Tracking the visual focus of attention for a varying number of wandering people.
We define and address the problem of finding the visual focus of attention for a varying number of wandering people (VFOA-W), determining where the people's movement is unconstrained. VFOA-W estimation is a new and important problem with mplications for behavior understanding and cognitive science, as well as real-world applications. One such application, which we present in this article, monitors the attention passers-by pay to an outdoor advertisement. Our approach to the VFOA-W problem proposes a multi-person tracking solution based on a dynamic Bayesian network that simultaneously infers the (variable) number of people in a scene, their body locations, their head locations, and their head pose. For efficient inference in the resulting large variable-dimensional state-space we propose a Reversible Jump Markov Chain Monte Carlo (RJMCMC) sampling scheme, as well as a novel global observation model which determines the number of people in the scene and localizes them. We propose a Gaussian Mixture Model (GMM) and Hidden Markov Model (HMM)-based VFOA-W model which use head pose and location information to determine people's focus state. Our models are evaluated for tracking performance and ability to recognize people looking at an outdoor advertisement, with results indicating good performance on sequences where a moderate number of people pass in front of an advertisement.


Polysaccharide-rich fraction of Agaricus brasiliensis enhances the candidacidal activity of murine macrophages.
A polysaccharide-rich fraction (ATF) of medicinal mushroom Agaricus brasiliensis was evaluated on the candidacidal activity, H2O2 and nitric oxide (NO) production, and expression of mannose receptors by murine peritoneal macrophages. Mice received three intraperitoneal (i.p.) injections of ATF and after 48 h their peritoneal resident macrophages were assayed against Candida albicans yeast forms. The treatment increased fungicidal activity and it was associated with higher levels of H2O2, whereas NO production was not affected. We also found that the treatment enhances mannose receptor expression by peritoneal macrophages, which are involved in the attachment and phagocytosis of non-opsonized microorganisms. Treatment of animals with ATF was able to enhance the clearance of C. albicans during the first 6 h after the experimental i.p. infection. Our results suggest that this extract can increase host resistance against some infectious agents through the stimulation of microbicidal activity of macrophages.


Notch Inhibitor PF-03084014 Inhibits Hepatocellular Carcinoma Growth and Metastasis via Suppression of Cancer Stemness due to Reduced Activation of Notch1-Stat3.
Aberrant activation of the Notch signaling pathway is implicated in many solid tumors, including hepatocellular carcinoma, indicating a potential use of Notch inhibitors for treatment. In this study, we investigated the antitumor and antimetastasis efficacy of the novel Notch inhibitor (γ-secretase inhibitor) PF-03084014 in hepatocellular carcinoma. Hepatocellular carcinoma spherical cells (stem-like cancer cells), a sphere-derived orthotopic tumor model and one patient-derived xenograft (PDX) model were used in our experiment. We demonstrated that PF-03084014 inhibited the self-renewal and proliferation of cancer stem cells. PF-03084014 reduced the hepatocellular carcinoma sphere-derived orthotopic tumor and blocked the hepatocellular carcinoma tumor liver to lung metastasis. We further tested the PF-03084014 in PDX models and confirmed the inhibition tumor growth effect. In addition, a low dose of PF-03084014 induced hepatocellular carcinoma sphere differentiation, resulting in chemosensitization. Antitumor activity was associated with PF-03084014-induced suppression of Notch1 activity, decreased Stat3 activation and phosphorylation of the Akt signaling pathway, and reduced epithelial-mesenchymal transition. These are the key contributors to the maintenance of cancer stemness and the promotion of cancer metastasis. Moreover, the Notch-Stat3 association was implicated in the clinical hepatocellular carcinoma prognosis. Collectively, PF-03084014 revealed antitumor and antimetastatic effects in hepatocellular carcinoma, providing evidence for the potential use of gamma-secretase inhibitors as a therapeutic option for the treatment of hepatocellular carcinoma. Mol Cancer Ther; 16(8); 1531-43. ©2017 AACR.


Polarization temporal dynamics in a dye microlaser.
We have investigated the temporal dynamics of the output polarization of a dye microlaser operating in a transient regime, i.e., under femtosecond excitation. In these conditions the dipole angular diffusion has an important role in the formation of the microlaser pulse, as is shown in a theoretical model. By performing the experiment for different values of the microcavity length and varying the polarization angle, we measured threshold and buildup time of the microlaser and compared the results obtained with two dye solutions of different viscosity. The agreement between theory and experimental results is adequate.


Cryptic Biological Invasions: a General Model of Hybridization.
The dispersal of non-native genes due to hybridization is a form of cryptic invasion with growing concern in evolution and conservation. This includes the spread of transgenic genes and antibiotic resistance. To investigate how genes and phenotypes are transmitted, we developed a general model that, for the first time, considers concurrently: multiple loci, quantitative and qualitative gene expression, assortative mating, dominance/recessivity inheritance and density-dependent demographic effects. Selection acting on alleles or genotypes can also be incorporated. Our results reveal that the conclusions about how hybridization threatens a species can be biased if they are based on single-gene models, while considering two or more genes can correct this bias. We also show that demography can amplify or balance the genetic effects, evidencing the need of jointly incorporating both processes. By implementing our model in a real case, we show that mallard ducks introduced in New Zealand benefit from hybridization to replace native grey-ducks. Total displacement can take a few generations and occurs by interspecific competition and by competition between hybrids and natives, demonstrating how hybridization may facilitate biological invasions. We argue that our general model represents a powerful tool for the study of a wide range of biological and societal questions.


The contribution of cannabis use to variation in the incidence of psychotic disorder across Europe (EU-GEI): a multicentre case-control study.
Cannabis use is associated with increased risk of later psychotic disorder but whether it affects incidence of the disorder remains unclear. We aimed to identify patterns of cannabis use with the strongest effect on odds of psychotic disorder across Europe and explore whether differences in such patterns contribute to variations in the incidence rates of psychotic disorder. We included patients aged 18-64 years who presented to psychiatric services in 11 sites across Europe and Brazil with first-episode psychosis and recruited controls representative of the local populations. We applied adjusted logistic regression models to the data to estimate which patterns of cannabis use carried the highest odds for psychotic disorder. Using Europe-wide and national data on the expected concentration of Δ9-tetrahydrocannabinol (THC) in the different types of cannabis available across the sites, we divided the types of cannabis used by participants into two categories: low potency (THC <10%) and high potency (THC ≥10%). Assuming causality, we calculated the population attributable fractions (PAFs) for the patterns of cannabis use associated with the highest odds of psychosis and the correlation between such patterns and the incidence rates for psychotic disorder across the study sites. Between May 1, 2010, and April 1, 2015, we obtained data from 901 patients with first-episode psychosis across 11 sites and 1237 population controls from those same sites. Daily cannabis use was associated with increased odds of psychotic disorder compared with never users (adjusted odds ratio [OR] 3·2, 95% CI 2·2-4·1), increasing to nearly five-times increased odds for daily use of high-potency types of cannabis (4·8, 2·5-6·3). The PAFs calculated indicated that if high-potency cannabis were no longer available, 12·2% (95% CI 3·0-16·1) of cases of first-episode psychosis could be prevented across the 11 sites, rising to 30·3% (15·2-40·0) in London and 50·3% (27·4-66·0) in Amsterdam. The adjusted incident rates for psychotic disorder were positively correlated with the prevalence in controls across the 11 sites of use of high-potency cannabis (r = 0·7; p=0·0286) and daily use (r = 0·8; p=0·0109). Differences in frequency of daily cannabis use and in use of high-potency cannabis contributed to the striking variation in the incidence of psychotic disorder across the 11 studied sites. Given the increasing availability of high-potency cannabis, this has important implications for public health. Medical Research Council, the European Community's Seventh Framework Program grant, São Paulo Research Foundation, National Institute for Health Research (NIHR) Biomedical Research Centre (BRC) at South London and Maudsley NHS Foundation Trust and King's College London and the NIHR BRC at University College London, Wellcome Trust.


Successful long-term thalidomide therapy for discoid lupus erythematosus-lichen planus overlap syndrome.
Although thalidomide is a U.S. Food and Drug Admistration (FDA) approved medication for erythema nodosum leprosum and multiple myeloma, it has many off-label uses, including for discoid lupus erythematosus (DLE), Behçet's disease, apththous ulcers in HIV patients, and prurigo nodularis. Herein, we present a patient with an overlap of discoid lupus erythematosus and lichen planus who was successfully treated with thalidomide for over 19 years without significant side effects. We propose that some of the most common side effects, including peripheral neuropathy, numbness, parasthesias, sedation, and constipation, can be avoided at lower doses, typically less than 100mg/day.


Aspirin-intolerance as a precipitating factor of life-threatening attacks of asthma requiring mechanical ventilation.
The records of 92 asthmatics who underwent mechanical ventilation were reviewed. In seven patients (8%) the severe attack was precipitated by a non-steroidal anti-inflammatory drug (NSAID); one of these patients died. Five of the cases had a history of asthmatic attacks provoked by NSAIDs; whilst in two the severe attack requiring mechanical ventilation was the first manifestation of NSAID-intolerance. In two patients the NSAID had been prescribed by their physicians. Another aspirin-intolerant patient, a general practitioner, self-administered a NSAID. The sudden attack in another patient was precipitated by a preparation which contains aspirin and is usually recommended for indigestion. In the fatal case the attack was provoked by a capsule containing aspirin, which had been given by a herbalist. Unlike other reports, we found that NSAID-intolerance is a frequent provoking factor in severe acute asthma requiring mechanical ventilation. Inadequate investigation of precipitating factors in asthmatics with severe sudden attacks is a possible reason why this phenomenon is underreported.


Improving coronary artery bypass grafting: a systematic review and meta-analysis on the impact of adopting transit-time flow measurement.
Despite there being numerous studies of intraoperative graft flow assessment by transit-time flow measurement (TTFM) on outcomes after coronary artery bypass grafting (CABG), the adoption of contemporary TTFM is low. Therefore, on 31 January 2018, a systematic literature search was performed to identify articles that reported (i) the amount of grafts classified as abnormal or which were revised or (ii) an association between TTFM and outcomes during follow-up. Random-effects models were used to create pooled estimates with 95% confidence intervals (CI) of (i) the rate of graft revision per patient, (ii) the rate of graft revision per graft and (iii) the rate of graft revision among grafts deemed abnormal based on TTFM parameters. The search yielded 242 articles, and 66 original articles were included in the systematic review. Of those articles, 35 studies reported on abnormal grafts or graft revisions (8943 patients, 15 673 grafts) and were included in the meta-analysis. In 4.3% of patients (95% CI 3.3-5.7%, I2 = 73.9) a revision was required and 2.0% of grafts (95% CI 1.5-2.5%; I2 = 66.0) were revised. The pooled rate of graft revisions among abnormal grafts was 25.1% (95% CI 15.5-37.9%; I2 = 80.2). Studies reported sensitivity ranging from 0.250 to 0.457 and the specificity from 0.939 to 0.984. Reported negative predictive values ranged from 0.719 to 0.980 and reported positive predictive values ranged from 0.100 to 0.840. This systematic review and meta-analysis showed that TTFM could improve CABG procedures. However, due to heterogeneous data, drawing uniform conclusions appeared challenging. Future studies should focus on determining the optimal use of TTFM and assessing its diagnostic accuracy.


HowNutsAreTheDutch (HoeGekIsNL): A crowdsourcing study of mental symptoms and strengths.
HowNutsAreTheDutch (Dutch: HoeGekIsNL) is a national crowdsourcing study designed to investigate multiple continuous mental health dimensions in a sample from the general population (n = 12,503). Its main objective is to create an empirically based representation of mental strengths and vulnerabilities, accounting for (i) dimensionality and heterogeneity, (ii) interactivity between symptoms and strengths, and (iii) intra-individual variability. To do so, HowNutsAreTheDutch (HND) makes use of an internet platform that allows participants to (a) compare themselves to other participants via cross-sectional questionnaires and (b) to monitor themselves three times a day for 30 days with an intensive longitudinal diary study via their smartphone. These data enable for personalized feedback to participants, a study of profiles of mental strengths and weaknesses, and zooming into the fine-grained level of dynamic relationships between variables over time. Measuring both psychiatric symptomatology and mental strengths and resources enables for an investigation of their interactions, which may underlie the wide variety of observed mental states in the population. The present paper describes the applied methods and technology, and presents the sample characteristics. Copyright © 2015 John Wiley & Sons, Ltd.


Basic composition and enriched integration in idiom processing: An EEG study.
We investigated the extent to which the literal meanings of the words forming literally plausible idioms (e.g., break the ice) are semantically composed and how the idiomatic meaning is integrated in the unfolding sentence representation. Participants read ambiguous idiom strings embedded in highly predictable, literal, and idiomatic contexts while their EEG was recorded. Control sentences only contained the idiom-final word whose cloze values were as high as in literal and idiomatic contexts. Event-related potentials data showed that differences in the amplitude of a frontal positivity (PNP) emerged at the beginning and at the end of the idiom strings, with the idiomatic context condition associated with more positive voltages. The time frequency analysis of the EEG showed an increase in power of the middle gamma frequency band only in the literal context condition. These findings suggest that sentence revision mechanisms, associated with the frontal PNP, are involved in idiom meaning integration, and that the literal semantic composition of the idiomatic constituents, associated with changes in gamma frequency, is not carried out after idiom recognition. (PsycINFO Database Record


Endogenous cytosine damage products alter the site selectivity of human DNA maintenance methyltransferase DNMT1.
Alterations in cytosine methylation patterns are usually observed in human tumors. The consequences of altered cytosine methylation patterns include both inappropriate activation of transforming genes and silencing of tumor suppressor genes. Despite the biological effect of methylation changes, little is known about how such changes are caused. The heritability of cytosine methylation patterns from parent to progeny cells is attributed to the fidelity of the methylation-sensitive human maintenance methyltransferase DNMT1, which methylates with high specificity the unmethylated strand of a hemimethylated CpG sequence following DNA replication. We have been studying DNA damage that might alter the specificity of DNMT1, either inhibiting the methylation of hemimethylated sites or triggering the inappropriate methylation of previously unmethylated sites. Here, we show that known forms of endogenous DNA damage can cause either hypermethylation or hypomethylation. Inflammation-induced 5-halogenated cytosine damage products, including 5-chlorocytosine, mimic 5-methylcytosine and induce inappropriate DNMT1 methylation within a CpG sequence. In contrast, oxidation damage of the methyl group of 5-methylcytosine, with the formation of 5-hydroxymethylcytosine, prevents DNMT1 methylation of the target cytosine. We propose that reduced DNMT1 selectivity resulting from DNA damage could cause heritable changes in cytosine methylation patterns, resulting in human tumor formation. These data may provide a mechanistic link for the associations documented between inflammation and cancer.


Structural Barriers to Pre-exposure Prophylaxis Use Among Young Sexual Minority Men: The P18 Cohort Study.
Despite decreasing rates of HIV among many populations, HIV-related health disparities among gay, bisexual and other men who have sex with men persist, with disproportional percentages of new HIV diagnoses among racial and ethnic minority men. Despite increasing awareness of HIV pre-exposure prophylaxis (PrEP), PrEP use remains low. In addition to exploring individual-level factors for this slow uptake, structural drivers of PrEP use must also be identified in order to maximize the effectiveness of biomedical HIV prevention strategies. Using cross-sectional data from an ongoing cohort study of young sexual minority men (N=492), we examine the extent to which structural-level barriers, including access to health care, medication logistics, counseling support, and stigma are related to PrEP use. While almost all participants indicated awareness of PrEP, only 14% had ever used PrEP. PrEP use was associated with lower concerns about health care access, particularly paying for PrEP. Those with greater concerns talking with their provider about their sexual behaviors were less likely to use PrEP. Paying for PrEP and talking to one's provider about sexual behaviors are concerns for young sexual minority men. In particular, stigma from healthcare providers poses a significant barrier to PrEP use in this population. Providers need not only to increase their own awareness of and advocacy for PrEP as an effective risk-management strategy for HIV prevention, but also must work to create open and non-judgmental spaces in which patients can discuss sexual behaviors without the fear of stigma.


Hominid humeral fragment from early Pleistocene of northwestern Kenya.
The distal end of a hominoid humerus was recovered from early Pleistocene sediments in the Kanapoi drainage near the southern end of Lake Rudolf. Lava capping the sediments yielded a potassium/argon date of 2.5 million years. The fragment can be distinguished on inspection from gorilla and orangutan; discriminate analysis of humeri of Homo and Pan assigns it as hominid. From other evidence we consider it more likely to represent Australopithecus s.s. than Paranthropus.


[Antilipemic drug-induced skin manifestations].
Systemically administered lipid-lowering drugs can induce eczema, ichthyosis, or psoriasis as side effects. Common abnormalities in these diseases are a rough, scaly skin, a disturbed permeability barrier and disturbed epidermal proliferation and differentiation. The disturbed epidermal differentiation is accompanied by changes in the keratin composition and the cornified envelope proteins as well as by changes in the lipid composition. Lipid-lowering drugs do not necessarily all cause the same diseases, because they inhibit different steps in the cholesterol synthetic pathway. The lipid-lowering drugs lovastatin (Mevacor), simvastatin (Zocor) and pravastatin (Selipram) can cause eczema; these drugs inhibit an early step of cholesterol biosynthesis, viz. HMG CoA reductase activity. The lipid-lowering drugs triparanol and diazacholesterol inhibit a late step in cholesterol biosynthesis, delta-24-sterol reductase, and they can induce ichthyosis or palmoplantar hyperkeratosis. In contrast, systemically applied gemfibrozil, which mainly lowers triglycerides, can cause an exacerbation of psoriasis. These observations show the importance of the lipid metabolism in the pathogenesis of eczema, ichthyosis, and psoriasis.


Excellent Semiconductors Based on Tetracenotetracene and Pentacenopentacene: From Stable Closed-Shell to Singlet Open-Shell.
Designing stable open-shell organic materials through the modifications of the π-topology of molecular organic semiconductors has recently attracted considerable attention. However, their uses as an active layer in organic field-effect transistors (OFETs) are very limited, and the obtained hole and electron charge mobilities are around 10-3 cm2 V-1 s-1. Herein, we disclose the synthesis of two peri-fused materials, so-called tetracenotetracene (TT) and pentacenopentacene (PP), which have low band gaps of 1.79 and 1.42 eV, respectively. Their ground state natures have been investigated by different experiments including steady state absorption, electron spin resonance, superconducting quantum interfering device, and variable-temperature NMR along with DFT calculations. TT and PP have closed-shell and singlet open-shell structures in their ground state, respectively, and possess high stability. Their biradical characteristics were found to be 0.50 and 0.64. The origin of the open-shell character of PP is related to the concomitant opening of two tetracenes with the recovering of two extra aromatic sextets and a small HOMO-LUMO energy gap (gap <1.5 eV). Thanks to the high stability, thin film OFET devices could be fabricated. In TG-BC configuration PP shows a remarkably high hole mobility of 1.4 cm2 V-1 s-1, while TT exhibits a hole mobility of 0.77 cm2 V-1 s-1. In the configuration of BG-TC, ambipolar behaviors for both were obtained with hole and electron mobilities of 0.21 and 0.01 cm2 V-1 s-1 for PP and 0.14 and 0.006 cm2 V-1 s-1 for TT.


The safety assessment of Pythium irregulare as a producer of biomass and eicosapentaenoic acid for use in dietary supplements and food ingredients.
Polyunsaturated fatty acids, docosahexaenoic acid (DHA, 22:6, n-3), eicosapentaenoic acid (EPA, 20:5, n-3), and arachidonic acid (ARA, 20:4 n-6), have multiple beneficial effects on human health and can be used as an important ingredient in dietary supplements, food, feed and pharmaceuticals. A variety of microorganisms has been used for commercial production of these fatty acids. The microorganisms in the Pythium family, particularly Pythium irregulare, are potential EPA producers. The aim of this work is to provide a safety assessment of P. irregulare so that the EPA derived from this species can be potentially used in various commercial applications. The genus Pythium has been widely recognized as a plant pathogen by infecting roots and colonizing the vascular tissues of various plants such as soybeans, corn and various vegetables. However, the majority of the Pythium species (including P. irregulare) have not been reported to infect mammals including humans. The only species among the Pythium family that infects mammals is P. insidiosum. There also have been no reports showing P. irregulare to contain mycotoxins or cause potentially allergenic responses in humans. Based on the safety assessment, we conclude that P. irregulare can be considered a safe source of biomass and EPA-containing oil for use as ingredients in dietary supplements, food, feed and pharmaceuticals.


Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region.
The Ras-related nuclear protein, Ran, has been implicated in nuclear transport. By screening a HeLa cell lambda expression library with Ran-GTP and sequencing overlapping cDNA clones, we have obtained the derived primary structure of a protein with a calculated molecular mass of 358 kDa. Using antibodies raised against an expressed segment of this protein, we obtained punctate nuclear surface staining by immunofluorescence microscopy that is characteristic for nucleoporins. Electron microscopy of immunogold-decorated rat liver nuclear envelopes sublocalized the 358-kDa protein at (or near) the tip of the cytoplasmic fibers of the nuclear pore complex (NPC). In agreement with current convention, this protein was therefore termed Nup358 (for nucleoporin of 358 kDa). Nup358 contains a leucine-rich region, four potential Ran binding sites (i.e. Ran binding protein 1 homologous domains) flanked by nucleoporin-characteristic FXFG or FG repeats, eight zinc finger motifs, and a C-terminal cyclophilin A homologous domain. Consistent with the location of Nup358 at the cytoplasmic fibers of the NPC, we found decoration with Ran-gold at only the cytoplasmic side of the NPC. Thus, Nup358 is the first nucleoporin shown to contain binding sites for two of three soluble nuclear transport factors so far isolated, namely karyopherin and Ran-GTP.


Skeletal scintigraphy with technetium-99m-tetraphenyl porphyrin sulfonate for the detection and determination of osteomyelitis in an animal model.
This article explores the accumulation of 99mTc-tetraphenyl porphyrin sulfonate (TPPS4) at inflammatory sites, especially osteomyelitis, and compares the results with 111In Cl3 and 111In-WBC in an animal model. Osteomyelitis was induced in 12 New Zealand white rabbits by injecting staphylococcus aureus in the left tibia. Three weeks later, radiographs confirmed the disease. Two hours later, after injection of 74 MBq 99mTc-TPPS4, scintiphotos of the lower extremities were acquired and repeat scintiphotos were obtained 24 hr after injection of 5.55 MBq 111In Cl3. After these studies, 24- and 48-hr scintiphotos of the lower extremities were acquired after injecting 5.55 MBq 111In-labeled WBC. The left tibia averaged three times the uptake with 99mTc-TPPS4 compared with right tibia; with 111In Cl3 and 111In WBC the ratios are two times. These three radiopharmaceuticals reveal positive images, but the image quality using 99mTc-TPPS4 is better, as would be expected from the more favorable physical characteristics of 99mTc and the higher uptake. The traditional combination of three-phase bone and 67Ga-citrate scintigraphy can be replaced by a single injection of 99mTc-TPPS4 with imaging as early as 2 hr. Finally, the use 99mTc-TPPS4 should result in a substantial reduction in radiopharmaceutical cost.


A novel measure of corporate carbon emission disclosure, the effect of capital expenditures and corporate governance.
The UK's 2050 net-zero emission target is one of the most ambitious goals in the world. Organisations should play a vital role by communicating a sufficient level of carbon emission information with the stakeholders. Motivated by the necessity of measuring the level of carbon disclosure, this paper provides a unique carbon emission disclosure measurement based on a sample of UK firms from 2013 to 2019. We apply different methods to assess the validity and reliability of our developed measurement. The results suggest that our measurement captures the actual CO2 emission, including scope 1, scope 2, and also captures the environmental, social and governance (ESG) score. Additionally, we explore the association between capital expenditure, corporate governance and the level of carbon emission disclosure. Further, the results show a positive relationship between capital expenditure and carbon emission disclosure. Also, there is a significant positive relationship between internal governance and carbon emission disclosure. Moreover, the analysis suggests that internal governance strengthen the relationship between capital expenditure and carbon emission disclosure. We also use quantile regression, and the findings show that capital expenditure and internal governance have a positive impact on carbon emission disclosure under all quantiles. Our data suggest that capital expenditure declines within the UK by around 53% over the last six years. Following the reduction in capital expenditure, the results demonstrate 39% decline in the CO2 emission level. The results also indicate that for every $1 million capital expenditure, approximately 2.75 Metric tons of carbon dioxide (MtCO2) emissions increase. Business investment is around 70% of the UK's total investment. Therefore, the reduction in capital expenditure is one of the primary reasons that might explain the decrease in the UK's overall CO2 emission level. The unique findings of this paper are relevant to the government, management and standard-setters.


Circadian rhythmicity persists through the Polar night and midnight sun in Svalbard reindeer.
Studies of locomotor activity in Svalbard reindeer reported the temporary absence of diel rhythms under Arctic photic conditions. However, using Lomb-Scargle periodogram analyses with high statistical power we found diel or circadian rhythmicity throughout the entire year in measures of behaviour, temperature in the rumen and heart rate in free-living Svalbard reindeer. Significant diel rhythmicity was only lacking during some of the 15-day intervals analysed in the less frequently measured heart rate. During Polar Night these rhythms were free-running and attenuated. During continual daylight in summer, rhythms where entrained to 24 hours corresponding with the daily variation in the intensity of solar radiation, but attenuated when continuous daylight coincided with the period of growing forage. Diel rhythmicity was reduced during this short period of peak foraging activity, which coincided with peak heart rate and temperature in the rumen, most likely to facilitate fattening when food is abundant. For the rest of the year, heart rate and temperature showed the most pronounced and long-lasting suppression ever found in ungulates. The profound seasonal changes in foraging, metabolic activity, and power of diel and circadian rhythmicity of Svalbard reindeer can be viewed as adaptations to the extreme living conditions in the High Arctic.


Gut AstA mediates sleep deprivation-induced energy wasting in Drosophila.
Severe sleep deprivation (SD) has been highly associated with systemic energy wasting, such as lipid loss and glycogen depletion. Despite immune dysregulation and neurotoxicity observed in SD animals, whether and how the gut-secreted hormones participate in SD-induced disruption of energy homeostasis remains largely unknown. Using Drosophila as a conserved model organism, we characterize that production of intestinal Allatostatin A (AstA), a major gut-peptide hormone, is robustly increased in adult flies bearing severe SD. Interestingly, the removal of AstA production in the gut using specific drivers significantly improves lipid loss and glycogen depletion in SD flies without affecting sleep homeostasis. We reveal the molecular mechanisms whereby gut AstA promotes the release of an adipokinetic hormone (Akh), an insulin counter-regulatory hormone functionally equivalent to mammalian glucagon, to mobilize systemic energy reserves by remotely targeting its receptor AstA-R2 in Akh-producing cells. Similar regulation of glucagon secretion and energy wasting by AstA/galanin is also observed in SD mice. Further, integrating single-cell RNA sequencing and genetic validation, we uncover that severe SD results in ROS accumulation in the gut to augment AstA production via TrpA1. Altogether, our results demonstrate the essential roles of the gut-peptide hormone AstA in mediating SD-associated energy wasting.


Penetrating head injury in planned and repetitive deliberate self-harm.
44-year-old man presented to his local emergency department wearing a baseball cap and complaining of headaches that had progressively worsened over the preceding 11 weeks. After we provided generous analgesia and performed simple investigations that failed to identify a diagnosis, the patient removed his cap to reveal an assortment of metallic objects embedded in his scalp. Plain radiographs showed 11 nails penetrating into his brain. A detailed history revealed a diagnosis of paranoid schizophrenia, and the patient confirmed that he had hammered a nail into his head each week for the past 11 weeks to rid him of evil. The nails were removed with the patient under general anesthesia, and he made an uncomplicated recovery with no neurological deficits.


Potential of geographical variation analysis for realigning providers to value-based care. ECHO case study on lower-value indications of C-section in five European countries.
Although C-section is a highly effective procedure, literature abounds with evidence of overuse and particularly misuse, in lower-value indications such as low-risk deliveries. This study aims to quantify utilization of C-section in low-risk cases, mapping out areas showing excess-usage in each country and to estimate excess-expenditure as a proxy of the opportunity cost borne by healthcare systems. Observational, ecologic study on deliveries in 913 sub-national administrative areas of five European countries (Denmark, England, Portugal, Slovenia and Spain) from 2002 to 2009. The study includes a cross-section analysis with 2009 data and a time-trend analysis for the whole period. Main endpoints: age-standardized utilization rates of C-section in low-risk pregnancies and deliveries per 100 deliveries. Secondary endpoints: Estimated excess-cases per geographical unit of analysis in two scenarios of minimized utilization. C-section is widely used in all examined countries (ranging from 19% of Slovenian deliveries to 33% of deliveries in Portugal). With the exception of Portugal, there are no systematic variations in intensity of use across areas in the same country. Cross-country comparison of lower-value C-section leaves Denmark with 10% and Portugal with 2%, the highest and lowest. Such behaviour was stable over the period of analysis. Within each country, the scattered geographical patterns of use intensity speak for local drivers playing a major role within the national trend. The analysis conducted suggests plenty of room for enhancing value in obstetric care and equity in women's access to such within the countries studied. The analysis of geographical variations in lower-value care can constitute a powerful screening tool.


Differences between "classical" risk factors for infections caused by methicillin-resistant Staphylococcus aureus (MRSA) and risk factors for nosocomial bloodstream infections caused by multiple clones of the staphylococcal cassette chromosome mec type IV MRSA strain.
To identify risk factors associated with nosocomial bloodstream infections caused by multiple clones of the staphylococcal cassette chromosome mec (SCCmec) type IV strain of methicillin-resistant Staphylococcus aureus (MRSA). An unmatched case-control study (at a ratio of 1:2) performed during the period from October 2002 through September 2003. A 2,000-bed tertiary care teaching hospital affiliated with the University of São Paulo in São Paulo, Brazil. Case patients (n=30) were defined either as patients who had a bloodstream infection due to SCCmec type IV MRSA diagnosed at least 48 hours after hospital admission or as neonates with the infection who were born in the hospital. Control patients (n=60) were defined as patients with SCCmec type III MRSA infection diagnosed at least 48 hours after hospital admission. Genes encoding virulence factors were studied in the isolates recovered from case patients, and molecular typing of the SCCmec type IV MRSA isolates was also done by pulsed-field gel electrophoresis and multilocus sequence typing. In multivariate analysis, the following 3 variables were significantly associated with having a nosocomial bloodstream infection caused by SCCmec type IV strains of MRSA: an age of less than 1 year, less frequent use of a central venous catheter (odds ratio [OR], 0.07 [95% confidence interval {CI}, 0.02-0.28]; p= .025), and female sex. A second analysis was performed that excluded the case and control patients from the neonatal unit, and, in multivariate analysis, the following variables were significantly associated with having a nosocomial bloodstream infection caused by SCCmec type IV strains of MRSA: less frequent use of a central venous catheter (OR, 0.12 [95% CI, 0.03-0.55]; p= .007), lower Acute Physiology and Chronic Health Evaluation II score on admission (OR, 0.14 [95% CI, 0.03-0.61]; p= .009), less frequent surgery (OR, 0.21 [95% CI, 0.06-0.83]; p= .025), and female sex (OR, 5.70 [95% CI, 1.32-24.66]; p= .020). Of the 29 SCCmec type IV MRSA isolates recovered from case patients, none contained the Panton-Valentine leukocidin, gamma-hemolysin, enterotoxin B or C, or toxic shock syndrome toxin-1. All of the isolates contained genes for the LukE-LukD leukocidin and alpha-hemolysin. Genes for enterotoxin A were present in 1 isolate, and genes for beta-hemolysin were present in 3 isolates. "Classical" risk factors do not apply to patients infected with the SCCmec type IV strain of MRSA, which is an important cause of nosocomial bacteremia. This strain infects a patient population that is less ill and has had less frequent invasive procedures than a patient population infected with the multidrug-resistant strain of SCCmec type III MRSA. We found that virulence factors were rare and that Panton-Valentine leukocidin was absent. There were multiple clones of the SCCmec type IV strain in our hospital. Children under 1 year of age were at a higher risk. There was a predominant clone (sequence type 5) in this patient population.


Impact of left ventricular assist device (LVAD) support on the cardiac reverse remodeling process.
With improved technology and expanding indications for use, left ventricular assist devices (LVADs) are assuming a greater role in the care of patients with end-stage heart failure. Following LVAD implantation with the intention of bridge to transplant, it became evident that some patients exhibit substantial recovery of ventricular function. This prompted explantation of some devices in lieu of transplantation, the so-called bridge-to-recovery (BTR) therapy. However, clinical outcomes following these experiences are not always successful. Patients treated in this fashion have often progressed rapidly back to heart failure. Special knowledge has emerged from studies of hearts supported by LVADs that provides insights into the basic mechanisms of ventricular remodeling and possible limits of ventricular recovery. In general, it was these studies that spawned the concept of reverse remodeling now recognized as an important goal of many heart failure treatments. Important examples of myocardial and/or ventricular properties that do not regress towards normal during LVAD support include abnormal extracellular matrix metabolism, increased tissue angiotensin levels, myocardial stiffening and partial recovery of gene expression involved with metabolism. Nevertheless, studies of LVAD-heart interactions have led to the understanding that although we once considered the end-stage failing heart of patients near death to be irreversibly diseased, an unprecedented degree of myocardial recovery is possible, when given sufficient mechanical unloading and restoration of more normal neurohormonal milieu. Evidence supporting and unsupporting the notion of reverse remodeling and clinical implications of this process will be reviewed.


Systemic vascular effects during acute rejection of lung allografts.
Circulating leukocytes activated during rejection of organ allografts could potentially have generalized effects on systemic blood vessels of the transplant recipient. Experiments were designed, therefore, to determine the function of the endothelium and smooth muscle of arteries from nontransplanted organs in dogs who received single lung transplants. Dogs underwent single lung allotransplantation and were immunosuppressed for 5 days. Immunosuppression was then withheld for 3 days, allowing rejection to occur. Dogs were studied at this time (rejecting) or following treatment for rejection for an additional 6-8 days (treated). Arteries from unoperated, untreated dogs also were studied to provide baseline responses of healthy tissue. Rings cut from left circumflex coronary, nonoperated native pulmonary, and renal arteries were suspended in organ chambers for measurement of isometric force. Endothelium-dependent relaxations to the calcium ionophore A23187 were not affected by rejection in any of the arteries. Contractions to angiotensin I were reduced significantly only in native pulmonary arteries. Contractions to KCI and endothelin-1 increased in renal arteries with endothelium during rejection. These contractions in renal arteries were reduced following treatment of rejection. None of the responses of the coronary arteries were affected significantly by rejection of the lung allograft. These results demonstrate that contractions of arteries in the transplant recipient's native organs are altered during rejection of lung allografts. The effects are organ specific, may include production of endothelium-derived contractile factors in renal arteries, and can be partially reversed by treatment of rejection.


Development of a screening method for the determination of PCBs in water using QuEChERS extraction and gas chromatography-triple quadrupole mass spectrometry.
A new method has been developed and validated for the simultaneous analysis of 11 polychlorinated biphenyls (PCBs), in water samples at trace levels by gas chromatography coupled to triple quadrupole mass spectrometry (GC-QqQ-MS/MS). Water samples were extracted by the QuEChERS (Quick Easy Cheap Effective Rugged and Safe) method. The QqQ analyzer acquired data in multiple reactions monitoring (MRM), permitting both quantification and confirmation in a single injection with a running time reduced up to 11.0 min. The effect of matrix interferences in extracts on analyte quantification and the identification of PCBs in water samples was deeply studied. The results showed that PCBs were prone to strong matrix interactions in water samples, and the quantification and identification of PCBs were highly affected by a matrix enhancement effect. To evaluate the performance of the method, validation experiments were carried out on water samples at three spiking levels (1.6, 8.0, 40.0 µg L(-1)). Recovery was in the range of 95 - 109% at 1.6 µg L(-1), 90 - 95% at 8.0 µg L(-1) and 97 - 102% at 40.0 µg L(-1), respectively. Precision values expressed as relative standard deviation (RSD) were lower than 15%. Linearity in the range of 0.5 - 50.0 µg L(-1) provided determination coefficients (R(2)) higher than 0.999 for all compounds. The limits of detection (LODs) for PCBs were ≤0.1 µg L(-1) and the limits of quantification (LOQs) ranged from 0.04 to 0.3 µg L(-1). The applicability of the proposed method to detect and quantify PCBs has been demonstrated in analyse of 15 real samples.


A stochastic analysis of distance estimation approaches in single molecule microscopy - quantifying the resolution limits of photon-limited imaging systems.
Optical microscopy is an invaluable tool to visualize biological processes at the cellular scale. In the recent past, there has been significant interest in studying these processes at the single molecule level. An important question that arises in single molecule experiments concerns the estimation of the distance of separation between two closely spaced molecules. Presently, there exists different experimental approaches to estimate the distance between two single molecules. However, it is not clear as to which of these approaches provides the best accuracy for estimating the distance. Here, we address this problem rigorously by using tools of statistical estimation theory. We derive formulations of the Fisher information matrix for the underlying estimation problem of determining the distance of separation from the acquired data for the different approaches. Through the Cramer-Rao inequality, we derive a lower bound to the accuracy with which the distance of separation can be estimated. We show through Monte-Carlo simulations that the bound can be attained by the maximum likelihood estimator. Our analysis shows that the distance estimation problem is in fact related to the localization accuracy problem, the latter being a distinct problem that deals with how accurately the location of an object can be determined. We have carried out a detailed investigation of the relationship between the Fisher information matrices of the two problems for the different experimental approaches considered here. The paper also addresses the issue of a singular Fisher information matrix, which presents a significant complication when calculating the Cramer-Rao lower bound. Here, we show how experimental design can overcome the singularity. Throughout the paper, we illustrate our results by considering a specific image profile that describe the image of a single molecule.


Management of the asymptomatic primary in the palliative treatment of metastatic colorectal cancer.
The management of the asymptomatic primary in stage IV colorectal cancer presents a dilemma. There is an increased morbidity and mortality from surgical resection. Nonresectional management of the primary is associated with the risks of obstruction, perforation or haemorrhage. Our practice in patients with stage IV disease is palliative chemotherapy and symptom control. We reviewed our nonoperatively managed patients with colorectal liver metastases in order to identify the percentage of patients requiring urgent operative interventions for symptoms related to the primary. SUBJECTS/PATIENTS AND METHOD: A retrospective review of all patients treated for stage IV disease at our institution from 2003-2006 was undertaken. Patients were identified from multidisciplinary team (MDT) records. Demographic detail, treatment, and follow-up data were extracted from hospital records. These were analysed with Microsoft Excel. Thirty-seven patients were identified. 26 Male:11 Female. Median age 63 years (range 38-78). The median survival from diagnosis was 14 months. Three (8%) patients developed obstruction whilst having palliative chemotherapy. Two required a defunctioning stoma, and one was treated by means of a stent. There were no similarities between these three patients in terms of age, sex, site or stage of primary, volume of liver metastases, and alkaline phosphatase (ALP) or carcinoembryonic antigen (CEA) levels. Of 37 patients initially treated palliatively for stage IV colorectal cancer, 92% required no surgical treatment of their primary. Therefore it is the experience of this MDT that it is acceptable to treat such patients in an expectant manner. It is not possible to predict those patients, likely to require surgical intervention.


The protective effect of community factors on childhood asthma.
Asthma burden in the US is not evenly distributed. Although asthma prevalence varies widely across urban neighborhoods, little attention has been paid to the community as a key contributor. To determine the effect of positive socio-environmental community factors on childhood asthma prevalence in Chicago. From 2003 to 2005, an asthma screening survey was conducted among children attending Chicago Public/Catholic schools from kindergarten through eighth grade. One hundred five schools participated, yielding a stratified representation of 4 race-income groups. Positive community factors, such as social capital, economic potential, and community amenities, were assessed by using the Metro Chicago Information Center's Community Vitality Index. Of the surveys returned, 45,177 (92%) were geocoded into 287 neighborhoods. Neighborhoods were divided into quartile groups by asthma prevalence (mean, 8%, 12%, 17%, 25%). Community vitality (54% vs 44%; P < .0001) and economic potential (64% vs 38%; P < .0001) were significantly higher in neighborhoods with low asthma prevalence. Neighborhood interaction (36% vs 73%; P < .0001) and stability (40% vs 53%; P < .0001) were significantly higher in neighborhoods with high asthma prevalence. Overall, positive factors explained 21% of asthma variation. Childhood asthma increased as the black population increased in a community (P < .0001). Accordingly, race/ethnicity was controlled. In black neighborhoods, these factors remained significantly higher in neighborhoods with low asthma prevalence. When considered alongside socio-demographic/individual characteristics, overall community vitality as well as social capital continued to contribute significantly to asthma variation. Asthma prevalence in Chicago is strongly associated with socio-environmental factors thought to enrich a community. A deeper understanding of this impact may lend insight into interventions to reduce childhood asthma.


Human pregnancy after transfer of intact frozen-thawed embryos.
Since the birth of the first baby as a result of in vitro fertilisation (IVF) in 1978, many clinics around the world have achieved pregnancies and births for their patients by using IVF and gamete intrafallopian transfer procedures. With the storage of excess embryos, multiple laparoscopies can be avoided; this has favoured the development of better cryopreservation techniques. In our clinic 8-cell human embryos are frozen in a 1.5M dimethyl sulphoxide solution as cryoprotectant using the slow freeze-thaw method. Sixteen thawed embryos were replaced in 8 patients, resulting in 1 pregnancy. Of the thawed embryos 51.6% survived the freezing process in that they had 50% or more of the original number of blastomeres and also the zona pellucida intact.


[Effects of glycine on auditory evoked potentials among diabetic patients with auditory pathway neuropathy].
Glycine inhibits the formation of advanced glycation end products that may cause central and peripheral neuronal damage, affecting also the auditory nerve. To evaluate the effect of glycine on auditory nerve conduction and hearing level among patients with type 2 diabetes mellitus and auditory neuropathy. Twenty grams of oral glycine per day were administered during 6 months to 28 type 2 diabetic patients aged 58 ± 6 years, with auditory pathway neuropathy. Hearing tests and evoked otoacustic potentials were performed regularly. Fifteen diabetic patients aged 49 ± 8 years, without auditory nerve neuropathy did not receive glycine and were followed as a control group. Among patients receiving glycine, a significant improvement in left ear audiometry at 125, 250 and 500 Hz and right ear audiometry at 500 Hz, was observed. Waves I, III and V (p= 0.02) of evoked otoacustic potentials improved significantly in the left ear and wave I in the right ear. Among controls, waves V and III of evoked otoacoustic potentials had a significant impairment in the left ear. There was an improvement in auditory evoked potentials in patients receiving glycine and an impairment in untreated control patients.


The region of phage T4 genes 34, 33 and 59: primary structures and organization on the genome.
The product of gene 33 is essential for the regulation of late transcription and gene product 59 is required in recombination, DNA repair and replication. The exact functions of both proteins are not known. Restriction fragments spanning the genomic area of genes 33 and 59 have been cloned into phage M13 and a 4.9 kb nucleotide sequence has been determined. Translation of the DNA sequence predicted that gp33 contains 112 amino acids with a mol.wt. of 12.816 kd while gp59 is composed of 217 amino acids adding up to a mol.wt. of 25.967 kd. The genomic area studied here also contains 3 open reading frames of genes not identified to date and it is thought to include the NH2-terminal part of g34. One of the open reading frames seems to code for the 10 kd protein, probably involved in the regulation of transcription of bacteriophage T4. This protein is predicted to consist of 89 amino acid residues with a mol.wt. of 10.376 kd. Gene 33 and the gene for the 10 kd protein were cloned separately on high expression vectors resulting in over-production of the two proteins.


A systematic gene-gene and gene-environment interaction analysis of DNA repair genes XRCC1, XRCC2, XRCC3, XRCC4, and oral cancer risk.
Oral cancer is the sixth most common cancer worldwide with a high mortality rate. Biomarkers that anticipate susceptibility, prognosis, or response to treatments are much needed. Oral cancer is a polygenic disease involving complex interactions among genetic and environmental factors, which require multifaceted analyses. Here, we examined in a dataset of 103 oral cancer cases and 98 controls from Taiwan the association between oral cancer risk and the DNA repair genes X-ray repair cross-complementing group (XRCCs) 1-4, and the environmental factors of smoking, alcohol drinking, and betel quid (BQ) chewing. We employed logistic regression, multifactor dimensionality reduction (MDR), and hierarchical interaction graphs for analyzing gene-gene (G×G) and gene-environment (G×E) interactions. We identified a significantly elevated risk of the XRCC2 rs2040639 heterozygous variant among smokers [adjusted odds ratio (OR) 3.7, 95% confidence interval (CI)=1.1-12.1] and alcohol drinkers [adjusted OR=5.7, 95% CI=1.4-23.2]. The best two-factor based G×G interaction of oral cancer included the XRCC1 rs1799782 and XRCC2 rs2040639 [OR=3.13, 95% CI=1.66-6.13]. For the G×E interaction, the estimated OR of oral cancer for two (drinking-BQ chewing), three (XRCC1-XRCC2-BQ chewing), four (XRCC1-XRCC2-age-BQ chewing), and five factors (XRCC1-XRCC2-age-drinking-BQ chewing) were 32.9 [95% CI=14.1-76.9], 31.0 [95% CI=14.0-64.7], 49.8 [95% CI=21.0-117.7] and 82.9 [95% CI=31.0-221.5], respectively. Taken together, the genotypes of XRCC1 rs1799782 and XRCC2 rs2040639 DNA repair genes appear to be significantly associated with oral cancer. These were enhanced by exposure to certain environmental factors. The observations presented here warrant further research in larger study samples to examine their relevance for routine clinical care in oncology.


Melatonin reduces dimethylnitrosamine-induced liver fibrosis in rats.
Increased deposition of the extracellular matrix components, particularly collagen, is a central phenomenon in liver fibrosis. Stellate cells, the central mediators in the pathogenesis of fibrosis are activated by free radicals, and synthesize collagen. Melatonin is a potent physiological scavenger of hydroxyl radicals. Melatonin has also been shown to be involved in the inhibitory regulation of collagen content in tissues. At present, no effective treatment of liver fibrosis is available for clinical use. We aimed to test the effects of melatonin on dimethylnitrosamine (DMN)-induced liver damage in rats. Wistar albino rats were injected with DMN intraperitoneally. Following a single dose of 40 mg/kg DMN, either saline (DMN) or 100 mg/kg daily melatonin was administered for 14 days. In other rats, physiologic saline or melatonin were injected for 14 days, following a single injection of saline as control. Hepatic fibrotic changes were evaluated biochemically by measuring tissue hydroxyproline levels and histopathogical examination. Malondialdehyde (MDA), an end product of lipid peroxidation, and glutathione (GSH) and superoxide dismutase (SOD) levels were evaluated in blood and tissue homogenates. DMN caused hepatic fibrotic changes, whereas melatonin suppressed these changes in five of 14 rats (P < 0.05). DMN administration resulted in increased hydroxyproline and MDA levels, and decreased GSH and SOD levels, whereas melatonin reversed these effects. When melatonin was administered alone, no significant changes in biochemical parameters were noted. In conclusion, the present study suggests that melatonin functions as a potent fibrosuppressant and antioxidant, and may be a therapeutic choice.


Relative Scale Estimation and 3D Registration of Multi-Modal Geometry Using Growing Least Squares.
The advent of low cost scanning devices and the improvement of multi-view stereo techniques have made the acquisition of 3D geometry ubiquitous. Data gathered from different devices, however, result in large variations in detail, scale, and coverage. Registration of such data is essential before visualizing, comparing and archiving them. However, state-of-the-art methods for geometry registration cannot be directly applied due to intrinsic differences between the models, e.g., sampling, scale, noise. In this paper we present a method for the automatic registration of multi-modal geometric data, i.e., acquired by devices with different properties (e.g., resolution, noise, data scaling). The method uses a descriptor based on Growing Least Squares, and is robust to noise, variation in sampling density, details, and enables scale-invariant matching. It allows not only the measurement of the similarity between the geometry surrounding two points, but also the estimation of their relative scale. As it is computed locally, it can be used to analyze large point clouds composed of millions of points. We implemented our approach in two registration procedures (assisted and automatic) and applied them successfully on a number of synthetic and real cases. We show that using our method, multi-modal models can be automatically registered, regardless of their differences in noise, detail, scale, and unknown relative coverage.


Characterisation of long-term non-progression of HIV-1 infection after seroconversion: a cohort study.
Some individuals remain AIDS-free with a high and stable CD4 cell count without antiretroviral therapy (ART) for many years. We estimated long-term progression-free survival after HIV seroconversion and aimed to identify factors associated with loss of long-term non-progression (LTNP) status. For this cohort study, we used data for individuals with well-estimated dates of HIV-1 seroconversion from the CASCADE Collaboration a network of 28 HIV seroconverter cohort studies in Europe, Australia, Canada, and sub-Saharan Africa. The first cohort began enrolling patients in 1979, and for this analysis we used data pooled in May 1, 2011. We defined non-progression as being HIV-positive without AIDS, ART-naive, and with CD4 counts of 500 cells per μL or higher. We defined LTNP as non-progression during the first 10 years after seroconversion. We used longitudinal methods to characterise LTNP. Of the 4979 HIV seroconverters in our dataset, 3708 (75%) were men. Median time to progression was 2·07 years (95% CI 1·96-2·17), giving estimated progression-free survivals of 18·4% (17·2-19·6) 5 years, 4·0% (3·6-4·5) 10 years, and 1·4% (0·9-1·5) 15 years after seroconversion. The rate of progression did not change beyond 10 years after seroconversion (0·28 [95%CI 0·26-0·31] per person-year at 10 years after seroconversion, 0·24 [0·19-0·29] per person-year at 15 years, and 0·18 [0·10-0·33] per person-year at 20 years). At 10 years since HIV seroconversion, 283 individuals had LTNP, of whom 202 subsequently lost this status (median time to loss of status 2·49 years [2·05-2·92]). In univariable analyses, loss of LTNP status was associated with CD4 cell count at 10 years after seroconversion (p < 0·0001) and HIV RNA load at 10 years after seroconversion (p = 0·005), but not age (p = 0·544), mode of infection (p = 0·621), sex (p = 0·676), or calendar year of seroconversion (p = 0·397). In the multivariable analyses, loss of LTNP status was associated with lower CD4 counts at 10 years after seroconversion (p < 0·0001). After exclusion of CD4 cell counts from the model, higher HIV RNA load at 10 years after seroconversion was independently associated with loss of LTNP status (p = 0·009). Progression-free survival is rare. Most individuals with LTNP eventually lose immunological and clinical control of HIV infection eventually. European Union Seventh Framework Programme.


Proteomic analysis and molecular modelling characterize the iron-regulatory protein haemojuvelin/repulsive guidance molecule c.
HJV (haemojuvelin) plays a key role in iron metabolism in mammals by regulating expression of the liver-derived hormone hepcidin, which controls systemic iron uptake and release. Mutations in HJV cause juvenile haemochromatosis, a rapidly progressing iron overload disorder in humans. HJV, also known as RGMc (repulsive guidance molecule c), is a member of the three-protein RGM family. RGMs are GPI (glycosylphosphatidylinositol)-linked glycoproteins that share ~50% amino acid identity and several structural motifs, including the presence of 14 cysteine residues in analogous locations. Unlike RGMa and RGMb, HJV/RGMc is composed of both single-chain and two-chain isoforms. To date there is no structural information for any member of the RGM family. In the present study we have mapped the disulfide bonds in mouse HJV/RGMc using a proteomics strategy combining sequential MS steps composed of ETD (electron transfer dissociation) and CID (collision-induced dissociation), in which ETD induces cleavage of disulfide linkages, and CID establishes disulfide bond assignments between liberated peptides. The results of the present study identified an HJV/RGMc molecular species containing four disulfide linkages. We predict using ab initio modelling that this molecule is a single-chain HJV/RGMc isoform. Our observations outline a general approach using tandem MS and ab initio molecular modelling to define unknown structural features in proteins.


Rab7B/42 Is Functionally Involved in Protein Degradation on Melanosomes in Keratinocytes.
Keratinocytes uptake melanosomes from melanocytes and retain them in the perinuclear region, where they form melanin caps. Although these processes are crucial to protecting nuclear DNA against ultraviolet injury, the molecular basis of melanosome uptake and decomposition in keratinocytes is poorly understood. One of the major reasons for its being poorly understood is the lack of a specific marker protein that can be used to visualize or monitor melanosomes (or melanosome-containing compartments) that have been incorporated into keratinocytes. In this study, we performed a comprehensive localization screening for mammalian Rab family small GTPases (Rab1-45) and succeeded in identifying 11 Rabs that were enriched around melanosomes that had been incorporated into keratinocytes. We also established a new assay by using a recently developed melanosome probe (called M-INK) as a means of quantitatively assessing the degradation of proteins on incorporated melanosomes in control and each of a series of Rab-knockdown keratinocytes. The results showed that knockdown or CRISPR/Cas9-mediated knockout of Rab7B (also identified as Rab42) in keratinocytes caused strong inhibition of protein degradation on melanosomes. Our findings indicated that Rab7B/42 is recruited to melanosome-containing compartments and that it promotes protein degradation on melanosomes in keratinocytes.Key words: degradation, keratinocytes, melanocytes, melanosome, Rab small GTPase.


Discriminating primary clinical states in bipolar disorder with a comprehensive symptom scale.
We assessed the spectrum and severity of bipolar symptoms that differentiated bipolar disorder (BD) clinical states, employing the Bipolar Inventory of Symptoms Scale (BISS) which provides a broader item range of traditional depression and mania rating scales. We addressed symptoms differentiating mixed states from depression or mania/hypomania. One hundred and sixteen subjects who met DSM-IV-TR criteria for BD and were currently in a depressed, manic/hypomanic, mixed episode, or recovered state were interviewed using the BISS. A subset of manic items differed between mixed episodes and mania/hypomania or depression. Most anxiety items were more severe in mixed subjects. BISS Depression and Manic subscales differentiated episodes from recovered status. The majority of depression and manic symptoms differentiated mood states in the predicted direction. Mixed episodes had overall greater mood severity than manic/hypomanic episodes or depressed episodes. These results indicate that a small subset of symptoms, several of which are absent in DSM-IV-TR criteria and traditional rating scales for bipolar studies, aid in distinguishing mixed episodes from depressive or manic/hypomanic episodes. The results also support the utility of a comprehensive BD symptom scale in distinguishing primary clinical states of BD.


Role of two-component sensory systems of Salmonella enterica serovar Dublin in the pathogenesis of systemic salmonellosis in cattle.
Salmonella enterica serovar Dublin (S. Dublin) is associated with enteritis, typhoid and abortion in cattle. Infections are acquired by the oral route, and the bacteria transit through varied anatomical and cellular niches to elicit systemic disease. S. Dublin must therefore sense and respond to diverse extrinsic stimuli to control gene expression in a spatial and temporal manner. Two-component systems (TCSs) play key roles in such processes, and typically contain a membrane-associated sensor kinase (SK) that modifies a cognate response regulator. Analysis of the genome sequence of S. Dublin identified 31 conserved SK genes. Each SK gene was separately disrupted by lambda Red recombinase-mediated insertion of transposons harbouring unique sequence tags. Calves were challenged with a pool of the mutants together with control strains of defined virulence by the oral and intravenous routes. Quantification of tagged mutants in output pools derived from various tissues and cannulated lymphatic vessels allowed the assignment of spatial roles for each SK following oral inoculation or when the intestinal barrier was bypassed by intravenous delivery. Mutant phenotypes were also assigned in cultured intestinal epithelial cells. Mutants with insertions in barA, envZ, phoQ, ssrA or qseC were significantly negatively selected at all enteric and systemic sites sampled after oral dosing. Mutants lacking baeS, dpiB or citA were negatively selected at some but not all sites. After intravenous inoculation, only barA and phoQ mutants were significantly under-represented at systemic sites. The novel role of baeS in intestinal colonization was confirmed by oral co-infection studies, with a mutant exhibiting modest but significant attenuation at a number of enteric sites. This is the first systematic analysis of the role of all Salmonella TCSs in a highly relevant model of enteric fever. Spatial roles were assigned to eight S. Dublin SKs, but most were not essential for intestinal or systemic infection of the target host.


Lymphocyte proteomics of Parkinson's disease patients reveals cytoskeletal protein dysregulation and oxidative stress.
There is increasing evidence of biochemical alterations in peripheral blood lymphocytes of Parkinson's disease (PD) patients. In this work, we describe the changes in protein levels in peripheral lymphocytes of PD patients in order to identify potential peripheral biomarkers. By means of 2D electrophoresis and mass spectrometry protein identification, we compared patients under L-3,4-dihydroxyphenylalanine (L-DOPA) treatment, patients under subthalamic nucleus deep-brain stimulation and healthy controls. Statistical analysis of the results demonstrated that cofilin-1, tropomyosin, and a specific actin isoform vary significantly in patients, regardless of the therapy. Two different isoforms of gamma-fibrinogen either correlate with the disease state or with the disease duration. Eventually, specific changes associated with the different therapies allowed to highlight oxidative stress conditions in lymphocytes in patients treated with higher doses of L-DOPA. As a whole, peripheral blood lymphocytes are sensitive reporters of PD over inter-individual variability, and allow the identification of specific alterations that could be further exploited for diagnostic purposes.


Mitochondrial oxidative phosphorylation system is recruited to detergent-resistant lipid rafts during myogenesis.
Since detergent-resistant lipid rafts play important roles in the signal transduction for myogenesis, their comprehensive proteomic analysis could provide new insights to understand their function in myotubes. Here, the detergent-resistant lipid rafts were isolated from C2C12 myotubes and analyzed by capillary RPLC/MS/MS. Among the 327 proteins (or protein groups) identified, 28% were categorized to the plasma membrane or raft proteins, 29% to mitochondria, 20% to microsomal proteins, 10% to other proteins, and 13% to unknown proteins. The localization of oxidative phosphorylation (OXPHOS) complexes in the sarcolemma lipid rafts was further confirmed from C2C12 myotubes by cellular fractionation, surface-biotin labeling, immunofluorescence, and lipid raft fractionation. After adding exogenous cytochrome c, the sarcolemma isolated from myotubes had an ability to consume oxygen in the presence of NADH or succinate. The generation of NADH-dependent extracellular superoxide was increased by inhibiting or downregulating OXPHOS I, III, and IV in myotubes, indicating that OXPHOS proteins are major sources for extracellular ROS in skeletal muscle. With all these data, we can conclude that OXPHOS proteins are associated with the sarcolemma lipid rafts during C2C12 myogenesis to generate extracellular ROS.


Survey of medical students to assess their knowledge and attitudes toward organ transplantation and donation.
Kidney transplantation is the most preferred treatment modality for patients with end-stage renal disease (ESRD). This study aims at understanding the awareness, attitudes, and beliefs among the medical and nonmedical students. The study population consisted of 500 medical students and 39 nonmedical controls, who were surveyed using a reliable questionnaire that examined their knowledge and attitudes. A 24-item self-administered questionnaire, which assessed the levels of knowledge, attitude regarding organ donation with dichotomous scale and demographic data were used. Of the 500 medical students who received the questionnaire, 376 (75.2%) with a mean age of 22.1 ± 2.5 years responded; 39% were male, 43.6% were Bahraini, and 32.2% were from Saudi Arabia, 51.3% were in Grades-5 and 6 and 58.8% resided in big cities. The medical students had a highly positive attitude and great willingness toward organ donation. Majority of them (75.3%) knew the treatment of ESRD, and 70.7% recognized correctly that kidney transplantation is the optimal treatment for ESRD. However, only 10.4% knew that it is performed in Bahrain since 1995. A total of 241 participants (64.3%) reported positive attitude toward living kidney donation and 71.8% expressed their agreement to donation after death. More than half (66.8%) believed that their religion permits organ donation, although 56.6% of the participants believed that there is a danger after donating a kidney. When compared to nonmedical students, there was no statistically significant difference in the attitudes toward living kidney donation (P = 0.823) or organ donation after death (P = 0.066).


Human airway monohydroxyeicosatetraenoic acid generation and mucus release.
The effects of 5-, 8-, 9-, 11-, 12-, and 15-monohydroxyeicosatetraenoic acid (HETE) (0.1-100 nM) on mucous glycoprotein release from cultured human airways were determined. Each of the HETE was an active secretagogue of mucus at concentrations greater than 1-10 nM with 12- and 15-HETE, the most active. Both 5- and 9-hydroperoxyeicosatetraenoic acid (HPETE) were also active as secretagogues at 100 nM, although of somewhat lower potency. As cultured airways were capable of responding to HETE with mucous glycoprotein release, it was of interest to identify and quantitate airway HETE formation. Accordingly, airways were incubated with tracer quantities of [14C]arachidonate for 16-48 h, and the spontaneous formation of 5-, 12- and 11- and/or 15-HETE was measured by high-pressure liquid chromatography. Indeed, sizeable quantities of 11- and/or 15- greater than 5- greater than 12-HETE were generated. This HETE generation was increased by the addition of 25 micrograms/ml of arachidonate and was reduced somewhat after 18-21 d in continuous tissue culture. Reversed anaphylaxis of human airways using anti-human IgE markedly increased the HETE formation, resulting in the production of micromolar concentrations of 5- and 11- and/or 15-HETE. Thus, human airways not only are capable of responding to the presence of HETE with mucous glycoprotein release, but also generate (both spontaneously and in response to anaphylaxis) at least three species of HETE, and do so in quantities capable of acting as mucus secretagogues.


Scoring system for differentiating perforated and non-perforated pediatric appendicitis.
Appendicitis is the most common indication for emergency pediatric surgery and its most significant complication is perforation. Perforated appendicitis (PA) may be managed conservatively, whereas non-perforated appendicitis (NP) is managed surgically. Recent studies have shown that ultrasound (US) is effective for differentiating between PA and NP, and does not expose pediatric patients to ionizing radiation. The purpose of this study is to enhance the accuracy of differentiation with a novel scoring system based on clinical, laboratory, and US findings. This retrospective study included 243 patients aged 2-17 years who presented between 2006 and 2013 with surgically proven appendicitis, of whom 60 had perforation. Clinical and laboratory data were collected and US images evaluated by a pediatric radiologist. To create the scoring system, point values were assigned to each parameter. A randomly selected training sample of 137 subjects was used to create a scoring prediction model. The model was tested on the remaining 106 patients. Scores of ≥6, ≥11, and ≥15 yielded specificities of 64, 91, and 99%, and sensitivities of 96, 61, and 29%, respectively (p < 0.001). We have designed a scoring system incorporating clinical, laboratory, and sonographic findings which can differentiate PA from NP with high specificity.


Evaluating sex as a biological variable in preclinical research: the devil in the details.
Translating policy into action is a complex task, with much debate surrounding the process whereby US and Canadian health funding agencies intend to integrate sex and gender science as an integral component of methodological rigor and reporting in health research. Effective January 25, 2016, the US National Institutes of Health implemented a policy that expects scientists to account for the possible role of sex as a biological variable (SABV) in vertebrate animal and human studies. Applicants for NIH-funded research and career development awards will be asked to explain how they plan to factor consideration of SABV into their research design, analysis, and reporting; strong justification will be required for proposing single-sex studies. The Canadian Institutes of Health Research is revising their peer review accreditation process to ensure that peer reviewers are skilled in applying a critical lens to protocols that should be incorporating sex and gender science. The current paper outlines the components that peer reviewers in North America will be asked to assess when considering whether SABV is appropriately integrated into research designs, analyses, and reporting. Consensus argues against narrowly defining rules of engagement in applying SABV, with criteria provided for reviewers as guidance only. Scores will not be given for each criterion; applications will be judged on the overall merit of scientific innovation, rigor, reproducibility, and potential impact.


Impact of New Cardiovascular Events on Quality of Life and Hospital Costs in People With Cardiovascular Disease in the United Kingdom and United States.
Background Despite optimized risk factor control, people with prior cardiovascular disease remain at high cardiovascular disease risk. We assess the immediate- and longer-term impacts of new vascular and nonvascular events on quality of life (QoL) and hospital costs among participants in the REVEAL (Randomized Evaluation of the Effects of Anacetrapib Through Lipid Modification) trial in secondary prevention. Methods and Results Data on demographic and clinical characteristics, health-related quality of life (QoL: EuroQoL 5-Dimension-5-Level), adverse events, and hospital admissions during the 4-year follow-up of the 21 820 participants recruited in Europe and North America informed assessments of the impacts of new adverse events on QoL and hospital costs from the UK and US health systems' perspectives using generalized linear regression models. Reductions in QoL were estimated in the years of event occurrence for nonhemorrhagic stroke (-0.067 [United Kingdom], -0.069 [US]), heart failure admission (-0.072 [United Kingdom], -0.103 [US]), incident cancer (-0.064 [United Kingdom], -0.068 [US]), and noncoronary revascularization (-0.071 [United Kingdom], -0.061 [US]), as well as in subsequent years following these events. Myocardial infarction and coronary revascularization (CRV) procedures were not found to affect QoL. All adverse events were associated with additional hospital costs in the years of events and in subsequent years, with the highest additional costs in the years of noncoronary revascularization (£5830 [United Kingdom], $14 133 [US Medicare]), of myocardial infarction with urgent CRV procedure (£5614, $24722), and of urgent/nonurgent CRV procedure without myocardial infarction (£4674/£4651 and $15 251/$17 539). Conclusions Stroke, heart failure, and noncoronary revascularization procedures substantially reduce QoL, and all cardiovascular disease events increase hospital costs. These estimates are useful in informing cost-effectiveness of interventions to reduce cardiovascular disease risk in secondary prevention. Registration URL: https://www.clinicaltrials.gov; Unique identifier: NCT01252953; https://www.Isrctn.com. Unique identifier: ISRCTN48678192; https://www.clinicaltrialsregister.eu. Unique identifier: 2010-023467-18.


Colonic mucosal and exfoliome transcriptomic profiling and fecal microbiome response to a flaxseed lignan extract intervention in humans.
Microbial metabolism of lignans from high-fiber plant foods produces bioactive enterolignans, such as enterolactone (ENL) and enterodiol (END). Enterolignan exposure influences cellular pathways important to cancer risk and is associated with reduced colon tumorigenesis in animal models and lower colorectal cancer risk in humans. The aim of this study was to test the effects of a flaxseed lignan supplement (50 mg secoisolariciresinol diglucoside/d) compared with placebo on host gene expression in colon biopsies and exfoliated colonocyte RNA in feces and fecal microbial community composition, and to compare responses in relation to ENL excretion. We conducted a 2-period randomized, crossover intervention in 42 healthy men and women (20-45 y). We used RNA-seq to measure differentially expressed (DE) genes in colonic mucosa and fecal exfoliated cells through the use of edgeR and functional analysis with Ingenuity Pathway Analysis. We used 16S ribosomal RNA gene (V1-V3) analysis to characterize the fecal microbiome, and measured END and ENL in 24-h urine samples by gas chromatography-mass spectrometry. We detected 32 DE genes (false discovery rate <0.05) in the exfoliome, but none in the mucosal biopsies, in response to 60 d of lignan supplement compared with placebo. Statistically significant associations were detected between ENL excretion and fecal microbiome measured at baseline and at the end of the intervention periods. Further, we detected DE genes in colonic mucosa and exfoliome between low- and high-ENL excreters. Analysis of biopsy samples indicated that several anti-inflammatory upstream regulators, including transforming growth factor β and interleukin 10 receptor, were suppressed in low-ENL excreters. Complementary analyses in exfoliated cells also suggested that low-ENL excreters may be predisposed to proinflammatory cellular events due to upregulation of nuclear transcription factor κB and NOS2, and an inhibition of the peroxisome proliferator-activated receptor γ network. These results suggest that ENL or other activities of the associated gut microbial consortia may modulate response to a dietary lignan intervention. This has important implications for dietary recommendations and chemoprevention strategies. This study was registered at clinicaltrials.gov as NCT01619020.


Monitoring trends in HIV prevalence among young people, aged 15 to 24 years, in Manicaland, Zimbabwe.
In June 2001, the United Nations General Assembly Special Session (UNGASS) set a target of reducing HIV prevalence among young women and men, aged 15 to 24 years, by 25% in the worst-affected countries by 2005, and by 25% globally by 2010. We assessed progress toward this target in Manicaland, Zimbabwe, using repeated household-based population serosurvey data. We also validated the representativeness of surveillance data from young pregnant women, aged 15 to 24 years, attending antenatal care (ANC) clinics, which UNAIDS recommends for monitoring population HIV prevalence trends in this age group. Changes in socio-demographic characteristics and reported sexual behaviour are investigated. Progress towards the UNGASS target was measured by calculating the proportional change in HIV prevalence among youth and young ANC attendees over three survey periods (round 1: 1998-2000; round 2: 2001-2003; and round 3: 2003-2005). The Z-score test was used to compare differences in trends between the two data sources. Characteristics of participants and trends in sexual risk behaviour were analyzed using Student's and two-tailed Z-score tests. HIV prevalence among youth in the general population declined by 50.7% (from 12.2% to 6.0%) from round 1 to 3. Intermediary trends showed a large decline from round 1 to 2 of 60.9% (from 12.2% to 4.8%), offset by an increase from round 2 to 3 of 26.0% (from 4.8% to 6.0%). Among young ANC attendees, the proportional decline in prevalence of 43.5% (from 17.9% to 10.1%) was similar to that in the population (test for differences in trend: p value=0.488) although ANC data significantly underestimated the population prevalence decline from round 1 to 2 (test for difference in trend: p value=0.003) and underestimated the increase from round 2 to 3 (test for difference in trend: p value=0.012). Reductions in risk behaviour between rounds 1 and 2 may have been responsible for general population prevalence declines. In Manicaland, Zimbabwe, the 2005 UNGASS target to reduce HIV prevalence by 25% was achieved. However, most prevention gains occurred before 2003. ANC surveillance trends overall were an adequate indicator of trends in the population, although lags were observed. Behaviour data and socio-demographic characteristics of participants are needed to interpret ANC trends.


Growth-restricting effects of siRNA transfections: a largely deterministic combination of off-target binding and hybridization-independent competition.
Perturbation of gene expression by means of synthetic small interfering RNAs (siRNAs) is a powerful way to uncover gene function. However, siRNA technology suffers from sequence-specific off-target effects and from limitations in knock-down efficiency. In this study, we assess a further problem: unintended effects of siRNA transfections on cellular fitness/proliferation. We show that the nucleotide compositions of siRNAs at specific positions have reproducible growth-restricting effects on mammalian cells in culture. This is likely distinct from hybridization-dependent off-target effects, since each nucleotide residue is seen to be acting independently and additively. The effect is robust and reproducible across different siRNA libraries and also across various cell lines, including human and mouse cells. Analyzing the growth inhibition patterns in correlation to the nucleotide sequence of the siRNAs allowed us to build a predictor that can estimate growth-restricting effects for any arbitrary siRNA sequence. Competition experiments with co-transfected siRNAs further suggest that the growth-restricting effects might be linked to an oversaturation of the cellular miRNA machinery, thus disrupting endogenous miRNA functions at large. We caution that competition between siRNA molecules could complicate the interpretation of double-knockdown or epistasis experiments, and potential interactions with endogenous miRNAs can be a factor when assaying cell growth or viability phenotypes.


Mechanisms of adenosine triphosphate-, thrombin-, and trypsin-induced relaxation of rat thoracic aorta.
The mechanisms by which adenosine triphosphate, thrombin, and trypsin cause relaxation of vascular smooth muscle were investigated. Relaxation of the rat thoracic aorta with adenosine triphosphate, thrombin, and/or trypsin was associated with increased levels of cyclic guanosine monophosphate in both time- and concentration-dependent manners. Thrombin and trypsin did not alter cyclic adenosine monophosphate levels, whereas adenosine triphosphate increased cyclic adenosine monophosphate levels after significant relaxation occurred. Removal of the endothelium abolished adenosine triphosphate-, thrombin-, and trypsin-induced relaxation and the associated increased levels of cyclic nucleotides. Relaxation due to these agents was also inhibited by exposure to nordihydroguaiaretic acid, a lipoxygenase inhibitor, and eicosatetraynoic acid, a lipoxygenase and cyclooxygenase inhibitor. Indomethacin, a cyclooxygenase inhibitor, potentiated relaxation to these agents, whereas the increased levels of cyclic nucleotides due to adenosine triphosphate were unaltered. Bromophenacyl bromide, a phospholipase A2 inhibitor, decreased relaxation due to adenosine triphosphate, thrombin, and trypsin and the associated increased levels of cyclic nucleotides. Removal of extracellular calcium, which also presumably inhibits phospholipase A2, prevented the elevated levels of cyclic nucleotides and the inhibitory effects of adenosine triphosphate and trypsin on contraction. In contrast, sodium nitroprusside-induced relaxation and/or increased levels of cyclic guanosine monophosphate were unaltered by nordihydroguaiaretic acid, eicosatetraynoic acid, bromophenacyl bromide, and removal of extracellular calcium. After incubation of intact tissue with 32P-orthophosphate, the patterns of protein phosphorylation caused by adenosine triphosphate, thrombin, and trypsin were indistinguishable from those of acetylcholine, sodium nitroprusside and 8-bromo cyclic guanosine monophosphate. All these agents dephosphorylated myosin light chain. Thus, the present study supports the hypothesis that relaxation induced by adenosine triphosphate, thrombin, and trypsin is mediated through the formation of an endothelial factor which elevates cyclic guanosine monophosphate levels and causes cyclic guanosine monophosphate-dependent protein phosphorylation and dephosphorylation of myosin light chain.


Thalamic field potentials in chronic central pain treated by periventricular gray stimulation -- a series of eight cases.
Chronic deep brain stimulation (DBS) of the periventricular gray (PVG) has been used for the treatment of chronic central pain for decades. In recent years motor cortex stimulation (MCS) has largely supplanted DBS in the surgical management of intractable neuropathic pain of central origin. However, MCS provides satisfactory pain relief in about 50-75% of cases, a range comparable to that reported for DBS (none of the reports are in placebo-controlled studies and hence the further need for caution in evaluating and comparing these results). Our experience also suggests that there is still a role for DBS in the control of central pain. Here we present a series of eight consecutive cases of intractable chronic pain of central origin treated with PVG DBS with an average follow-up of 9 months. In each case, two electrodes were implanted in the PVG and the ventroposterolateral thalamic nucleus, respectively, under guidance of corneal topography/magnetic resonance imaging image fusion. The PVG was stimulated in the frequency range of 2-100 Hz in alert patients while pain was assessed using the McGill-Melzack visual analogue scale. In addition, local field potentials (FPs) were recorded from the sensory thalamus during PVG stimulation. Maximum pain relief was obtained with 5-35 Hz stimulation while 50-100 Hz made the pain worse. This suggests that pain suppression was frequency dependent. Interestingly, we detected low frequency thalamic FPs at 0.2-0.4 Hz closely associated with the pain. During 5-35 Hz PVG stimulation the amplitude of this potential was significantly reduced and this was associated with marked pain relief. At the higher frequencies (50-100 Hz), however, there was no reduction in the FPs and no pain suppression. We have found an interesting and consistent correlation between thalamic electrical activity and chronic pain. This low frequency potential may provide an objective index for quantifying chronic pain, and may hold further clues to the mechanism of action of PVG stimulation. It may be possible to use the presence of these slow FPs and the effect of trial PVG DBS on both the clinical status and the FPs to predict the probable success of future pain control in individual patients.


Mechanistic study of hydrazine decomposition on Ir(111).
Hydrogen transport and storage technology remain one of the critical challenges of the hydrogen economy. Hydrazine (N2H4) is a carbon-free hydrogen carrier which has been widely used as fuel in the field of space exploration. We have combined experiments and computer simulations in order to gain a better understanding of the N2H4 decomposition on Ir catalyst, the most efficient catalyst for hydrazine decomposition up to date. We have identified metallic Ir rather than IrO2 as the active phase for hydrazine decomposition and carried out density functional theory (DFT) calculations to systematically investigate the changes in the electronic structure along with the catalytic decomposition mechanisms. Three catalytic mechanisms to hydrazine decomposition over Ir(111) have been found: (i) intramolecular reaction between hydrazine molecules, (ii) intramolecular reaction between co-adsorbed amino groups, and (iii) hydrazine dehydrogenation assisted by co-adsorbed amino groups. These mechanisms follow five different pathways for which transition states and intermediates have been identified. The results show that hydrazine decomposition on Ir(111) starts preferentially with an initial N-N bond scission followed by hydrazine dehydrogenation assisted by the amino group produced, eventually leading to ammonia and nitrogen production. The preference for N-N scission mechanisms was rationalized by analyzing the electronic structure. This analysis showed that upon hydrazine adsorption, the π bond between nitrogen atoms becomes weaker.


Himalayan fossils of the oldest known pantherine establish ancient origin of big cats.
Pantherine felids ('big cats') include the largest living cats, apex predators in their respective ecosystems. They are also the earliest diverging living cat lineage, and thus are important for understanding the evolution of all subsequent felid groups. Although the oldest pantherine fossils occur in Africa, molecular phylogenies point to Asia as their region of origin. This paradox cannot be reconciled using current knowledge, mainly because early big cat fossils are exceedingly rare and fragmentary. Here, we report the discovery of a fossil pantherine from the Tibetan Himalaya, with an age of Late Miocene-Early Pliocene, replacing African records as the oldest pantherine. A 'total evidence' phylogenetic analysis of pantherines indicates that the new cat is closely related to the snow leopard and exhibits intermediate characteristics on the evolutionary line to the largest cats. Historical biogeographic models provide robust support for the Asian origin of pantherines. The combined analyses indicate that 75% of the divergence events in the pantherine lineage extended back to the Miocene, up to 7 Myr earlier than previously estimated. The deeper evolutionary origin of big cats revealed by the new fossils and analyses indicate a close association between Tibetan Plateau uplift and diversification of the earliest living cats.


Identification of neonatal hearing impairment: infants with hearing loss.
This article describes the audiologic findings and medical status of infants who were found to have hearing loss, detected as part of the Identification of Neonatal Hearing Impairment (INHI) project. In addition, the neonatal and maternal health variables for the group of infants who could not be tested with visual reinforcement audiometry (VRA) due to developmental and visual disability are presented. The overall goal of the INHI project was to evaluate the test performance of auditory brain stem response and evoked otoacoustic emission (OAE) tests given in the newborn period. These tools were evaluated on the basis of the infants' hearing when tested behaviorally with VRA at 8 to 12 mo corrected age. The neonatal test results, VRA results, medical history information and a record of intercurrent events occurring between the neonatal period and the time of VRA were collated and reviewed. The purpose of this article is to review the characteristics of those infants who were found to have hearing loss. Of 2995 infants who had VRA tests judged to be of good or fair reliability, 168 had a finding of hearing loss for at least one ear, an incidence of 5.6%. Sixty-six infants had bilateral losses, an incidence of 2%, and 22 infants had bilateral hearing losses in the moderate to profound range, an incidence 0.7%. The prevalence of middle ear problems was greater than 50% among these infants with hearing loss. From the larger group of 168 infants with hearing loss, a group of 56 infants (86 ears) was chosen as those with a low probability that the hearing loss was due to transient middle ear pathology and was more likely hearing loss of a permanent nature. These were the infants used for the analyses of neonatal test performance (Norton et al., 2000). In this selected group there were 30 infants with bilateral impairment of at least mild degree, which is an incidence of 1%. There were approximately equal numbers of ears in the mild, moderate, severe and profound range of hearing loss. Risk factors associated with hearing loss were reviewed for the total sample of infants tested with VRA and for those infants with hearing loss. A history of treatment with aminoglycosides was the risk factor most often reported in the entire sample; however, there was no difference in prevalence of this risk factor for the normal-hearing and hearing-impaired groups. The risk factor associated with the highest incidence of hearing loss was stigmata of syndromes associated with sensorineural hearing loss and other neurosensory disorders. Sixty-seven infants who returned for follow-up could not be tested with VRA due to severe developmental delay or visual disability. Many of these infants had medical histories indicating the sequelae of extreme prematurity and/or very low birthweight. Most of the hearing losses found in this study were mild and, based on clinical history and tympanometry tests, many of the mild and some of the moderate impairments may have been acquired in early infancy due to middle ear effusion. In the group of infants used for determination of neonatal test performance there were approximately equal numbers of mild, moderate, severe and profound losses. Only a small percentage of infants with a conventional risk indicator for hearing loss actually had a hearing loss, and there were a significant number of infants with hearing loss who did not have a risk indicator. These findings support the need for an early identification program based on universal neonatal hearing screening rather than by targeted testing of those with risk indicators.


An optimized transformation protocol for Anthoceros agrestis and three more hornwort species.
Land plants comprise two large monophyletic lineages, the vascular plants and the bryophytes, which diverged from their most recent common ancestor approximately 480 million years ago. Of the three lineages of bryophytes, only the mosses and the liverworts are systematically investigated, while the hornworts are understudied. Despite their importance for understanding fundamental questions of land plant evolution, they only recently became amenable to experimental investigation, with Anthoceros agrestis being developed as a hornwort model system. Availability of a high-quality genome assembly and a recently developed genetic transformation technique makes A. agrestis an attractive model species for hornworts. Here we describe an updated and optimized transformation protocol for A. agrestis, which can be successfully used to genetically modify one more strain of A. agrestis and three more hornwort species, Anthoceros punctatus, Leiosporoceros dussii, and Phaeoceros carolinianus. The new transformation method is less laborious, faster, and results in the generation of greatly increased numbers of transformants compared with the previous method. We have also developed a new selection marker for transformation. Finally, we report the development of a set of different cellular localization signal peptides for hornworts providing new tools to better understand the hornwort cell biology.