the research collaboration amongst European HIV cure groups and the community and facilitate interactions with other academic and community cure consortia, private parties, and policy makers. Our main aim is to create a European research agenda, data sharing, and development of best practice for clinical and translational science to achieve breakthroughs with clinically feasible HIV cure strategies. This consortium should also enable setting up collaborative studies accessible to a broader group of people living with HIV. Besides reservoir studies, we have identified three overlapping scientific interests in the consortium that provide a starting point for further research within a European network: developing "shock and kill" cure strategies, defining HIV cure biomarkers, and connecting cure cohorts. This strategy should aid stakeholders to sustain progress in HIV cure research regardless of coincidental global health or political crises.


Anti-bacterial activities and phytochemical screening of extracts of different parts of Thalictrum rhynchocarpum.
Parts of the plant Thalictrum rhyncocarpum are used in herbal medicine in Kenya to treat various infections. The aim of this study was to evaluate in-vitro anti-bacteria activities and phytochemical profiles of solvent extracts of the leaves, stem bark and root of Thalictrum rhyncocarpum against Bacillus subtilis-6633, Staphylococcus aures-SG 511, Escherichia coli SG 458, Pseudomonus aeruginosa-K799/61 and Mycobacterium vaccae-10670. Anti-bacterial activity tests were carried out using disc diffusion assay and tube dilution technique, and phytochemical screening was carried out through Thin Layer Chromatography. The crude extracts showed antibacterial effects on M. vaccae, P. aeruginosa and B. subtilis. M. vaccae was most sensitive, particularly to the methanol root extract. Phytochemical screening of the extracts suggested the presence of glycosides and alkaloids in the stem bark and root extracts, and flavonoids and triterpenes in the leaf extracts. The study showed interesting levels of activities of solvent extracts of different parts of T. rhyncocarpum against some of the bacteria tested (M. vaccae, P. aeruginosa and B. subtilis). The results provide some scientific rationale for the traditional use of the plant in Kenya to treat different microbial infections.


Impact of gastrointestinal tract variability on oral drug absorption and pharmacokinetics: An UNGAP review.
The absorption of oral drugs is frequently plagued by significant variability with potentially serious therapeutic consequences. The source of variability can be traced back to interindividual variability in physiology, differences in special populations (age- and disease-dependent), drug and formulation properties, or food-drug interactions. Clinical evidence for the impact of some of these factors on drug pharmacokinetic variability is mounting: e.g. gastric pH and emptying time, small intestinal fluid properties, differences in pediatrics and the elderly, and surgical changes in gastrointestinal anatomy. However, the link of colonic factors variability (transit time, fluid composition, microbiome), sex differences (male vs. female) and gut-related diseases (chronic constipation, anorexia and cachexia) to drug absorption variability has not been firmly established yet. At the same time, a way to decrease oral drug pharmacokinetic variability is provided by the pharmaceutical industry: clinical evidence suggests that formulation approaches employed during drug development can decrease the variability in oral exposure. This review outlines the main drivers of oral drug exposure variability and potential approaches to overcome them, while highlighting existing knowledge gaps and guiding future studies in this area.


Developmental regulation of ganglioside antigens recognized by the JONES antibody.
The JONES monoclonal antibody has been immunocytochemically associated with regions of the developing rat brain where cell and axon migrations are occurring (Mendez-Otero et al., 1986, 1988). In the present study the antigens recognized by the JONES antibody were analyzed in a variety of brain regions and at developmental ages selected to correspond to the preceding immunocytochemical observations. In accordance with earlier results from retina, JONES binding could not be detected in SDS gels from developing brain. Binding of the antibody was, however, prominent in chloroform/methanol extracts of the same tissues, and it was completely removed from tissue sections by brief chloroform/methanol form/methanol treatment. Enzymatic analyses of chloroform/methanol extracts indicated that the JONES epitope was sensitive to neuraminidase but insensitive to proteases. Overlay assays on developed high-performance thin-layer chromatographic plates (HPTLC) indicate that in all regions the JONES epitope resides on 2 or 3 ganglioside bands, depending on the age examined. These bands migrate between ganglioside standards GD 1a and GM2 on HPTLC plates and have been designated GJ1, GJ2, and GJ3, with the higher number designating the more rapidly migrating species. Occasionally, additional bands migrating in the range of polysialogangliosides were observed. The pattern of expression of GJ species was studied in forebrain, retina, and cerebellar tissue taken from embryonic day 18 (E18), postnatal day 0 (P0), P7, P14, and adult animals. Both region-specific differences in the relative prominence of each band and stage-specific differences in the total amount of the JONES gangliosides were detected. The stage-specific differences in the amount of JONES antigens are well correlated with the developmental periods of maximal cell migration in each region. While the JONES gangliosides are most prominent in forebrain before birth, in they are most prominent during the first 2 postnatal weeks. In cerebellum, JONES antigen expression is more pronounced during the 2 periods of cell migration in this tissue. In retina, the more rapidly migrating GJ3 band was the most prominent band at all stages examined, and this same band is retained in the adult. In cerebellum and forebrain GJ3 is also the most pronounced band during development. However, in contrast to the retina, the more slowly migrating GJ1 band is retained in adult forebrain and cerebellum. A variety of non-brain tissues have also been examined for the presence of the JONES antigens.(ABSTRACT TRUNCATED AT 400 WORDS)


PI3K regulation of RAC1 is required for KRAS-induced pancreatic tumorigenesis in mice.
New drug targets are urgently needed for the treatment of patients with pancreatic ductal adenocarcinoma (PDA). Nearly all PDAs contain oncogenic mutations in the KRAS gene. Pharmacological inhibition of KRAS has been unsuccessful, leading to a focus on downstream effectors that are more easily targeted with small molecule inhibitors. We investigated the contributions of phosphoinositide 3-kinase (PI3K) to KRAS-initiated tumorigenesis. Tumorigenesis was measured in the Kras(G12D/+);Ptf1a(Cre/+) mouse model of PDA; these mice were crossed with mice with pancreas-specific disruption of genes encoding PI3K p110α (Pik3ca), p110β (Pik3cb), or RAC1 (Rac1). Pancreatitis was induced with 5 daily intraperitoneal injections of cerulein. Pancreata and primary acinar cells were isolated; acinar cells were incubated with an inhibitor of p110α (PIK75) followed by a broad-spectrum PI3K inhibitor (GDC0941). PDA cell lines (NB490 and MiaPaCa2) were incubated with PIK75 followed by GDC0941. Tissues and cells were analyzed by histology, immunohistochemistry, quantitative reverse-transcription polymerase chain reaction, and immunofluorescence analyses for factors involved in the PI3K signaling pathway. We also examined human pancreas tissue microarrays for levels of p110α and other PI3K pathway components. Pancreas-specific disruption of Pik3ca or Rac1, but not Pik3cb, prevented the development of pancreatic tumors in Kras(G12D/+);Ptf1a(Cre/+) mice. Loss of transformation was independent of AKT regulation. Preneoplastic ductal metaplasia developed in mice lacking pancreatic p110α but regressed. Levels of activated and total RAC1 were higher in pancreatic tissues from Kras(G12D/+);Ptf1a(Cre/+) mice compared with controls. Loss of p110α reduced RAC1 activity and expression in these tissues. p110α was required for the up-regulation and activity of RAC guanine exchange factors during tumorigenesis. Levels of p110α and RAC1 were increased in human pancreatic intraepithelial neoplasias and PDAs compared with healthy pancreata. KRAS signaling, via p110α to activate RAC1, is required for transformation in Kras(G12D/+);Ptf1a(Cre/+) mice.


Nitric oxide is the primary mediator of cytotoxicity induced by GSH depletion in neuronal cells.
Glutathione (GSH) levels progressively decline during aging and in neurodegenerative disorders. However, the contribution of such event in mediating neuronal cell death is still uncertain. In this report, we show that, in neuroblastoma cells as well as in primary mouse cortical neurons, GSH decrease, induced by buthionine sulfoximine (BSO), causes protein nitration, S-nitrosylation and DNA strand breaks. Such alterations are also associated with inhibition of cytochrome c oxidase activity and microtubule network disassembly, which are considered hallmarks of nitric oxide (NO) toxicity. In neuroblastoma cells, BSO treatment also induces cell proliferation arrest through the ERK1/2-p53 pathway that finally results in caspase-independent apoptosis, as evident from the translocation of apoptosis-inducing factor from mitochondria towards nuclei. A deeper analysis of the signaling processes indicates that the NO-cGMP pathway is involved in cell proliferation arrest and death. In fact, these events are completely reversed by L-NAME, a specific NO synthase inhibitor, indicating that NO, rather than the depletion of GSH per se, is the primary mediator of cell damage. In addition, the guanylate cyclase (GC) inhibitor LY83583 is able to completely block activation of ERK1/2 and counteract BSO toxicity. In cortical neurons, NMDA (N-methyl-D-aspartic acid) treatment results in GSH decrease and BSO-mediated NO cytotoxicity is enhanced by either epidermal growth factor (EGF) or NMDA. These findings support the idea that GSH might represent the most important buffer of NO toxicity in neuronal cells, and indicate that the disruption of cellular redox buffering controlled by GSH makes neuronal cells susceptible to endogenous physiological flux of NO.


Apoptosis controls the speed of looping morphogenesis in Drosophila male terminalia.
In metazoan development, the precise mechanisms that regulate the completion of morphogenesis according to a developmental timetable remain elusive. The Drosophila male terminalia is an asymmetric looping organ; the internal genitalia (spermiduct) loops dextrally around the hindgut. Mutants for apoptotic signaling have an orientation defect of their male terminalia, indicating that apoptosis contributes to the looping morphogenesis. However, the physiological roles of apoptosis in the looping morphogenesis of male terminalia have been unclear. Here, we show the role of apoptosis in the organogenesis of male terminalia using time-lapse imaging. In normal flies, genitalia rotation accelerated as development proceeded, and completed a full 360° rotation. This acceleration was impaired when the activity of caspases or JNK or PVF/PVR signaling was reduced. Acceleration was induced by two distinct subcompartments of the A8 segment that formed a ring shape and surrounded the male genitalia: the inner ring rotated with the genitalia and the outer ring rotated later, functioning as a 'moving walkway' to accelerate the inner ring rotation. A quantitative analysis combining the use of a FRET-based indicator for caspase activation with single-cell tracking showed that the timing and degree of apoptosis correlated with the movement of the outer ring, and upregulation of the apoptotic signal increased the speed of genital rotation. Therefore, apoptosis coordinates the outer ring movement that drives the acceleration of genitalia rotation, thereby enabling the complete morphogenesis of male genitalia within a limited developmental time frame.


Pulmonary lymphangitic carcinomatosis from metastatic gastric adenocarcinoma: case report.
Pulmonary lymphangitic carcinomatosis (PLC) is the diffuse infltration of lymphatic systems by adenocarcinomas. Because of its vague clinical symptoms and absence of malignant features, confrmatory diagnosis requires transbronchial or open-lung biopsy. Tumor spread is postulated to occur by means of retrograde lymphatic permeation with tumor growth along the lymphatic channels and involvement of vascular endothelial growth factor-C in a signaling pathway. The authors describe the case of a 45-year-old woman whose presentation with cough and dyspnea led to diagnosis of pulmonary lymphangitic carcinomatosis with superimposed pulmonary hypertension, tumor thrombotic microangiopathy, and ultimately metastatic signet ring cell gastric adenocarcinoma. Because of the patient's poor prognosis, she was given palliative care and died 29 days after admission. This case emphasizes the importance of the osteopathic principle of treating the whole patient and evaluating all organ systems. Physicians should be aware of PLC from gastric adenocarcinoma and consider it in patients with pulmonary symptoms who are unresponsive to antibiotics.


Speaking of risk, managing uncertainty: decision-making about cholesterol-reducing treatment in general practice.
Decision making regarding treatment options and risk communication between health professionals and patients have become key areas of research and quality improvement. Decisions are directed towards applying evidence, exploring individual patient concerns, and addressing medical uncertainty. Compared with research on health professionals' views on higher stakes decisions, relatively little attention has been paid to improving decision making and risk communication for lifestyle-related risk conditions involving lower stakes or longer term treatment options, such as medication to reduce risk of future disease. To examine general practitioners (GPs) who experienced difficulties with decision making regarding treatment options and risk communication with asymptomatic patients with high cholesterol and risk of cardiovascular disease. An exploratory qualitative and ethnographically informed approach was used. Danish GPs (six male, six female, average age 48 years), were interviewed in three groups. Interviews were transcribed and coded, while analytical concepts about medical uncertainty were identified. The study identified two modalities of medical uncertainty: epistemological uncertainty about scientific knowledge and evidence-based medicine; and situational uncertainty produced in the one-to-one relationship between the GP and the patient during the consultation. The study also stressed that the decision making about cholesterol-reducing treatment is interpreted by the GPs as reversible and provisional. These modalities of medical uncertainty can be addressed proactively when developing concepts, tools and training interventions to optimise communication about treatment options in primary care.


Escape of a Small Molecule from Inside T4 Lysozyme by Multiple Pathways.
The T4 lysozyme L99A mutant is often used as a model system to study small-molecule binding to proteins, but pathways for ligand entry and exit from the buried binding site and the associated protein conformational changes have not been fully resolved. Here, molecular dynamics simulations were employed to model benzene exit from its binding cavity using the weighted ensemble (WE) approach to enhance sampling of low-probability unbinding trajectories. Independent WE simulations revealed four pathways for benzene exit, which correspond to transient tunnels spontaneously formed in previous simulations of apo T4 lysozyme. Thus, benzene unbinding occurs through multiple pathways partially created by intrinsic protein structural fluctuations. Motions of several α-helices and side chains were involved in ligand escape from metastable microstates. WE simulations also provided preliminary estimates of rate constants for each exit pathway. These results complement previous works and provide a semiquantitative characterization of pathway heterogeneity for binding of small molecules to proteins.


Multiple rare alleles contribute to low plasma levels of HDL cholesterol.
Heritable variation in complex traits is generally considered to be conferred by common DNA sequence polymorphisms. We tested whether rare DNA sequence variants collectively contribute to variation in plasma levels of high density lipoprotein cholesterol (HDL-C). We sequenced three candidate genes (ABCA1, APOA1, and LCAT) that cause Mendelian forms of low HDL-C levels in individuals from a population-based study. Nonsynonymous sequence variants were significantly more common (16% versus 2%) in individuals with low HDL-C (<fifth percentile) than in those with high HDL-C (>95th percentile). Similar findings were obtained in an independent population, and biochemical studies indicated that most sequence variants in the low HDL-C group were functionally important. Thus, rare alleles with major phenotypic effects contribute significantly to low plasma HDL-C levels in the general population.


Thrombotic microangiopathy mimicking membranoproliferative glomerulonephritis.
A 4-year-old boy presented with proteinuria and developed progressive renal failure over 6 years. In the patient's family, five individuals were affected with atypical haemolytic uraemic syndrome (aHUS) but not the patient. Renal biopsies (n = 3) showed glomerular basement membrane thickening with double contours, endothelial swelling and deposits of C3 and C1q. Electron microscopy revealed mesangial and subendothelial electron-dense deposits. Complement mutations in membrane cofactor protein (Y155D) and C3 (R713W and G1094R) were detected in all affected family members. The patient also had transient autoantibodies to factor H. The findings suggest that aHUS and glomerulopathy resembling membranoproliferative glomerulonephritis may have a common molecular background.


Does off-pump coronary artery bypass surgery reduce mortality in high risk patients?
The aim of this retrospective study was to compare outcome in two groups of patients who were classified according to their risk groups and underwent coronary revascularization with or without cardiopulmonary bypass. Between January 1996 and July 2000, 480 cases that underwent coronary artery bypass surgery (CABG) were included in a retrospective nonrandomized manner for study. Group 1 included 210 patients who were revascularized using off-pump techniques. Octopus 2 and 3 (Medtronic, Inc., Minneapolis, MN) were used for tissue stabilization. Group 2 included 270 cases who underwent CABG using CPB. Emergency cases, combined operations, reoperations, and patients in cardiogenic shock were excluded. Demographic variables were comparable between two the groups. Using the Allegheny Clinic Risk Scoring Scale [Magovern 1996], patients in both groups were scored as low, moderate, and high risk. In Group 1, 37 % of patients consisted of high risk patients while Group 2 had 14% (p < 0.05). Student's t-test and chi-square test were used for statistical analysis and alfa < 0.05 was considered significant. Mortality was 1.4% in Group 1 and 1.5% in Group 2 (p = ns). Mean anastomosis per patient was 2.6 +/- 0.6 in Group 1 and 3.2 +/- 0.5 in Group 2 (p < 0.05). Patients in Group 1 needed less blood transfusions and less inotropic support postoperatively (p < 0.05). There were also fewer minor neurological events (p < 0.05) and pulmonary complications (Type 2) in Group 1. Atrial fibrillation rate, infection, and major neurological deficit (Type 1) were similar in both groups. Mortality was less among Group 1 high risk patients (3.9 %) in comparison to Group 2 high risk patients (7.9 %), but this did not reach statistical significance. In low or moderate risk patients, CABG can be performed safely with or without CPB. In high risk patients with several comorbidities, off-pump CABG seems to be a safe and efficient method that can improve outcome.


A cross-sectional survey of the nature and correlates of sleep disturbance in people with psoriasis.
Research suggests that sleep disturbance is common in psoriasis. While several sleep investigations have been conducted in psoriasis populations, many have methodological shortcomings, and no study has examined multiple dimensions of sleep-wake functioning. Moreover, research has yet to be performed comprehensively examining the range of physical and psychological factors that may affect sleep in people with psoriasis. To characterize sleep disturbance using validated measures and to identify physical and psychological predictors of sleep quality in people with psoriasis. An online survey was conducted (186 respondents; mean age 39·2 years) comprising validated measures assessing sleep [Pittsburgh Sleep; Quality Index (PSQI), Berlin Questionnaire, Pre-Sleep Arousal Scale]; chronotype (Morningness-Eveningness Questionnaire); mood (Hospital Anxiety and Depression Scale); itch (5-D Itch Scale); and psoriasis severity (Simplified Psoriasis Index). Group comparisons and regression analyses were used to examine predictors of poor sleep. The mean PSQI score was 9·2 ± 4·3, with 76·3% scoring above the threshold for poor sleep (≥ 6 on the PSQI) and 32·5% scoring 'positive' for probable obstructive sleep apnoea (OSA). Poor sleep and high likelihood of OSA were associated with more severe psoriasis (P < 0·05; η = 0·07; η2 = 0·005). Cognitive arousal (β = 0·26, P = 0·001), itch (β = 0·26, P < 0·001) and depression (β = 0·24, P = 0·001) were the most robust predictors of poor sleep quality, which, together with somatic arousal (β = 0·17, P = 0·022), accounted for 43% of variance in PSQI scores. Poor sleep is common in psoriasis and associated with psychological and physical factors. Rates of probable OSA are also high. Given the importance of restorative sleep for health, sleep complaints should receive greater clinical attention in the management of psoriasis.


Association of serum sex steroid levels and bone mineral density with CYP17 and CYP19 gene polymorphisms in postmenopausal women in Turkey.
Many clinical conditions, including osteoporosis, are associated with serum levels of sex steroids. Enzymes that regulate rate-limiting steps of steroidogenic pathways, such as CYP17 and CYP19, are also regarded as significant factors that may cause the development of these conditions. We investigated the association of two common polymorphisms, in the promoter region (T→C substitution) of CYP17 and exon 3 (G→A) of CYP19, with bone mineral density (BMD) in the lumbar spine and femoral neck and serum androgen/estradiol, in a case-control study of 172 postmenopausal women aged 62.3 ± 9.6 years (mean ± SD). The CYP17 TC genotype was significantly overrepresented in patients compared to controls, and TC genotype neck T-score and lumbar T-score values were significantly higher in patients compared to controls. CYP17 TC and TT genotype testosterone and DHEA-SO(4) levels were lower in patients compared to controls. All three genotypes of CYP19 had almost the same distribution among patients. The CYP19 AG genotype, however, was most frequent among controls. CYP19 lumbar BMD levels were close to each other among the different genotypes; however, AA and AG genotypes were significantly lower in patients. Testosterone and DHEA-SO(4) levels in the CYP19 GG genotype were higher compared to those of the other genotypes in patients but not in controls. CYP19 GA individuals had lower E(2) levels and lower BMD in controls and patients. Femoral neck BMD and lumbar T-score were also diminished with GA transition. In conclusion, CYP17 and CYP19 gene polymorphisms were found to be associated with osteoporosis in postmenopausal women in Turkey.


Pathology of the liver sinusoids.
The hepatic sinusoids comprise a complex of vascular conduits to transport blood from the porta hepatis to the inferior vena cava through the liver. Under normal conditions, portal venous and hepatic artery pressures are equalized within the sinusoids, oxygen and nutrients from the systemic circulation are delivered to the parenchymal cells and differentially distributed throughout the liver acini, and proteins of liver derivation are carried into the cardiac/systemic circulation. Liver sinusoid structures are lined by endothelial cells unique to their location, and Kupffer cells. Multifunctional hepatic stellate cells and various immune active cells are localized within the space of Disse between the sinusoid and the adjacent hepatocytes. Flow within the sinusoids can be compromised by physical or pressure blockage in their lumina as well as obstructive processes within the space of Disse. The intimate relationship of the liver sinusoids to neighbouring hepatocytes is a significant factor affecting the health of hepatocytes, or transmission of the effects of injury within the sinusoidal space. Pathologists should recognize several patterns of injury involving the sinusoids and surrounding hepatocytes. In this review, injury, alterations and accumulations within the liver sinusoids are illustrated and discussed.


Mitochondrial DNA control region analysis of three ethnic populations in lower Northern part of Thailand.
The lower northern part of Thailand contains various genetically diverse ethnic populations. The sequences of the mitochondrial DNA hypervariable region were studied in three ethnic populations inhabiting Phitsanulok Province. One hundred and nine nucleotide sequences - 53, 29, and 27 from Hmongs (Hill tribe), Lao Songs, and Thai-Siams, respectively - were collected. The haplotypes were generated from 1130 nucleotides of the entire control region. Eighty-six haplotypes were found in the three ethnic populations, and no shared haplotypes were found between populations. Point heteroplasmy was noted at position 311 (C→Y). Haplotypes with ACAC-insertion at position 512 were observed in immigrant individuals from the Lao Song population. The Thai-Siam population showed higher genetic diversity than the other populations. The Hmong and Lao Song populations showed less genetic diversity than those living in their original area. The neutrality testing suggested that the result might be influenced by genetic drift.


Comparison of BD GeneOhm Cdiff real-time PCR assay with a two-step algorithm and a toxin A/B enzyme-linked immunosorbent assay for diagnosis of toxigenic Clostridium difficile infection.
The BD GeneOhm Cdiff assay, a real-time PCR assay for the detection of the Clostridium difficile toxin B (tcdB) gene, was compared with the toxin A/B (Tox A/B) II enzyme-linked immunosorbent assay (ELISA) and a two-step algorithm which includes a C. Diff Chek-60 glutamate dehydrogenase (GDH) antigen assay followed by cytotoxin neutralization. Four hundred liquid or semisolid stool samples submitted for diagnostic C. difficile testing, 200 GDH antigen positive and 200 GDH antigen negative, were selected for analysis. All samples were tested by the C. Diff Chek-60 GDH antigen and cytotoxin neutralization assays, the Tox A/B II ELISA, and the BD GeneOhm Cdiff assay. Specimens with discrepant results were tested by toxigenic culture as an independent "gold standard." Of 200 GDH-positive samples, 71 were positive by the Tox A/B II ELISA, 88 were positive by the two-step method, 93 were positive by PCR, and 96 were positive by the GDH antigen assay only. Of 200 GDH-negative samples, 3 were positive by PCR only. Toxigenic culture was performed for 41 samples with discrepant results, and 39 were culture positive. Culture resolution of discrepant results showed the Tox A/B II assay to have detected 70 (66.7%), the two-step method to have detected 87 (82.9%), and PCR to have detected 96 (91.4%) of 105 true positives. The BD GeneOhm Cdiff assay was more sensitive in detecting toxigenic C. difficile than the Tox A/B II assay (P < 0.0001); however, the difference between PCR and the two-step method was not significant (P = 0.1237). Enhanced sensitivity and rapid turnaround time make the BD GeneOhm Cdiff assay an important advance in the diagnosis of toxigenic C. difficile infection.


Phytochrome-mediated bioelectric potentials in mung bean seedlings.
Excised root tips from dark-grown mung bean seedlings (Phaseolus aureus) that adhere to a negatively charged glass surface when irradiated with red light and release when irradiated by far-red light develop a positive bioelectric potential (about 1.0 millivolt) at the tip in red light and a negative bioelectric potential in far-red light. The sign of the bioelectric potential was repeatedly photo-reversible, and the adhesion and release kinetics were similar to those of the development of the bioelectric potentials. Photoconversion of the phytochrome holochrome perhaps changes permeability characteristics of the cell membrane, resulting in an induced localized electrochemical gradient manifested as a bioelectric potential. This supports the view that phytochrome in situ is membrane bound.


Challenge of liver disease in systemic lupus erythematosus: Clues for diagnosis and hints for pathogenesis.
Systemic lupus erythematosus (SLE) encompass a broad spectrum of liver diseases. We propose here to classify them as follows: (1) immunological comorbilities (overlap syndromes); (2) non-immunological comorbilities associated to SLE; and (3) a putative liver damage induced by SLE itself, referred to as "lupus hepatitis". In the first group, liver injury can be ascribed to overlapping hepatopathies triggered by autoimmune mechanisms other than SLE occurring with higher incidence in the context of lupus (e.g., autoimmune hepatitis, primary biliary cirrhosis). The second group includes non-autoimmune liver diseases, such as esteatosis, hepatitis C, hypercoagulation state-related liver lesions, hyperplasic parenchymal and vascular lesions, porphyria cutanea tarda, and drug-induced hepatotoxicity. Finally, the data in the literature to support the existence of a hepatic disease produced by SLE itself, or the occurrence of a SLE-associated prone condition that increases susceptibility to acquire other liver diseases, is critically discussed. The pathological mechanisms underlying each of these liver disorders are also reviewed. Despite the high heterogeneity in the literature regarding the prevalence of SLE-associated liver diseases and, in most cases, lack of histopathological evidence or clinical studies large enough to support their existence, it is becoming increasingly apparent that liver is an important target of SLE. Consequently, biochemical liver tests should be routinely carried out in SLE patients to discard liver disorders, particularly in those patients chronically exposed to potentially hepatotoxic drugs. Diagnosing liver disease in SLE patients is always challenging, and the systematization of the current information carried out in this review is expected to be of help both to attain a better understanding of pathogenesis and to build an appropriate work-up for diagnosis.


Optimal suppression of quantization noise with pseudoperiodic multilevel phase gratings.
A comprehensive two-step approach to design staircase-type multilevel diffractive phase elements (DPEs) that generate arbitrary desired diffraction patterns with the highest possible accuracy is presented. First a preliminary periodic grating with an unconstrained phase delay and an optimized nonuniform amplitude profile is designed by means of a customized iterative Fourier-transform algorithm. Then this preliminary grating is subjected to a phase quantization in which strict periodicity is forgone in favour of the best possible preservation of the shape of the power spectrum yielding a final phase only DPE with only rudimentary periodicity. An arbitrarily high similarity between the diffraction patterns of the final DPE and the preliminary grating can be achieved independently of the number D of discrete phase delay levels as long as D > or = 3. The signal-to-noise ratio of the final DPE is close to the theoretical upper limit. These properties are confirmed in computer simulations and demonstrated in optical experiments. Pseudoperiodic DPEs may have applications in optical computing, optical communication and networking, optical authentication, or coherent laser coupling.


Personal Fixations-Based Object Segmentation With Object Localization and Boundary Preservation.
As a natural way for human-computer interaction, fixation provides a promising solution for interactive image segmentation. In this paper, we focus on Personal Fixations-based Object Segmentation (PFOS) to address issues in previous studies, such as the lack of appropriate dataset and the ambiguity in fixations-based interaction. In particular, we first construct a new PFOS dataset by carefully collecting pixel-level binary annotation data over an existing fixation prediction dataset, such dataset is expected to greatly facilitate the study along the line. Then, considering characteristics of personal fixations, we propose a novel network based on Object Localization and Boundary Preservation (OLBP) to segment the gazed objects. Specifically, the OLBP network utilizes an Object Localization Module (OLM) to analyze personal fixations and locates the gazed objects based on the interpretation. Then, a Boundary Preservation Module (BPM) is designed to introduce additional boundary information to guard the completeness of the gazed objects. Moreover, OLBP is organized in the mixed bottom-up and top-down manner with multiple types of deep supervision. Extensive experiments on the constructed PFOS dataset show the superiority of the proposed OLBP network over 17 state-of-the-art methods, and demonstrate the effectiveness of the proposed OLM and BPM components. The constructed PFOS dataset and the proposed OLBP network are available at https://github.com/MathLee/OLBPNet4PFOS.


Estrous cycle in the rat: effects on self-stimulation behavior.
The performance of female rats, in pressing a bar for electrical stimulation of the hypothalamus, changes during the estrous cycle. Highest barpressing rates accompany the appearance of vaginal cornification. This increase is not an artifact of increased spontaneous activity at estrus, although the factors underlying these changes in activity may also mediate the changes in self-stimulation behavior.


Hormonal regulation of gap junction differentiation.
Thin-section, tracer, and freeze-cleave experiments on hypophysectomized Rana pipiens larvae reveal that gap junctions form between differentiating ependymoglial cells in response to thyroid hormone. These junctions assemble in large particle-free areas of the plasma membrane known as formation plaques. Between 20 and 40 h after hormone application, formation plaque area increases approximately 26-fold while gap junction area rises about 20-fold. The differentiation of these junctions requires the synthesis of new protein and probably RNA as well. On the basis of inhibitor experiments, it can be reported that formation plaques develop at about 16-20 h after hormone treatment and stages in the construction of gap junctions appear 4-8 h later. These studies suggest that gap junction subunits are synthesized and inserted into formation plaque membrane during the differentiation of the anuran ependymoglial cells.


Do body mass index trajectories affect the risk of type 2 diabetes? A case-control study.
Although obesity is a well-studied risk factor for diabetes, there remains an interest in whether "increasing body mass index (BMI)," "high BMI per se," or both are the actual risk factors for diabetes. The present study aimed to retrospectively compare BMI trajectories of individuals with and without diabetes in a case-control design and to assess whether increasing BMI alone would be a risk factor. Using comprehensive health check-up data measured over ten years, we conducted a case-control study and graphically drew the trajectories of BMIs among diabetic patients and healthy subjects, based on coefficients in fitted linear mixed-effects models. Patient group was matched with healthy control group at the onset of diabetes with an optimal matching method in a 1:10 ratio. Simple fixed-effects models assessed the differences in increasing BMIs over 10 years between patient and control groups. At the time of matching, the mean ages in male patients and controls were 59.3 years [standard deviation (SD) = 9.2] and 57.7 years (SD = 11.2), whereas the mean BMIs were 25.0 kg/m(2) (SD = 3.1) and 25.2 kg/m(2) (SD = 2.9), respectively. In female patients and controls, the mean ages were 61.4 years (SD = 7.9) and 60.1 years (SD = 9.6), whereas the mean BMIs were 24.8 kg/m(2) (SD = 3.5) and 24.9 kg/m(2) (SD = 3.4), respectively. The simple fixed-effects models detected no statistical significance for the differences of increasing BMIs between patient and control groups in males (P = 0.19) and females (P = 0.67). Sudden increases in BMI were observed in both male and female patients when compared with BMIs 1 year prior to diabetes onset. The present study suggested that the pace of increasing BMIs is similar between Japanese diabetic patients and healthy individuals. The increasing BMI was not detected to independently affect the onset of type 2 diabetes.


Host cell transcriptional profiling during malaria liver stage infection reveals a coordinated and sequential set of biological events.
Plasmodium sporozoites migrate to the liver where they traverse several hepatocytes before invading the one inside which they will develop and multiply into thousands of merozoites. Although this constitutes an essential step of malaria infection, the requirements of Plasmodium parasites in liver cells and how they use the host cell for their own survival and development are poorly understood. To gain new insights into the molecular host-parasite interactions that take place during malaria liver infection, we have used high-throughput microarray technology to determine the transcriptional profile of P. berghei-infected hepatoma cells. The data analysis shows differential expression patterns for 1064 host genes starting at 6 h and up to 24 h post infection, with the largest proportion correlating specifically with the early stages of the infection process. A considerable proportion of those genes were also found to be modulated in liver cells collected from P. yoelii-infected mice 24 and 40 h after infection, strengthening the data obtained with the in vitro model and highlighting genes and pathways involved in the host response to rodent Plasmodium parasites. Our data reveal that host cell infection by Plasmodium sporozoites leads to a coordinated and sequential set of biological events, ranging from the initial stage of stress response up to the engagement of host metabolic processes and the maintenance of cell viability throughout infection.


A Novel and Generic Workflow of Indocyanine Green Perfusion Assessment Integrating Standardization and Quantification Toward Clinical Implementation.
This study aims to generate a reproducible and generalizable Workflow model of ICG-angiography integrating Standardization and Quantification (WISQ) that can be applied uniformly within the surgical innovation realm independent of the user. Tissue perfusion based on indocyanine green (ICG)-angiography is a rapidly growing application in surgical innovation. Interpretation of results has been subjective and error-prone due to the lack of a standardized and quantitative ICG-workflow and analytical methodology. There is a clinical need for a more generic, reproducible, and quantitative ICG perfusion model for objective assessment of tissue perfusion. In this multicenter, proof-of-concept study, we present a generic and reproducible ICG-workflow integrating standardization and quantification for perfusion assessment. To evaluate our model's clinical feasibility and reproducibility, we assessed the viability of parathyroid glands after performing thyroidectomy. Biochemical hypoparathyroidism was used as the postoperative endpoint and its correlation with ICG quantification intraoperatively. Parathyroid gland is an ideal model as parathyroid function post-surgery is only affected by perfusion. We show that visual subjective interpretation of ICG-angiography by experienced surgeons on parathyroid perfusion cannot reliably predict organ function impairment postoperatively, emphasizing the importance of an ICG quantification model. WISQ was able to standardize and quantify ICG-angiography and provided a robust and reproducible perfusion curve analysis. A low ingress slope of the perfusion curve combined with a compromised egress slope was indicative for parathyroid organ dysfunction in 100% of the cases. WISQ needs prospective validation in larger series and may eventually support clinical decision-making to predict and prevent postoperative organ function impairment in a large and varied surgical population.


Management of heatstroke and heat exhaustion.
Heat exhaustion and heatstroke are part of a continuum of heat-related illness. Both are common and preventable conditions affecting diverse patients. Recent research has identified a cascade of inflammatory pathologic events that begins with mild heat exhaustion and, if uninterrupted, can lead eventually to multiorgan failure and death. Heat exhaustion is characterized by nonspecific symptoms such as malaise, headache, and nausea. Treatment involves monitoring the patient in a cool, shady environment and ensuring adequate hydration. Untreated heat exhaustion can progress to heatstroke, a much more serious illness involving central nervous system dysfunction such as delirium and coma. Other systemic effects, including rhabdomyolysis, hepatic failure, arrhythmias, disseminated intravascular coagulation, and even death, are not uncommon. Prompt recognition and immediate cooling through evaporation or full-body ice-water immersion are crucial. Physicians also must monitor electrolyte abnormalities, be alert to signs of renal or hepatic failure, and replace fluids in patients with heatstroke. Most experts believe that physicians and public health officials should focus greater attention on prevention. Programs involving identification of vulnerable individuals, dissemination of information about dangerous heat waves, and use of heat shelters may help prevent heat-related illness. These preventive measures, when paired with astute recognition of the early signs of heat-related illness, can allow physicians in the ambulatory setting to avert much of the morbidity and mortality associated with heat exhaustion and heatstroke.


Role of the human kidney in glucose counterregulation.
Animal experiments indicate that the kidney may play an important role in glucose counterregulation. Because the human kidney normally takes up and releases glucose, and since patients with end-stage renal disease are prone to hypoglycemia, we examined whether the kidney is also involved in human glucose counterregulation. Accordingly, we compared renal glucose release (RGR) and uptake (RGU) during 4-h hyperinsulinemic-hypoglycemic (approximately 3.2 mmol/l, n = 9) and -euglycemic (approximately 5 mmol/l, n = 10) control clamp experiments in normal postabsorptive subjects. A combination of renal balance and isotopic ([3H]glucose, [14C]glutamine) techniques was used, which permitted hepatic glucose release (HGR) and glutamine gluconeogenesis to be calculated as the difference between systemic (overall) and renal values. In both experiments, infusion of insulin increased plasma insulin comparably (approximately 210 pmol/l). In euglycemic control experiments, RGR and HGR decreased more than 50% (both P<0.001) and RGU increased approximately 35% (P = 0.02). In hypoglycemic experiments, both HGR (P = 0.034) and RGR (P<0.001) increased to a comparable extent (1.69+/-0.47 and 1.67+/-0.15 pmol x kg-(-1) x min(-1), respectively, P = 0.96) above rates observed in control experiments; hepatic and renal glutamine gluconeogenesis increased by 0.19+/-0.06 (P<0.008) and 0.30+/-0.07 pmol x kg(-1) x min(-1) (P< 0.001), respectively. RGU decreased by 65% compared with control experiments (P<0.001), so that renal glucose balance changed from a net uptake of 80+/-19 micromol/min to a net release of 130+/-9 micromol/min, P< 0.001. These observations provide evidence that the kidney may play an important role in human glucose counterregulation.


Trainee Occupational Therapists Scoring the Barthel ADL.
Within medical applications there are two main types of information design; paper-based and digital information [1]. As technology is constantly changing, information within healthcare management and delivery is continually being transitioned from traditional paper documents to digital and online resources. Activity of Daily Living (ADL) charts are still predominantly paper based and are therefore prone to "human error" [2]. In light of this, an investigation has taken place into the design for reducing the amount of human error, between a paper based ADL, specifically the Barthel Index, and the same ADL created digitally. The digital ADL was developed as an online platform as this offers the best method of data capture for a large group of participants all together [3]. The aim of the study was to evaluate the usability of the Barthel Index ADL in paper format and then reproduce the same ADL digitally. This paper presents the findings of a study involving 26 participants who were familiar with ADL charts, and used three scenarios requiring them to complete both a paper ADL and a digital ADL. An evaluation was undertaken to ascertain if there were any 'human errors' in completing the paper ADL and also to find similarities/differences through using the digital ADL. The results from the study indicated that 22/26 participants agreed that the digital ADL was better, if not the same as a paper based ADL. Further results indicated that participants rate highly the added benefit of the digital ADL being easy to use and also that calculation of assessment scores were performed automatically. Statistically the digital BI offered a 100 % correction rate in the total calculation, in comparison to the paper based BI where it is more common for users to make mathematical calculation errors. Therefore in order to minimise handwriting and calculation errors, the digital BI proved superior than the traditional paper based method.


Presentation, natural history, and management of carotid cavernous aneurysms.
We present the largest reported cohort of carotid cavernous aneurysms (CCA), comparing the neuro-ophthalmic presentation, complications, and outcome with and without endovascular treatment. Retrospective review of 185 patients with 206 CCAs examined between 1980 and 2001 at a tertiary neuro-ophthalmology and neurovascular service. Patients' symptoms and findings at presentation were recorded and compared with those at outcome. The effect of treatment on outcome and on complication rate was analyzed using the chi test, multivariate analysis of covariance, model-selection log-linear analysis, and multinomial logistic regression. Long-term follow-up was available for 189 of 206 CCAs. Seventy-four CCAs underwent treatment (endovascular, 67 [91%]; surgical treatment, 6 [9%]), and 115 were followed for an average of 4 years, two of which required later treatment. Treatment reduced the incidence and severity of pain, even after adjusting for the severity of initial pain (F(1,192 = 9.59, P = 0.002). Treatment did not significantly affect the patient's final diplopia after adjusting for their initial diplopia (F(1, 182 = 2.01, P = 0.158). Statistical examination revealed that the treated group had a higher proportion of neurological and visual complications than people who were not treated (2(2). = 25.26, P = 0.0003). Endovascular treatment of carotid cavernous aneurysms leads to a significantly higher rate of pain resolution compared with untreated patients, even after adjusting for initial pain severity. Diplopia may not resolve after treatment. The results of this study underscore our approach indicating treatment only in cases of debilitating pain, visual loss from compression, or diplopia in primary gaze or in patients with risk factors for major complications such as pre-existing coagulopathy or sphenoid sinus erosion.


Prevalence of depression in granted and refused requests for euthanasia and assisted suicide: a systematic review.
There is an established link between depression and interest in hastened death in patients who are seriously ill. Concern exists over the extent of depression in patients who actively request euthanasia/physician-assisted suicide (PAS) and those who have their requests granted. To estimate the prevalence of depression in refused and granted requests for euthanasia/PAS and discuss these findings. Methods A systematic review was performed in MEDLINE and PsycINFO in July 2010, identifying studies reporting rates of depression in requests for and cases of euthanasia/PAS. One author critically appraised the strength of the data using published criteria. 21 studies were included covering four countries. There was considerable heterogeneity in methods of assessing depression and selecting patients. In the highest quality studies, in the Netherlands and Oregon, 8-47% of patients requesting euthanasia/PAS had depressive symptoms and 2-17% of completed euthanasia/PAS cases had depressive symptoms. In the Netherlands, depression was significantly higher in refused than granted requests, and there was no significant difference in the rate of depression between euthanasia cases and similar patients who had not made a request for euthanasia. It is unclear whether depression increases the probability of making a request for euthanasia/PAS, but in the Netherlands most requests in depressed patients are rejected, leaving a depression rate in cases that is similar to the surrounding population. Less evidence is available elsewhere, but some level of depression has been identified in patients undergoing euthanasia/PAS in all the countries studied. Whether the presence of depression is ever compatible with an ethical decision on euthanasia/PAS is discussed.


Gender Differences in Intimate Partner Violence in Current and Prior Relationships.
Although much available research indicates that intimate partner violence (IPV) is male perpetrated, growing recent evidence suggests a gender symmetry model of family violence. This article examines gender differences in IPV in current and prior relationships reported by young adults. Data comprised 2,060 young adults (62.1% females) who participated in the 30-year follow-up of the Mater Hospital and University of Queensland Study of Pregnancy (MUSP) in Brisbane, Australia. The Composite Abuse Scale was used to measure IPV during the last 12 months in the respondents' most recent relationship. Similar proportions of males and females reported leaving their prior relationships. Both males and females who were not currently in a relationship reported experiencing much higher rates of IPV than those who were in a relationship. There were no differences in the past experience of IPV between males and females who were not currently in a relationship, but males in a current relationship reported they experienced most forms of IPV more often than did females. IPV typically involves both male and female perpetrators and victims. It does appear that the majority of relationships involving higher rates of IPV were dissolved. IPV was more likely to have occurred in relationships that ended than in relationships that persisted. Males more often remain in an abusive relationship and report experiencing higher rates of IPV in their current relationships compared with females.


Spatial organization and dynamics of the association of Rec102 and Rec104 with meiotic chromosomes.
Meiotic double-strand breaks (DSBs) are formed by Spo11 in conjunction with at least nine other proteins whose roles are not well understood. We find that two of these proteins, Rec102 and Rec104, interact physically, are mutually dependent for proper subcellular localization, and share a requirement for Spo11 and Ski8 for their recruitment to meiotic chromosomes, suggesting that they work together as a functional unit. Rec102 associated extensively with chromatin loops during leptotene and zygotene and showed preferential binding in the vicinity at least of most DSB sites, consistent with a direct role in DSB formation. However, Rec102 was associated with both DSB-hot and DSB-cold regions, ruling out a simple model in which sites of DSB formation are dictated by where Rec102/104 complexes load. Both proteins persisted on chromatin until pachytene before abruptly disappearing, indicating that they remain on chromosomes well after DSB formation. These studies reveal unexpected behaviors for Rec102 and Rec104, and point to distinct roles and subcomplexes among the DSB proteins.


Ionotropic glutamate receptor AMPA 1 is associated with ovulation rate.
Ionotropic glutamate receptors mediate most excitatory neurotransmission in the central nervous system by opening ion channels upon the binding of glutamate. Despite the essential roles of glutamate in the control of reproduction and anterior pituitary hormone secretion, there is a limited understanding of how glutamate receptors control ovulation. Here we reveal the function of the ionotropic glutamate receptor AMPA-1 (GRIA1) in ovulation. Based on a genome-wide association study in Bos taurus, we found that ovulation rate is influenced by a variation in the N-terminal leucine/isoleucine/valine-binding protein (LIVBP) domain of GRIA1, in which serine is replaced by asparagine. GRIA1(Asn) has a weaker affinity to glutamate than GRIA1(Ser), both in Xenopus oocytes and in the membrane fraction of bovine brain. This single amino acid substitution leads to the decreased release of gonadotropin-releasing hormone (GnRH) in immortalized hypothalamic GT1-7 cells. Cows with GRIA1(Asn) have a slower luteinizing hormone (LH) surge than cows with GRIA1(Ser). In addition, cows with GRIA1(Asn) possess fewer immature ovarian follicles before superovulation and have a lower response to hormone treatment than cows with GRIA1(Ser). Our work identified that GRIA1 is a critical mediator of ovulation and that GRIA1 might be a useful target for reproductive therapy.


Histone methylation by NUE, a novel nuclear effector of the intracellular pathogen Chlamydia trachomatis.
Sequence analysis of the genome of the strict intracellular pathogen Chlamydia trachomatis revealed the presence of a SET domain containing protein, proteins that primarily function as histone methyltransferases. In these studies, we demonstrated secretion of this protein via a type III secretion mechanism. During infection, the protein is translocated to the host cell nucleus and associates with chromatin. We therefore named the protein nuclear effector (NUE). Expression of NUE in mammalian cells by transfection reconstituted nuclear targeting and chromatin association. In vitro methylation assays confirmed NUE is a histone methyltransferase that targets histones H2B, H3 and H4 and itself (automethylation). Mutants deficient in automethylation demonstrated diminished activity towards histones suggesting automethylation functions to enhance enzymatic activity. Thus, NUE is secreted by Chlamydia, translocates to the host cell nucleus and has enzymatic activity towards eukaryotic substrates. This work is the first description of a bacterial effector that directly targets mammalian histones.


The T790M mutation in EGFR kinase causes drug resistance by increasing the affinity for ATP.
Lung cancers caused by activating mutations in the epidermal growth factor receptor (EGFR) are initially responsive to small molecule tyrosine kinase inhibitors (TKIs), but the efficacy of these agents is often limited because of the emergence of drug resistance conferred by a second mutation, T790M. Threonine 790 is the "gatekeeper" residue, an important determinant of inhibitor specificity in the ATP binding pocket. The T790M mutation has been thought to cause resistance by sterically blocking binding of TKIs such as gefitinib and erlotinib, but this explanation is difficult to reconcile with the fact that it remains sensitive to structurally similar irreversible inhibitors. Here, we show by using a direct binding assay that T790M mutants retain low-nanomolar affinity for gefitinib. Furthermore, we show that the T790M mutation activates WT EGFR and that introduction of the T790M mutation increases the ATP affinity of the oncogenic L858R mutant by more than an order of magnitude. The increased ATP affinity is the primary mechanism by which the T790M mutation confers drug resistance. Crystallographic analysis of the T790M mutant shows how it can adapt to accommodate tight binding of diverse inhibitors, including the irreversible inhibitor HKI-272, and also suggests a structural mechanism for catalytic activation. We conclude that the T790M mutation is a "generic" resistance mutation that will reduce the potency of any ATP-competitive kinase inhibitor and that irreversible inhibitors overcome this resistance simply through covalent binding, not as a result of an alternative binding mode.


Coupled enhancer and coding sequence evolution of a homeobox gene shaped leaf diversity.
Here we investigate mechanisms underlying the diversification of biological forms using crucifer leaf shape as an example. We show that evolution of an enhancer element in the homeobox gene REDUCED COMPLEXITY (RCO) altered leaf shape by changing gene expression from the distal leaf blade to its base. A single amino acid substitution evolved together with this regulatory change, which reduced RCO protein stability, preventing pleiotropic effects caused by its altered gene expression. We detected hallmarks of positive selection in these evolved regulatory and coding sequence variants and showed that modulating RCO activity can improve plant physiological performance. Therefore, interplay between enhancer and coding sequence evolution created a potentially adaptive path for morphological evolution.


Selective vasodilators redistribute intestinal blood flow and depress oxygen uptake.
In this paper we report the effects of adenosine and isoproterenol on intestinal oxygen consumption. Using the recently developed technique of laser-Doppler velocimetry (LDV), we also studied the effects of these two agents on regional blood flow in the canine intestinal mucosa and muscularis. In isolated gut loops perfused at constant blood flow, both vasodilators depressed O2 uptake, but isoproterenol increased rubidium extraction while adenosine depressed rubidium extraction. Under constant pressure perfusion both dilators depressed O2 uptake but increased total flow. Isoproterenol increased the capillary filtration coefficient (Kf), but adenosine depressed Kf. LDV flowmetry showed adenosine increased muscularis flow and depressed mucosal flow. Both LDV flowmetry and radioactive microspheres showed isoproterenol increased mucosal but depressed muscularis flow. Our data indicate that the deleterious effects of isoproterenol and adenosine in depressing intestinal oxygen uptake can be explained by the ability of these agents to selectively vasodilate one of the regional circulations within the gut wall and by a "vascular steal" mechanism to reduce blood flow in the other vascular bed not as sensitive to the agent in question. The reduction in the regional blood flow thus depresses the O2 consumption of the gut as a whole. By comparing our regional laser-Doppler blood flow data with a simple analog model, we show that the seemingly paradoxical effect of vasodilators in depressing a regional blood flow while increasing total perfusion is explicable by a "vascular steal" mechanism.


Is the Life Space Assessment applicable to a palliative care population? Its relationship to measures of performance and quality of life.
The spatial environments that palliative care patients frequent for business and leisure constrict as their disease progresses and their physical functioning deteriorates. Measuring a person's movement within his or her own environment is a clinically relevant and patient-centered outcome because it measures function in a way that reflects actual and not theoretical participation. This exploratory study set out to test whether the Life-Space Assessment (LSA) would correlate with other commonly used palliative care outcome measures of function and quality of life. The baseline LSA, Australia-modified Karnofsky Performance Status Scale (AKPS), and the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-Core 15-Palliative (EORTC QLQ-C15-PAL) scores from two large clinical trials were used to calculate correlation coefficients between the measures. Convergent validity analysis was undertaken by comparing LSA scores between participants with higher (≥70) and lower (≤60) AKPS scores. The LSA was correlated significantly and positively with the AKPS, with a moderate correlation coefficient of 0.54 (P<0.001). There was a significant weak negative correlation between the LSA and the EORTC QLQ-C15-PAL, with a small coefficient of -0.22 (P=0.027), but a strong correlation between the LSA and the EORTC QLQ-C15-PAL item related to independent activities of daily living (r=-0.654, P<0.01). A significant difference in the LSA score between participants with higher (≥70) and lower (≤60) AKPS scores t(97)=-4.35, P<0.001) was found. The LSA appears applicable to palliative care populations given the convergent validity and capacity of this instrument to differentiate a person's ability to move through life-space zones by performance status. Further research is required to validate and apply the LSA within community palliative care populations.


Changes in terminology for childhood overweight and obesity.
A variety of different terms, metrics, and cut-off values have been used to describe and assess overweight and obesity in children. Body mass index (BMI) calculated as weight in kilograms divided by height in meters squared can be used to express weight adjusted for height. In order to account for variability by sex and age, BMI in children is compared to sex- and age-specific reference values. In the United States, the Centers for Disease Control and Prevention (CDC) 2000 growth charts serve as reference values. The terminology used for high BMI-for-age in children in the United States to date has been based on the recommendation of an expert committee convened by federal agencies. This committee recommended the use of BMI and defined overweight as a BMI-forage at or above the 95th percentile of a specified reference population and the designation of "at risk for overweight" for BMI values between the 85th and the 95th percentiles of BMI for age. More recently, although the cut-off values and the interpretation have not changed, changes in terminology were proposed. An American Medical Association expert committee report retained the two cut-off values of the 85th and 95th percentiles of BMI-for-age but used different terminology, referring to BMI-for-age from the 85th up to the 95th percentile as "overweight" and to BMI-for-age at or above the 95th percentile as "obesity." The National Center for Health Statistics (NCHS) and other CDC publications will continue to include prevalence estimates at the 85th and 95th percentiles as before but will change the terminology to use the term "overweight" for a BMI-for-age between the 85th and 95th percentile (formerly called "at risk for overweight") and the term "obesity" for a BMI-for-age at or above the 95th percentile (formerly called "overweight").


Aortic valve replacement for endocarditis: determinants of early and late outcome.
The study aim was to determine risk factors for operative mortality, recurrent infection, reoperation and long-term survival following aortic valve replacement (AVR) for infective endocarditis. Between 1973 and 1997, 109 patients (91 male, 18 female, mean age 52.6 years) underwent isolated AVR for infective endocarditis in our unit. Native valve endocarditis was present in 89 (81.6%) and prosthetic valve endocarditis in 20 (18.4%). Active culture-positive endocarditis was present in 53 (48.6%). Preoperatively, 99 patients (90.8%) were in NYHA classes III and IV. Indications for surgery included cardiac failure in 41 patients, valvular dysfunction in 38, vegetations in 18, sepsis in seven, abscess in six and embolism in four. Mechanical valves were implanted in 69 patients (63.3%) and bioprostheses in 40 (36.7%), including a homograft in 19 (17.4%). Follow up was complete (mean 5.8 years; range: 0-23.8 years; total 633.5 patient-years). The operative mortality was 10.1% (11 deaths). At ten years, freedom from recurrent infection was 94.2%, and freedom from reoperation 83.6%. Biological valve and younger age were significant adverse parameters for freedom from reoperation (p = 0.01 and p = 0.01). There have been 21 late deaths, 15 due to cardiac causes. Kaplan-Meier survival, including operative mortality, at five and ten years was 77.4% and 68.0%, respectively. On Cox proportional hazards regression, Staphylococcus aureus infection (p = 0.008) and older age (p = 0.04) were independent adverse predictors of survival. AVR for endocarditis carries a relatively high operative mortality, but can result in a satisfactory freedom from recurrent infection, reoperation and long-term survival. Analysis of our series demonstrates that implantation of a biological valve limits the freedom from reoperation and that infection by Staph. aureus reduces the probability of long-term survival.


Endothelin-1 as a putative modulator of erectile dysfunction: I. Characteristics of contraction of isolated corporal tissue strips.
Normal erectile physiology is heavily dependent on a delicate balance between the effects of endogenous vasoconstricting and vasorelaxing hormones on the tone of the corporal smooth muscle. Recent studies indicate that endothelin-1 (ET-1) is present and physiologically active in the human corpora. The primary goal of the present investigation was to further define the role of ET-1 in corporal physiology and to ascertain whether it might play a role in augmenting corporal tone in vivo, as reported in other vascular tissues. Thus, we conducted pharmacological studies of ET-1-induced steady-state contractions in isolated human corporal smooth muscle strips to determine if there were any detectable age- or diabetes-related alterations in ET-1-induced contractions. For statistical analysis, the patient population was divided into 2 age groups, A (< or = 59 years of age; = 11 patients) and B (> or = 60 years of age; n = 7 patients), and further subdivided into 2 diagnostic categories, diabetic (n = 7 patients) and nondiabetic (n = 11 patients). Construction of cumulative concentration response curves (CRCs) for ET-1-induced contractions revealed characteristically slow onset and long-lasting responses. Endothelin-1 CRC data were computer fit to the logistic equation to derive Emax (calculated maximal response), pEC50 (negative logarithm of the concentration that elicits one-half of the calculated maximal response) and slope factor (n) values. Two-factor analysis of variance revealed no detectable age- or diabetes-related alterations, nor any age-diabetes interaction in any of the logistic parameters. Furthermore, logistic analysis of ET-1 CRC data on 14 isolated corporal tissue strips derived from 3 potent patients with documented spontaneous erections revealed no differences in ET-1 contractility from that observed for patients with organic erectile dysfunction. Importantly, despite an apparent absence of age- or diabetes-related alterations in ET-1-induced steady-state contractions, preliminary studies demonstrated that coadministration of the alpha 1-adrenergic agonist phenylephrine and ET-1 produce much greater contractile responses than those observed in the presence of phenylephrine (PE) alone. Moreover, the magnitude of the augmentation was precisely that predicted by a model for simple additivity of agonist effects. Such observations suggest that the physiological relevance of ET-1 to corporal physiology may be related to its ability to augment the contractile responses of other vasomodulators present in the human corpora, in particular, perhaps modulating the contractile responses to sympathetic activity.


Sympathetic modulation of muscle spindle afferent sensitivity to stretch in rabbit jaw closing muscles.
Previous reports showed that sympathetic stimulation affects the activity of muscle spindle afferents (MSAs). The aim of the present work is to study the characteristics of sympathetic modulation of MSA response to stretch: (i) on the dynamic and static components of the stretch response, and (ii) on group Ia and II MSAs to evaluate potentially different effects. In anaesthetised rabbits, the peripheral stump of the cervical sympathetic nerve (CSN) was stimulated at 10 impulses s(-1) for 45-90 s. The responses of single MSAs to trapezoidal displacement of the mandible were recorded from the mesencephalic trigeminal nucleus. The following characteristic parameters were determined from averaged trapezoidal responses: initial frequency (IF), peak frequency at the end of the ramp (PF), and static index (SI). From these, other parameters were derived: dynamic index (DI = PF - SI), dynamic difference (DD = PF - IF) and static difference (SD = SI - IF). The effects of CSN stimulation were also evaluated during changes in the state of intrafusal muscle fibre contraction induced by succinylcholine and curare. In a population of 124 MSAs, 106 units (85.4 %) were affected by sympathetic stimulation. In general, while changes in resting discharge varied among different units (Ia vs. II) and experimental conditions (curarised vs. non-curarised), ranging from enhancement to strong depression of firing, the amplitude of the response to muscle stretches consistently decreased. This was confirmed and detailed in a quantitative analysis performed on 49 muscle spindle afferents. In both the non-curarised (23 units) and curarised (26 units) condition, stimulation of the CSN reduced the response amplitude in terms of DD and SD, but hardly affected DI. The effects were equally present in both Ia and II units; they were shown to be independent from gamma drive and intrafusal muscle tone and not secondary to muscle hypoxia. Sympathetic action on the resting discharge (IF) was less consistent. In the non-curarised condition, IF decreased in most Ia units, while in II units decreases and increases occurred equally often. In the curarised condition, IF in group II units mostly increased. The results have important functional implications on the control of motor function in a state of 'high' sympathetic activity, like excessive stress, as well as in certain pathological conditions such as sympathetically maintained pain.


Long-term training-induced changes in sprinting speed and sprint momentum in elite rugby union players.
Speed and sprint momentum are considered to be important physical qualities for rugby. The purpose of the study was to understand the development of these qualities in senior and junior international rugby players. In part 1 of the study, a group of senior (n = 38) and junior (n = 31) players were tested for speed over 40 m. Initial sprint velocity (ISV), maximal sprint velocity (MSV), initial sprint momentum (ISM), and maximal sprint momentum (MSM) were calculated using 10-m splits. In part 2 of the study, a group of junior (n = 12) and senior (n = 15) players were tracked over a 2-year period for body mass, ISV, MSV, ISM, and MSM. In part 1, senior backs and forwards were not found to have significantly greater ISV and MSV than junior players but were found to have greater ISM and MSM. Forwards were found to have significantly greater ISM and MSM than backs but significantly lower ISV and MSV than backs. In part 2, no significant differences were found over the 2 years between senior and junior players, but greater effect sizes for juniors were generally found when compared with seniors for improvements in ISV (d = 0.73 vs. 0.79), MSV (d = 1.09 vs. 0.68), ISM (d = 0.96 vs. 0.54), and MSM (d = 1.15 vs. 0.50). Sprint momentum is a key discriminator between senior and junior players, and large changes can be made by junior players as they transition into senior rugby. Speed appears to peak for players in their early 20s but sprint momentum appears to be more trainable.


Global public-private partnerships: Part II--What are the health issues for global governance?
This is the second of a two-part review of global public-private partnerships (GPPPs) for health development. Part I was published in the April issue of the Bulletin (Vol. 78, No. 4). The recent emergence of GPPPs is rapidly reconfiguring the international health landscape. While most multilateral and bilateral agencies are currently grappling with how to proceed, there is little information in the public domain concerning how individual partnerships work and to date very little consideration of the many implications of this trend. This paper differentiates between product-based, product development-based and issues/systems-based GPPPs and describes a number of examples of each type in the health sector. The benefits of these initiatives, not least the major resources which they harness for specific health problems, are identified. The final section of the paper explores the implications and dilemmas posed by GPPPs. It discusses whether or not shared goals can transcend conflicting values and mandates and how governance of partnership arrangements may transform and undermine certain attributes of multilateral organizations. The paper concludes that the current climate of goodwill between public and private sectors offers an opportunity that should not be missed: it can be used not only to foster new partnership but to ensure that partnership is truly in the interests of international public health.


Gel-on-a-chip: continuous, velocity-dependent DNA separation using nanoscale lateral displacement.
We studied the trajectories of polymers being advected while diffusing in a pressure driven flow along a periodic pillar nanostructure known as nanoscale deterministic lateral displacement (nanoDLD) array. We found that polymers follow different trajectories depending on their length, flow velocity and pillar array geometry, demonstrating that nanoDLD devices can be used as a continuous polymer fractionation tool. As a model system, we used double-stranded DNA (dsDNA) with various contour lengths and demonstrated that dsDNA in the range of 100-10 000 base pairs (bp) can be separated with a size-selective resolution of 200 bp. In contrast to spherical colloids, a polymer elongates by shear flow and the angle of polymer trajectories with respect to the mean flow direction decreases as the mean flow velocity increases. We developed a phenomenological model that explains the qualitative dependence of the polymer trajectories on the gap size and on the flow velocity. Using this model, we found the optimal separation conditions for dsDNA of different sizes and demonstrated the separation and extraction of dsDNA fragments with over 75% recovery and 3-fold concentration. Importantly, this velocity dependence provides a means of fine-tuning the separation efficiency and resolution, independent of the nanoDLD pillar geometry.


The Urokinase Plasminogen Activation System in Pancreatic Cancer: Prospective Diagnostic and Therapeutic Targets.
Pancreatic cancer is a highly aggressive malignancy that features high recurrence rates and the poorest prognosis of all solid cancers. The urokinase plasminogen activation system (uPAS) is strongly implicated in the pathophysiology and clinical outcomes of patients with pancreatic ductal adenocarcinoma (PDAC), which accounts for more than 90% of all pancreatic cancers. Overexpression of the urokinase-type plasminogen activator (uPA) or its cell surface receptor uPAR is a key step in the acquisition of a metastatic phenotype via multiple mechanisms, including the increased activation of cell surface localised plasminogen which generates the serine protease plasmin. This triggers multiple downstream processes that promote tumour cell migration and invasion. Increasing clinical evidence shows that the overexpression of uPA, uPAR, or of both is strongly associated with worse clinicopathological features and poor prognosis in PDAC patients. This review provides an overview of the current understanding of the uPAS in the pathogenesis and progression of pancreatic cancer, with a focus on PDAC, and summarises the substantial body of evidence that supports the role of uPAS components, including plasminogen receptors, in this disease. The review further outlines the clinical utility of uPAS components as prospective diagnostic and prognostic biomarkers for PDAC, as well as a rationale for the development of novel uPAS-targeted therapeutics.


Testing the efficacy of a molecular surveillance network: methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecium (VREF) genotypes in six hospitals in the metropolitan New York City area. The BARG Initiative Pilot Study Group. Bacterial Antibiotic Resistance Group.
Molecular fingerprinting techniques are rapidly becoming indispensable tools for hospital epidemiology. On the other hand, the relative complexity and unfamiliarity of these techniques to most hospital diagnostic laboratories limit their usefulness. In an attempt to provide a solution for this dilemma, we tested the feasibility and efficacy of a cooperative venture in which molecular typing of isolates recovered from patients in six hospitals was performed at two microbiology research laboratories with expertise in these techniques. In a small preliminary study, 30 methicillin-resistant Staphylococcus aureus (MRSA) and 30 vancomycin-resistant Enterococcus faecium (VREF) isolates were collected over a 3-week period from six hospitals in the metropolitan New York area and transported to the Laboratory of Microbiology at The Rockefeller University during the summer months of 1994. Nineteen of the 27 confirmed MRSA isolates were closely related strains carrying the same mecA and the same Tn554 polymorphs in a pulsed-field gel electrophoresis (PFGE) background represented by closely related subtypes of a single pattern, indicating the wide distribution of this MRSA clone among the participating hospitals. Typing of the same 27 MRSA isolates was also performed at the Tuberculosis Center of the Public Health Research Institute and identical results were obtained. The 29 confirmed VREF isolates were highly heterogeneous and belonged to as many as 23 distinct clonal types as defined by PFGE patterns and probing with vanA. Characterization of the 60 isolates by these methods was completed in one month of full-time effort by a single experienced laboratory assistant guided by a doctoral-level expert in molecular fingerprinting techniques. The collection of samples for both MRSA and VREF was not intended to address epidemiological questions but to determine the feasibility of a multicenter study. On the basis of our preliminary findings we are encouraged that a larger cooperative effort is possible and with the correct sampling method we believe that epidemiological and surveillance studies could be accomplished that would provide a tracking system to assist hospitals, clinics, and chronic care facilities in controlling the spread of multidrug-resistant pathogens.