The multiple-wing-hairs gene encodes a novel GBD-FH3 domain-containing protein that functions both prior to and after wing hair initiation.
The frizzled signaling/signal transduction pathway controls planar cell polarity (PCP) in both vertebrates and invertebrates. Epistasis experiments argue that in the Drosophila epidermis multiple wing hairs (mwh) acts as a downstream component of the pathway. The PCP proteins accumulate asymmetrically in pupal wing cells where they are thought to form distinct protein complexes. One is located on the distal side of wing cells and a second on the proximal side. This asymmetric protein accumulation is thought to lead to the activation of the cytoskeleton on the distal side, which in turn leads to each cell forming a single distally pointing hair. We identified mwh as CG13913, which encodes a novel G protein binding domain-formin homology 3 (GBD-FH3) domain protein. The Mwh protein accumulated on the proximal side of wing cells prior to hair formation. Unlike planar polarity proteins such as Frizzled or Inturned, Mwh also accumulated in growing hairs. This suggested that mwh had two temporally separate functions in wing development. Evidence for these two functions also came from temperature-shift experiments with a temperature-sensitive allele. Overexpression of Mwh inhibited hair initiation, thus Mwh acts as a negative regulator of the cytoskeleton. Our data argued early proximal Mwh accumulation restricts hair initiation to the distal side of wing cells and the later hair accumulation of Mwh prevents the formation of ectopic secondary hairs. This later function appears to be a feedback mechanism that limits cytoskeleton activation to ensure a single hair is formed.


[Disseminated pulmonary tuberculosis: significance of high-resolution computerized tomography].
Clinical and X-Ray studies were performed in 85 patients with disseminated pulmonary tuberculosis. All the patients underwent routine computerized tomography (CT) and high-resolution CT. According to the pathogenetic process, the authors identified hematogenic (n = 38), lymphogenic (n = 19), bronchogenic (n = 18) and mixed (n = 10) disseminations. High-resolution CT was found to have great advantages in detecting various types of tuberculous disseminations and in assessing the pattern of pulmonary abnormalities. Disseminated tuberculosis was revealed in 7 patients who had no pathological changes on routine lung X-ray films. The specific signs of hematogenic, lymphogenic disseminations and bronchgenic inoculations were identified in other forms of pulmonary tuberculosis. CT symptomatology is shown to be determined by the pathogenetic variant of its development and the stage of the process. Small focal changes in the lung were prevalent in patients with acute and subacute hematogenic forms of the disease. Infiltrates with decay cavities, thin-wall caverns, emphysema and bronchoectases were detected over the chronic course. Lymphogenic disseminations were characterized by the predominance of interstitial changes along with multiple minor foci. High-resolution CT had advantages in identifying decay cavities, signs of fibrosis and in evaluating mediastinal lymph nodes. CT data are of great significance for differential diagnosis of disseminated tuberculosis with lung metastases and diffuse interstitial diseases.


Airborne Raman lidar.
We designed and tested an airborne lidar system using Raman scattering to make simultaneous measurements of methane, water vapor, and temperature in a series of flights on a NASA-operated C-130 aircraft. We present the results for methane detection, which show that the instrument has the requisite sensitivity to atmospheric trace gases. Ultimately these measurements can be used to examine the transport of chemically processed air from within the polar vortex to mid-latitudinal regions and the exchange of stratospheric air between tropical and mid-latitudinal regions.


Focused ion beam milling and MicroED structure determination of metal-organic framework crystals.
We report new advancements in the determination and high-resolution structural analysis of beam-sensitive metal organic frameworks (MOFs) using microcrystal electron diffraction (MicroED) coupled with focused ion beam milling at cryogenic temperatures (cryo-FIB). A microcrystal of the beam-sensitive MOF, ZIF-8, was ion-beam milled in a thin lamella approximately 150 nm thick. MicroED data were collected from this thin lamella using an energy filter and a direct electron detector operating in counting mode. Using this approach, we achieved a greatly improved resolution of 0.59 Å with a minimal total exposure of only 0.64 e-/A2. These innovations not only improve model statistics but also further demonstrate that ion-beam milling is compatible with beam-sensitive materials, augmenting the capabilities of electron diffraction in MOF research.


The alpha-ketoglutarate-dehydrogenase complex: a mediator between mitochondria and oxidative stress in neurodegeneration.
Damage from oxidative stress and mitochondrial dysfunction occur together in many common neurodegenerative diseases. The enzymes that form the mitochondrial alpha-ketoglutarate- dehydrogenase complex (KGDHC), a key and arguably rate-limiting enzyme system of the tricarboxylic acid cycle, might mediate the interaction of these processes. KGDHC activity is reduced in numerous age-related neurodegenerative diseases and is diminished by oxidative stress. In Alzheimer's disease (AD), the reduction correlates highly to diminished mental performance. Thus, research has focused on the mechanisms by which select oxidants reduce KGDHC and the consequences of such a reduction. Diminished KGDHC in cells is associated with apoptosis without changes in the mitochondrial membrane potential. Studies of isolated mitochondria and of animal models suggest that a reduction in KGDHC can predispose to damage by other toxins that promote neurodegeneration. Diminished oxidative metabolism can be plausibly linked to pathological features of neurodegenerative diseases (e.g., reduced mental function, the plaques and tangles in AD). Thus, reductions in KGDHC might be central to the pathophysiology of these diseases. Studies of proteins, cells, animal models, and humans suggest that treatments to diminish, or bypass, the reduction in KGDHC might be beneficial in age-related neurodegenerative disorders.


Mineralocorticoid specificity of renal type I receptors: in vivo binding studies.
We have injected rats with [3H]aldosterone or [3H]corticosterone, plus 100-fold excess of the highly specific glucocorticoid RU 28362, with or without excess unlabeled aldosterone or corticosterone and compared type I receptor occupancy in kidney and hippocampus. Thirty minutes after subcutaneous injection [3H]aldosterone was well retained in renal papilla-inner medulla, renal cortex-outer medulla, and hippocampus; in contrast, [3H]corticosterone was well retained only in hippocampus. Competition studies for [3H]aldosterone binding sites showed corticosterone to be a poor competitor in the kidney compared with hippocampus. Time-course studies, with rats killed 10-180 min after tracer administration, showed very low uptake/retention of [3H]corticosterone by kidney; in hippocampus [3H]corticosterone retention was similar to that of [3H]aldosterone in kidney, and retention of [3H]aldosterone by hippocampus was much more prolonged than of either tracer in any other tissue. Studies in 10-day-old rats, with very low levels of corticosteroid binding globulin (CBG), showed a high degree of aldosterone selectivity in both zones of the kidney, whereas [3H]aldosterone and [3H]corticosterone were equivalently bound in hippocampus. We interpret these data as evidence for a mechanism unrelated to extravascular CBG conferring mineralocorticoid specificity on renal type I receptors and propose two models derived from our findings consistent with such differential selectivity.


Effect of dexamethasone treatment on maturational changes in the NMDA receptor in sheep brain.
The objective of the present study was to examine the effect of antenatal or postnatal treatment with corticosteroids on the NMDA receptor, one of the mediators of both normal brain development and hypoxic-ischemic injury, by determining the characteristics of the receptor MK-801 binding site in untreated and corticosteroid-treated fetal and newborn lambs. (3)H-MK-801 binding was performed in cerebral cortical cell membranes from fetal sheep at 88, 120, and 136 d gestation (term = 150 d), and from 5-d-old lambs and adult ewes. Animals were randomized to receive dexamethasone [fetuses: 6 mg, i.m. every 12 hr for four doses to mother; lambs: 0.01 mg/kg (low dose) or 0.25 mg/kg (high dose) every 12 hr for four doses] or placebo. During development, B(max) (apparent number of receptors) increased, reaching a maximum in 5-d-old lambs (p < 0.05) and decreasing in the adult brain. K(d) (dissociation constant) did not change, suggesting that receptor affinity was not altered during maturation. Dexamethasone treatment had no effect on MK-801 binding in the fetus or adult, but in lambs was associated with a significant decrease in B(max) from 2.17 +/- 0.18 pmol/mg protein in placebo-treated animals to 1.65 +/- 0.8 and 1.62 +/- 0.07 pmol/mg protein in low-dose and high-dose animals, respectively. Affinity for (3)H-MK-801 decreased 20% after dexamethasone treatment in lambs only (p < 0.05). Thus, dexamethasone treatment appears to modify the NMDA receptor only during a specific period of brain development.


Fuzzy robust credibility-constrained programming for environmental management and planning.
In this study, a fuzzy robust credibility-constrained programming (FRCCP) is developed and applied to the planning for waste management systems. It incorporates the concepts of credibility-based chance-constrained programming and robust programming within an optimization framework. The developed method can reflect uncertainties presented as possibility-density by fuzzy-membership functions. Fuzzy credibility constraints are transformed to the crisp equivalents with different credibility levels, and ordinary fuzzy inclusion constraints are determined by their robust deterministic constraints by setting a-cut levels. The FRCCP method can provide different system costs under different credibility levels (lambda). From the results of sensitivity analyses, the operation cost of the landfill is a critical parameter. For the management, any factors that would induce cost fluctuation during landfilling operation would deserve serious observation and analysis. By FRCCP, useful solutions can be obtained to provide decision-making support for long-term planning of solid waste management systems. It could be further enhanced through incorporating methods of inexact analysis into its framework. It can also be applied to other environmental management problems.


Maternal Phytosterol Supplementation during Pregnancy and Lactation Modulates Lipid and Lipoprotein Response in Offspring of apoE-Deficient Mice.
In utero exposure to excessive cholesterol has been shown to increase fetal plasma cholesterol concentration and predispose adult offspring to cardiovascular disease (CVD) risk. Because lipid-lowering drugs are contraindicated during pregnancy, natural cholesterol-lowering compounds may be a safe and effective alternative to reduce CVD risk in offspring born to hypercholesterolemic mothers. This study used the hypercholesterolemic apolipoprotein E-deficient (apoE(-/-)) mouse model to test the hypothesis that mothers supplemented with phytosterols during gestation and lactation would produce offspring with a more favorable lipid profile than offspring from unsupplemented mothers, despite having a genetic predisposition toward hypercholesterolemia. Sixteen female apoE(-/-) mice were randomly assigned to 2 diets fed throughout the gestation and lactation periods: a cholesterol-enriched diet (CH) (0.15%) or the cholesterol-enriched diet supplemented with phytosterols (CH/PS) (2%). Serum lipids and lipoproteins were measured by enzyme assay and nuclear magnetic resonance spectroscopy, respectively, and liver cholesterol was analyzed by GC. Compared with the CH-fed dams at the end of lactation, phytosterol-supplemented dams displayed lower (P < 0.05) serum total cholesterol (-55%), non-HDL cholesterol (-56%), and LDL cholesterol (-47%), but no change (P > 0.05) in HDL cholesterol and triacylglycerol (TG) concentrations. Pups from phytosterol-fed dams demonstrated lower (P < 0.05) total cholesterol (-25%), non-HDL cholesterol (-25%), LDL cholesterol (-47%), and TGs (-41%), without any change (P > 0.05) in HDL cholesterol compared with pups from CH-fed dams. Furthermore, compared with pups from CH-fed dams, pups from phytosterol-supplemented dams displayed a lower (P < 0.05) number of total LDL particles (-34%), VLDL particles (-31%), and HDL particles (-30%). Our results in apoE(-/-) mice suggest that even under strong genetic predisposition to hypercholesterolemia, pups born to mothers supplemented with phytosterols during gestation and lactation exhibit favorable liver and serum lipid responses compared with pups from unsupplemented mothers.


Multiple members of the TNF superfamily contribute to IFN-gamma-mediated inhibition of erythropoiesis.
IFN-gamma inhibits the growth and differentiation of erythroid precursor cells and mediates hemopoietic suppression through mechanisms that are not completely understood. We found that treatment of human erythroid precursor cells with IFN-gamma up-regulates the expression of multiple members of the TNF family, including TRAIL and the recently characterized protein TWEAK. TWEAK and its receptor fibroblast growth factor-inducible 14 (Fn14) were expressed by purified erythroblasts at all the stages of maturation. Exposure to recombinant TWEAK or agonist anti-Fn14 Abs was able to inhibit erythroid cell growth and differentiation through caspase activation. Because other members of the TNF family such as TRAIL and CD95 ligand (CD95L) are known to interfere with erythroblast growth and differentiation, we investigated the role of different TNF/TNFR family proteins as potential effectors of IFN-gamma in the immature hemopoietic compartment. Treatment of erythroid precursor cells with agents that blocked either TRAIL, CD95L, or TWEAK activity was partially able to revert the effect of IFN-gamma on erythroid proliferation and differentiation. However, the simultaneous inhibition of TRAIL, TWEAK, and CD95L resulted in a complete abrogation of IFN-gamma inhibitory effects, indicating the requirement of different receptor-mediated signals in IFN-gamma-mediated hemopoietic suppression. These results establish a new role for TWEAK and its receptor in normal and IFN-gamma-mediated regulation of hematopoiesis and show that the effects of IFN-gamma on immature erythroid cells depend on multiple interactions between TNF family members and their receptors.


Protein wasting due to acidosis of prolonged fasting.
During a total fast in obese subjects, the daily rate of nitrogen excretion undergoes only a small further decline after 2 wk, the excretion rate being about 5 g N/day. At this time, ammonium and urea each constitute about one-half of this excretion. The purpose of this study was to consider two alternative hypotheses: first, that the near plateau in nitrogen excretion represents an irreducible minimum rate of net protein breakdown in order to supply essential organs with calories in the form of glucose; second, that protein breakdown could be further reduced by minimizing the requirement to provide nitrogen for ammonium excretion during the ketoacidosis of fasting. Because ammonium excretion is largely controlled by acid-base balance, 150 mmol of sodium bicarbonate plus 60 mmol of potassium chloride were administered daily to decrease ammonium excretion in eight obese subjects who were totally fasting for more than 14 days. Urine ammonium nitrogen fell with this treatment (from 3.8 +/- 0.4 to 2.0 +/- 0.4 g N/g creatinine). In addition, there was a smaller fall in the rate of urea excretion (from 2.5 +/- 0.2 to 2.1 +/- 0.3 g N/g creatinine) together with a fall in the blood urea nitrogen. Therefore, it appears that ammonium excretion contributes to the negative nitrogen balance of a prolonged total fast, as assessed over a 3-day period of observation, is responsible for about one-third of the net lean body mass loss.


A multimodal brain-based feedback and communication system.
The Thought Translation Device (TTD) is a brain-computer interface based on the self-regulation of slow cortical potentials (SCPs) and enables completely paralyzed patients to communicate using their brain potentials. Here, an extended version of the TTD is presented that has an auditory and a combined visual and auditory feedback modality added to the standard visual feedback. This feature is necessary for locked-in patients who are no longer able to focus their gaze. In order to test performance of physiological regulation with auditory feedback 54 healthy participants were randomly assigned to visual, auditory or combined visual-auditory feedback of slow cortical potentials. The training consisted of three sessions with 500 trials per session with random assignment of required cortical positivity or negativity in half of the trials. The data show that physiological regulation of SCPs can be learned with auditory and combined auditory and visual feedback although the performance of auditory feedback alone was significantly worse than with visual feedback alone.


Haplotype data for 23 Y-chromosome markers in a reference sample from Bosnia and Herzegovina.
To detect polymorphisms of 23 Y-chromosomal short tandem repeat (STR) loci, including 6 new loci, in a reference database of male population of Bosnia and Herzegovina, as well as to assess the importance of increasing the number of Y-STR loci utilized in forensic DNA analysis. The reference sample consisted of 100 healthy, unrelated men originating from Bosnia and Herzegovina. Sample collection using buccal swabs was performed in all geographical regions of Bosnia and Herzegovina in the period from 2010 to 2011. DNA samples were typed for 23 Y STR loci, including 6 new loci: DYS576, DYS481, DYS549, DYS533, DYS570, and DYS643, which are included in the new PowerPlex® Y 23 amplification kit. The absolute frequency of generated haplotypes was calculated and results showed that 98 samples had unique Y 23 haplotypes, and that only two samples shared the same haplotype. The most polymorphic locus was DYS418, with 14 detected alleles and the least polymorphic loci were DYS389I, DYS391, DYS437, and DYS393. This study showed that by increasing the number of highly polymorphic Y STR markers, to include those tested in our analysis, leads to a reduction of repeating haplotypes, which is very important in the application of forensic DNA analysis.


High-resolution genotyping of Listeria monocytogenes by fluorescent amplified fragment length polymorphism analysis compared to pulsed-field gel electrophoresis, random amplified polymorphic DNA analysis, ribotyping, and PCR-restriction fragment length polymorphism analysis.
The purpose of this study was to evaluate fluorescent amplified fragment length polymorphism (AFLP) analysis for the inter- and intraspecies differentiation of a collection of 96 strains of Listeria monocytogenes and 10 non-L. monocytogenes strains representing six other Listeria species of different origin. The AFLP technique was compared with three other molecular typing methods--ribotyping, random amplified polymorphic DNA analysis (RAPD), and pulsed-field gel electrophoresis (PFGE)--in terms of discriminatory ability. PCR-restriction fragment length polymorphism was included for virulence gene allele characterization. The 96 L. monocytogenes strains were divided into two major clusters by AFLP fingerprinting at a similarity level of 82% in concordance with the results of PFGE, RAPD, and ribotyping. One main cluster consisted of all of the 24 L. monocytogenes hly allele 1 strains, while another main cluster consisted of all of the 72 L. monocytogenes hly allele 2 strains. This indicates the existence of two distinct phylogenetic divisions. Isolates of the remaining Listeria species were not included in the clusters. AFLP, PFGE, and RAPD typing were highly discriminatory methods, with discrimination (D) indices of 0.974, 0.969, and 0.954, respectively, whereas ribotyping had a lower D index of 0.874. AFLP, PFGE, and RAPD typing showed some level of agreement in terms of strain grouping and differentiation. However, all three methods subdivided types of strains grouped by the other methods. Isolates with identical DNA profiles were distributed across the spectrum of origin. It was not possible to associate certain types with specific food sectors or clinical cases, which is indicative of the spread of L. monocytogenes clones across species. Overall, AFLP fingerprinting was suitable for the high-resolution genotyping of L. monocytogenes and had an equally high or higher differentiation power compared to PFGE or RAPD typing.


Comparison of pro re nata versus Bimonthly Injection of Intravitreal Aflibercept for Typical Neovascular Age-Related Macular Degeneration.
The aim of this study was to clarify the 1-year outcomes of pro re nata (PRN) and bimonthly intravitreal injections of aflibercept (IVA) for typical neovascular age-related macular degeneration (tAMD) after the initial 3 monthly IVA. We conducted a prospective, interventional study. Fifty-eight treatment-naïve patients with tAMD were randomly assigned to the PRN (30 patients) or the bimonthly (28 patients) treatment group. Both groups initially received 3 monthly IVA. Visual acuity, central macular retinal thickness (CRT), and central choroidal thickness (CCT) were evaluated at 12 months. Subanalysis was performed to identify factors associated with the best-corrected visual acuity (BCVA). BCVA was significantly improved only in the bimonthly group at 12 months. CRT and CCT were significantly decreased in both groups. Subanalysis showed that the only factor associated with BCVA improvement at 12 months was the existence of pigment epithelial detachment at baseline. BCVA showed significant improvement only in the bimonthly group but not in the PRN group at 12 months.


Physical mapping and complementation analysis of transposon Tn5 mutations in Caulobacter crescentus: organization of transcriptional units in the hook gene cluster.
Using the cloned DNA from the hook protein gene region of Caulobacter crescentus ( Ohta et al., Proc. Natl. Acad. Sci. U.S.A. 79:4863-4867, 1982), we have identified and physically mapped 19 Tn5-induced and 2 spontaneous insertion mutations to this region of the chromosome. These nonmotile mutants define a major cluster of fla genes that covers approximately 17 kilobases on the chromosome (hook gene cluster). Complementation analysis of the mutants using DNA fragments from the region subcloned in the broad host range plasmid pRK290 has shown that these fla genes are organized into at least five transcriptional units (I to V). Transcriptional unit II contains at least one gene in addition to the hook protein gene, which makes this the first operon described in C. crescentus. Expression of the hook protein gene and the genetically unlinked flagellin A and B genes by this set of mutants also furnishes additional insights into the hierarchial regulation of flagellar genes. We have found that the spontaneous insertion mutant ( SC511 ) of the hook protein gene ( flaK ) makes no flagellin A or B and that genes downstream from the hook protein gene are required in trans for expression of the hook protein operon and the flagellin A and B genes. Recombination and complementation results thus place flaK , flaJ , flaN , and flaO (R. C. Johnson and B. Ely , J. Bacteriol . 137:627-634, 1979) in the hook gene cluster, identify at least three new genes ( flbD , flbG , and flbF ), and suggest that this cluster may contain several additional, as yet unidentified, fla genes.


Exploring design requirements for repurposing dental virtual patients from the web to second life: a focus group study.
Since their inception, virtual patients have provided health care educators with a way to engage learners in an experience simulating the clinician's environment without danger to learners and patients. This has led this learning modality to be accepted as an essential component of medical education. With the advent of the visually and audio-rich 3-dimensional multi-user virtual environment (MUVE), a new deployment platform has emerged for educational content. Immersive, highly interactive, multimedia-rich, MUVEs that seamlessly foster collaboration provide a new hotbed for the deployment of medical education content. This work aims to assess the suitability of the Second Life MUVE as a virtual patient deployment platform for undergraduate dental education, and to explore the requirements and specifications needed to meaningfully repurpose Web-based virtual patients in MUVEs. Through the scripting capabilities and available art assets in Second Life, we repurposed an existing Web-based periodontology virtual patient into Second Life. Through a series of point-and-click interactions and multiple-choice queries, the user experienced a specific periodontology case and was asked to provide the optimal responses for each of the challenges of the case. A focus group of 9 undergraduate dentistry students experienced both the Web-based and the Second Life version of this virtual patient. The group convened 3 times and discussed relevant issues such as the group's computer literacy, the assessment of Second Life as a virtual patient deployment platform, and compared the Web-based and MUVE-deployed virtual patients. A comparison between the Web-based and the Second Life virtual patient revealed the inherent advantages of the more experiential and immersive Second Life virtual environment. However, several challenges for the successful repurposing of virtual patients from the Web to the MUVE were identified. The identified challenges for repurposing of Web virtual patients to the MUVE platform from the focus group study were (1) increased case complexity to facilitate the user's gaming preconception in a MUVE, (2) necessity to decrease textual narration and provide the pertinent information in a more immersive sensory way, and (3) requirement to allow the user to actuate the solutions of problems instead of describing them through narration. For a successful systematic repurposing effort of virtual patients to MUVEs such as Second Life, the best practices of experiential and immersive game design should be organically incorporated in the repurposing workflow (automated or not). These findings are pivotal in an era in which open educational content is transferred to and shared among users, learners, and educators of various open repositories/environments.


Development of a Decision Support System for the Management of Mummy Berry Disease in Northwestern Washington.
Mummy berry, caused by Monilinia vaccinii-corymbosi, is the most important disease of the northern highbush blueberry (Vaccinium corymbosum L.) in North America and can cause up to 70% yield losses in affected fields. A key event in the mummy berry disease cycle is the primary infection phase where ascospores are released by apothecia that infect emerging floral and vegetative tissues. Current management of mummy berry disease in northwestern Washington is predominantly reliant on the prevention of primary infections through prophylactic, calendar-based fungicide spray applications early in the growing season. To improve the understanding of risk during these periods and to help tailor management strategies, we developed a decision support system (DSS) based on field records spanning over five seasons and four locations in northwestern Washington. Environmental conditions across the region were highly uniform but different dynamics of apothecial development were observed under high- and low-management regimes. Based on our analysis, we suggest basing the initial iteration of the DSS on two sub-models. The first sub-model predicts the onset of apothecia based on chill-unit accumulation under high- and low-management regimes, and the second predicts primary infection risk, which provides opportunities to improve the timing of fungicide applications. The synoptic DSS proposed here is based on the current biological knowledge of the pathosystem and available data for the northwestern Washington region. We provide the analysis and the DSS implementation and evaluation as an open-source repository, providing opportunities for further improvements. Finally, we provide suggestions for future research and the operational efforts needed for improving the utility and accuracy of the mummy berry DSS.


[Potentiation of the antiviral activity of human interferons by virazole in in vitro experiments].
Combination of interferon and virazole results in potentiation of their antiviral activity. This was demonstrated with all three types of natural interferons and the recombinant RL-2-interferon. This effect is retained at a temperature of 37 degrees C and increases when the chemical drug is combined with a mixture of alpha- and gamma-interferon.


Patient's Perspectives of Experimental HCV-Positive to HCV-Negative Renal Transplantation: Report from a Single Site.
Background: With growing transplant wait times, clinical trials are evaluating the safety and efficacy of transplanting HCV-infected donor (HCV+) organs into HCV-noninfected recipients (HCV D+/R-). Such transplants raise ethical questions about safety, consent, and access to organs. Methods: We interviewed eight of the ten total HCV D+/R- transplant recipients enrolled in a pilot clinical trial examining the safety and feasibility of these novel transplants regarding their experiences in the trial, including their decision-making and perceptions of the informed consent process. Results: All interviewees reported positive experiences and expressed confidence regarding their decision to participate. Participants accepted an HCV + organ based on their assessments of the risks and potential benefits of HCV D+/R- transplants. For many, the risks of HCV were minimal compared to the risks of not receiving a transplant. All participants recalled providing informed consent, reporting that the process was thorough and that all their questions were addressed. Participants did not regret receiving an HCV D+/R- transplant and did not report experiencing stigma. However, given their understanding of HCV cure rates in the general population and the survival benefit associated with kidney transplantation, participants may have conflated research regarding HCV D+/R- transplantation with clinical care, suggesting a potential therapeutic misconception. Conclusions: Recipients of experimental HCV D+/R- transplants generally seemed to recognize the risks and benefits of these novel transplants and did not regret participating. Such salutary reported experiences are important in assessing the appropriateness of further research into the feasibility of HCV D+/R- transplants.


Promoter hypermethylation of BRCA1 correlates with absence of expression in hereditary breast cancer tumors.
Germline mutations in BRCA1 account for a low proportion of hereditary cases in diverse populations. Several efforts have been made to find new genes involved in the inheritance of breast cancer with no success until today. The participation of BRCA1 in the development of breast cancer has been proposed in several studies where hypermethylation of its promoter and a decrease in expression has been reported for sporadic cases and one study on familial cases. To explore the participation of BRCA1 in hereditary carcinogenesis through a different mechanism than the inheritance of germline mutations, we studied the methylation status of its promoter in breast tumors, from patients previously screened for BRCA1/BRCA2 germline mutations. We also determined the presence of the BRCA1 protein in these tumors and correlated both events with tumor grade, hormone receptors and ERBB2 presence. Promoter hypermethylation of the BRCA1 gene was detected in 51% of our biopsies, among which 67% did not express the respective protein. This result leads us to suggest that hypermethylation could be considered as an inactivating mechanism for BRCA1 expression, either as a first or second hit. Moreover, a number of biopsies with absence of expression on BRCA1 showed negative detection of estrogen and progesterone receptors, a similar phenotype to BRCA1 mutated breast tumors.


p53 mutations define the chromatin landscape to confer drug tolerance in pancreatic cancer.
Somatic inactivation of p53 (TP53) mainly occurs as missense mutations that lead to the acquisition of neomorphic mutant protein forms. p53 mutants have been postulated to exert gain-of-function (GOF) effects, including promotion of metastasis and drug tolerance, which generally contribute to the acquisition of the lethal phenotype. Here, by integrating a p53R270H -dependent transcriptomic analysis with chromatin accessibility (ATAC-seq) profiling, we shed light on the molecular basis of a p53 mutant-dependent drug-tolerant phenotype in pancreatic cancer. p53R270H finely tunes chromatin accessibility in specific genomic loci, orchestrating a transcriptional programme that participates in phenotypic evolution of the cancer. We specifically focused on the p53R270H -dependent regulation of the tyrosine kinase receptor macrophage-stimulating protein receptor (MST1r). MST1r deregulation substantially impinged on drug response in the experimental model, recapitulating the p53R270H -dependent phenotype, and strongly correlated with p53 mutant and aggressive phenotype in pancreatic cancer patients. As cellular plasticity in the final stages of the evolution of pancreatic cancer seems to predominantly originate from epigenetic mechanisms, we propose that mutant p53 participates in the acquisition of a lethal phenotype by fine-tuning the chromatin landscape.


Natural variation in female reproductive hormones does not affect contrast sensitivity.
Evidence suggests that females experience adaptive shifts in facial preferences across the menstrual cycle. However, recent discussions and meta-analyses suggest that these findings are equivocal. A previously unexplored question is the extent to which shifts in female preferences are modulated by hormone-dependent changes occurring in low-level vision, such as visual sensitivity. This mechanistic approach has been a novel method for investigating the extent to which complex perceptual phenomena are driven by low-level versus higher-level perceptual processes. We investigated whether the contrast sensitivity function-an early dimension of vision-is also influenced by variation in female reproductive hormones. Visual contrast thresholds were measured for 1, 4 and 16 cycles/degree gratings during the ovulatory, luteal and menstrual phases of the menstrual cycle in naturally cycling women, and women using oral contraceptives. Male participants were tested at similar time intervals. Results showed that visual contrast sensitivity does not differ according to sex, or use of oral contraception, nor does it vary relative to hormonal shifts across the menstrual cycle. These findings suggest that shifts in female preferences are not driven by changes in visual sensitivity, and are therefore likely attributable to changes in higher-level perception or cognition.


Prevalence, determinants, and effects of violence during pregnancy: A maternity-based cross-sectional study in Luanda, Angola.
Information on the extent of violence against women is scarce in Angola. This study aims to estimate the prevalence of violence against pregnant women in Angola and to identify its sociodemographic determinants and effects on pregnancy outcomes. A cross-sectional study was conducted between December 2012 and February 2013, involving 995 women who delivered at a Maternity in Luanda, Angola. Information was collected through questionnaires administered by interviewers. The prevalence of violence during pregnancy was 13.0%. Exclusively physical, psychological or sexual violence was reported by 4.3%, 7.7% and 0.2% of the women, respectively. After adjustment, the occurrence of physical violence decreased with increasing age and education, and was more common among women who consumed alcohol during pregnancy, while psychological violence was significantly more frequent among women aged 20 to 24 years and those who had their first sexual intercourse before the age of 15, and less frequent among those who were married or in cohabitation. This first study describing violence against pregnant Angolan women showed that violence is a frequent event, supporting that violence assessment should be considered in antenatal care.


[Revascularization of the limbs in patients of elderly and senile age with chronic arterial ischemia].
The results of examination and treatment of about 3000 patients having chronic arterial ischemia of extremities allowed to define three main types of arterial diseases appearing in arterial ischemia of extremities of different degrees of manifestation: spastic arterial diseases, obliterating diseases and mixed types, combining spasms and arterial obliteration. In case of spastic arterial diseases ischemia is to be treated conservatively. Biosurgical revascularization of extremities having practically no contraindications is recommended in case of obliterating arterial diseases with critical arterial ischemia of extremities, when surgical generally accepted methods have a short-term treatment effect or their application is impossible, in case of peripheral form of arterial affection and also for acute associated diseases of middle-aged and old patients. Mixed arterial ischemia is revealed as a rule for middle-aged and old patients. Biosurgical revascularization and conservative treatment methods were applied for their treatment. The paper is devoted to the treatment of middle-aged and old patients having obliterating arterial diseases of extremities with a clinical presentation of chronic arterial ischemia.


Interaction of dietary fatty acids with tumour necrosis factor family cytokines during colon inflammation and cancer.
Intestinal homeostasis is precisely regulated by a number of endogenous regulatory molecules but significantly influenced by dietary compounds. Malfunction of this system may result in chronic inflammation and cancer. Dietary essential n-3 polyunsaturated fatty acids (PUFAs) and short-chain fatty acid butyrate produced from fibre display anti-inflammatory and anticancer activities. Both compounds were shown to modulate the production and activities of TNF family cytokines. Cytokines from the TNF family (TNF- α, TRAIL, and FasL) have potent inflammatory activities and can also regulate apoptosis, which plays an important role in cancer development. The results of our own research showed enhancement of apoptosis in colon cancer cells by a combination of either docosahexaenoic acid (DHA) or butyrate with TNF family cytokines, especially by promotion of the mitochondrial apoptotic pathway and modulation of NF κ B activity. This review is focused mainly on the interaction of dietary PUFAs and butyrate with these cytokines during colon inflammation and cancer development. We summarised recent knowledge about the cellular and molecular mechanisms involved in such effects and outcomes for intestinal cell behaviour and pathologies. Finally, the possible application for the prevention and therapy of colon inflammation and cancer is also outlined.


Interaction between residues in the Mg2+-binding site regulates BK channel activation.
As a unique member of the voltage-gated potassium channel family, a large conductance, voltage- and Ca(2+)-activated K(+) (BK) channel has a large cytosolic domain that serves as the Ca(2+) sensor, in addition to a membrane-spanning domain that contains the voltage-sensing (VSD) and pore-gate domains. The conformational changes of the cytosolic domain induced by Ca(2+) binding and the conformational changes of the VSD induced by membrane voltage changes trigger the opening of the pore-gate domain. Although some structural information of these individual functional domains is available, how the interactions among these domains, especially the noncovalent interactions, control the dynamic gating process of BK channels is still not clear. Previous studies discovered that intracellular Mg(2+) binds to an interdomain binding site consisting of D99 and N172 from the membrane-spanning domain and E374 and E399 from the cytosolic domain. The bound Mg(2+) at this narrow interdomain interface activates the BK channel through an electrostatic interaction with a positively charged residue in the VSD. In this study, we investigated the potential interdomain interactions between the Mg(2+)-coordination residues and their effects on channel gating. By introducing different charges to these residues, we discovered a native interdomain interaction between D99 and E374 that can affect BK channel activation. To understand the underlying mechanism of the interdomain interactions between the Mg(2+)-coordination residues, we introduced artificial electrostatic interactions between residues 172 and 399 from two different domains. We found that the interdomain interactions between these two positions not only alter the local conformations near the Mg(2+)-binding site but also change distant conformations including the pore-gate domain, thereby affecting the voltage- and Ca(2+)-dependent activation of the BK channel. These results illustrate the importance of interdomain interactions to the allosteric gating mechanisms of BK channels.


Selective androgen receptor modulators: the future of androgen therapy?
Selective androgen receptor modulators (SARMs) are small molecule drugs that function as either androgen receptor (AR) agonists or antagonists. Variability in AR regulatory proteins in target tissues permits SARMs to selectively elicit anabolic benefits while eschewing the pitfalls of traditional androgen therapy. SARMs have few side effects and excellent oral and transdermal bioavailability and may, therefore, represent viable alternatives to current androgen therapies. SARMs have been studied as possible therapies for many conditions, including osteoporosis, Alzheimer's disease, breast cancer, stress urinary incontinence (SUI), prostate cancer (PCa), benign prostatic hyperplasia (BPH), male contraception, hypogonadism, Duchenne muscular dystrophy (DMD), and sarcopenia/muscle wasting/cancer cachexia. While there are no indications for SARMs currently approved by the Food and Drug Administration (FDA), many potential applications are still being explored, and results are promising. In this review, we examine the literature assessing the use of SARMS for a number of indications.


Subversion of NK-cell and TNFα Immune Surveillance Drives Tumor Recurrence.
Understanding how incompletely cleared primary tumors transition from minimal residual disease (MRD) into treatment-resistant, immune-invisible recurrences has major clinical significance. We show here that this transition is mediated through the subversion of two key elements of innate immunosurveillance. In the first, the role of TNFα changes from an antitumor effector against primary tumors into a growth promoter for MRD. Second, whereas primary tumors induced a natural killer (NK)-mediated cytokine response characterized by low IL6 and elevated IFNγ, PD-L1hi MRD cells promoted the secretion of IL6 but minimal IFNγ, inhibiting both NK-cell and T-cell surveillance. Tumor recurrence was promoted by trauma- or infection-like stimuli inducing VEGF and TNFα, which stimulated the growth of MRD tumors. Finally, therapies that blocked PD-1, TNFα, or NK cells delayed or prevented recurrence. These data show how innate immunosurveillance mechanisms, which control infection and growth of primary tumors, are exploited by recurrent, competent tumors and identify therapeutic targets in patients with MRD known to be at high risk of relapse. Cancer Immunol Res; 5(11); 1029-45. ©2017 AACR.


[Analysis of the sequences of internal transcribed spacers ITS1, ITS2 and the 5.8S ribosomal gene of species of the Amaranthus genus].
Analysis of the sequence ITS1-5.8S-ITS2 in 11 samples of the amaranth species (Amaranthus caudatus, A. cruentus, A. hybridus, A. tricolor, A. paniculatus, A. hypohondriacus) was performed. It has been shown that the variability of the sequences of the intergenic spacers ITS1, ITS2 and 5.8S rRNA gene of the amaranth species analyzed is extremely low. A possible secondary structure of the 5.8S rRNA molecule was determined for the first time; three conservative motifs were identified. A single nucleotide substitution found in A. hybridus did not change the loop topology. In the sample of Celosia cristata taken as an external group, a four-nucleotide insertion in the 5'-end of the gene and a one-nucleotide deletion in the fourth hairpin not affecting the general topology of the 5.8S rRNA molecule were found.


[Usefulness of electromyographic techniques of the bulbocavernosus muscle in the diagnosis of neurogenic impotence].
To analyze the utility of electromyographic study of the bulbocavernosus muscle. 126 impotent patients were evaluated by physical examination and neuroandrologic profile. The physical examination consisted in an exploration of the anal tone and the bulbocavernosus reflex. The neuroandrologic profile consisted in selective electromyography of the bulbocavernosus muscle, the determination of the S2-S4 evoked potentials, somatosensory potential of the pudenal nerve, electromyography of cavernous smooth muscle (SPACE) sympathetic skin response and cystometry. The results of the bulbocavernosus electromyography and S2-S4 evoked potentials were different from the data obtained from the physical examination of the anal tone and the bulbocavernosus reflex. Selective bulbocavernosus electromyography showed a sensitivity of 57% and a specificity of 84%. The S2-S4 evoked potentials showed a sensitivity of 61% and a specificity of 100%. The diagnostic value of the S2-S4 evoked potentials increased at 39 msec cutoff (sensitivity = 66%; specificity = 95%). Exploration of the anal tone is very specific for neurologic lesion in impotence (93%), but has a low sensitivity (30%). The clinical bulbocavernosus reflex showed a good specificity (85%) and sensitivity (75%) in the diagnosis of pudendal efferent lesion, but is less useful in the diagnosis of neurogenic impotence. The highest diagnostic value was obtained with the S2-S4 evoked potentials. Selective electromyography of the bulbocavernosus muscle showed a moderate diagnostic value. Physical examination of peripheral pudendal innervation does not appear to be useful for screening for neurologic lesion in impotence because of its low sensitivity, which may be due to existing incomplete nervous lesions.


[Dietary intake and energy balance in the adult population in Niterói, Rio de Janeiro State, Brazil: the Nutrition, Physical Activity, and Health Survey (PNAFS)].
This study aimed to assess typical daily dietary intake and energy expenditure in a probabilistic sample of adults in the city of Niterói, Rio de Janeiro State, Brazil. Dietary intake was measured by 24-hour dietary recall, body mass and height were measured in the household, and energy expenditure was estimated by the Flex-heart rate method. The results showed mean energy intake of 1,570.9 (24.1) and 2,188.8 (46.1) kcal.day-1 for women and men, respectively. Energy expenditure was higher in obese women and men (1,511.5+/-19.5 and 2,222.3+/-68.8 kcal.day-1, respectively) due to their higher body mass. Measured energy balance (intake - expenditure) decreased with increasing body mass index (BMI) in both women and men and was negative (-3.2; 131 kcal.day-1) in obese men. In conclusion, energy intake among adults in Niterói falls within the recommended values, despite high prevalence of overweight and obesity in women and men. Energy intake decreased with increasing body mass, indicating possible overestimation of energy expenditure. It is important to develop better estimates of energy expenditure in this population.


Poor Subjective Sleep Quality Predicts Symptoms in Irritable Bowel Syndrome Using the Experience Sampling Method.
Sleep quality may affect symptom experience in irritable bowel syndrome (IBS). Our aim was to investigate the relationship between sleep quality and gastrointestinal (GI) symptoms using actigraphy and the experience sampling method. Patients with IBS were recruited from a tertiary Neurogastroenterology clinic and the community. GI symptoms and mood were recorded on a smartphone application, 10 times per day, over 7 consecutive days. Subjective sleep quality was recorded every morning to reflect the night before. Objective measures of sleep quality were estimated from wrist-worn actigraphy. Cross-lagged structural equation models were built to assess the directionality of sleep-symptom relationships over time. Eighty patients with IBS completed the study (mean age: 37 years [range 20-68], 89% female, 78% community). Approximately 66% had a Pittsburgh Sleep Quality Index score ≥ 8, indicating a clinically significant sleep disturbance. Approximately 82% (95% CI: 72-90) screened positive for a sleep disorder, most commonly insomnia. In cross-lagged analysis, poor subjective sleep quality predicted next-day abdominal pain (0.036 < P < 0.040) and lower GI symptoms (0.030 < P < 0.032), but not vice versa. No significant relationship with GI symptoms was found for any objective sleep measure using actigraphy. Poor subjective sleep quality was associated with higher next-day lower GI symptom levels, but not vice versa. Objective sleep measures did not predict next-day abdominal symptoms, potentially supporting the conclusion that it is the perception of sleep quality that is most influential. This study may be used to guide future research into the effect of sleep interventions on GI symptoms.


Clinicians on the board: what difference does it make?
Around the world clinical professionals have increased their involvement in the management of health services. However the evidence to suggest that these changes will lead to improved performance remains fragmented. In this paper we address this matter focussing on the impact of clinicians appointed to the boards of directors of English NHS hospital trusts. Although the number of clinicians involved in the strategic governance of hospital trusts is relatively low by international standards, they do appear to have an impact on overall performance. Drawing on published information from hospital trust annual reports, publicly available performance measures from the Healthcare Commission and data gathered by Dr Foster over a three year period (2006-9), the paper reports two main findings. First, the analysis reveals a significant and positive association between a higher percentage of clinicians on boards and the quality ratings of service providers, especially where doctors are concerned. This positive influence is also confirmed in relation to lower morbidity rates and tests to exclude the possibility of reverse causality (doctors joining boards of already successful organisations). Second, we do not find the same level of support for clinical professions such as nurses and other allied health professions turned directors.


Phase I Clinical Study of Irinotecan Plus S-1 in Patients With Advanced or Recurrent Cervical Cancer Previously Treated With Platinum-Based Chemotherapy.
This study aimed to determine the maximum tolerated dose and acute dose-limiting toxicities (DLTs) of intravenous irinotecan plus oral S-1 in patients with advanced or recurrent uterine cervical cancer. Irinotecan was administered intravenously over the course of 90 minutes on day 1, and S-1 was given orally in 2 divided doses from days 1 to 14 of a 21-day cycle. The dose of S-1 was escalated in a stepwise fashion from 40 (level 1) to 60 mg/m (level 2) and then 80 mg/m (level 3), whereas the dosage of irinotecan remained the same (150 mg/m). The primary end point for the escalation study was acute DLT that occurred within 2 cycles of chemotherapy. Twelve patients were enrolled and treated over 3 dose levels. Their median age was 47 years (range, 28-48 years). At level 1, one episode of grade 3 anemia and a grade 3 fatigue were observed, but no DLT developed. At level 2, the first patient experienced febrile neutropenia, which was considered to be a DLT. To evaluate the toxicity of this dose level, 5 more patients were evaluated. However, no DLT developed in these patients. At level 3, although grade 1 to 2 hematological and nonhematological toxicities developed, no DLT occurred. In women with advanced or recurrent cervical cancer previously treated with platinum-based chemotherapy, S-1 plus irinotecan in a triweekly setting is a reasonable treatment regimen with an acceptable toxicity profile. The recommended doses of S-1 and irinotecan for this regimen are 80 and 150 mg/m, respectively.


Molecular analysis of the maize wx-B3 allele indicates that precise excision of the transposable Ac element is rare.
The somatic and germinal behavior of the maize wx-B3 mutation indicates that this Ac allele rarely reverts. Endosperms containing wx-B3 display tiny and infrequent Wx revertant sectors while no significant reversion is detected when wx-B3 pollen is stained with I/KI. Previous studies of other transposable element alleles that revert infrequently have implicated low levels of element excision. Unlike these other alleles, the wx-B3 Ac element is indistinguishable from fully active Ac elements with respect to its structure, and its ability to transpose from the Wx gene or to trans-activate a Ds element. Characterization of somatic and germinal excision events lead us to conclude that excision of the wx-B3 Ac element almost always produces null alleles. Furthermore, the excellent correlation between the position of the wx-B3 mutation on the physical and genetic maps indicates that the Ac insertion is the only lesion of wx-B3. As a result, precise excision of this Ac should restore Wx function. The fact that revertant sectors and pollen grains are rare indicates that precise excision of Ac is also rare. The finding that the wx-B3 reversion frequency is comparable whether wx-B3 is hemizygous or over a wx allele with a wild-type insertion site illustrates a fundamental difference between the excision mechanisms of Ac and Drosophila P elements.


Dental and dental hygiene students' attitudes in a joint local anesthesia course.
In 1995, the local anesthesia course was combined for dental and dental hygiene students. It was also the first time that dental hygiene students were instructed clinically in local anesthetic techniques. Faculty felt it important to evaluate student attitudes regarding these curriculum changes. This study assessed students' perceptions of the adequacy of background preparation courses, concerns about administering local anesthetics and attitudes about its administration by dental hygienists. A pre/post survey was completed by dental and dental hygiene students. T-tests were used to determine if differences in attitudes existed between groups and if changes in attitudes occurred within groups after taking the course. Results indicated that both groups believed they had adequate background preparation. Apprehensions about administering local anesthetics were similar, but the dental hygiene students felt significantly less anxiety (p < 0.01). Results also differed significantly on the issue of hygienists administering local anesthetics, with hygiene students in support of it and dental students opposed (p < 0.01). Results confirmed that no changes were needed in the existing prerequisite courses for both groups. A need to address student anxiety about local anesthetic administration has been identified. In addition, it may be beneficial to expand dental students' appreciation of dental hygiene education to foster mutually successful working relationships in the future.


Moral distress and burnout in internal medicine residents.
Residents frequently encounter situations in their workplace that may induce moral distress or burnout. The objective of this study was to measure overall and rotation-specific moral distress and burnout in medical residents, and the relationship between demographics and moral distress and burnout. The revised Moral Distress Scale and the Maslach Burnout Inventory (Human Service version) were administered to Internal Medicine residents in the 2013-2014 academic year at the University of British Columbia. Of the 88 residents, 45 completed the surveys. Participants (mean age 30+/-3; 46% male) reported a median moral distress score (interquartile range) of 77 (50-96). Twenty-six percent of residents had considered quitting because of moral distress, 21% had a high level of burnout, and only 5% had a low level of burnout. Moral distress scores were highest during Intensive Care Unit (ICU) and Clinical Teaching Unit (CTU) rotations, and lowest during elective rotations (p<0.0001). Women reported higher emotional exhaustion. Moral distress was associated with depersonalization (p=0.01), and both moral distress and burnout were associated with intention to leave the job. Internal Medicine residents report moral distress that is greatest during ICU and CTU rotations, and is associated with burnout and intention to leave the job.


Overexpression of hydroxymethylglutaryl CoA synthase 2 and 2,4-dienoyl-CoA reductase in rat pancreas following chronic alcohol consumption.
The mechanism of alcohol-induced pancreatic damage is unclear. The aim of this study was to clarify the effects of chronic alcohol intake on the pancreatic proteome. Rats were fed an alcohol-containing Lieber-DeCarli liquid diet, and the pancreatic proteome was compared with that of pair-fed control rats using agarose 2-dimensional gel electrophoresis followed by liquid chromatography-tandem mass spectrometry. The expression of 3 proteins was consistently altered in alcohol-fed rats: 1 protein was down-regulated, and 2 proteins were up-regulated. The 2 up-regulated proteins were identified as 2,4-dienoyl-CoA reductase and hydroxymethylglutaryl-CoA synthase (HMGCS2). The combined concentration of malondialdehyde and 4-hydroxyalkenals was significantly greater in alcohol-fed rats. It is noteworthy that the reactivity of anti-4-hydroxy-2-nonenal antibody was significantly higher toward HMGCS2 isolated from alcohol-fed rats. The activity of HMGCS2 was higher in alcohol-fed rats, but the relative increase in enzyme activity in alcohol-fed rats was less than the relative increase in HMGCS2 expression. Chronic alcohol consumption results in distinct alterations in the expression of 3 pancreatic proteins. The reactivity of 4-hydroxy-2-nonenal toward one of the up-regulated proteins, HMGCS2, increased markedly following chronic alcohol intake, suggesting that up-regulation of HMGCS2 is connected with alterations of lipid peroxidation induced by alcohol.


New insights into HIV epidemic in South Africa: key findings from the National HIV Prevalence, Incidence and Behaviour Survey, 2012.
This article presents key findings from the 2012 HIV prevalence, incidence and behaviour survey conducted in South Africa and explores trends in the HIV epidemic. A representative household based survey collected behavioural and biomedical data among people of all ages. Chi-squared test for association and formal trend tests (2002, 2005, 2008 and 2012) were used to test for associations and trends in the HIV epidemic across the four surveys. In 2012 a total of 38 431 respondents were interviewed from 11 079 households; 28 997 (67.5%) of 42 950 eligible individuals provided blood specimens. HIV prevalence was 12.2% [95% CI: 11.4-13.1] in 2012 with prevalence higher among females 14.4% than males 9.9%. Adults aged 25-49 years were most affected, 25.2% [95% CI: 23.2-27.3]. HIV prevalence increased from 10.6% [95%CI: 9.8-11.6] in 2008 to 12.2% [95% CI: 11.4-13.1] in 2012 (p < 0.001). Antiretroviral treatment (ART) exposure doubled from 16.6% in 2008 to 31.2% in 2012 (p < 0.001). HIV incidence in 2012 among persons 2 years and older was 1.07% [95% CI: 0.87-1.27], with the highest incidence among Black African females aged 20-34 years at 4.5%. Sexual debut before 15 years was reported by 10.7% of respondents aged 15-24 years, and was significantly higher among male youth than female (16.7% vs. 5.0% respectively, p < 0.001). Reporting of multiple sexual partners in the previous 12 months increased from 11.5% in 2002 to 18.3% in 2012 (p < 0.001). Condom use at last sex dropped from 45.1% in 2008 to 36.2% in 2012 (p < 0.001). Levels of accurate HIV knowledge about transmission and prevention were low and had decreased between 2008 and 2012 from 31.5% to 26.8%. South Africa is on the right track with scaling up ART. However, there have been worrying increases in most HIV-related risk behaviours. These findings suggest that there is a need to scale up prevention methods that integrate biomedical, behavioural, social and structural prevention interventions to reverse the tide in the fight against HIV.


Renal responses to acute volume expansion in spontaneously hypertensive rats is related to the baseline sodium excretion.
Essential hypertension is associated with an exaggerated natriuresis in response to intravenous infusion of isotonic saline. We examined proximal tubular fluid output and segmental tubular handling of sodium in conscious spontaneously hypertensive rats (SHR), their normotensive counterparts Wistar-Kyoto rats (WKY), and ordinary Wistar rats using servo-controlled sodium and fluid balance and Li clearance technique. Sodium (Na) excretion rose to 2.72 ± 0.75 (by a factor of 8) and 1.73 ± 0.68 μmol/min (by a factor of 6.3) (p < 0.05) in SHR and WKY, respectively, thus confirming the presence of exaggerated natriuresis in SHR. FE(Li) rose to 34 ± 4 and 29 ± 2 (p < 0.05) and C(Na)/C(Li) rose to 3.0 ± 0.8 and 2.0 ± 0.6 (p < 0.05) in SHR and WKY, respectively, demonstrating that Na reabsorption in both the proximal and the distal nephron was involved. Additional experiments showed that giving the rats saline instead of water to drink for four days prior to the clearance measurement led to a remarkable increase in the natriuretic response to volume expansion. There was a close correlation between the peak increase in FE(Na) and the logarithmic values of the baseline FE(Na) values. In conclusion, the study confirms the presence of an exaggerated natriuresis in response to volume expansion in conscious SHR rats compared to WKY rats, and that the renal response to acute volume expansion is related to the baseline sodium excretion.


Microalgal luxury uptake of phosphorus in waste stabilization ponds - frequency of occurrence and high performing genera.
Microalgae commonly found in waste stabilization ponds (WSPs) are able to accumulate elevated phosphorus levels within their cells in a process known as luxury uptake. However, there are few studies focused on luxury uptake in full scale WSPs. In order to comprehensively quantify the occurrence of this phenomenon, eight different WSP sites comprising seven primary facultative, six maturation and two high rate algal ponds (HRAPs) spread over several climatic regions were monitored over four seasons. Of the 15 ponds studied, 13 of these exhibited elevated levels of biomass phosphorus content at some point; however, the occurrence in HRAPs was limited. More than half of the samples tested had elevated phosphorus contents and this occurred in all climatic zones surveyed. The phosphorus content of the biomass was significantly correlated to decreasing rainfall and increasing total dissolved phosphorus. Microscopic analysis revealed that nearly all the 17 microalgal and five cyanobacterial genera identified performed luxury uptake, but at varying frequencies. This is the first time that the genera of algae responsible for luxury uptake in full scale WSPs has been studied. Chlamydomonas/Cryptomonas, Micractinium/Microcystis and Scenedesmus were the only microalgal genera found to both commonly occur in WSPs and consistently perform luxury uptake.


Whole body elemental composition in patients with essential hypertension.
The whole body content of sodium, chlorine and potassium has been measured in 30 patients with essential hypertension, using the techniques of in vivo neutron activation analysis and whole body counting. Total exchangeable sodium and potassium were also measured, and found to be well correlated with the total body amounts of these elements. Comparable measurements on normotensive subjects could not be obtained, but results for both elements were similar to those expected on the basis of published values for healthy normal body composition. Similarly, no abnormality was found in the average body content of the other major elements (chlorine, calcium, phosphorus and nitrogen). We therefore have no evidence that essential hypertension is associated with any abnormality in the mean body content of these elements. However, there was some evidence of a relationship between body sodium and blood pressure in this study group.


Effect of immune-enhancing enteral nutrition formula enriched with plant-derived n-3 fatty acids on natural killer cell activity in rehabilitation patients.
Enteral nutrition formulas with immune-enhancing nutrients, such as n-3 fatty acids, may manage patients' nutritional status and pathophysiological processes. The aim of our study was to investigate natural killer (NK) cell activity alterations and related cytokine changes resulting from feeding with soybean oil-containing enteral nutrition formula (control group) and plant-derived n-3 fatty acid-enriched enteral nutrition formula. Subjects participated for 14 consecutive days and consumed enteral formula containing canola and flaxseed oil (n3EN, test group) in nonsurgical patients hospitalized for rehabilitation. Blood samples were collected on the first day and 14 days after the consumption of each formula daily, and anthropometric parameters were collected. Hematology and biochemical values were analyzed, and NK cell activities and serum cytokine concentration were measured. A total of sixty subjects were included in the analysis, excluding dropouts. No significant differences were found in biochemical parameters. The n3EN group's NK cell activities at effector:tumor cell ratios of 10:1, 5:1, 2.5:1 and 0.625:1 were significantly higher than those of the control group after two weeks (P < 0.05). However, there were no statistically significant differences in serum cytokine interleukin (IL)-12, interferon-γ, IL-1β, IL-6 and tumor necrosis factor-α values between the two groups. In conclusion, this study elucidates the beneficial effects of plant-derived n-3 fatty acid supplementation in enteral formula on NK cell activity.


The search for standards.
The International Organization for Standardization (ISO) assigned the responsibility for developing valve standards to its technical committee on surgical implants. A sub-committee on cardiovascular implants and the working group for heart valves were formed more than 20 years ago. The working group has learned a great deal, not only from its experience with ISO, but most of all from the collaboration of its international experts representing science, engineering and manufacturing. The first valve standard was published in 1984, revised in 1989, and the next revision will probably be published in 1995. After the formation of this ISO working group in 1972, the U.S. Food and Drug Administration (FDA) developed regulations for the premarket approval of heart valves in the United States. The European Committee for Standardization (CEN) was subsequently formed and appointed a working group to develop heart valve standards for Europe. It rapidly became apparent that the lack of co-ordination and communication of three separate organizations writing documents with different requirements and tests would lead to inconsistent and conflicting requirements. The ultimate outcome could have been a threat to public welfare by limiting the availability of medical devices. The commitment to the harmonization of standards by CEN and ISO, and the recent FDA policy to participate actively in standards development, have greatly improved this situation. Past errors, omissions and inappropriate attitudes have often slowed the standards process or have resulted in incomplete standards. This learning experience, while painful, should point the way to more timely and complete standards development in the future.


Molecular typing of Borrelia burgdorferi sensu lato: taxonomic, epidemiological, and clinical implications.
Borrelia burgdorferi sensu lato, the spirochete that causes human Lyme borreliosis (LB), is a genetically and phenotypically divergent species. In the past several years, various molecular approaches have been developed and used to determine the phenotypic and genetic heterogeneity within the LB-related spirochetes and their potential association with distinct clinical syndromes. These methods include serotyping, multilocus enzyme electrophoresis, DNA-DNA reassociation analysis, rRNA gene restriction analysis (ribotyping), pulsed-field gel electrophoresis, plasmid fingerprinting, randomly amplified polymorphic DNA fingerprinting analysis, species-specific PCR and PCR-based restriction fragment length polymorphism (RFLP) analysis, and sequence analysis of 16S rRNA and other conserved genes. On the basis of DNA-DNA reassociation analysis, 10 different Borrelia species have been described within the B. burgdorferi sensu lato complex: B. burgdorferi sensu stricto, Borrelia garinii, Borrelia afzelii, Borrelia japonica, Borrelia andersonii, Borrelia valaisiana, Borrelia lusitaniae, Borrelia tanukii, Borrelia turdi, and Borrelia bissettii sp. nov. To date, only B. burgdorferi sensu stricto, B. garinii, and B. afzelii are well known to be responsible for causing human disease. Different Borrelia species have been associated with distinct clinical manifestations of LB. In addition, Borrelia species are differentially distributed worldwide and may be maintained through different transmission cycles in nature. In this paper, the molecular methods used for typing of B. burgdorferi sensu lato are reviewed. The current taxonomic status of B. burgdorferi sensu lato and its epidemiological and clinical implications, especiallly correlation between the variable clinical presentations and the infecting Borrelia species, are discussed in detail.


Distinct contributions of functional and deep neural network features to representational similarity of scenes in human brain and behavior.
Inherent correlations between visual and semantic features in real-world scenes make it difficult to determine how different scene properties contribute to neural representations. Here, we assessed the contributions of multiple properties to scene representation by partitioning the variance explained in human behavioral and brain measurements by three feature models whose inter-correlations were minimized a priori through stimulus preselection. Behavioral assessments of scene similarity reflected unique contributions from a functional feature model indicating potential actions in scenes as well as high-level visual features from a deep neural network (DNN). In contrast, similarity of cortical responses in scene-selective areas was uniquely explained by mid- and high-level DNN features only, while an object label model did not contribute uniquely to either domain. The striking dissociation between functional and DNN features in their contribution to behavioral and brain representations of scenes indicates that scene-selective cortex represents only a subset of behaviorally relevant scene information.


X-ray diffraction studies of the composition of gallstones from English and Australian patients.
The crystalline composition of two collections of gallstones from patients in England and Australia have been determined by the x-ray powder diffraction technique. Twelve substances have been identified including a form of cholesterol which hitherto has not been reported. The weighted percentage composition averaged over each collection shows that cholesterol is the major constituent of the Australian stones and cholesterol and cholesterol monohydrate the major constituents of the English calculi. The cholesterol is possibly a decomposition product of the monohydrate. The calcium carbonates-calcite, aragonite, and vaterite-constitute most of the remainder of the calculi. Although their percentage composition is much smaller than that of the cholesterols, they are nevertheless present in a high proportion of stones. Small traces of apatite, whitlockite, sodium chloride, calcium stearate and palmitic acid (or other long-chain compounds having closely related spacings) have been found. Small spheroids scattered throughout some stones appear to be mainly calcium stearate, although the total quantity available is too small and too impure for a definite identification.


8-hydroxydeguanosine and nitrotyrosine are prognostic factors in urinary bladder carcinoma.
Oxidative stress markers and peroxiredoxins are connected to cancer. A large set of urinary bladder carcinomas were studied for the expression of nitrotyrosine and 8-hydroxydeguanosine (8OHdG), two markers indicating oxidative damage. Serum and urine 8-OHdG were assessed in a subset of patients. We also analysed immunohisto-chemically the expression of nrf2, keap1, all six peroxiredoxins (prx) and thioredoxin (trx) in these tumors. 15 % of the cases showed 8OHdG and 36 % nitrotyrosine positivity. Expression of nitrotyrosine and 8OHdG associated with a poor prognosis (p=0.050, p=0.011, respectively). Peroxiredoxin positivity ranged from 39 % to 84 % lowest expression being for prx 4 and highest for prx 3. Prx 4 expression associated with a poor prognosis (p=0.025) with high grade (p=0.044) and larger tumors (p=0.023). Cytoplasmic trx positivity was seen in 91 % and nuclear in 59 % of tumors. Nuclear and cytoplasmic trx associated with each other (p<0.001), and nuclear trx associated with prx 6 (p=0.001), prx 2 (p<0.001), and prx 5 (p<0.001). 8OHdG associated with nuclear trx positivity (p=0.002), inversely with prx 1 (p=0.025) and with keap1 (p=0.020). Nuclear nrf2 was associated with nitrotyrosine (p=0.042). The results show that the amount of oxidative stress in urinary bladder tumors affects the prognosis of the patients. Of antioxidative enzymes, prx4 associated with an unfavourable prognosis. Selective inhibition of prx4 expression might then be one additional option of treatment of bladder cancer.


Metabolic control theory: a graph-theoretic approach.
The regulatory properties of metabolic pathways are investigated using a graph-theoretic approach. Applying concepts from classical graph theory, analytical expressions are derived for the flux control coefficients of a linear pathway subject to feedback inhibition. It is shown that the relative importance of the various enzymes in the control of flux can be easily established with the aid of this graphical procedure. Another advantage of this technique is that it can be implemented in a step-by-step fashion for simplifying the pathway structure. The graph-theoretic method can also be used for studying the cause-effect relationships of the individual enzymes in a given pathway.


Targeting ligand-induced binding sites on GPIIb/IIIa via single-chain antibody allows effective anticoagulation without bleeding time prolongation.
Therapeutic anticoagulation is widely used, but limitations in efficacy and bleeding complications cause an ongoing search for new agents. However, with new agents developed it seems to be an inherent problem that increased efficiency is accompanied by an increase in bleeding complications. We investigate whether targeting of anticoagulants to activated platelets provides a means to overcome this association of potency and bleeding. Ligand-induced binding sites (LIBS) on fibrinogen/fibrin-binding GPIIb/IIIa represent an abundant clot-specific target. We cloned an anti-LIBS single-chain antibody (scFv(anti-LIBS)) and genetically fused it with a potent, direct factor Xa (fXa) inhibitor, tick anticoagulant peptide (TAP). Specific antibody binding of fusion molecule scFv(anti-LIBS)-TAP was proven in flow cytometry; anti-fXa activity was demonstrated in chromogenic assays. In vivo anticoagulative efficiency was determined by Doppler-flow in a ferric chloride-induced carotid artery thrombosis model in mice. ScFv(anti-LIBS)-TAP prolonged occlusion time comparable to enoxaparine, recombinant TAP, and nontargeted mutant-scFv-TAP. ScFv(anti-LIBS)-TAP revealed antithrombotic effects at low doses at which the nontargeted mutant-scFv-TAP failed. In contrast to the other anticoagulants tested, bleeding times were not prolonged by scFv(anti-LIBS)-TAP. The novel clot-targeting approach of anticoagulants via single-chain antibody directed against a LIBS-epitope on GPIIb/IIIa promises effective anticoagulation with reduced bleeding risk.


Increase in the number of atrial natriuretic hormone receptors in regenerating rat liver.
Forty-eight hours after partial (approximately 67%) hepatectomy the activity of the particulate guanylate cyclase was increased by 2-fold in the regenerating rat liver. This increase was not an artifact of membrane isolation procedures, and as determined by 125I-labeled Tyr-28 atrial natriuretic hormone-(1-28) ANF binding, was accompanied by a 2-fold increase in the number of ANF receptors. The Kd of the receptors in membranes of regenerating livers was not significantly different from the Kd of the receptors in livers of sham-operated rats. The linear synthetic descysteine analog of ANF, analog I, which binds only to the 66-kDa receptors, displaced approximately 40% of the specifically bound 125I-ANF in liver membranes from both hepatectomized and sham-operated (control) animals. Affinity cross-linking studies with 125I-ANF confirmed the increase in the 116-kDa ANF receptor in membranes of regenerating livers. In perfused livers derived from control and hepatectomized animals, the basal rates of cGMP production were not significantly different. However, atriopeptin II-stimulated cGMP production was twice as great in regenerating livers as compared with controls. These data demonstrate that the increase in particulate guanylate cyclase activity observed during liver regeneration is due to an increase in the 116-kDa ANF receptor-associated activity. Additionally, our data demonstrate that the regenerating rat liver may be a valuable model with which to study the role of the hepatic ANF receptor/particulate guanylate cyclase.


Protection of BALB/c mice against experimental Helicobacter pylori infection by oral immunisation with H pylori heparan sulphate-binding proteins coupled to cholera toxin beta-subunit.
The presence of Helicobacter pylori in the gastroduodenal mucosae is associated with chronic active gastritis, peptic ulcers and gastric cancers such as adenocarcinoma and low-grade gastric B-cell lymphoma. In response to the presence of antibiotic-resistant strains, the use of vaccines to combat this infection has become an attractive alternative. The present study used a murine model of infection by a mouse-adapted H. pylori strain to determine whether infection in BALB/c mice can be successfully eradicated by intragastric vaccination with H. pylori heparan sulphate-binding proteins (HSBP) covalently coupled to the beta-subunit of cholera toxin (CTB). It was shown that vaccination confers protection against exposure of BALB/c mice to the pathogen, as revealed by microbiological, histopathological and molecular methods.


[Monosynaptic excitation of motor neurons of the nucleus of the accessory nerve by reticulofugal impulses].
Responses of accessory motoneurons to electrical stimulation of the ipsilateral nucleus gigantocellularis of medullary reticular formation were recorded intracellularly in cats under chloraloze-nembutal anesthesia. The stimulation evoked in the accessory motoneurons monosynaptic EPSPs with latency from 0.6 to 1.0 ms, maximal amplitude 4.0 mV and duration 10 ms. Weak facilitation of EPSPs and absence of action potentials were demonstrated using paired shocks and rhythmic stimulation of the nucleus.


Crystal structures of human SIRT3 displaying substrate-induced conformational changes.
SIRT3 is a major mitochondrial NAD(+)-dependent protein deacetylase playing important roles in regulating mitochondrial metabolism and energy production and has been linked to the beneficial effects of exercise and caloric restriction. SIRT3 is emerging as a potential therapeutic target to treat metabolic and neurological diseases. We report the first sets of crystal structures of human SIRT3, an apo-structure with no substrate, a structure with a peptide containing acetyl lysine of its natural substrate acetyl-CoA synthetase 2, a reaction intermediate structure trapped by a thioacetyl peptide, and a structure with the dethioacetylated peptide bound. These structures provide insights into the conformational changes induced by the two substrates required for the reaction, the acetylated substrate peptide and NAD(+). In addition, the binding study by isothermal titration calorimetry suggests that the acetylated peptide is the first substrate to bind to SIRT3, before NAD(+). These structures and biophysical studies provide key insight into the structural and functional relationship of the SIRT3 deacetylation activity.


Early Evaluation of Response Using 18F-FDG PET Influences Management in Gastrointestinal Stromal Tumor Patients Treated with Neoadjuvant Imatinib.
18F-FDG PET has previously been proven effective as an early way to evaluate the response of gastrointestinal stromal tumors (GISTs) to imatinib treatment. However, it is unclear whether early evaluation of response affects treatment decisions in GIST patients treated with neoadjuvant intent. Methods: We retrospectively scored changes in management based on early evaluation of response by 18F-FDG PET in patients in the Dutch GIST registry treated with neoadjuvant imatinib. Results: Seventy 18F-FDG PET scans were obtained for 63 GIST patients to evaluate for an early response to neoadjuvant imatinib. The scans led to a change in management in 27.1% of the patients. Change in management correlated strongly with lack of metabolic response (P < 0.001) and non-KIT exon 11-mutated GISTs (P < 0.001). Conclusion: Performing 18F-FDG PET for early evaluation of response often results in a change of management in GIST patients harboring the non-KIT exon 11 mutation and should be considered the standard of care in GIST patients treated with neoadjuvant intent.


Hyperglycaemic siblings of Type II (non-insulin-dependent) diabetic patients have increased PAI-1, central obesity and insulin resistance compared with their paired normoglycaemic sibling.
First-degree relatives of Type II (non-insulin-dependent) diabetic patients in cross-sectional studies have increased insulin resistance, associated cardiovascular risk factors and abnormalities of fibrinolysis and coagulation. To minimise between-family genetic and environmental confounders, we investigated within-family relationships between early hyperglycaemia and risk factors. Thirteen age and gender matched sibling pairs of Type II (non-insulin-dependent) diabetic patients, one hyperglycaemic, one normoglycaemic (fasting plasma glucose at screening 6.0-7.7 mmol.l(-1) and < 6.0 mmol.l(-1), respectively) were assessed for plasminogen activator inhibitor antigen (PAI-1), tissue plasminogen activator antigen (t-PA), fibrinogen, Factor VII and Factor VIII/von Willebrand factor antigen. Fasting lipid profiles, blood pressure and HOMA insulin sensitivity (%S) were also measured in siblings and in matched subjects without family history of diabetes. Hyperglycaemic and normoglycaemic siblings (7 female, 6 male) were aged, mean (SD) 56.8 (8.7) and 55.8 (8.4) years. Hyperglycaemic siblings had increased PAI-1 antigen, geometric mean (i.q.r.): 26.3 (15.1-45.6) vs 11.1 (2.1-23.3) ng/ml, p=0.0002, similar t-PA antigen, mean (SD) 9.5 (4.3) vs 7.4 (2.5) ng/ml, p=0.2 and fibrinogen 2.2 (0.3) vs 2.3 (0.6) g/l, p=0.5, and reduced %S 66.3 (30.5) vs 82.9 (25), p=0.04. PAI-1 correlated negatively with %S ( r=-0.55, p=0.005). No significant differences were found in blood pressure or fasting lipids. A minor increase in plasma glucose in non-diabetic sibling pairs of Type II (non-insulin-dependent) diabetic patients was associated with reduced insulin sensitivity, increased central adiposity and a doubling of PAI-1 antigen concentration, suggesting impaired fibrinolysis. It is possible that this could contribute to increased cardiovascular risk in these subjects.


Visual outcome in ocular sarcoidosis: retrospective evaluation of risk factors.
To analyze risk factors associated with poor visual outcome in patients with ocular sarcoidosis. In this retrospective study, charts of 44 patients with uveitis and biopsy-proven sarcoidosis were reviewed. Ocular parameters evaluated were as follows: location, type of uveitis, visual acuity, presence of posterior synechia, iris nodules, vitritis, snowballs, chorioretinal lesions, retinal vasculitis, papillitis, macular edema, cataract, and glaucoma. Final visual acuity of the worst-seeing eye at last follow-up was the outcome considered in univariable and multivariable analyses. Visual acuity of the worst-seeing eye was stratified into 2 categories according to the threshold 20/50 (=20/50 and >20/50). A total of 44 patients with bilateral uveitis were studied. The majority of patients presented with panuveitis (52%), granulomatous type (61%), posterior synechia (62%). The most frequent vision-threatening complications were cystoid macular edema (56%) and cataract (56%). The median best-corrected visual acuity in the worst-seeing eye at presentation and at end of follow-up was respectively 0.4 (interquartile range [IQR] 0.26-0.80) and 0.63 (IQR 0.36-1.00). At univariable analysis, the presence of iris nodules (p=0.049), cystoid macular edema (p=0.007), and cataract (p=0.007) were clinically significant conditions for a visual outcome of 20/50 or worse in the worst-seeing eye. In multivariable analysis, cystoid macular edema (p=0.034) was the only statistically significant predictor associated with unfavorable visual outcome. In this study, we attempted to find risk factors related to poor visual outcome in patients with ocular sarcoidosis. The results suggest that only the presence of cystoid macular edema was significantly associated with worst visual outcome.


Alterations in the reproductive patterns of female mice exposed to xenobiotics.
Chemicals, by virtue of their varied interactions with biological molecules, are expected to differ in the way they may alter female reproduction. Reproductive toxicity may reflect effects either on the female germ cells or on various maternal processes such as ovulation, implantation, pregnancy, and parturition. In either case, the ultimate manifestation of chemical toxicity on female reproduction is a decrease in the number of normal young born. Very little information is available on the effects of chemicals that are nonhormonal in nature on the long-term ability of treated females to produce offspring. This report presents the results of long-term female total reproductive capacity (TRC) tests on 29 chemicals, including pharmaceuticals, pesticides, and alkylating and industrial agents. For each chemical, the minimum test involved an evaluation of the maximum tolerated dose administered as a single intraperitoneal injection. Females were single-pair mated with an untreated male for most of the female's reproductive life span (a minimum of 347 days posttreatment) and scored for the number of live births produced during this period. Confirmatory dominant lethal experiments or histological examinations for numbers of small follicles were carried out when mutagenic effects or cytotoxicity, respectively, were suspected as the basis for reduced fertility. Of the 29 chemicals studied, 17 had reproductive effects which may be grouped into one of three classes: (1) those that reduced the total number of young and litters per female, (2) those that reduced the total number of young but not of litters, and (3) those that had no significant effect on the total number of young produced but reduced the size of the first and/or second litters. The TRC provides a capacity for detecting a range of toxic insults upon female reproduction. Many of the chemicals were indeed shown to affect the reproductive performance of females through mutagenic and/or cytotoxic effects on follicles. In some cases, however, no causative mechanism could be identified for the observed reduction in reproductive performance. Nevertheless, with this report the number of chemicals tested by this TRC procedure has been quadrupled and the categories of chemicals tested have been substantially broadened.


Guidelines for the management of dermatomycosis (2019).
The "Guidelines for the management of dermatomycosis" of the Japanese Dermatological Association were first published in Japanese in 2009 and the Guidelines Committee of the Japanese Dermatological Association revised it in 2019. The first guidelines was prepared according to the opinions of the Guidelines Committee members and it was of educational value. The revised version is composed of introductory descriptions of the disease concepts, diagnosis, medical mycology and recent advances in treatment, along with clinical questions (CQ), which is intended to help in general practice for dermatologists. The CQ are limited to those involved in therapy but include some of the recently launched antifungal agents. The level of evidence and the degree of recommendation for each item were reviewed by the committee based on clinical studies published by 2018. For rare dermatomycoses, recommendations by the committee are described in the guidelines. In this field, there are still few good quality studies on treatment. Periodic revision in line with new evidence is necessary.


Effect of pneumococcal conjugate vaccination on serotype-specific carriage and invasive disease in England: a cross-sectional study.
We investigated the effect of the 7-valent pneumococcal conjugate vaccine (PCV7) programme in England on serotype-specific carriage and invasive disease to help understand its role in serotype replacement and predict the impact of higher valency vaccines. Nasopharyngeal swabs were taken from children <5 y old and family members (n=400) 2 y after introduction of PCV7 into routine immunization programs. Proportions carrying Streptococcus pneumoniae and serotype distribution among carried isolates were compared with a similar population prior to PCV7 introduction. Serotype-specific case carrier ratios (CCRs) were estimated using national data on invasive disease. In vaccinated children and their contacts vaccine-type (VT) carriage decreased, but was offset by an increase in non-VT carriage, with no significant overall change in carriage prevalence, odds ratio 1.06 (95% confidence interval 0.76-1.49). The lower CCRs of the replacing serotypes resulted in a net reduction in invasive disease in children. The additional serotypes covered by higher valency vaccines had low carriage but high disease prevalence. Serotype 11C emerged as predominant in carriage but caused no invasive disease whereas 8, 12F, and 22F emerged in disease but had very low carriage prevalence. Because the additional serotypes included in PCV10/13 have high CCRs but low carriage prevalence, vaccinating against them is likely to significantly reduce invasive disease with less risk of serotype replacement. However, a few serotypes with high CCRs could mitigate the benefits of higher valency vaccines. Assessment of the effect of PCV on carriage as well as invasive disease should be part of enhanced surveillance activities for PCVs. Please see later in the article for the Editors' Summary.


Cell-mediated immunity in patients positive for hepatitis-associated antigen.
Cell-mediated immunity to antigens prepared from both serum and liver of patients positive for hepatitis-associated antigen (H.A.A.) was measured by using the leucocyte migration test. Altogether, 43 patients with H.A.A.-positive acute and chronic liver disease, eight with serum antibody to H.A.A., and 13 controls were studied. The cell-mediated immunity detected was specific for H.A.A. or other antigenic determinants of the associated infective agent and could be found only in patients with evidence of previous contact with H.A.A.Cell-mediated immunity to the H.A.A.-positive test antigens was found in all but one of the patients with acute hepatitis, in about half of the patients with chronic aggressive hepatitis or cirrhosis, rarely in those with chronic persistent hepatitis, and in none of the apparently healthy carriers.Our results support the hypothesis that the cellular immune response plays an important part in the clearance of the infective agent from H.A.A.-positive patients and in the pathogenesis of the associated liver cell injury.


An integrative in-silico approach for therapeutic target identification in the human pathogen Corynebacterium diphtheriae.
Corynebacterium diphtheriae (Cd) is a Gram-positive human pathogen responsible for diphtheria infection and once regarded for high mortalities worldwide. The fatality gradually decreased with improved living standards and further alleviated when many immunization programs were introduced. However, numerous drug-resistant strains emerged recently that consequently decreased the efficacy of current therapeutics and vaccines, thereby obliging the scientific community to start investigating new therapeutic targets in pathogenic microorganisms. In this study, our contributions include the prediction of modelome of 13 C. diphtheriae strains, using the MHOLline workflow. A set of 463 conserved proteins were identified by combining the results of pangenomics based core-genome and core-modelome analyses. Further, using subtractive proteomics and modelomics approaches for target identification, a set of 23 proteins was selected as essential for the bacteria. Considering human as a host, eight of these proteins (glpX, nusB, rpsH, hisE, smpB, bioB, DIP1084, and DIP0983) were considered as essential and non-host homologs, and have been subjected to virtual screening using four different compound libraries (extracted from the ZINC database, plant-derived natural compounds and Di-terpenoid Iso-steviol derivatives). The proposed ligand molecules showed favorable interactions, lowered energy values and high complementarity with the predicted targets. Our proposed approach expedites the selection of C. diphtheriae putative proteins for broad-spectrum development of novel drugs and vaccines, owing to the fact that some of these targets have already been identified and validated in other organisms.


Development of a qualitative sequential immunoassay for characterizing the intrinsic properties of circulating cardiac troponin I.
With myocardial infarction (MI), cardiac troponin is released from the heart into circulation, where it can be detected with immunoassays independently quantifying cardiac troponin I (cTnI) or cTnT. There is, however, no single immunoassay that sequentially probes the posttranslational modification status of cTnI or directly characterizes whether circulating cTnI is bound to cTnC and/or cTnT. Here we describe the development of a qualitative immunoassay to directly probe the primary and ternary structure of circulating cTnI through diffractive optics technology (dotLab System, Axela). Anti-cTnI antibody 8I-7 was immobilized on a patterned sensor to capture cTnI. One or more detector antibodies were sequentially introduced to probe for amino acid sequence integrity or phosphorylation status of cTnI, or its association with cTnC and/or cTnT. Respective immunocaptures were recorded as real-time diffractive intensities (DIs), and the DI differences were analyzed. Each immunodetection was independent of the others but was done in a single sequential assay. This diffraction-based immunoassay successfully characterized cTnI. The unamplified assay determined whether cTnI was degraded at N-terminus and/or C-terminus or phosphorylated. Sequential application of multiple detector antibodies without an antibody-stripping step enables real-time interrogation of 5 different epitopes of cTnI, or direct detection of the cTn complex (cTnI-cTnC-cTnT) in a single sequential assay. Finally, this assay was optimized with amplification to directly detect circulating cTnI bound to cTnC and cTnT in serum from an MI patient. The dot Immunoassay is the first qualitative sequential immunoassay to address the direct interactions of the troponin subunits and various modified forms of cTnI.


Role of the electrochemical proton gradient in genetic transformation of Haemophilus influenzae.
The uptake of homologous DNA by Haemophilus influenzae was studied as a function of the proton motive force in completely competent cultures in the pH range of 6 to 8. The composition and magnitude of the proton motive force were varied by using the ionophores valinomycin and nigericin (in the presence of various potassium ion concentrations) and by using protonophores. No interaction of the ionophores with the DNA transformation system itself was observed. Either component of the proton motive force, the electrical potential or the pH gradient, can drive the uptake of DNA, and the extent of the uptake of DNA is ultimately determined by the total proton motive force. The transformation frequency increases with the proton motive force, which reaches a maximum value at around -130 mV. These results are consistent with an electrogenic proton-DNA symport mechanism, but direct evidence for such a system is not available. The proton motive force was followed during competence development of H. influenzae at pH 8. In the initial phase (up to 50 min), the proton motive force remained constant at about -90 mV, whereas the transformation frequency rose steeply. In the second phase, the proton motive force increased. The transformation frequency in this phase increased with the proton motive force, as in completely competent cultures. These observations and the observed inhibition by NAD of both the proton motive force and the transformation frequency indicate that structural components of the competent state are formed in the initial phase of competence development, whereas the second phase is characterized by an increase of the proton motive force.