Transient characteristics and stability analysis of standing wave thermoacoustic-piezoelectric harvesters.
Standing wave thermoacoustic-piezoelectric (TAP) energy harvesters convert thermal energy, such as solar or waste heat energy, directly into electrical energy without the need for any moving components. The input thermal energy generates a steep temperature gradient along a porous medium called "stack." At a critical threshold of the temperature gradient, self-sustained acoustic waves are developed inside an acoustic resonator. The associated pressure fluctuations impinge on a piezoelectric diaphragm, placed at the end of the resonator, to generate electricity. The behavior of this multi-field system is modeled using the electrical analogy approach. The developed model combines the descriptions of the acoustic resonator and the stack with the characteristics of the piezoelectric diaphragm. The equivalent electric network is analyzed to determine the system's stability and predict the temperature gradient necessary to developing self-sustained oscillations inside the harvester. The developed network is utilized also to investigate the transient performance of the harvester by employing the network theory and Simulation Program with Integrated Circuit Emphasis software package. The established stability boundaries are validated against the predictions of the root locus technique. Furthermore, the obtained results are compared with experimental results extracted from testing a prototype of the harvester. The developed approach presents an innovative tool for the design of TAP energy harvesters.


Prevalence of distal renal tubular acidosis in primary Sjögren's syndrome.
Our objectives were to analyse the prevalence of distal renal tubular acidosis (dRTA) in primary SS (pSS) and to compare a novel urinary acidification test with furosemide and fludrocortisone (FF) with the gold standard ammonium chloride (NH4Cl) to detect dRTA. Urinary acidification was assessed in 57 pSS patients using NH4Cl and FF. A urinary acidification defect was defined as an inability to reach a urinary pH of <5.3 after NH4Cl. The prevalence of complete dRTA (urinary acidification defect with acidosis) was 5% (3/57). All three patients had positive SSA/Ro and SSB/La autoantibodies and impaired kidney function. The prevalence of incomplete dRTA (urinary acidification defect without acidosis) was 25% (14/57). Compared with patients without dRTA, patients with incomplete dRTA had significantly lower venous pH and serum bicarbonate and higher urinary pH. SSB/La antibodies were more prevalent in the dRTA groups (P < 0.05). Compared with NH4Cl, the positive and negative predictive values of FF were 46% and 82%, respectively. Vomiting occurred more often during the urinary acidification test with NH4Cl than with FF (9 vs 0, P < 0.05). Incomplete dRTA is common in pSS and causes mild acidaemia and higher urinary pH, which may contribute to bone demineralization and kidney stone formation. FF cannot replace NH4Cl in testing urinary acidification in pSS, but may be considered as a screening tool, given its reasonable negative predictive value and better tolerability.


Modified vastus lateralis flap in treating a difficult hip infection.
Difficult-to-treat hip infections are typically a reconstructive challenge to orthopedic surgeons. Treatment regimens must be tailored to each patient on the basis of an overall evaluation of systemic host factors and local wound factors. To treat a medically ill patient with compromised local wounds in the hip area, a modified vastus lateralis flap transposition was developed. This flap transposition is of a simple design and effective as both a soft-tissue coverage and a dead space filler. From January 2002 to June 2003, 10 patients (5 male patients and 5 female patients) suffering difficult hip infections were referred to Chang Gung Memorial Hospital at Chia-Yi for further treatment. Before visiting the clinic, each patient had undergone repeated failed operations, including debridement, introduction of antibiotic-loaded cement spacers, and fasciocutaneous flap transposition. After being diagnosed with difficult hip infections, these patients were treated with the modified vastus lateralis flap transposition immediately after radical debridement. The average follow-up period was 17.4 months (range, 9-27 months). The hip infections in all 10 patients were controlled and the wounds healed uneventfully. The need for another soft-tissue procedure was not indicated. The C-reactive protein returned to a near-normal and stable level within 1 month. The only complication occurred when case 1 developed a contralateral iliopsoas abscess and secondary septic hip 9 months after the procedure. There was no morbidity or mortality related to the technique. For medically ill patients with compromised local wounds, the modified vastus lateralis muscle flap transposition after radical debridement is a simple and effective way of resolving refractory and limb-threatening hip infections.


Effects of malleable kinetochore morphology on measurements of intrakinetochore tension.
The distance between fluorescent spots formed by various kinetochore proteins (delta) is commonly interpreted as a manifestation of intrakinetochore tension (IKT) caused by microtubule-mediated forces. However, large-scale changes of the kinetochore architecture (such as its shape or dimensions) may also contribute to the value of delta. To assess contributions of these non-elastic changes, we compare behaviour of delta values in human kinetochores with small yet mechanically malleable kinetochores against compound kinetochores in Indian muntjac (IM) cells whose architecture remains constant. Due to the micrometre-scale length of kinetochore plates in IM, their shape and orientation are discernible in conventional light microscopy, which enables precise measurements of IKT independent of contributions from changes in overall architecture of the organelle. We find that delta in IM kinetochores remains relatively constant when microtubule-mediated forces are suppressed by Taxol, but it prominently decreases upon detachment of microtubules. By contrast, large decreases of delta observed in Taxol-treated human cells coincide with prominent changes in length and curvature of the kinetochore plate. These observations, supported by computational modelling, suggest that at least 50% of the decrease in delta in human cells reflects malleable reorganization of kinetochore architecture rather than elastic recoil due to IKT.


Analytical Transmission Electron Microscopy of Amphibole Fibers From the Lungs of Quebec Miners.
The objective of this study is to describe the morphology, molecular structure, and chemistry of amphibole fibers from lung samples from workers in the chrysotile mines at Asbestos and Thetford Mines, Quebec. A fibrous tremolite-actinolite contaminant in an asbestos ore sample from the deposit at Asbestos was used for comparison. Lattice imaging was performed using high-resolution transmission electron microscopy (HRTEM). Silica-rich amorphous coatings (SIRA) that may be related to carcinogenesis are noted on all of the HRTEM photographs of fibers retained in lung, but not on fiber surfaces of the bulk comparison sample. Fibers found in lung samples and in a bulk comparison sample are produced primarily by splitting of thicker crystals and, as such, might not be considered asbestos fibers on the basis of certain mineralogical criteria. Implications of SIRA coatings with respect to carcinogenesis are worthy of further study.


A multigene expression panel for the molecular diagnosis of Barrett's esophagus and Barrett's adenocarcinoma of the esophagus.
In order to identify genes or combination of genes that have the power to discriminate between premalignant Barrett's esophagus and Barrett's associated adenocarcinoma, we analysed a panel of 23 genes using quantitative real-time RT-PCR (qRT-PCR, Taqman and bioinformatic tools. The genes chosen were either known to be associated with Barrett's carcinogenesis or were filtered from a previous cDNA microarray study on Barrett's adenocarcinoma. A total of 98 tissues, obtained from 19 patients with Barrett's esophagus (BE group) and 20 patients with Barrett's associated esophageal adenocarcinoma (EA group), were studied. Triplicate analysis for the full 23 gene of interest panel, and analysis of an internal control gene, was performed for all samples, for a total of more than 9016 single PCR reactions. We found distinct classes of gene expression patterns in the different types of tissues. The most informative genes clustered in six different classes and had significantly different expression levels in Barrett's esophagus tissues compared to adenocarcinoma tissues. Linear discriminant analysis (LDA) distinguished four genetically different groups. The normal squamous esophagus tissues from patients with BE or EA were not distinguishable from one another, but Barrett's esophagus tissues could be distinguished from adenocarcinoma tissues. Using the most informative genes, obtained from a logistic regression analysis, we were able to completely distinguish between benign Barrett's and Barrett's adenocarcinomas. This study provides the first non-array parallel mRNA quantitation analysis of a panel of genes in the Barrett's esophagus model of multistage carcinogenesis. Our results suggest that mRNA expression quantitation of a panel of genes can discriminate between premalignant and malignant Barrett's disease. Logistic regression and LDAs can be used to further identify, from the complete panel, gene subsets with the power to make these diagnostic distinctions. Expression analysis of a limited number of highly selected genes may have clinical usefulness for the treatment of patients with this disease.


Impact of PRRSV infection and dietary soybean meal on ileal amino acid digestibility and endogenous amino acid losses in growing pigs.
Porcine reproductive and respiratory syndrome virus (PRRSV) is a significant disease in the swine industry, and increasing soybean meal (SBM) consumption during this disease challenge may improve performance. Our objectives were to determine the impact of SBM level on apparent total tract (ATTD) and ileal (AID) digestibility during PRRSV infection and to determine ileal basal endogenous losses (BEL) during PRRSV infection. Forty PRRSV negative gilts were fitted with a T-cannula in the distal ileum. Treatments were arranged in a 2 × 2 factorial with high and low SBM (HSBM, 29% vs. LSBM, 10%), with and without PRRSV (n = 6/treatment). The remaining pigs (n = 8/challenge status) were fed a N-free diet. Chromic oxide was used as an indigestible marker. On day post inoculation (dpi) 0, at 47.7 ± 0.57 kg BW, 20 pigs were inoculated with live PRRSV; 20 control pigs were sham inoculated. Infection was confirmed by serum PCR. Feces were collected at dpi 5 to 6 and dpi 16 to 17, and ileal digesta collected at dpi 7 to 8 and dpi 18 to 19. Feed, feces, and digesta were analyzed for DM, N, and GE. Digesta and feed were analyzed for AA. Data were analyzed in a 2 × 2 + 2 factorial design to determine main effects of diet and PRRSV and their interaction. Data from N-free fed pigs were analyzed separately to determine BEL and hindgut disappearance due to PRRSV infection. All control pigs remained PRRSV negative. There were no interactions for AID of AA; however, HSBM reduced DM, GE, Lys, and Met AID and increased Arg and Gly AID during both collection periods (P < 0.05). At dpi 7 to 8 only, PRRSV reduced DM and GE AID (P < 0.05). At 7 to 8 dpi, BEL of Arg, Ala, and Pro were reduced (P < 0.05) due to PRRSV by 64%, 39%, and 94%, respectively. At dpi 18 to 19, BEL of Thr tended (P = 0.06) to be increased in PRRSV-infected pigs; however, no other differences were observed. Pigs fed LSBM had increased Lys, Met, Thr, Trp, and Pro standardized ileal digestibility (SID), primarily at 7 to 8 dpi. At 7 to 8 dpi, PRRSV reduced Arg, Gly, and Pro SID (P < 0.01), and SID Pro continued to be reduced by 17% at dpi 18 to 19. Compared with HSBM pigs, LSBM reduced hindgut disappearance of DM and GE at dpi 5 to 8 and dpi 16 to 19, while N disappearance was reduced at dpi 5 to 8. There were no differences between control and PRRSV N-free fed pigs. Altogether, SBM inclusion impacts SID of AA and hindgut disappearance of nutrients, regardless of PRRSV. In contrast, there is minimal impact of PRRSV on BEL, and therefore, SID of most AA are not different.


Relaxed molecular clock provides evidence for long-distance dispersal of Nothofagus (southern beech).
Nothofagus (southern beech), with an 80-million-year-old fossil record, has become iconic as a plant genus whose ancient Gondwanan relationships reach back into the Cretaceous era. Closely associated with Wegener's theory of "Kontinentaldrift", Nothofagus has been regarded as the "key genus in plant biogeography". This paradigm has the New Zealand species as passengers on a Moa's Ark that rafted away from other landmasses following the breakup of Gondwana. An alternative explanation for the current transoceanic distribution of species seems almost inconceivable given that Nothofagus seeds are generally thought to be poorly suited for dispersal across large distances or oceans. Here we test the Moa's Ark hypothesis using relaxed molecular clock methods in the analysis of a 7.2-kb fragment of the chloroplast genome. Our analyses provide the first unequivocal molecular clock evidence that, whilst some Nothofagus transoceanic distributions are consistent with vicariance, trans-Tasman Sea distributions can only be explained by long-distance dispersal. Thus, our analyses support the interpretation of an absence of Lophozonia and Fuscospora pollen types in the New Zealand Cretaceous fossil record as evidence for Tertiary dispersals of Nothofagus to New Zealand. Our findings contradict those from recent cladistic analyses of biogeographic data that have concluded transoceanic Nothofagus distributions can only be explained by vicariance events and subsequent extinction. They indicate that the biogeographic history of Nothofagus is more complex than envisaged under opposing polarised views expressed in the ongoing controversy over the relevance of dispersal and vicariance for explaining plant biodiversity. They provide motivation and justification for developing more complex hypotheses that seek to explain the origins of Southern Hemisphere biota.


Isolated plasma membranes regulate neurotransmitter expression and facilitate effects of a soluble brain cholinergic factor.
The choice of which transmitter will be expressed by a neuron is influenced by diffusible differentiating factors produced by a variety of nonneuronal cells. In this study we show that human, bovine, and rat brains contain a soluble heparin-binding factor that stimulates cholinergic and peptidergic expression. We also show that neuronal contact with other cell membranes influences neurotransmitter development and acts synergistically with the effects of the soluble brain factor. Exposure of cultured sympathetic neurons to purified plasma membranes derived either from cultured Schwann cells or from cultured sympathetic neurons promoted expression of choline acetyltransferase (CAT), a cholinergic trait, and of substance P (SP). CAT activity and SP were also stimulated by a 50-kDa soluble protein that was purified 14,000-fold from human, calf, and rat brain by heparin-Sepharose affinity chromatography. CAT activity after concurrent treatment with plasma membranes and the soluble factor far exceeded the sum of the enzyme activities after the individual treatments, suggesting that membrane molecules and the factor facilitated each other. Thus, cell-surface molecules, which have been shown previously to influence neuronal morphogenesis and neurite elongation, may also help determine the transmitter phenotype of the neuron. Moreover, cell-surface molecules may modulate the effects of diffusible differentiating factors.


Sub-wavelength patterning of the optical near-field.
We report the sub-wavelength patterning of the optical near-field by total internal reflection illumination of a regular array of resonant gold nano-particles. Under appropriate conditions, the in-plane coupling between Localized Surface Plasmon (LSP) fields gives rise to sub-wavelength light spots between the structures. Measurements performed with an Apertureless Scanning Near-Field Optical Microscope (ASNOM) show a good agreement with theoretical predictions based on the Green dyadic method. This concept might offer a convenient way to elaborate extended optical trap landscapes for manipulation of sub-micrometer systems.


Association between respiratory tract methicillin-resistant Staphylococcus aureus and survival in cystic fibrosis.
The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in the respiratory tract of individuals with cystic fibrosis (CF) has increased dramatically; however, its impact on outcomes in CF is unclear. Because the time between infection with bacteria in CF and death can be decades, observational studies with long periods of follow-up are well suited to address the current gap in knowledge. To determine whether isolation of MRSA from the respiratory tract of CF patients is associated with worse survival compared with patients who never have a culture positive for MRSA. Cohort study of 19,833 CF patients aged 6 to 45 years seen at centers accredited by the Cystic Fibrosis Foundation in the United States. Patients entered between January 1996 and December 2006 and were followed up through December 2008. Cox regression models with time-varying covariates were used to compare survival between CF patients with and without respiratory tract MRSA. Time from age at entry until age at death from any cause. In 137,819 patient-years of observation (median, 7.3 years/patient), 2537 CF patients died and 5759 patients had MRSA detected. The mortality rate was 18.3 deaths (95% confidence interval [CI], 17.5-19.1) per 1000 patient-years in patients without MRSA and 27.7 deaths (95% CI, 25.3-30.4) per 1000 patient-years in those with MRSA. Among those with MRSA, the attributable risk percentage of death associated with MRSA was 34.0% (95% CI, 26.7%-40.4%). The unadjusted hazard ratio associated with MRSA was 1.47 (95% CI, 1.32-1.62). After adjustment for time-varying covariates associated with severity of illness, MRSA remained associated with a higher risk of death (1.27; 95% CI, 1.11-1.45). Detection of MRSA in the respiratory tract of CF patients was associated with worse survival.


Microarray meta-analysis identifies acute lung injury biomarkers in donor lungs that predict development of primary graft failure in recipients.
To perform a meta-analysis of gene expression microarray data from animal studies of lung injury, and to identify an injury-specific gene expression signature capable of predicting the development of lung injury in humans. We performed a microarray meta-analysis using 77 microarray chips across six platforms, two species and different animal lung injury models exposed to lung injury with or/and without mechanical ventilation. Individual gene chips were classified and grouped based on the strategy used to induce lung injury. Effect size (change in gene expression) was calculated between non-injurious and injurious conditions comparing two main strategies to pool chips: (1) one-hit and (2) two-hit lung injury models. A random effects model was used to integrate individual effect sizes calculated from each experiment. Classification models were built using the gene expression signatures generated by the meta-analysis to predict the development of lung injury in human lung transplant recipients. Two injury-specific lists of differentially expressed genes generated from our meta-analysis of lung injury models were validated using external data sets and prospective data from animal models of ventilator-induced lung injury (VILI). Pathway analysis of gene sets revealed that both new and previously implicated VILI-related pathways are enriched with differentially regulated genes. Classification model based on gene expression signatures identified in animal models of lung injury predicted development of primary graft failure (PGF) in lung transplant recipients with larger than 80% accuracy based upon injury profiles from transplant donors. We also found that better classifier performance can be achieved by using meta-analysis to identify differentially-expressed genes than using single study-based differential analysis. Taken together, our data suggests that microarray analysis of gene expression data allows for the detection of "injury" gene predictors that can classify lung injury samples and identify patients at risk for clinically relevant lung injury complications.


Rectal examination in patients with pain in the right lower quadrant of the abdomen.
To determine whether rectal examination provides any diagnostic information in patients admitted to hospital with pain in the right lower quadrant of the abdomen. Casualty officer or surgical registrar recorded symptoms and signs on admission on detailed forms. Final diagnosis was noted on discharge from hospital. District general hospital. 1204 Consecutive patients admitted to hospital with pain in the right lower quadrant of the abdomen as their major complaint; 1028 had a rectal examination on admission. Odds ratio for each symptom and sign related to final diagnosis. Results of multiple logistic regression analysis for acute appendicitis. Right sided rectal tenderness, present in 309 of those examined, was more common in patients with acute appendicitis (odds ratio 1.34, p less than 0.05). This odds ratio was considerably less than that for other clinical signs--namely, tenderness in the right lower quadrant (odds ratio 5.09), rebound tenderness (3.34), guarding (3.07), and muscular rigidity in the abdomen (5.03). In the logistic regression analysis of patients with acute appendicitis, when allowance was made for the presence or absence of rebound tenderness, rectal tenderness on the right lost its significance. Six patients had masses palpable rectally, of which three were palpable on abdominal examination; the other three patients had acute appendicitis. No other unexpected diagnoses were established, and no useful additional diagnostic information was obtained by routine rectal examination. If patients presenting with pain in the right lower quadrant of the abdomen are tested for rebound tenderness then rectal examination does not give any further diagnostic information.


Not all sequence tags are created equal: designing and validating sequence identification tags robust to indels.
Ligating adapters with unique synthetic oligonucleotide sequences (sequence tags) onto individual DNA samples before massively parallel sequencing is a popular and efficient way to obtain sequence data from many individual samples. Tag sequences should be numerous and sufficiently different to ensure sequencing, replication, and oligonucleotide synthesis errors do not cause tags to be unrecoverable or confused. However, many design approaches only protect against substitution errors during sequencing and extant tag sets contain too few tag sequences. We developed an open-source software package to validate sequence tags for conformance to two distance metrics and design sequence tags robust to indel and substitution errors. We use this software package to evaluate several commercial and non-commercial sequence tag sets, design several large sets (max(count) = 7,198) of edit metric sequence tags having different lengths and degrees of error correction, and integrate a subset of these edit metric tags to polymerase chain reaction (PCR) primers and sequencing adapters. We validate a subset of these edit metric tagged PCR primers and sequencing adapters by sequencing on several platforms and subsequent comparison to commercially available alternatives. We find that several commonly used sets of sequence tags or design methodologies used to produce sequence tags do not meet the minimum expectations of their underlying distance metric, and we find that PCR primers and sequencing adapters incorporating edit metric sequence tags designed by our software package perform as well as their commercial counterparts. We suggest that researchers evaluate sequence tags prior to use or evaluate tags that they have been using. The sequence tag sets we design improve on extant sets because they are large, valid across the set, and robust to the suite of substitution, insertion, and deletion errors affecting massively parallel sequencing workflows on all currently used platforms.


One cannot hear the density of a drum (and further aspects of isospectrality).
It is well known that certain pairs of planar domains have the same spectra of the Laplacian operator. We prove that these domains are still isospectral for a wider class of physical problems, including the cases of heterogeneous drums and quantum billiards in an external field. In particular we show that the isospectrality is preserved when the density or the potential is symmetric under reflections along the folding lines of the domain. These results are also confirmed numerically using the finite-difference method: We find that the pairs of numerical matrices obtained in the discretization are exactly isospectral up to machine precision.


SCOUR: a stepwise machine learning framework for predicting metabolite-dependent regulatory interactions.
The topology of metabolic networks is both well-studied and remarkably well-conserved across many species. The regulation of these networks, however, is much more poorly characterized, though it is known to be divergent across organisms-two characteristics that make it difficult to model metabolic networks accurately. While many computational methods have been built to unravel transcriptional regulation, there have been few approaches developed for systems-scale analysis and study of metabolic regulation. Here, we present a stepwise machine learning framework that applies established algorithms to identify regulatory interactions in metabolic systems based on metabolic data: stepwise classification of unknown regulation, or SCOUR. We evaluated our framework on both noiseless and noisy data, using several models of varying sizes and topologies to show that our approach is generalizable. We found that, when testing on data under the most realistic conditions (low sampling frequency and high noise), SCOUR could identify reaction fluxes controlled only by the concentration of a single metabolite (its primary substrate) with high accuracy. The positive predictive value (PPV) for identifying reactions controlled by the concentration of two metabolites ranged from 32 to 88% for noiseless data, 9.2 to 49% for either low sampling frequency/low noise or high sampling frequency/high noise data, and 6.6-27% for low sampling frequency/high noise data, with results typically sufficiently high for lab validation to be a practical endeavor. While the PPVs for reactions controlled by three metabolites were lower, they were still in most cases significantly better than random classification. SCOUR uses a novel approach to synthetically generate the training data needed to identify regulators of reaction fluxes in a given metabolic system, enabling metabolomics and fluxomics data to be leveraged for regulatory structure inference. By identifying and triaging the most likely candidate regulatory interactions, SCOUR can drastically reduce the amount of time needed to identify and experimentally validate metabolic regulatory interactions. As high-throughput experimental methods for testing these interactions are further developed, SCOUR will provide critical impact in the development of predictive metabolic models in new organisms and pathways.


Fibronectin modulates macrophage adhesion and FBGC formation: the role of RGD, PHSRN, and PRRARV domains.
To probe the role of human plasma fibronectin in modulating human blood-derived macrophage adhesion and fusion to form multinucleated foreign-body giant cells (FBGC), a series of biomimetic oligopeptides based on the functional structure of fibronectin was designed and synthesized. Peptides incorporated the RGD and PHSRN integrin-binding sequences from FIII-10 and FIII-9 modules, respectively, and the PRRARV sequence from the C-terminal heparin-binding domain, either alone or in combination. Peptides were immobilized onto a polyethyleneglycol-based polymer substrate. The following conclusions were reached. Fibronectin modulated macrophage adhesion and the extent (i.e., size) of FBGC formation on control surfaces in the presence of serum proteins. Macrophages adhered to all substrates with relatively subtle differences between adhesion mediated by RGD, PHSRN, PRRARV, or combinations thereof. beta1-integrin subunit was essential in macrophage adhesion to peptide-grafted networks in a receptor-peptide specific manner, whereas beta3-integrin subunit was less important. Macrophage adhesion to PRRARV was mediated primarily by the direct interaction with integrins. RGD or PHSRN alone did not provide an adequate substrate for macrophage fusion to form FBGCs. However, the PHSRN synergistic site and the RGD site in a single oligopeptide provided a substrate for FBGC formation that was statistically comparable to that on the positive control material in the presence of serum proteins. This response was highly dependent upon the relative orientation between RGD and PHSRN. PRRARV did not support FBGC formation. These results demonstrate the importance of fibronectin and, specifically, the synergy between RGD and PHSRN domains, in supporting macrophage fusion to form FBGCs.


Characterization of the mycobacterial acyl-CoA carboxylase holo complexes reveals their functional expansion into amino acid catabolism.
Biotin-mediated carboxylation of short-chain fatty acid coenzyme A esters is a key step in lipid biosynthesis that is carried out by multienzyme complexes to extend fatty acids by one methylene group. Pathogenic mycobacteria have an unusually high redundancy of carboxyltransferase genes and biotin carboxylase genes, creating multiple combinations of protein/protein complexes of unknown overall composition and functional readout. By combining pull-down assays with mass spectrometry, we identified nine binary protein/protein interactions and four validated holo acyl-coenzyme A carboxylase complexes. We investigated one of these--the AccD1-AccA1 complex from Mycobacterium tuberculosis with hitherto unknown physiological function. Using genetics, metabolomics and biochemistry we found that this complex is involved in branched amino-acid catabolism with methylcrotonyl coenzyme A as the substrate. We then determined its overall architecture by electron microscopy and found it to be a four-layered dodecameric arrangement that matches the overall dimensions of a distantly related methylcrotonyl coenzyme A holo complex. Our data argue in favor of distinct structural requirements for biotin-mediated γ-carboxylation of α-β unsaturated acid esters and will advance the categorization of acyl-coenzyme A carboxylase complexes. Knowledge about the underlying structural/functional relationships will be crucial to make the target category amenable for future biomedical applications.


A pure population of ectodermal cells derived from human embryonic stem cells.
Embryonic stem (ES) cells represent a unique cellular model to recapitulate in vitro early steps of embryonic development and an unlimited cellular source in therapy for many diseases, as well as targets for drug discovery and toxicology screens. Although previous studies have reported epidermal differentiation of mouse and human embryonic stem (huES) cells, the heterogeneity of the resulting cell culture impairs the evaluation of differentiated cells for cell therapy. We report here the reproducible isolation of a homogenous ectodermal cell population, IT1, from human ES cells. Like primary cells, IT1 cells remain homogenous over 15 passages, expand up to 60 population doublings, and then die through senescence. Accordingly, IT1 cells display a normal karyotype and a somatic cell cycle kinetics and do not produce teratoma in nude mice. The production of K14-expressing epithelial cells driven by p63 expression strengthens the ectodermal nature of IT1 cells. Since IT1 can be isolated from different huES cell lines, it may provide a ready source of ectodermal progenitors for the development of a toxicology cell model, new-drug-screening strategies, and cell therapy transplantation.


Conjectures on some curious connections among social status, calorie restriction, hunger, fatness, and longevity.
Many animal and human studies show counterintuitive effects of environmental influences on energy balance and life span. Relatively low social and/or economic status seems to be associated with and produce greater adiposity, and reduced provision (e.g., caloric restriction) of food produces greater longevity. We suggest that a unifying factor may be perceptions of the environment as "energetically insecure" and inhospitable to reproduction, which may in turn provoke adiposity-increasing and longevity-extending mechanisms. We elaborate on two main aspects of resources (or the perceptions thereof) on body weight and longevity. We first discuss the effects of social dominance on body weight regulation in human and animal models. Second, we examine models of the interactions between caloric restriction, body composition, and longevity. Finally, we put forth a relational model of the influences of differing environmental cues on body composition and longevity.


Coexistence of behavioural types in an aquatic top predator: a response to resource limitation?
Intra-population variation in behaviour unrelated to sex, size or age exists in a variety of species. The mechanisms behind behavioural diversification have only been partly understood, but density-dependent resource availability may play a crucial role. To explore the potential coexistence of different behavioural types within a natural fish population, we conducted a radio telemetry study, measuring habitat use and swimming activity patterns of pike (Esox lucius), a sit-and-wait predatory fish. Three behavioural types co-occurred in the study lake. While two types of fish only selected vegetated littoral habitats, the third type opportunistically used all habitats and increased its pelagic occurrence in response to decreasing resource biomasses. There were no differences in size, age or lifetime growth between the three behavioural types. However, habitat-opportunistic pike were substantially more active than the other two behavioural types, which is energetically costly. The identical growth rates exhibited by all behavioural types indicate that these higher activity costs of opportunistic behaviour were compensated for by increased prey consumption in the less favourable pelagic habitat resulting in approximately equal fitness of all pike groups. We conclude that behavioural diversification in habitat use and activity reduces intraspecific competition in preferred littoral habitats. This may facilitate the emergence of an ideal free distribution of pike along resource gradients.


Correlating Flavivirus virulence and levels of intrinsic disorder in shell proteins: protective roles vs. immune evasion.
Computational analyses revealed correlations between the intrinsic disorder propensity of shell proteins and case fatality rates (CFRs) among Flaviviruses and within at least two Flavivirus species, such as tick-borne encephalitis virus (TBEV) and dengue virus (DENV). The shell proteins analyzed in this study are capsid (C) and membrane (PrM, Pr, and M) proteins. The highest correlations can be found when regression analyses were conducted using Pr (Flavivirus: r(2) = 0.78, p < 0.01) or M (Flavivirus: r(2) = 0.91, p < 0.01) as an independent variable with C and CFR as co-explanatory and dependent variables, respectively. Interestingly, while predicted intrinsic disorder levels (PIDs) of both C and M are positively correlated with the virulence, the PIDs of Pr and CFR are negatively correlated. This is likely due to the fact that the Pr portion of PrM plays various roles in protecting the virion from damage, whereas M and C are assisted by greater potential in binding promiscuity as a result of greater disorder. The C protein of yellow fever virus (YFV), which is the most virulent virus in the sample, has the highest PID levels, whereas the second most virulent TBEV FE subtype has the second highest PID score due to its C protein, and the least virulent West Nile virus (WNV) has the least disordered C protein. This knowledge can be used while working on the development and identification of attenuated strains for vaccine. Curiously, unlike Flaviviruses, a disordered outer shell was described for hepatitis C virus (HCV), human immunodeficiency virus (HIV), and human simplex virus 2 (HSV-2), which currently have no effective vaccine.


The influence of eyelid morphology on normal corneal shape.
To investigate associations between the topography of the cornea and the morphology of the eyelids in a population of young adult subjects with a range of refractive errors. Corneal topography data and digital images of the anterior eye were acquired for 100 young adult subjects. The corneal topography data and palpebral fissure images were analyzed to determine a range of parameters describing the shape of the cornea and the morphology of the palpebral fissure. Correlation analysis was carried out to investigate for significant associations between the parameters describing the topography of the cornea and the parameters describing the morphology of the palpebral fissure. A number of highly significant correlations were found between the best-fitting corneal spherocylinder and the eyelid morphology parameters. The corneal best-fit sphere (M) was significantly correlated with the horizontal palpebral fissure width (r = -0.428; P < 0.001). Corneal astigmatism power vector J45 was significantly correlated with the angle of the palpebral fissure (r = 0.392; P < 0.001). The axis of corneal astigmatism was also found to be significantly correlated with the angle of the palpebral fissure (r = 0.317; P = 0.005). A number of significant associations exist between the corneal spherocylinder and the morphology of the eyelids in a normal adult population.


Alteration of drug metabolizing enzymes in sulphite oxidase deficiency.
The aim of this study was to investigate the possible effects of sulphite oxidase (SOX, E.C. 126.96.36.199) deficiency on xenobiotic metabolism. For this purpose, SOX deficiency was produced in rats by the administration of a low molybdenum diet with concurrent addition of 200 ppm tungsten to their drinking water. First, hepatic SOX activity in deficient groups was measured to confirm SOX deficiency. Then, aminopyrine N-demethylase, aniline 4-hydroxylase, aromatase, caffeine N-demethylase, cytochrome b5 reductase, erythromycin N-demethylase, ethoxyresorufin O-deethylase, glutathione S-transferase, N-nitrosodimethylamine N-demethylase and penthoxyresorufin O-deethylase activities were determined to follow changes in the activity of drug metabolizing enzymes in SOX-deficient rats. Our results clearly demonstrated that SOX deficiency significantly elevated A4H, caffeine N-demethylase, erythromycin N-demethylase and N-nitrosodimethylamine N-demethylase activities while decreasing ethoxyresorufin O-deethylase and aromatase activities. These alterations in drug metabolizing enzymes can contribute to the varying susceptibility and response of sulphite-sensitive individuals to different drugs and/or therapeutics used for treatments.


High beta rhythm amplitude in olfactory learning signs a well-consolidated and non-flexible behavioral state.
Beta rhythm (15-30 Hz) is a major candidate underlying long-range communication in the brain. In olfactory tasks, beta activity is strongly modulated by learning but its condition of expression and the network(s) responsible for its generation are unclear. Here we analyzed the emergence of beta activity in local field potentials recorded from olfactory, sensorimotor and limbic structures of rats performing an olfactory task. Rats performed successively simple discrimination, rule transfer, memory recall tests and contingency reversal. Beta rhythm amplitude progressively increased over learning in most recorded areas. Beta amplitude reduced to baseline when new odors were introduced, but remained high during memory recall. Intra-session analysis showed that even expert rats required several trials to reach a good performance level, with beta rhythm amplitude increasing in parallel. Notably, at the beginning of the reversal task, beta amplitude remained high while performance was low and, in all tested animals, beta amplitude decreased before rats were able to learn the new contingencies. Connectivity analysis showed that beta activity was highly coherent between all structures where it was expressed. Overall, our results suggest that beta rhythm is expressed in a highly coherent network when context learning - including both odors and reward - is consolidated and signals behavioral inflexibility.


Staphylococcus aureus sortase A contributes to the Trojan horse mechanism of immune defense evasion with its intrinsic resistance to Cys184 oxidation.
Staphylococcus aureus is a Gram-positive bacterial pathogen that causes serious infections which have become increasingly difficult to treat due to antimicrobial resistance and natural virulence strategies. Bacterial sortase enzymes are important virulence factors and good targets for future antibiotic development. It has recently been shown that sortase enzymes are integral to bacterial survival of phagocytosis, an underappreciated, but vital, step in S. aureus pathogenesis. Of note, the reaction mechanism of sortases relies on a solvent-accessible cysteine for transpeptidation. Because of the common strategy of oxidative damage employed by professional phagocytes to kill pathogens, it is possible that this cysteine may be oxidized inside the phagosome, thereby inhibiting the enzyme. This study addresses this apparent paradox by assessing the ability of physiological reactive oxygen species, hydrogen peroxide and hypochlorite, to inhibit sortase A (SrtA) from S. aureus. Surprisingly, we found that SrtA is highly resistant to oxidative inhibition, both in vitro and in vivo. The mechanism of resistance to oxidative damage is likely mediated by maintaining a high reduction potential of the catalytic cysteine residue, Cys184. This is due to the unusual active site utilized by S. aureus SrtA, which employs a reverse protonation mechanism for transpeptidation, resulting in a high pK(a) as well as reduction potential for Cys184. The results of this study suggest that S. aureus SrtA is able to withstand the extreme conditions encountered in the phagosome and maintain function, contributing to survival of phagocytotic killing.


Comparative analysis of carotenoid accumulation in two goji (Lycium barbarum L. and L. ruthenicum Murr.) fruits.
The traditional Chinese medicinal plants Lycium barbarum L. and L. ruthenicum Murr. are valued for the abundance of bioactive carotenoids and anthocyanins in their fruits, respectively. However, the cellular and molecular mechanisms contributing to their species-specific bioactive profiles remain poorly understood. In this study, the red fruit (RF) of L. barbarum was found to accumulate high levels of carotenoids (primarily zeaxanthin), while they were undetectable in the black fruit (BF) of L. ruthenicum. Cytological and gene transcriptional analyses revealed that the chromoplast differentiation that occurs in the chloroplast during fruit ripening only occurs in RF, indicating that the lack of chromoplast biogenesis in BF leads to no sink for carotenoid storage and the failure to synthesize carotenoids. Similar enzyme activities of phytoene synthase 1 (PSY1), chromoplast-specific lycopene β-cyclase (CYC-B) and β-carotene hydroxylase 2 (CRTR-B2) were observed in both L. ruthenicum and L. barbarum, suggesting that the undetectable carotenoid levels in BF were not due to the inactivation of carotenoid biosynthetic enzymes. The transcript levels of the carotenoid biosynthetic genes, particularly PSY1, phytoene desaturase (PDS), ζ-carotene desaturase (ZDS), CYC-B and CRTR-B2, were greatly increased during RF ripening, indicating increased zeaxanthin biosynthesis. Additionally, carotenoid cleavage dioxygenase 4 (CCD4) was expressed at much higher levels in BF than in RF, suggesting continuous carotenoid degradation in BF. The failure of the chromoplast development in BF causes low carotenoid biosynthesis levels and continuous carotenoid degradation, which ultimately leads to undetectable carotenoid levels in ripe BF. In contrast, the successful chromoplast biogenesis in RF furnishes the sink necessary for carotenoid storage. Based on this observation, the abundant zeaxanthin accumulation in RF is primarily determined via both the large carotenoid biosynthesis levels and the lack of carotenoid degradation, which are regulated at the transcriptional level.


MR in temporal lobe epilepsy: analysis with pathologic confirmation.
We evaluated the MR findings in patients with temporal lobe epilepsy to determine the predictive value of MR imaging in assessing patient outcome. MR studies from 186 of 274 consecutive patients who underwent temporal lobectomy for intractable epilepsy were reviewed retrospectively. Images were interpreted by an experienced neuroradiologist, who was blinded to the side of seizure activity and to pathologic findings. MR imaging exhibited 93% sensitivity and 83% specificity in detecting hippocampal/amygdalar abnormalities (n = 121), and 97% sensitivity and 97% specificity in detecting abnormalities in the rest of the temporal lobe (n = 60). Abnormal high signal of the hippocampus on T2-weighted images had a sensitivity of 93% and specificity of 74% in predicting mesial temporal sclerosis (n = 115). The presence of hippocampal atrophy on MR correlated with the duration of seizures. Sensitivity and specificity of MR imaging in detecting temporal lobe tumors (n = 42) were 83% and 97%, respectively, based on abnormal signal and mass effect. After surgery, 63% of patients were seizure free and 28% had a significant reduction of seizure frequency at an average of 24 months (range, 12 to 78 months) after surgery. Patients with a single lesion in the anterior temporal lobe or hippocampus/amygdala had a better outcome than patients with multiple lesions (n = 22). Interrater agreement varied from 0.4 to 0.93, with best agreement for tumors or abnormal hippocampal signal on T2-weighted images. MR imaging is highly sensitive in detecting and locating abnormalities in the temporal lobe and the hippocampus/amygdala in patients with temporal lobe epilepsy. Hippocampal atrophy appears to correspond to the duration of seizure disorder.


[Tension attributes of the family caregiver of frail older adults].
The objective of this study was to assess both theoretically and empirically the tension attributes of the family caregiver of frail older adults. The theoretical phase consisted of analyzing 52 studies addressing this issue. The analysis showed that the phenomenon is evidenced by the caregiver through physical and emotional changes, unbalance between activity and rest, and compromised individual coping. In the field phase, an assessment was made of 30 women who, during the nursing consultation, showed evidence of tension due to their role as caregiver. Data collection was performed through interviews at home, which were recorded and followed a script created based on the theoretical findings. Every time it was considered adequate, excerpts from the caregivers' discourses were added to improve understanding. The empirical assessment confirmed the tension attributes revealed in the theoretical analysis.


Pharmacogenomics Study for Raloxifene in Postmenopausal Female with Osteoporosis.
Osteoporosis is characterized by decreased bone mineral density and increased risk of fracture. Raloxifene is one of the treatments of osteoporosis. However, the responses were variable among patients. Previous studies revealed that the genetic variants are involved in the regulation of treatment outcomes. To date, studies that evaluate the influence of genes across all genome on the raloxifene treatment response are still limited. In this study, a total of 41 postmenopausal osteoporosis patients under regular raloxifene treatment were included. Gene-based analysis using MAGMA was applied to investigate the genetic association with the bone mineral density response to raloxifene at the lumbar spine or femoral neck site. Results from gene-based analysis indicated several genes (GHRHR, ABHD8, and TMPRSS6) related to the responses of raloxifene. Besides, the pathways of iron ion homeostasis, osteoblast differentiation, and platelet morphogenesis were enriched which implies that these pathways might be relatively susceptible to raloxifene treatment outcome. Our study provided a novel insight into the response to raloxifene.


Outcomes of haploidentical vs matched sibling transplantation for acute myeloid leukemia in first complete remission.
HLA-haploidentical hematopoietic cell transplantation (Haplo-HCT) using posttransplantation cyclophosphamide (PT-Cy) has improved donor availability. However, a matched sibling donor (MSD) is still considered the optimal donor. Using the Center for International Blood and Marrow Transplant Research database, we compared outcomes after Haplo-HCT vs MSD in patients with acute myeloid leukemia (AML) in first complete remission (CR1). Data from 1205 adult CR1 AML patients (2008-2015) were analyzed. A total of 336 patients underwent PT-Cy-based Haplo-HCT and 869 underwent MSD using calcineurin inhibitor-based graft-versus-host disease (GVHD) prophylaxis. The Haplo-HCT group included more reduced-intensity conditioning (65% vs 30%) and bone marrow grafts (62% vs 7%), consistent with current practice. In multivariable analysis, Haplo-HCT and MSD groups were not different with regard to overall survival (P = .15), leukemia-free survival (P = .50), nonrelapse mortality (P = .16), relapse (P = .90), or grade II-IV acute GVHD (P = .98). However, the Haplo-HCT group had a significantly lower rate of chronic GVHD (hazard ratio, 0.38; 95% confidence interval, 0.30-0.48; P < .001). Results of subgroup analyses by conditioning intensity and graft source suggested that the reduced incidence of chronic GVHD in Haplo-HCT is not limited to a specific graft source or conditioning intensity. Center effect and minimal residual disease-donor type interaction were not predictors of outcome. Our results indicate a lower rate of chronic GVHD after PT-Cy-based Haplo-HCT vs MSD using calcineurin inhibitor-based GVHD prophylaxis, but similar other outcomes, in patients with AML in CR1. Haplo-HCT is a viable alternative to MSD in these patients.


The petroleum ether extract of Nigella sativa exerts lipid-lowering and insulin-sensitizing actions in the rat.
We studied the effect of a 4-week intragastric gavage with a petroleum ether extract of Nigella sativa seeds on blood glucose, insulin and lipids in the normal rat. Petroleum ether extract caused a 25% reduction in food intake that translated into a transient weight loss. No sign of toxicity of the plant could be seen in vivo or in vitro. Fasting plasma glucose remained stable throughout Nigella sativa treatment. At the end of the 4-week treatment, Nigella sativa-treated rats had lower fasting plasma levels of insulin and triglycerides, and higher HDL-cholesterol as compared to pair-fed controls. Response to insulin was evaluated in hepatocytes isolated from animals of all groups by Western blot analysis of phosphorylated MAPK p44/42erk and PKB. In vivo Nigella sativa treatment resulted in greater dose-dependent activation of MAPK and PKB in response to insulin. These results suggest that the petroleum ether extract of Nigella sativa has a slight anorexic effect, and that it contains the hypolipidemic activity previously obtained with the plant. More significantly, our data demonstrate that in vivo treatment with the petroleum ether extract exerts an insulin-sensitizing action by enhancing the activity of the two major intracellular signal transduction pathways of the hormone's receptor.


Overhauser Dynamic Nuclear Polarization-Enhanced NMR Relaxometry.
We present a new methodological basis for selectively illuminating a dilute population of fluid within a porous medium. Specifically, transport in porous materials can be analyzed by now-standard nuclear magnetic resonance (NMR) relaxometry and NMR pulsed field gradient (PFG) diffusometry methods in combination with with the prominent NMR signal amplification tool, dynamic nuclear polarization (DNP). The key components of the approach introduced here are (1) to selectively place intrinsic or extrinsic paramagnetic probes at the site or local volume of interest within the sample, (2) to amplify the signal from the local solvent around the paramagnetic probes with Overhauser DNP, which is performed in situ and under ambient conditions, and (3) to observe the ODNP-enhanced solvent signal with 1D or 2D NMR relaxometry methods, thus selectively amplifying only the relaxation dynamics of the fluid that resides in or percolates through the local porous volume that contains the paramagnetic probe. Here, we demonstrate the proof of principle of this approach by selectively amplifying the NMR signal of only one solvent population, which is in contact with a paramagnetic probe and occluded from a second solvent population. An apparent one-component T2 relaxation decay is shown to actually contain two distinct solvent populations. The approach outlined here should be universally applicable to a wide range of other 1D and 2D relaxometry and PFG diffusometry measurements, including T1-T2 or T1-D correlation maps, where the occluded population containing the paramagnetic probes can be selectively amplified for its enhanced characterization.


[Incidence and nature of thoracic pains in outpatients (author's transl)].
The incidence of different types of thoracic pain in outpatients at the Medical University Policlinic, Basel, was obtained for comparison from a random sample of the Swiss population. The data from the Rose questionnaire which was used were compared with the case histories. Of 515 patients, 233 admitted having thoracic pains. These complaints were the reason for consulting the doctor in 71 patients. Of all patients who admitted thoracic pains, 14% complained of typical ischemic pains and 60% of unspecific thoracic pains. Perusal of the case histories showed that the doctor was only able to distinguish between coronary heart disease, other organic causes and functional complaints after selective history taking and clinical examination in 89% angina pectoris and 74% unspecific thoracic pains.


Attention Deficit Hyperactivity Disorder (ADHD) among longer-term prison inmates is a prevalent, persistent and disabling disorder.
ADHD is a common and disabling disorder, with an increased risk for coexisting disorders, substance abuse and delinquency. In the present study, we aimed at exploring ADHD and criminality. We estimated the prevalence of ADHD among longer-term prison inmates, described symptoms and cognitive functioning, and compared findings with ADHD among psychiatric outpatients and healthy controls. At Norrtälje Prison, we approached 315 male inmates for screening of childhood ADHD by the Wender Utah Rating Scale (WURS-25) and for present ADHD by the Adult ADHD Self-Report Screener (ASRS-Screener). The response rate was 62%. Further, we assessed 34 inmates for ADHD and coexisting disorders. Finally, we compared findings with 20 adult males with ADHD, assessed at a psychiatric outpatient clinic and 18 healthy controls. The estimated prevalence of adult ADHD among longer-term inmates was 40%. Only 2 out of 30 prison inmates confirmed with ADHD had received a diagnosis of ADHD during childhood, despite most needed health services and educational support. All subjects reported lifetime substance use disorder (SUD) where amphetamine was the most common drug. Mood and anxiety disorders were present among half of subjects; autism spectrum disorder (ASD) among one fourth and psychopathy among one tenth. Personality disorders were common; almost all inmates presented conduct disorder (CD) before antisocial personality disorder (APD). Prison inmates reported more ADHD symptoms during both childhood and adulthood, compared with ADHD psychiatric outpatients. Further, analysis of executive functions after controlling for IQ showed both ADHD groups performed poorer than controls on working memory tests. Besides, on a continuous performance test, the ADHD prison group displayed poorer results compared with both other groups. This study suggested ADHD to be present among 40% of adult male longer-term prison inmates. Further, ADHD and coexisting disorders, such as SUD, ASD, personality disorders, mood- and anxiety disorders, severely affected prison inmates with ADHD. Besides, inmates showed poorer executive functions also when controlling for estimated IQ compared with ADHD among psychiatric outpatients and controls. Our findings imply the need for considering these severities when designing treatment programmes for prison inmates with ADHD.


Conversion, correction, and International Scale standardization: results From a Multicenter External Quality Assessment Study for BCR-ABL1 testing.
Monitoring BCR-ABL1 expression levels relative to clinically validated response criteria on the International Scale (IS) is vital in the optimal management of patients with chronic myeloid leukemia, yet significant variability remains across laboratories worldwide. To assess method performance, interlaboratory precision, and different IS standardization modalities in representative laboratories performing routine BCR-ABL1 testing. Fifteen blinded test specimens with 5-level nominal BCR-ABL1 to ABL1 IS percentage ratios ranging from 5% to 0.0005% and 4-level secondary IS reference panels, the ARQ IS Calibrator Panels, were tested by relative quantitative polymerase chain reaction in 15 laboratories in 5 countries. Both raw and IS percentage ratios calculated by using local conversion factors (CFs) or analytic correction parameters (CPs) were collected and analyzed. A total of 670 valid positive results were generated. BCR-ABL1 detection was associated with variable ABL1 quality metric passing rates (P < .001) and reached at least 0.01% in 13 laboratories. Intralaboratory precision was within 2.5-fold for all sample levels combined with a relative mean difference greater than 5-fold across laboratories. International Scale accuracy was increased by using both the CF and CP standardization methods. Classification agreement for major molecular response status was 90% after CF conversion and 93% after CP correction, with precision improved by 3-fold for the CP method. Despite preanalytic and analytic differences between laboratories, conversion and correction are effective IS standardization methods. Validated secondary reference materials can facilitate global diffusion of the IS without the need to perform sample exchange and improve the accuracy and precision of BCR-ABL1 quantitative measurements, including at low levels of residual disease.


Chronic ITP: analysis of various factors at presentation which predict failure to first line treatment and their response to second line therapy.
To observe the significance of various factors in chronic idiopathic thrombocytopenic purpura (ITP) which predict the response of first line (corticosteroids) and second line therapy (splenectomy) and to evaluate their response to second line therapy. This was a descriptive, prospective study conducted from August 2004 till January 2006. Patients of all age groups and both genders with diagnosis of chronic ITP were included. Treatment protocol and criteria for response assessment was explained. During 17 months period, 86 patients with chronic ITP were analyzed. Non-responders to first line therapy were 74 patients who ultimately required splenectomy. Complete response (CR) was had in 37 (50.7%) patients, 10 (13.7%) and 27 (36.5%) had partial response (PR) and no response (NR) respectively. Analysis of variables like younger age, sex and low platelet count at presentation failed to show any significant influence on response to first line treatment. However response to splenectomy was found to be higher in patients who had initial complete or partial response with steroids and later relapsed and the platelet count was more than 300x10(9)/L on day 14 of surgery. Splenectomy remains the most effective treatment of chronic ITP. No significant factor was identified which predicted initial response to first line treatment. However patients who initially responded to steroids and had platelet counts above 300 X109/L about a fortnight after splenectomy showed promising results post-operatively (p=0.003 and p=0.001).


Effect of childhood victimization on occupational prestige and income trajectories.
Violence toward children (childhood victimization) is a major public health problem, with long-term consequences on economic well-being. The purpose of this study was to determine whether childhood victimization affects occupational prestige and income in young adulthood. We hypothesized that young adults who experienced more childhood victimizations would have less prestigious jobs and lower incomes relative to those with no victimization history. We also explored the pathways in which childhood victimization mediates the relationships between background variables, such as parent's educational impact on the socioeconomic transition into adulthood. A nationally representative sample of 8,901 young adults aged 18-28 surveyed between 1999-2009 from the National Longitudinal Survey of Youth 1997 (NLSY) were analyzed. Covariate-adjusted multivariate linear regression and path models were used to estimate the effects of victimization and covariates on income and prestige levels and on income and prestige trajectories. After each participant turned 18, their annual 2002 Census job code was assigned a yearly prestige score based on the 1989 General Social Survey, and their annual income was calculated via self-reports. Occupational prestige and annual income are time-varying variables measured from 1999-2009. Victimization effects were tested for moderation by sex, race, and ethnicity in the multivariate models. Approximately half of our sample reported at least one instance of childhood victimization before the age of 18. Major findings include 1) childhood victimization resulted in slower income and prestige growth over time, and 2) mediation analyses suggested that this slower prestige and earnings arose because victims did not get the same amount of education as non-victims. Results indicated that the consequences of victimization negatively affected economic success throughout young adulthood, primarily by slowing the growth in prosperity due to lower education levels.


The Atypical Antipsychotic Agent, Clozapine, Protects Against Corticosterone-Induced Death of PC12 Cells by Regulating the Akt/FoxO3a Signaling Pathway.
Schizophrenia is one of the most severe psychiatric disorders. Increasing evidence implicates that neurodegeneration is a component of schizophrenia pathology and some atypical antipsychotics are neuroprotective and successful in slowing the progressive morphological brain changes. As an antipsychotic agent, clozapine has superior and unique effects, but the intracellular signaling pathways that mediate clozapine action remain to be elucidated. The phosphatidylinositol-3-kinase/protein kinase B/Forkhead box O3 (PI3K/Akt/FoxO3a) pathway is crucial for neuronal survival. However, little information is available regarding this pathway with clozapine. In the present study, we investigated the protective effect of clozapine on the PC12 cells against corticosterone toxicity. Our results showed that corticosterone decreases the phosphorylation of Akt and FoxO3a, leading to the nuclear localization of FoxO3a and the apoptosis of PC12 cells, while clozapine concentration dependently protected PC12 cells against corticosterone insult. Pathway inhibitors studies displayed that the protective effect of clozapine was reversed by LY294002 and wortmannin, two PI3K inhibitors, or Akt inhibitor VIII although several other inhibitors had no effect. The shRNA knockdown results displayed that downregulated Akt1 or FoxO3a attenuated the protective effect of clozapine. Western blot analyses revealed that clozapine induced the phosphorylation of Akt and FoxO3a by the PI3K/Akt pathway and reversed the reduction of the phosphorylated Akt and FoxO3a and the nuclear translocation of FoxO3a evoked by corticosterone. Together, our data indicates that clozapine protects PC12 cells against corticosterone-induced cell death by modulating activity of the PI3K/Akt/FoxO3a pathway.


Same-day antiretroviral therapy initiation for people living with HIV who have tuberculosis symptoms: a systematic review.
Tuberculosis symptoms are very common among people living with HIV (PLHIV) initiating antiretroviral therapy (ART), are not specific for tuberculosis disease and may result in delayed ART start. The risks and benefits of same-day ART initiation in PLHIV with tuberculosis symptoms are unknown. We systematically reviewed nine databases on 12 March 2020 to identify studies that investigated same-day ART initiation among PLHIV with tuberculosis symptoms and reported both their approach to TB screening and clinical outcomes. We extracted and summarized data about TB screening, numbers of people starting same-day ART and outcomes. We included four studies. Two studies deferred ART for everyone with any tuberculosis symptoms (one or more of cough, fever, night sweats or weight loss) and substantial numbers of people had deferred ART start (28% and 39% did not start same-day ART). Two studies permitted some people with tuberculosis symptoms to start same-day ART, and fewer people deferred ART (2% and 16% did not start same-day). Two of the four studies were conducted sequentially; proven viral load suppression at 8 months was 31% when everyone with tuberculosis symptoms had ART deferred, and 44% when the algorithm was changed so that some people with tuberculosis symptoms could start same-day ART. Although tuberculosis symptoms are very common in people starting ART, there is insufficient evidence about whether presence of tuberculosis symptoms should lead to ART start being deferred or not. Research to inform clear guidelines would help to maximise the benefits of same-day ART.


Efficacy of Agnikarma over the padakanistakam (little toe) and Katibasti in Gridhrasi: A comparative study.
Gridhrasi (Sciatica) is one of the Vatavyadhi which is caused by aggravated Vata dosha. This disease is characterized by ruja (pain) in the waist, back, thigh, knee and calf regions along the course of sciatic nerve. In spite of the different types of treatment modalities mentioned in ancient and modern medical sciences, they have some or the other shortcomings and drawbacks. Considering all these, the present study was taken up with the objective of evaluating the efficacy of Agnikarma (treatment done with cauterization) over the padakanistakam (little toe) in the management of Gridhrasi. To consider the significance of the method of Agnikarma, the efficacy of Katibasti in the management of Gridhrasi which has been established in the previous work was also studied. The study was performed after obtaining Ethics Committee approval and patients' written informed consent. Forty cases presenting with classical features of Gridhrasi (Sciatica) due to lumbar intervertebral disc prolapse were selected. The management of Gridhrasi by Agnikarma and Katibasti was conducted by including the patients in two groups, namely Group A (study group) and Group B (control group). The data were collected and the observations were made before the treatment, on 8(th) day, 15(th) day and on 22(nd) day of the treatment. The data obtained from the results were subjected for statistical analysis and conclusions were drawn. There was a significant reduction in the parameters, pain (P < 0.01) and straight-leg raising (SLR) test (P < 0.01), of the study group compared to the control group (P < 0.01). Pain was assessed through Numerical Pain Analogue Scale. After the treatment with Agnikarma, the pain was totally relieved in 80% of cases. It was reduced to moderate degree in 20% of cases and in 95% of cases, SLR test became negative. After the treatment with Katibasti, the pain was totally relieved in 50% of cases. It was reduced to moderate degree in 20% and to mild degree in 25% of cases. In 60% of cases, SLR test became negative. However, changes in the radiological findings were not found in both the methods of management. Analysis of overall effect of treatment in the present study reveals that Agnikarma was Both the procedures were conducted in to that of Katibasti. The management of Gridhrasi by Agnikarma was more efficacious as compared with Katibasti in reducing pain. However, there were no radiological changes produced by both the methods of treatment. Further studies may be conducted by future scholars by taking more samples with more number of sittings.


Classification using belief functions: relationship between case-based and model-based approaches.
The transferable belief model (TBM) is a model to represent quantified uncertainties based on belief functions, unrelated to any underlying probability model. In this framework, two main approaches to pattern classification have been developed: the TBM model-based classifier, relying on the general Bayesian theorem (GBT), and the TBM case-based classifier, built on the concept of similarity of a pattern to be classified with training patterns. Until now, these two methods seemed unrelated, and their connection with standard classification methods was unclear. This paper shows that both methods actually proceed from the same underlying principle, i.e., the GBT, and that they essentially differ by the nature of the assumed available information. This paper also shows that both methods collapse to a kernel rule in the case of precise and categorical learning data and for certain initial assumptions, and a simple relationship between basic belief assignments produced by the two methods is exhibited in a special case. These results shed new light on the issues of classification and supervised learning in the TBM. They also suggest new research directions and may help users in selecting the most appropriate method for each particular application, depending on the nature of the information at hand.


Immunity and squamous cell carcinoma of the anus: epidemiological, clinical and therapeutic aspects.
Squamous cell carcinoma of the anus (SCCA) is a rare disease, but its incidence has been increasing dramatically since the 1970s. Men who have sex with men (MSM) and infection with human immunodeficiency virus (HIV) are the two main risk factors. Risk of developing SCCA is increased more than 100-fold in HIV-seropositive MSM. We review here how immunodeficiency could promote SCCA and how restoration of immunity since the advent of combined antiretroviral therapy can influence the natural history and incidence of SCCA. We also review the prognostic and therapeutic implications of immunosuppression in these patients. Finally, we show how, with anti-HPV vaccines, immunity is a target in the prevention of SCCA and could in the future be used in its treatment.


Fungal Contaminants in Energy Efficient Dwellings: Impact of Ventilation Type and Level of Urbanization.
The presence of growing fungi in the indoor environment has been associated with the development of respiratory problems such as asthma or allergic rhinitis, as well as the worsening of respiratory pathologies. Their proliferation indoors could be a result of water leakage or inadequate ventilation. Although the factors promoting mould growth have been widely investigated in traditional dwellings, little work has been done in energy efficient dwellings. Here, the effectiveness of ventilation type, i.e., natural or mechanical, in influencing mould development was estimated in 44 recent and 105 retrofitted energy efficient dwellings. Fungi growing on surfaces were investigated in the dwellings situated in rural, peri-urban, and urban regions of Switzerland. The presence of these fungi was also investigated in bedroom settled dust. Information on building characteristics and owners' lifestyle were collected. Significant associations were found with the level of urbanisation, the location of mouldy area in dwellings, and the diversity of fungal taxa. Dwellings in peri-urban zones showed the most frequent fungal contamination in the owners' bedroom and the highest diversity of fungal genera among dwellings. While the urbanisation level or the ventilation type favoured no specific genus, we found marked disparities in the diversity of fungi growing on surfaces in naturally ventilated versus mechanically ventilated dwellings. Aspergillus, in particular, was a frequent surface contaminant in bedrooms with natural ventilation, but not in those mechanically ventilated. We observed a strong association between fungal growth on surfaces and the number of fungal particles counted in the settled dust of owners' bedrooms. These results demonstrate the importance of ventilation systems in energy efficient dwellings in controlling fungal proliferation in living areas.


Dimensions of the endometrial cavity and intrauterine device expulsion or removal for displacement: a nested case-control study.
To evaluate the effect of the dimensions of the uterine cavity in relation to the expulsion, or removal for displacement, of intrauterine devices (IUDs) MLCu375 and TCu380A. A case-control study nested in a multicentre clinical trial. Eighteen family planning clinics in China. Forty-eight pairs of TCu380A users and 118 pairs of MLCu375 users. The women were classified as cases if IUD expulsion or displacement occurred during the first year of follow-up after insertion. One control was randomly selected for each case matched by IUD model, centre, age, service provider for insertion, and date of insertion. Axial length of the uterine cavity (LUC) and the largest transverse diameter of the coronal section of the uterine cavity (LTD) were measured using abdominal ultrasound. We used multivariate conditional logistic regressions to estimate the adjusted odds ratios (aORs) of expulsions or displacements among women with different sizes of uterine cavity according to the quartile of LUC and LTD, respectively. Among MLCu375 users, women with LTD ≥ 27 mm had a higher risk of expulsion or displacement (aOR 2.40; 95% confidence interval, 95% CI, 1.02-5.63), after adjusting for the volume of menstrual flow, dysmenorrhoea, parity, uterine position, MLCu375 type, and LUC. Among TCu380A users, the association between LTD ≥ 37 mm and expulsion or displacement (aOR 4.98; 95% CI 1.01-22.49) was statistically significant, after adjusting for LUC and potential confounding factors. Our study suggests that LTD should be considered when making the decision of which IUD model to use.


Accuracy of non-invasive measurement of haemoglobin concentration by pulse co-oximetry during steady-state and dynamic conditions in liver surgery.
The Masimo Radical 7 (Masimo Corp., Irvine, CA, USA) pulse co-oximeter(®) calculates haemoglobin concentration (SpHb) non-invasively using transcutaneous spectrophotometry. We compared SpHb with invasive satellite-lab haemoglobin monitoring (Hb(satlab)) during major hepatic resections both under steady-state conditions and in a dynamic phase with fluid administration of crystalloid and colloid solutions. Thirty patients undergoing major hepatic resection were included and randomized to receive a fluid bolus of 15 ml kg(-1) colloid (n=15) or crystalloid (n=15) solution over 30 min. SpHb was continuously measured on the index finger, and venous blood samples were analysed in both the steady-state phase (from induction until completion of parenchymal transection) and the dynamic phase (during fluid bolus). Correlation was significant between SpHb and Hb(satlab) (R(2)=0.50, n=543). The modified Bland-Altman analysis for repeated measurements showed a bias (precision) of -0.27 (1.06) and -0.02 (1.07) g dl(-1) for the steady-state and dynamic phases, respectively. SpHb accuracy increased when Hb(satlab) was <10 g dl(-1), with a bias (precision) of 0.41 (0.47) vs -0.26 (1.12) g dl(-1) for values >10 g dl(-1), but accuracy decreased after colloid administration (R(2)=0.25). SpHb correlated moderately with Hb(satlab) with a slight underestimation in both phases in patients undergoing major hepatic resection. Accuracy increased for lower Hb(satlab) values but decreased in the presence of colloid solution. Further improvements are necessary to improve device accuracy under these conditions, so that SpHb might become a sensitive screening device for clinically significant anaemia.


Expression of the LIM-homeodomain protein Isl1 in the developing and mature mouse retina.
The mammalian retina is comprised of six major neuronal cell types and is subdivided into more morphological and physiological subtypes. The transcriptional machinery underlying these subtype fate choices is largely unknown. The LIM-homeodomain protein, Isl1, plays an essential role in central nervous system (CNS) differentiation but its relationship to retinal neurogenesis remains unknown. We report here its dynamic spatiotemporal expression in the mouse retina. Among bipolar interneurons, Isl1 expression commences at postnatal day (P)5 and is later restricted to ON-bipolar cells. The intensity of Isl1 expression is found to segregate the pool of ON-bipolar cells into rod and ON-cone bipolar cells with higher expression in rod bipolar cells. As bipolar cell development proceeds from P5-10 the colocalization of Isl1 and the pan-bipolar cell marker Chx10 reveals the organization of ON-center bipolar cell nuclei to the upper portion of the inner nuclear layer. Further, whereas Isl1 is predominantly a ganglion cell marker prior to embryonic day (E)15.5, at E15.5 and later its expression in nonganglion cells expands. We demonstrate that these Isl1-positive, nonganglion cells acquire the expression of amacrine cell markers embryonically, likely representing nascent cholinergic amacrine cells. Taken together, Isl1 is expressed during the maturation of and is later maintained in retinal ganglion cells and subtypes of amacrine and bipolar cells where it may function in the maintenance of these cells into adulthood.


Learning before leaping: integration of an adaptive study design process prior to initiation of BetterBirth, a large-scale randomized controlled trial in Uttar Pradesh, India.
Pragmatic and adaptive trial designs are increasingly used in quality improvement (QI) interventions to provide the strongest evidence for effective implementation and impact prior to broader scale-up. We previously showed that an on-site coaching intervention focused on the World Health Organization Safe Childbirth Checklist (SCC) improved performance of essential birth practices (EBPs) in one facility in Karnataka, India. We report on the process and outcomes of adapting the intervention prior to larger-scale implementation in a randomized controlled trial in Uttar Pradesh (UP), India. Initially, we trained a local team of physicians and nurses to coach birth attendants in SCC use at two public facilities for 4-6 weeks. Trained observers evaluated adherence to EBPs before and after coaching. Using mixed methods and a systematic adaptation process, we modified and strengthened the intervention. The modified intervention was implemented in three additional facilities. Pre/post-change in EBP prevalence aggregated across facilities was analyzed. In the first two facilities, limited improvement was seen in EBPs with the exception of post-partum oxytocin. Checklists were used <25 % of observations. We identified challenges in physicians coaching nurses, need to engage district and facility leadership to address system gaps, and inadequate strategy for motivating SCC uptake. Revisions included change to peer-to-peer coaching (nurse to nurse, physician to physician); strengthened coach training on behavior and system change; adapted strategy for effective leadership engagement; and an explicit motivation strategy to enhance professional pride and effectiveness. These modifications resulted in improvement in multiple EBPs from baseline including taking maternal blood pressure (0 to 16 %), post-partum oxytocin (36 to 97 %), early breastfeeding initiation (3 to 64 %), as well as checklist use (range 32 to 88 %), all p < 0.01. Further adaptations were implemented to increase the effectiveness prior to full trial launch. The adaptive study design of implementation, evaluation, and feedback drove iterative redesign and successfully developed a SCC-focused coaching intervention that improved EBPs in UP facilities. This work was critical to develop a replicable BetterBirth package tailored to the local context. The multi-center pragmatic trial is underway measuring impact of the BetterBirth program on EBP and maternal-neonatal morbidity and mortality. NCT02148952 .


Practical disk diffusion method for detection of inducible clindamycin resistance in Staphylococcus aureus and coagulase-negative staphylococci.
Resistance to macrolides in staphylococci may be due to active efflux (encoded by msrA) or ribosomal target modification (macrolide-lincosamide-streptogramin B [MLSB] resistance; usually encoded by ermA or ermC). MLSB resistance is either constitutive or inducible following exposure to a macrolide. Induction tests utilize closely approximated erythromycin and clindamycin disks; the flattening of the clindamycin zone adjacent to the erythromycin disk indicates inducible MLSB resistance. The present study reassessed the reliability of placing erythromycin and clindamycin disks in adjacent positions (26 to 28 mm apart) in a standard disk dispenser, compared to distances of 15 or 20 mm. A group of 130 clinical isolates of Staphylococcus aureus and 100 isolates of erythromycin-resistant coagulase-negative staphylococci (CNS) were examined by disk approximation; all CNS isolates and a subset of S. aureus isolates were examined by PCR for ermA, ermC, and msrA. Of 114 erythromycin-resistant S. aureus isolates, 39 demonstrated constitutive resistance to clindamycin, while 33 showed inducible resistance by disk approximation at all three distances. Only one isolate failed to clearly demonstrate induction at 26 mm. Of 82 erythromycin-resistant CNS isolates that contained ermA or ermC, 57 demonstrated constitutive clindamycin resistance, and 25 demonstrated inducible resistance, at 20 and 26 mm. None of the 42 S. aureus isolates or 18 CNS isolates containing only msrA and none of the erythromycin-susceptible isolates yielded positive disk approximation tests. Simple placement of erythromycin and clindamycin disks at a distance achieved with a standard disk dispenser allowed detection of 97% of S. aureus strains and 100% of CNS strains with inducible MLSB resistance in this study.


Loss of adaptive variation during evolutionary responses to climate change.
The changes in species' geographical distribution demanded by climate change are often critically limited by the availability of key interacting species. In such cases, species' persistence will depend on the rapid evolution of biotic interactions. Understanding evolutionary limits to such adaptation is therefore crucial for predicting biological responses to environmental change. The recent poleward range expansion of the UK brown argus butterfly has been associated with a shift in female preference from its main host plant, rockrose (Cistaceae), onto Geraniaceae host plants throughout its new distribution. Using reciprocal transplants onto natural host plants across the UK range, we demonstrate reduced fitness of females from recently colonised Geraniaceae-dominated habitat when moved to ancestral rockrose habitats. By contrast, individuals from ancestral rockrose habitats show no reduction in fitness on Geraniaceae. Climate-driven range expansion in this species is therefore associated with the rapid evolution of biotic interactions and a significant loss of adaptive variation.


Intraocular light scatter, reflections, fluorescence and absorption: what we see in the slit lamp.
Much knowledge has been collected over the past 20 years about light scattering in the eye- in particular in the eye lens- and its visual effect, called straylight. It is the purpose of this review to discuss how these insights can be applied to understanding the slit lamp image. The slit lamp image mainly results from back scattering, whereas the effects on vision result mainly from forward scatter. Forward scatter originates from particles of about wavelength size distributed throughout the lens. Most of the slit lamp image originates from small particle scatter (Rayleigh scatter). For a population of middle aged lenses it will be shown that both these scatter components remove around 10% of the light from the direct beam. For slit lamp observation close to the reflection angles, zones of discontinuity (Wasserspalten) at anterior and posterior parts of the lens show up as rough surface reflections. All these light scatter effects increase with age, but the correlations with age, and also between the different components, are weak. For retro-illumination imaging it will be argued that the density or opacity seen in areas of cortical or posterior subcapsular cataract show up because of light scattering, not because of light loss. NOTES: (1) Light scatter must not be confused with aberrations. Light penetrating the eye is divided into two parts: a relatively small part is scattered, and removed from the direct beam. Most of the light is not scattered, but continues as the direct beam. This non-scattered part is the basis for functional imaging, but its quality is under the control of aberrations. Aberrations deflect light mainly over small angles (<1°), whereas light scatter is important because of the straylight effects over large angles (>1°), causing problems like glare and hazy vision. (2) The slit lamp image in older lenses and nuclear cataract is strongly influenced by absorption. However, this effect is greatly exaggerated by the light path lengths concerned. This obviates proper judgement of the functional importance of absorption, and hinders the appreciation of the Rayleigh nature of what is seen in the slit lamp image.


Prevalence and correlates of intimate partner violence among older Chinese couples in Hong Kong.
This study examined the prevalence and risk factors for intimate partner violence (IPV) among Chinese older couples in Hong Kong. A population representative sample was surveyed. The prevalence of IPV in older adults was found to be quite high in the present study, with a lifetime prevalence ranging from 1.4% to 53.6%, and a past year prevalence ranging from 0.4% to 36.1% for various forms of aggression. Results of logistic regression analyses showed that older persons who were younger among this "older" group, who were not employed, who had a substance abuse problem, who had witnessed parental violence during their childhood, who had a criminal history, who had a low level of assertiveness, who had an anger management problem, who experienced a low level of social support and/or experienced stressful conditions, were all more likely to fall victims of IPV. It is suggested that IPV in older couples is a complex phenomenon that is closely intertwined with other forms of domestic violence, including spousal violence, child abuse, in-law conflicts, and elderly adult abuse. Thus, before we have more definitive and concrete evidence that IPV in older couples should definitively come under the category of elder abuse or IPV, it is advisable to treat it under its own separate category of family violence.


Magenstrasse and Mill gastroplasty and sleeve gastrectomy as treatment for morbid obesity.
Laparoscopic nonbanded restrictive procedures are becoming more popular as staging and primary operations in bariatric surgery. The Magenstrasse and Mill (MM) procedure produces a restrictive gastric tubular pouch based along the lesser curvature; for the most part anatomy and physiology are preserved. In Sleeve Gastrectomy (SG), 80% of normal stomach is resected to produce restriction and to decrease ghrelin levels. This is a retrospective nonrandomized study evaluating the medical records of patients who had the laparoscopic MM (LMM) and laparoscopic SG (LSG) between January 2007 and October 2008. One bariatric surgeon performed the LMM and two bariatric surgeons performed the LSG. A total of 20 patients were identified: 13 SG and 7 MM. The mean age was 50 for the MM vs 42.9 for the SG. For the MM, the mean preoperative body mass index (BMI) was 65.4 +/- 11.1 kg/m2, with a mean excess weight of 282 +/- 73.7 kg. For the SG, the mean preoperative body mass index was 47.5 +/- 8.3 kg/m2, with a mean excess weight of 156.1 +/- 52.6 kg. The mean excess weight loss after six and 12 months for the M&M was 35 +/- 10.5% and 20.1 +/- 1.4%, vs 52.4 +/- 17.8% and 49% +/- 15.4% for the SG. Follow-up of one year was achieved in two M&M patients and three SG patients. Median follow-up of all patients was seven months (range 12-1). This is a short-term retrospective outcome study. The LMM patients were larger than LSG patients. Total weight loss was greater for the LMM patients. Operative time for the LMM is shorter. The percent excess weight loss in the short-term 12 month period was more in the LSG compared to the LMM. Long-term follow-up is needed.


Reconstitution of Rad53 activation by Mec1 through adaptor protein Mrc1.
Upon DNA replication stress, stalled DNA replication forks serve as a platform to recruit many signaling proteins, leading to the activation of the DNA replication checkpoint. Activation of Rad53, a key effector kinase in the budding yeast Saccharomyces cerevisiae, is essential for stabilizing DNA replication forks during replication stress. Using an activity-based assay for Rad53, we found that Mrc1, a replication fork-associated protein, cooperates with Mec1 to activate Rad53 directly. Reconstitution of Rad53 activation using purified Mec1 and Mrc1 showed that the addition of Mrc1 stimulated a more than 70-fold increase in the ability of Mec1 to activate Rad53. Instead of increasing the catalytic activity of Mec1, Mrc1 was found to facilitate the phosphorylation of Rad53 by Mec1 via promotion of a stronger enzyme-substrate interaction between them. Further, the conserved C-terminal domain of Mrc1 was found to be required for Rad53 activation. These results thus provide insights into the role of the adaptor protein Mrc1 in activating Rad53 in the DNA replication checkpoint.


An essential role for medullary thymic epithelial cells during the intrathymic development of invariant NKT cells.
In the thymus, interactions with both cortical and medullary microenvironments regulate the development of self-tolerant conventional CD4(+) and CD8(+) αβT cells expressing a wide range of αβTCR specificities. Additionally, the cortex is also required for the development of invariant NKT (iNKT) cells, a specialized subset of T cells that expresses a restricted αβTCR repertoire and is linked to the regulation of innate and adaptive immune responses. Although the role of the cortex in this process is to enable recognition of CD1d molecules expressed by CD4(+)CD8(+) thymocyte precursors, the requirements for additional thymus microenvironments during iNKT cell development are unknown. In this study, we reveal a role for medullary thymic epithelial cells (mTECs) during iNKT cell development in the mouse thymus. This requirement for mTECs correlates with their expression of genes required for IL-15 trans-presentation, and we show that soluble IL-15/IL-15Rα complexes restore iNKT cell development in the absence of mTECs. Furthermore, mTEC development is abnormal in iNKT cell-deficient mice, and early stages in iNKT cell development trigger receptor activator for NF-κB ligand-mediated mTEC development. Collectively, our findings demonstrate that intrathymic iNKT cell development requires stepwise interactions with both the cortex and the medulla, emphasizing the importance of thymus compartmentalization in the generation of both diverse and invariant αβT cells. Moreover, the identification of a novel requirement for iNKT cells in thymus medulla development further highlights the role of both innate and adaptive immune cells in thymus medulla formation.


A randomized clinical trial to compare plaque inhibition of a sodium fluoride/potassium nitrate dentifrice versus a stabilized stannous fluoride/sodium hexametaphosphate dentifrice.
To compare the plaque inhibition efficacy of a sodium fluoride/potassium nitrate (NaF/KNO3 with 1450 ppm F) test dentifrice to a 0.454% stannous fluoride/sodium hexametaphosphate/sodium fluoride positive control dentifrice (SnF2/SHMP with 1450 ppm F). Twenty-five subjects were randomized to a two-period, two-treatment, double blind crossover sequence using NaF/KNO3 (Sensodyne ProNamel dentifrice) and SnF2/SHMP (blend-a-med* EXPERT GUMS PROTECTION dentifrice). Each treatment was conducted with a standard manual toothbrush (Oral-B P35 Indicator). Digital plaque image analysis (DPIA) was used on three consecutive days to evaluate: (a) overnight plaque formation (A.M. pre-brushing); (b) following 40 seconds of brushing with the test product (A.M. post-brushing); and (c) mid-afternoon (P.M.). Images were analysed using an objective computer algorithm to calculate the total area of visible plaque. A four-day washout period was instituted for the crossover phase. All 25 subjects completed the study. The SnF2/SHMP positive control dentifrice provided statistically significantly lower levels of plaque area coverage versus the NaF/KNO3 test dentifrice at each timepoint. For the SnF2/SHMP dentifrice, plaque coverage was 23.0% lower (p< 0.0001) at A.M. pre-brushing, 17.3% (p= 0.0163) lower at A.M. post-brushing, and 22.6% (p= 0.0004) lower at the P.M. measure relative to the NaF/KNO3 dentifrice. The SnF2/SHMP dentifrice (blend-a-med EXPERT GUMS PROTECTION) inhibits plaque regrowth both overnight and during the day to a significantly greater degree than the NaF/KNO3 dentifrice (Sensodyne ProNamel). Dentists recommending an effective home use dentifrice for patients experiencing dentinal hypersensitivity and/or dental erosion may previously have needed to compromise on other key benefits, such as plaque control. blend-a-med EXPERT GUMS PROTECTION is a dentifrice when integrated into an oral hygiene routine can provide a proven treatment for hypersensitivity, dental erosion, and a reduction in the regrowth of plaque.


Lung function, radiological changes and exposure: analysis of ATSDR data from Libby, MT, USA.
In 2000, the Agency for Toxic Substances and Disease Registry (ATSDR; Atlanta, GA, USA) investigated lung disease in those exposed to the tremolite-contaminated vermiculite mine in Libby, MT, USA. Previously unreported spirometric results are presented here in relation to exposure and radiographic findings. 4,524 study participants were assigned to one of seven mutually exclusive exposure categories. Associations among radiographic findings, spirometric results and exposure were investigated, along with the effect of a reduction in exposure potential when production was moved to a wet process mill in the mid 1970s. Spirometry data for the total population by smoking status and age were within the normal range. Prevalence of pleural plaque increased with age, but was lowest in the environmentally exposed group (0.42-12.74%) and greatest in the W.R. Grace & Co. mineworkers (20-45.68%). For males, there was a significant (4.5%) effect of pleural plaques on forced vital capacity. For W.R. Grace & Co. workers and household contacts, a reduction in plaque (0.11 versus 1.64%) and in diffuse pleural thickening or costophrenic angle obliteration (1.94 and 0.13%) was noted for those exposed after 1976. These analyses do not support a clinically important reduction in spirometry of this cohort. The 1976 reductions in exposure have led to decrease in radiographic changes.


Neurosurgery residency and fellowship education in the United States: 2 decades of system development by the One Neurosurgery Summit organizations.
The purpose of this report is to chronicle a 2-decade period of educational innovation and improvement, as well as governance reform, across the specialty of neurological surgery. Neurological surgery educational and professional governance systems have evolved substantially over the past 2 decades with the goal of improving training outcomes, patient safety, and the quality of US neurosurgical care. Innovations during this period have included the following: creating a consensus national curriculum; standardizing the length and structure of neurosurgical training; introducing educational outcomes milestones and required case minimums; establishing national skills, safety, and professionalism courses; systematically accrediting subspecialty fellowships; expanding professional development for educators; promoting training in research; and coordinating policy and strategy through the cooperation of national stakeholder organizations. A series of education summits held between 2007 and 2009 restructured some aspects of neurosurgical residency training. Since 2010, ongoing meetings of the One Neurosurgery Summit have provided strategic coordination for specialty definition, neurosurgical education, public policy, and governance. The Summit now includes leadership representatives from the Society of Neurological Surgeons, the American Association of Neurological Surgeons, the Congress of Neurological Surgeons, the American Board of Neurological Surgery, the Review Committee for Neurological Surgery of the Accreditation Council for Graduate Medical Education, the American Academy of Neurological Surgery, and the AANS/CNS Joint Washington Committee. Together, these organizations have increased the effectiveness and efficiency of the specialty of neurosurgery in advancing educational best practices, aligning policymaking, and coordinating strategic planning in order to meet the highest standards of professionalism and promote public health.