EEG Reactivity Predicts Individual Efficacy of Vagal Nerve Stimulation in Intractable Epileptics.
Background: Chronic vagal nerve stimulation (VNS) is a well-established non-pharmacological treatment option for drug-resistant epilepsy. This study sought to develop a statistical model for prediction of VNS efficacy. We hypothesized that reactivity of the electroencephalogram (EEG) to external stimuli measured during routine preoperative evaluation differs between VNS responders and non-responders. Materials and Methods: Power spectral analyses were computed retrospectively on pre-operative EEG recordings from 60 epileptic patients with VNS. Thirty five responders and 25 non-responders were compared on the relative power values in four standard frequency bands and eight conditions of clinical assessment-eyes opening/closing, photic stimulation, and hyperventilation. Using logistic regression, groups of electrodes within anatomical areas identified as maximally discriminative by n leave-one-out iterations were used to classify patients. The reliability of the predictive model was verified with an independent data-set from 22 additional patients. Results: Power spectral analyses revealed significant differences in EEG reactivity between responders and non-responders; specifically, the dynamics of alpha and gamma activity strongly reflected VNS efficacy. Using individual EEG reactivity to develop and validate a predictive model, we discriminated between responders and non-responders with 86% accuracy, 83% sensitivity, and 90% specificity. Conclusion: We present a new statistical model with which EEG reactivity to external stimuli during routine presurgical evaluation can be seen as a promising avenue for the identification of patients with favorable VNS outcome. This novel method for the prediction of VNS efficacy might represent a breakthrough in the management of drug-resistant epilepsy, with wide-reaching medical and economic implications.


Flexible polyimide probes with microelectrodes and embedded microfluidic channels for simultaneous drug delivery and multi-channel monitoring of bioelectric activity.
The study of intracellular communication requires devices that can not only monitor the bioelectric activity, but also control and observe the biochemical environment at the cellular level. This paper reports on the development and characterisation of implantable polyimide microprobes that allow simultaneous, selective chemical delivery/probing and multi-channel recording/stimulation of bioelectric activity. The key component of the system is a flexible polyimide substrate with embedded microchannels that is batch-fabricated combining polyimide micromachining and a lamination technique. The devices provide platinum microelectrodes on both sides of the polyimide substrate with an active surface between 20 microm x 20 microm and 50 microm x 50 microm. The embedded microchannels permit highly localised drug delivery or probing at the tip of the device via channel outlets adjacent to the microelectrodes. The microelectrodes were characterised by electrical impedance spectroscopy and the microchannels were studied in microflow experiments. Two different fluid delivery schemes were explored in two different designs. The first device type consists of a simple combination of microchannels and microelectrodes on one substrate. Liquids are ejected at the tip of the device by pressure injection techniques. The second device was inspired by the so-called U-tube concept allowing for highly localised delivery of controlled amounts of liquids in the picoliters range. Thus, the influence of chemical compounds on the electrical activity of cells can be studied with high temporal and spatial resolution. The flexible, implantable devices can be used for studying the chemical and electrical information exchange and communication of cells in in vivo and in vitro experiments.


Outcome and long-term growth after extensive small bowel resection in the neonatal period: a survey of 87 children.
This retrospective study aims to analyze the outcome, the prognosis factors and the long-term growth of children after extensive small bowel (SB) resection in the neonatal period. 87 children, born between 1975 and 1991 who had undergone extensive neonatal small bowel resection, were followed up over a mean period of 15 years. Anatomical data influencing PN dependency and duration were analyzed. Data on height and weight were collected and compared using growth standards. Final heights were studied for patients who achieved their puberty and compared to predicted height based on Tanner's formula. Patients were analyzed according to PN weaning and growth: children still receiving PN (group A), patients weaned from initial PN but requiring PN once again or enteral feeding (group B), and children with permanent intestinal autonomy (group C). The overall survival is 89.7 %, depending on the date of birth. The duration of PN-dependency varies according to the intestinal length and the presence of the ileocaecal valve (ICV). All patients who remain PN dependent had less than 40 cm of small bowel and/or the absence of ICV. Patients in group B had a mean small bowel length of 35 +/- 19 cm, resection of the ICV in 50 % of cases, and a PN duration of 47.4 +/- 23.8 months. There was a significant decrease in height and weight gain within the 4 years after cessation of PN, requiring enteral or parenteral feeding. Patients in group C had a mean small bowel length of 57 +/- 19 cm, presence of ICV in 81 % of cases and a PN duration of 16.1 +/- 11.4 months. After PN weaning, they grow up normally with normal puberty and final height as predicted from genetic target height. PN duration is influenced by the length of residual SB and the absence of ICV. With good anatomic prognosis factors and short duration of initial PN, normal long-term growth may be predicted. Conversely, poor anatomical factors and protracted initial PN require careful monitoring of growth and may sometimes require nutritional support to be restarted. The last group, permanently dependent on PN, might be candidates for intestinal transplantation.


Arousal, working memory capacity, and sexual decision-making in men.
This study investigated whether working memory capacity (WMC) moderated the relationship between physiological arousal and sexual decision making. A total of 59 men viewed 20 consensual and 20 non-consensual images of heterosexual interaction while their physiological arousal levels were recorded using skin conductance response. Participants also completed an assessment of WMC and a date-rape analogue task for which they had to identify the point at which an average Australian male would cease all sexual advances in response to verbal and/or physical resistance from a female partner. Participants who were more physiologically aroused by and spent more time viewing the non-consensual sexual imagery nominated significantly later stopping points on the date-rape analogue task. Consistent with our predictions, the relationship between physiological arousal and nominated stopping point was strongest for participants with lower levels of WMC. For participants with high WMC, physiological arousal was unrelated to nominated stopping point. Thus, executive functioning ability (and WMC in particular) appears to play an important role in moderating men's decision making with regard to sexually aggressive behavior.


Oxidation of phenolic acid derivatives by soil and its relevance to allelopathic activity.
Previous studies have suggested that phenolic acids from legume green manures may contribute to weed control through allelopathy. The objectives of this study were to investigate the oxidation reactions of phenolic acids in soil and to determine the subsequent effects of oxidation upon phytotoxicity. Soils were reacted for 18 h with 0.25 mmol L(-1) benzoic and cinnamic acid derivative solutions and Mn release from the suspension was used as a marker for phenolic acid oxidation. The extent of oxidation in soil suspensions was in the order of 3,4dihydroxy- > 4-hydroxy-3-methoxy- > 4-hydroxy-approximately 2-hydroxy-substituted benzoic and cinnamic acids. The same ranking was observed for cyclic voltammetry peak currents of the cinnamic acid derivatives. This suggests that the oxidation of phenolic acids is controlled by the electron transfer step from the sorbed phenolic acid to the metal oxide. A bioassay experiment showed that the 4-hydroxy-, 4-hydroxy-3-methoxy-, and 3,4-dihydroxy-substituted cinnamic acids were bioactive at 0.25 mmol L(-1) concentration. Reaction with soil for 18 h resulted in the elimination of bioactivity of these three cinnamic acids at the 5% significance level. The oxidative reactivity of phenolic acids may limit the potential of allelopathy as a component of an integrated weed management system. However, the initial phytotoxicity after soil incorporation may coincide with the early, critical stage of weed emergence and establishment, so that allelopathic phenolic acids may still play a role in weed management despite their reactivity in soil systems.


Field-based video observations of wild barnacle cyprid behaviour in response to textural and chemical settlement cues.
Many marine invertebrate larvae respond behaviourally to environmental settlement cues, yet behaviours are often only inferred from settlement patterns or are limited to laboratory studies. The behaviour of wild cypris larvae of Semibalanus balanoides L. was filmed on settlement tiles in the field. Tiles were of five different textures with a nested treatment of crude conspecific adult extract (AE). The effects of texture and AE on eleven defined behaviours were analysed. Texture affected the gross and net exploratory distances, velocity, acceleration and time spent exploring. AE attracted more cyprids during the first minute of immersion and increased the time spent on surfaces. Relatively few arrivals that either travel far and fast, or exit the surface rapidly, may indicate a lower chance of settlement. An increase in time spent on a surface may increase the probability of being in contact with the surface when the sign stimulus to settle occurs.


Does oral alprazolam affect ventilation? A randomised, double-blind, placebo-controlled trial.
The respiratory effects of benzodiazepines have been controversial. This investigation aimed to study the effects of oral alprazolam on ventilation. In a randomised, double-blind cross-over protocol, 20 healthy men ingested 1 mg of alprazolam or placebo in random order, 1 week apart. Ventilation was unobtrusively monitored by inductance plethysmography along with end-tidal PCO(2) and pulse oximetry 60-160 min after drug intake. Subjects were encouraged to keep their eyes open. Mean +/- SD minute ventilation 120 min after alprazolam and placebo was similar (6.21 +/- 0.71 vs 6.41 +/- 1.12 L/min, P = NS). End-tidal PCO(2) and oxygen saturation did also not differ between treatments. However, coefficients of variation of minute ventilation after alprazolam exceeded those after placebo (43 +/- 23% vs 31 +/- 13%, P < 0.05). More encouragements to keep the eyes open were required after alprazolam than after placebo (5.2 +/- 5.7 vs 1.3 +/- 2.3 calls, P < 0.05). In a multiple regression analysis, higher coefficients of variation of minute ventilation after alprazolam were related to a greater number of calls. Oral alprazolam in a mildly sedative dose has no clinically relevant effect on ventilation in healthy, awake men. The increased variability of ventilation on alprazolam seems related to vigilance fluctuations rather than to a direct drug effect on ventilation.


Complete mitochondrial genome sequences of two extinct moas clarify ratite evolution.
The origin of the ratites, large flightless birds from the Southern Hemisphere, along with their flighted sister taxa, the South American tinamous, is central to understanding the role of plate tectonics in the distributions of modern birds and mammals. Defining the dates of ratite divergences is also critical for determining the age of modern avian orders. To resolve the ratite phylogeny and provide biogeographical data to examine these issues, we have here determined the first complete mitochondrial genome sequences of any extinct taxa--two New Zealand moa genera--along with a 1,000-base-pair sequence from an extinct Madagascan elephant-bird. For comparative data, we also generated 12 kilobases of contiguous sequence from the kiwi, cassowary, emu and two tinamou genera. This large dataset allows statistically precise estimates of molecular divergence dates and these support a Late Cretaceous vicariant speciation of ratite taxa, followed by the subsequent dispersal of the kiwi to New Zealand. This first molecular view of the break-up of Gondwana provides a new temporal framework for speciation events within other Gondwanan biota and can be used to evaluate competing biogeographical hypotheses.


Premarital HIV testing on prospective couples in a teaching hospital in sub Saharan Africa.
Most religious bodies insist on premarital screening for prospective couples. To determine the level of voluntary screening, prevalence and risk factors of HIV among premarital couples. A cross-sectional descriptive study using interviewer administered questionnaire and HIV screening to collect data. Systematic sampling of everythird premarital couples attending the General outpatient Clinic between November 2010 and October 2011. SPSS version 17 was used for data analysis of 386 subjects. A majority of respondents (83.4% or 322); X2 = 172.446, df = 1, p < .05) had knowledge of transmission of HIV. Mandatory HIV screening was 5.7% (22); X2 = 303.018, df = 1, p < .05), while voluntary testing was 30% (113). The sero-prevalence of HIV was 2.6%, which was statistically and significantly associated with cohabitation and upper social class. A majority (68.4% or 266); X2 = 272.166, df = 3, p < .05) would call off marriage plans if their spouses tested positive to HIV. CONCLUSION The knowledge of transmission of HIV was very high with low sero-prevalence (2.6%) among premarital couples. Voluntary HIV testing was higher than mandatory request, while cohabitation and upper social class are risk factors for HIV transmission.


Reasons for Reason-Giving: The Obamacare Debates.
The demanding and giving of reasons for their actions are core business for public policy makers in a democracy. But there are many different reasons for asking why questions, and correspondingly many different responses that might count as adequate answers. Seven different reasons for reason-giving are here distinguished and categorized along two dimensions: political moralism versus political realism, and high versus low politics. All of those were in play in the enactment and adjudication of the Affordable Care Act. The attempts at repealing it were characterized by low-politics and political-realist modes of reason-giving more exclusively.


Needs-appropriate services for people with young onset dementia: The perspectives of healthcare professionals.
Young onset dementia has become a growing challenge in national healthcare systems. Concerns about the situation have already been expressed; however, they mostly report the views of services end-users. Aimed at finding out useful further improvement, interviews documenting the working experiences of professionals addressing healthcare services to people with young onset dementia (YOD) were analysed qualitatively. The interviews resulted in four themes and twelve categories. The themes regarded services' complexity and responsiveness, the levels of education and knowledge on YOD, the impact that serving people with YOD reflects on staff working experience and quality of life, and a series of proposals aimed at improving services by giving people with YOD and families the provision of care they deserve. Although some of the themes recapped the ones reported in the previous literature, the mixture of internally and externally driven instances represented in the themes depict the complexity of care delivery in the services. Findings are discussed in the light of a pragmatical framework capable of suggesting what changes services should implement to be timely responsive.


[Colorectal cancer: controversial role of meat consumption].
Diet is supposed to influence the colorectal cancer etiology, but the precise causative factors are yet unknown. International ecological studies show a strong correlation between meat consumption and the colorectal cancer incidence. Most case-control studies (22 of 29) show an increased risk to develop a colorectal cancer for those eating higher amounts of meat. In contrast, only 2 out of the 5 best prospective cohort studies have shown this positive association for red meat. Two studies out of 4 show an association with processed meat. Consumption of white meat or of fish is not associated with a high risk, and might even reduce the occurrence of colorectal cancer. Several plausible hypotheses concerning the link between meat and colon carcinogenesis have been suggested. They involve saturated fat, protein, iron, heterocyclic amines produced by cooking, and N-nitroso compounds. High fat diets may promote cancer because they have a high caloric content, or because they lead to increased levels of bile acids in the colonic lumen. Six experimental studies are published on the effect of meat, or meat fractions, on the colon tumor incidence in rodents initiated with chemical carcinogens. Data from these studies do not support the belief that red meat (beef) has a specific effect on intestinal carcinogenesis. Instead, diets containing beef meat (cooked or raw) decrease carcinogenesis when compared to control diets containing similar amounts of fat and protein of vegetal origin. However, high fat or high protein diets often increase carcinogenesis when compared to low fat or low protein diets. Thus, one cannot state, nor exclude, that meat promotes colorectal cancer.


Measuring health-related quality of life in MSA: the MSA-QoL.
The objective of this study was to develop a new patient-reported outcome measure for patients with multiple system atrophy (MSA) and to test its psychometric properties. There were three stages. First, a pool of potential scale items was generated from in-depth patient interviews. Second, these items were administered, in the form of a questionnaire, to a sample of people with MSA and traditional psychometric methods used to develop a rating scale satisfying standard criteria for reliability and validity. Third, the psychometric properties of the rating scale were examined in a second sample. In stage one, a pool of 105 items was generated from 20 patient interviews. In stage two, a scale with three subscales (motor, 14 items; nonmotor, 12 items; emotional/social functioning, 14 items), satisfying standard criteria for reliability and validity, was developed from the response data of 317 patients with MSA (response rate 71%). In stage three, the scale was examined in 286 people with MSA. Missing data were low, scores in both subscales were evenly distributed, and floor and ceiling effects were small. Reliability was high (Cronbach's alpha 0.83-0.93; test-retest ICC 0.88-0.92). Validity was supported by the interscale correlations (r = 0.47-0.59), known group differences, and the magnitude and pattern of correlations with four other rating scales, disease severity, and disease duration. In conclusion, the patient-rated MSA health-related Quality of life scale (MSA-QoL) may be a suitable patient-reported scale for use in clinical trials and studies in MSA.


Incorporation of the purified human placental insulin receptor into phospholipid vesicles.
Purified human placental insulin receptors were incorporated into small unilamellar phospholipid vesicles by the addition of n-octyl beta-glucopyranoside solubilized phospholipids, followed by removal of the detergent on a Sephadex G-50 gel filtration column and extensive dialysis. The vesicles have an average diameter of 142 +/- 24 nm by Sephacryl S-1000 gel filtration chromatography and 119 +/- 20 nm by transmission electron microscopy. These vesicles are impermeant to small molecules as indicated by their ability to retain [gamma-32P]ATP, which could be released by the addition of 0.05% Triton X-100. Detergent permeabilization or freeze-thawing of the insulin receptor containing vesicles in the presence of 125I-insulin indicated that approximately 75% of the insulin binding sites were oriented right side out (extravesicularly). Sucrose gradient centrifugation of insulin receptors incorporated at various protein to phospholipid mole ratios demonstrated that the insulin receptors were inserted into the phospholipid bilayer structure in a concentration-dependent manner. Addition of [gamma-32P]ATP to the insulin receptor containing vesicles was relatively ineffective in promoting the autophosphorylation of the beta subunit in the absence or presence of insulin. Permeabilization of the vesicles with low detergent concentrations, however, stimulated the beta-subunit autophosphorylation approximately 2-fold in the absence and 10-fold in the presence of insulin. Insulin-stimulated beta-subunit autophosphorylation was also observed under conditions such that 94% of those vesicles containing insulin receptors had a single receptor per vesicle, suggesting that the initial beta-subunit autophosphorylating activity is intramolecular. Phospho amino acid analysis of the vesicle-incorporated insulin receptors demonstrated that the basal and insulin-stimulated beta-subunit autophosphorylation occurs exclusively on tyrosine residues. It is concluded that when purified insulin receptors are incorporated into a phospholipid bilayer, they insert into the vesicles primarily in the same orientation as occurs in the plasma membrane of intact cells and retain insulin binding as well as insulin-stimulated beta-subunit autophosphorylating activities.


DNA sequences homologous to the Drosophila opa repeat are present in murine mRNAs that are differentially expressed in fetuses and adult tissues.
A mouse embryonic cDNA containing two opa-like (CAX)n repeats was isolated on the basis of its cross-hybridization with a Drosophila K10 cDNA. Such repeated sequences were present in different murine mRNAs, some of which were specifically expressed during fetal life or in different adult tissues. This suggests that, as already described for Drosophila, opa-like sequences are parts of proteins involved in ontogenic or cell-type-specific functions in vertebrates. However, unlike Drosophila, such repeated sequences were not found within the murine homeo-boxes containing genes of the Hox-1 complex.


MHC class I expression in mice lacking the proteasome subunit LMP-7.
Proteasomes degrade endogenous proteins. Two subunits, LMP-2 and LMP-7, are encoded in a region of the major histocompatibility complex (MHC) that is critical for class I-restricted antigen presentation. Mice with a targeted deletion of the gene encoding LMP-7 have reduced levels of MHC class I cell-surface expression and present the endogenous antigen HY inefficiently; addition of peptides to splenocytes deficient in LMP-7 restores wild-type class I expression levels. This demonstrates the involvement of LMP-7 in the MHC class I presentation pathway and suggests that LMP-7 functions as an integral part of the peptide supply machinery.


The role of cdc25 in checkpoints and feedback controls in the eukaryotic cell cycle.
Major checkpoints that gate progression through the cell cycle function at the G1/S transition, entry into mitosis and exit from mitosis. Cells use feedback mechanisms to inhibit passage through these checkpoints in response to growth control signals, incomplete DNA replication or spindle assembly. In many organisms, transition points seem to involve regulation of the activity of cyclin-dependent kinases (cdks) not only through their interactions with various cyclins, but also by phosphorylation-dephosphorylation cycles acting on the kinase activity of the cdks. These phosphorylation cycles are modulated by the regulation of the opposing kinases and phosphatases that act on cdks and form feedback loops. In this article, we discuss the role of positive and negative feedback loops in cell cycle timing and checkpoints, focusing more specifically on the regulation of the dual specificity cdc25 phosphatase.


Less Is More: Oligomer Extraction and Hydrothermal Annealing Increase PDMS Adhesion Forces for Materials Studies and for Biology-Focused Microfluidic Applications.
Cues in the micro-environment are key determinants in the emergence of complex cellular morphologies and functions. Primary among these is the presence of neighboring cells that form networks. For high-resolution analysis, it is crucial to develop micro-environments that permit exquisite control of network formation. This is especially true in cell science, tissue engineering, and clinical biology. We introduce a new approach for assembling polydimethylsiloxane (PDMS)-based microfluidic environments that enhances cell network formation and analyses. We report that the combined processes of PDMS solvent-extraction and hydrothermal annealing create unique conditions that produce high-strength bonds between solvent-extracted PDMS (E-PDMS) and glass-properties not associated with conventional PDMS. Extraction followed by hydrothermal annealing removes unbound oligomers, promotes polymer cross-linking, facilitates covalent bond formation with glass, and retains the highest biocompatibility. Herein, our extraction protocol accelerates oligomer removal from 5 to 2 days. Resulting microfluidic platforms are uniquely suited for cell-network studies owing to high adhesion forces, effectively corralling cellular extensions and eliminating harmful oligomers. We demonstrate the simple, simultaneous actuation of multiple microfluidic domains for invoking ATP- and glutamate-induced Ca2+ signaling in glial-cell networks. These E-PDMS modifications and flow manipulations further enable microfluidic technologies for cell-signaling and network studies as well as novel applications.


Expected clinical competence from midwifery graduates during community service placement in Limpopo province, South Africa.
Community service nurses are placed in a hospital within the first year after qualifying to obtain clinical experience under the supervision of experienced professional nurses. When placed in clinical environments, new midwifery graduates are expected to be job ready, demonstrate competence in the provision of evidence-based care, practise independently and assume accountability and responsibility for their own actions. The study aimed at exploring the expectations of experienced midwives of clinical competence of newly graduated midwives during transition. The study was conducted at the training hospitals of the five districts in Limpopo province. The researcher used a qualitative approach which is explorative and descriptive in nature. The population comprised all the professional midwives with experience of 5 years and above working at the selected hospitals in Vhembe, Mopani, Capricorn, Waterberg and Sekhukhune districts of Limpopo province, South Africa. From each selected hospital, five experienced midwives were selected using non-probability, purposive sampling method. An in-depth individual face-to-face interview was used to collect data from the participants, until saturation was reached. The open-coding, Tesch's eight-step process was used to analyse data. Results revealed that newly graduated midwives failed to meet the perceived expectations by experienced midwives; this was reflected in sub-themes: limited sense of independence, limited participation in task sharing and commitment and competence versus incompetence to patient care. The experienced midwives expected newly graduated midwives to function as professionals; unfortunately, they expressed disappointment as graduates did not meet their expectations. Newly graduated midwives lacked sense of independence, commitment to patient care and did not display ability in task sharing.


Monitoring of pigeon paramyxovirus type-1 in organs of pigeons naturally infected with Salmonella Typhimurium.
An experimental pigeon paramyxovirus (pPMV-1) infection was followed by reverse transcription-nested polymerase chain reaction for 31 days after infection, in 16 organs of inoculated or contact pigeons naturally infected with Salmonella Typhimurium. With two exceptions, both groups presented similar results. Typical nervous signs and a green diarrhoea were observed. The spread of pPMV-1 was relatively quick, all organs being largely positive at 4 days after inoculation or contact. The lung, spleen, caecal tonsils, kidneys and brain, for which almost all tested samples remained positive during most of the experiment, seemed to be the principal targets for virus persistence. However, the virus was significantly recovered later in the brain parts and for longer in the trachea of the contact pigeons than of the inoculated ones.


Why do euploid embryos miscarry? A case-control study comparing the rate of aneuploidy within presumed euploid embryos that resulted in miscarriage or live birth using next-generation sequencing.
To determine whether undetected aneuploidy contributes to pregnancy loss after transfer of euploid embryos that have undergone array comparative genomic hybridization (aCGH). Case-control study. University-based fertility center. Cases included 38 patients who underwent frozen euploid ET as determined by aCGH, resulting in miscarriage. Controls included 38 patients who underwent frozen euploid ET as determined by aCGH, resulting in a live birth. Next-generation sequencing (NGS) protocols were internally validated. Saved amplified DNA samples from the blastocyst trophectoderm biopsies previously diagnosed as euploid by aCGH were reanalyzed using NGS. Cytogenetic reports of the products of conception for 20 of the pregnancies resulting in miscarriage were available for comparison. The incidence of aneuploidy and mosaicism using NGS within embryos resulting in miscarriage and live birth. Of euploid embryos analyzed by aCGH resulting in miscarriage, 31.6% were mosaic and 5.2% were polyploid by NGS. The rate of chromosomal abnormalities was significantly higher in embryos resulting in miscarriage (36.8%) than in those resulting in live births (15.8%). The rate of mosaicism was twice as high among embryos resulting in miscarriage than those resulting in live birth, but this was not statistically significant. Next-generation sequencing detected more cases of mosaicism than cytogenetic analysis of products of conception. Undetected aneuploidy may increase the risk of first trimester pregnancy loss. Next-generation sequencing may detect mosaicism and triploidy more frequently than aCGH, which could help to identify embryos at high risk of miscarriage. Mosaic embryos, however, should not be discarded as some can result in live births.


Nicotinamide deamidation by microorganisms in rat stomach.
We have extended our investigation of nicotinamide deamidation in the stomach of conventional rats. The bacterial species in the pars preventricularis were identified as Flavobacterium peregrinum, Escherichia coli, Streptococcus faecalis, and Lactobacillus acidophilus, listed in order of decreasing deamidase activity. Nicotinamide-7-(14)C ingested into rat stomach was rapidly deamidated to nicotinic acid. These results contribute to the accumulated evidence that microorganisms present in the pars preventricularis of rat stomach are responsible for the deamidation of nicotinamide to nicotinic acid, a known precursor of mammalian pyridine nucleotides.


Mutation patterns at dinucleotide microsatellite loci in humans.
Microsatellites are a major type of molecular markers in genetics studies. Their mutational dynamics are not clear. We investigated the patterns and characteristics of 97 mutation events unambiguously identified, from 53 multigenerational pedigrees with 630 subjects, at 362 autosomal dinucleotide microsatellite loci. A size-dependent mutation bias (in which long alleles are biased toward contraction, whereas short alleles are biased toward expansion) is observed. There is a statistically significant negative relationship between the magnitude (repeat numbers changed during mutation) and direction (contraction or expansion) of mutations and standardized allele size. Contrasting with earlier findings in humans, most mutation events (63%) in our study are multistep events that involve changes of more than one repeat unit. There was no correlation between mutation rate and recombination rate. Our data indicate that mutational dynamics at microsatellite loci are more complicated than the generalized stepwise mutation models.


Mapping a gene for familial hypertrophic cardiomyopathy to chromosome 14q1.
To identify the chromosomal location of a gene responsible for familial hypertrophic cardiomyopathy, we used clinical and molecular genetic techniques to evaluate the members of a large kindred. Twenty surviving and 24 deceased family members had hypertrophic cardiomyopathy; 58 surviving members were unaffected. Genetic-linkage analyses were performed with polymorphic DNA loci dispersed throughout the entire genome, to identify a locus that was inherited with hypertrophic cardiomyopathy in family members. The significance of the linkage detected between the disease locus and polymorphic loci was assessed by calculating a lod score (the logarithm of the probability of observing coinheritance of two loci, assuming that they are genetically linked, divided by the probability of detecting coinheritance if they are unlinked). A DNA locus (D14S26), previously mapped to chromosome 14 and of unknown function, was found to be coinherited with the disease in this family. No instances of recombination were observed between the locus for familial hypertrophic cardiomyopathy and D14S26, yielding a lod score of +9.37 (theta = 0). These data indicate that in this kindred, the odds are greater than 2,000,000,000:1 that the gene responsible for familial hypertrophic cardiomyopathy is located on chromosome 14 (band q1).


Medicinal values of fruit peels from Citrus sinensis, Punica granatum, and Musa paradisiaca with respect to alterations in tissue lipid peroxidation and serum concentration of glucose, insulin, and thyroid hormones.
Peel extracts from Citrus sinensis, Punica granatum, and Musa paradisiaca were investigated for their effects on tissue lipid peroxidation (LPO) and on the concentration of thyroid hormones, insulin, and glucose in male rats. In vitro inhibition of H(2)O(2)-induced LPO in red blood cells of rats by 0.25, 0.50, 1.0, and 2.0 microg/mL C. sinensis, P. granatum, and M. paradisiaca peel extracts was observed in a dose-specific manner. Maximum inhibition was observed at 0.50 microg/mL C. sinensis, 2.0 microg/mL P. granatum, and 1.0 microg/mL M. paradisiaca. In the in vivo investigation, out of four different concentrations of each peel extract, 25, 200, and 100 mg/kg C. sinensis, P. granatum, and M. paradisiaca, respectively, were found to maximally inhibit hepatic LPO. The most effective doses were further evaluated for effects on serum triiodothyronine (T(3)), thyroxine (T(4)), insulin, and glucose concentrations. C. sinensis exhibited antithyroidal, hypoglycemic, and insulin stimulatory activities, in addition to inhibition of LPO, as it significantly decreased the serum T(4) (P < .05) and glucose (P < .001) concentrations with a concomitant increase in insulin levels (P < .05). P. granatum decreased LPO in hepatic, cardiac, and renal tissues (P < .01, P < .001, and P < .05, respectively) and serum glucose concentration (P < .01). M. paradisiaca strongly inhibited the serum level of thyroid hormones (P < .01 for both T(3) and T(4)) but increased the level of glucose (P < .05). These findings reveal the hitherto unknown potential of the tested peel extracts in the regulation of thyroid function and glucose metabolism. Besides antiperoxidative activity, C. sinensis extract has antithyroidal, hypoglycemic, and insulin stimulatory properties, which suggest its potential to ameliorate both hyperthyroidism and diabetes mellitus.


Improving diets and nutrition through an integrated poultry value chain and nutrition intervention (SELEVER) in Burkina Faso: study protocol for a randomized trial.
The SELEVER study is designed to evaluate the impact of an integrated agriculture-nutrition package of interventions (including poultry value chain development, women's empowerment activities, and a behavior change communications strategy to promote improved diets and feeding, care, and hygiene practices) on the diets, health, and nutritional status of women and children in Burkina Faso. This paper presents the rationale and study design. The impact evaluation involves a cluster randomized controlled trial design that will be implemented in 120 rural communities/villages within 60 communes supported by SELEVER in the Boucle de Mouhoun, Centre-Ouest, and Haut-Bassins regions of Burkina Faso. Communities will be randomly assigned to one of three treatment arms, including: (1) SELEVER intervention group; (2) SELEVER with an intensive WASH component; and (3) control group without intervention. Primary outcomes include the mean probability of adequacy of diets for women and children (aged 2-4 years at baseline), infant and young child feeding practices of caregivers of children aged 0-2 years, and household poultry production and sales. Intermediate outcomes along the agriculture and nutrition pathways will also be measured, including child nutrition status and development. The evaluation will follow a mixed-methods approach, including a panel of child-, household-, community-, and market-level surveys, and data collection points during post-harvest and lean seasons, as well as one year after implementation completion to examine sustainability. To our knowledge, this study is the first to rigorously examine from a food systems perspective, the simultaneous impact of scaling-up nutrition-specific and nutrition-sensitive interventions through a livestock value-chain and community-intervention platform, across nutrition, health, and agriculture domains. The findings of this evaluation will provide evidence to support the design of market-based nutrition-sensitive interventions. ISRCTN registry, ISRCTN16686478 . Registered on 2 December 2016.


Block of intercellular communication: interaction of intracellular H+ and Ca2+.
The influence of elevated intracellular levels of H+ and Ca2+ on intercellular communication between cultured neonatal rat myocardial cells was examined by quantifying the percent of primary neighboring cells to which intracellularly injected Lucifer yellow had spread within 10 s of injection. Partial acidosis was induced by incubation in and then removal of NH4Cl. Intracellular Ca2+ was raised through the use of treatments that are standard in studies of heart muscle: reduction of the Na+ gradient, addition of caffeine, and combinations of these interventions. Under control conditions and during application of NH4Cl, cells exhibited spontaneous electrical and contractile activity and were well coupled (dye detectable in 100% of primary neighbors). Sustained intracellular acidosis without simultaneous elevation of intracellular Ca2+ (NH4Cl exposure followed by zero Na+, zero Ca2+) reduced the incidence of dye transfer to 90%. Elevation of intracellular Ca2+ (exposure to zero Na+, Ca2+-containing solution, with or without 10 mM caffeine) had no effect on coupling. These same interventions, when employed together, reduced the incidence of dye coupling to 18%. The results are consistent with a synergism of action of Ca2+ and H+ in the regulation of junctional permeability.


Evidence of high-risk sexual behaviors among injection drug users in the Kenya PLACE study.
Injection drug users (IDUs) in resource poor settings are at high risk for HIV transmission through unsafe needle-sharing and sexual practices. We report on the injecting and sexual behavior of a sample of IDUs from Malindi, Kenya. A Priority for Local AIDS Control Efforts (PLACE) study was conducted from April to May 2010 to identify areas where HIV transmission is most likely to occur and specific venues where people meet new sexual partners. Community informants (n=202) listed 157 unique venues from which 29 were randomly selected using a systematic fixed interval sampling strategy with probability of selection proportional to venue size. Twenty patrons and four workers were interviewed at each venue. Drug use practices were elicited in a staff-administered interview. Between 40% and 50% of IDUs reported needle-sharing, taking drugs from a common reservoir, using a ready-made solution without boiling, and not exchanging a used for a new syringe in the past month. Most could inconsistently or never get new syringes. In multivariate logistic regression models controlling for age, education, residence, and poverty status, IDUs were twice as likely as non-IDUs to report multiple partners in the past year (OR 1.94, 95% CI 1.26-3.00, p<.01) and multiple new partners in the past year (OR 2.11, 95% CI 1.30-3.42, p<.01). High prevalence of multiple sexual partnerships and risky injecting behaviors among IDUs and unavailability of new injecting needles are likely facilitating HIV transmission in Malindi, Kenya.


A high-throughput device for size based separation of C. elegans developmental stages.
Caenorhabditis elegans is a widely used model organism to study development, aging and behavior. Many of these biological studies require staging a large number of worms to assay a synchronized population of animals. Conventional synchronization techniques such as manual picking, gravity stratification and chemical bleaching are labor-intensive and could perturb animals' physiology. Thus, there is a need for a simple inexpensive technology to sort a mixed population of worms based on their developmental stages with minimal perturbation. Here we demonstrate a simple but accurate and high-throughput technique to sort based on animal size, which correlates well with developmental stages. The device consists of an array of geometrically optimized pillars that act as a sieve to allow worms of specific sizes to rapidly move through. With optimized chamber heights, pillar spacing and driving pressures, these binary separation devices are capable of independently separating a mixture of worms at two different stages at average efficiency of around 95%, and throughput of hundreds of worms per minute. In addition, when four devices are used sequentially, we demonstrate the ability to stratify a mixture of worms of all developmental stages with >85% overall efficiency.


Multiple ADH genes modulate risk for drug dependence in both African- and European-Americans.
Drug dependence (DD) is commonly co-morbid with alcohol dependence (AD). Many studies have also shown common genetic risk factors for these disorders. We previously reported associations of AD with seven alcohol dehydrogenase (ADH) genes. The present study examines the relationship between these genes and DD. We genotyped 16 markers within the ADH gene cluster and 38 unlinked ancestry-informative markers in a case-control sample of 718 individuals. All markers were consistent with Hardy-Weinberg equilibrium in controls, but some markers showed Hardy-Weinberg disequilibrium in cases (minimal P = 0.002). Genotypes of many markers were associated with DD, both before and after controlling for admixture effects (minimal P < 1.0 x 10(-6)). Diplotype trend regression analysis showed that ADH5 and ADH6 genotypes, and diplotypes at ADH1A, ADH1B, ADH1C and ADH7 (minimal P = 0.002), were associated with DD in European-Americans and/or African-Americans. This first report of an allelic association of these loci with DD provides new insight into the mechanism of genetic risk for DD. These findings, obtained using a series of powerful and reliable analytic methods, may also help to explain the high rate of co-morbidity between AD and DD.


The intense world theory - a unifying theory of the neurobiology of autism.
Autism covers a wide spectrum of disorders for which there are many views, hypotheses and theories. Here we propose a unifying theory of autism, the Intense World Theory. The proposed neuropathology is hyper-functioning of local neural microcircuits, best characterized by hyper-reactivity and hyper-plasticity. Such hyper-functional microcircuits are speculated to become autonomous and memory trapped leading to the core cognitive consequences of hyper-perception, hyper-attention, hyper-memory and hyper-emotionality. The theory is centered on the neocortex and the amygdala, but could potentially be applied to all brain regions. The severity on each axis depends on the severity of the molecular syndrome expressed in different brain regions, which could uniquely shape the repertoire of symptoms of an autistic child. The progression of the disorder is proposed to be driven by overly strong reactions to experiences that drive the brain to a hyper-preference and overly selective state, which becomes more extreme with each new experience and may be particularly accelerated by emotionally charged experiences and trauma. This may lead to obsessively detailed information processing of fragments of the world and an involuntarily and systematic decoupling of the autist from what becomes a painfully intense world. The autistic is proposed to become trapped in a limited, but highly secure internal world with minimal extremes and surprises. We present the key studies that support this theory of autism, show how this theory can better explain past findings, and how it could resolve apparently conflicting data and interpretations. The theory also makes further predictions from the molecular to the behavioral levels, provides a treatment strategy and presents its own falsifying hypothesis.


Photopic temporal processing in retinitis pigmentosa.
The relation between early changes in the photopic flicker electroretinogram (ERG) and photopic psychophysical changes in retinitis pigmentosa (RP) is poorly understood. Here, abnormalities in foveal and extrafoveal temporal contrast sensitivity functions (TCSFs) were studied in a group of carefully selected patients with RP who had relatively preserved macular function. The psychophysical results were compared with changes in the timing of the multifocal ERG. Subjects were patients with RP who had acuity > or =20/32 and no visual field defects within 6 degrees from the fovea. Maxwellian-view and direct-view optical systems were used to obtain foveal and extrafoveal TCSFs under a range of test conditions, including high retinal illuminances that yielded temporal contrast sensitivity independent of mean retinal illuminance. TCSFs were described using log sensitivity and corner frequency parameters. Foveal TCSFs in these patients showed overall reductions in sensitivity but no frequency-dependent defects. Also, no macular defects were found in the timing of the multifocal ERG. TCSFs from extrafoveal locations in moderate field defects, obtained at retinal illuminances that were sufficient to render flicker sensitivity independent of effective mean luminance, showed reductions in overall sensitivity as well as changes in temporal tuning. The multifocal ERGs from these extrafoveal locations showed signs of temporal slowing. Changes in temporal tuning (both psychophysical and electroretinographic) were found only within visual field scotomas, whereas changes of the log sensitivity parameter were found also in the relatively preserved foveas of this group of patients with early stage RP.


XIF3, a Xenopus peripherin gene, requires an inductive signal for enhanced expression in anterior neural tissue.
A full-length cDNA clone for the Xenopus intermediate filament gene XIF3 has been isolated. It is very similar in sequence to the rat intermediate filament cDNA clone 73 that is thought to encode the neuronal intermediate filament protein 'peripherin'. By analysing dissected embryos, we show that XIF3 is expressed predominantly in anterior and dorsal structures and most strongly in the brain of the tailbud (stage 26) embryo. In situ hybridization shows XIF3 transcripts to be localized in neural tissue and especially in regions that most probably correspond to the motor neurones of the neural tube and to some cranial nerve ganglia. New XIF3 transcripts are first found at the start of gastrulation at a low level throughout the ectoderm and are not localized to the presumptive neurectoderm. Expression subsequently increases by about 10-fold in neural tissue, and requires an interaction of the mesoderm with overlying ectoderm. Because new transcripts are found predominantly in neural tissue of the head, this response can be used as a marker of anterior neural induction.


Multiple activities of the plant pathogen type III effector proteins WtsE and AvrE require WxxxE motifs.
The broadly conserved AvrE-family of type III effectors from gram-negative plant-pathogenic bacteria includes important virulence factors, yet little is known about the mechanisms by which these effectors function inside plant cells to promote disease. We have identified two conserved motifs in AvrE-family effectors: a WxxxE motif and a putative C-terminal endoplasmic reticulum membrane retention/retrieval signal (ERMRS). The WxxxE and ERMRS motifs are both required for the virulence activities of WtsE and AvrE, which are major virulence factors of the corn pathogen Pantoea stewartii subsp. stewartii and the tomato or Arabidopsis pathogen Pseudomonas syringae pv. tomato, respectively. The WxxxE and the predicted ERMRS motifs are also required for other biological activities of WtsE, including elicitation of the hypersensitive response in nonhost plants and suppression of defense responses in Arabidopsis. A family of type III effectors from mammalian bacterial pathogens requires WxxxE and subcellular targeting motifs for virulence functions that involve their ability to mimic activated G-proteins. The conservation of related motifs and their necessity for the function of type III effectors from plant pathogens indicates that disturbing host pathways by mimicking activated host G-proteins may be a virulence mechanism employed by plant pathogens as well.


Micropulse water vapor differential absorption lidar: transmitter design and performance.
An all diode-laser-based micropulse differential absorption lidar (DIAL) laser transmitter for tropospheric water vapor and aerosol profiling is presented. The micropulse DIAL (MPD) transmitter utilizes two continuous wave (cw) external cavity diode lasers (ECDL) to seed an actively pulsed, overdriven tapered semiconductor optical amplifier (TSOA). The MPD laser produces up to 7 watts of peak power over a 1 µs pulse duration (7 µJ) and a 10 kHz pulse repetition frequency. Spectral switching between the online and offline seed lasers is achieved on a 1Hz basis using a fiber optic switch to allow for more accurate sampling of the atmospheric volume between the online and offline laser shots. The high laser spectral purity of greater than 0.9996 coupled with the broad tunability of the laser transmitter will allow for accurate measurements of tropospheric water vapor in a wide range of geographic locations under varying atmospheric conditions. This paper describes the design and performance characteristics of a third generation MPD laser transmitter with enhanced laser performance over the previous generation DIAL system.


Captopril, a specific inhibitor of angiotensin converting enzyme, enhances both trypsin and vitellogenin titers in the grey fleshfly Neobellieria bullata.
A strong and constitutive angiotensin converting enzyme- or ACE-like activity was demonstrated in the hemolymph of the adult grey fleshfly Neobellieria bullata. In a competition assay, the N. bullata trypsin modulating oostatic factor (Neb-TMOF) was confirmed to be an in vitro substrate for this circulating Neb-ACE. Oral uptake of captopril, a selective and specific inhibitor of ACE, resulted in a complete phenotypic knockout of circulating ACE activity. When compared with control animals, captopril-fed female flies showed an increase in the liver meal-induced trypsin peak in the midgut and elevated levels of protein meal-induced yolk polypeptides in the hemolymph. The latter effect was not due to a slower vitellogenin uptake by the ovaries, because oocyte growth was not affected by the captopril treatment. The apparent synergism between the demonstrated ACE functionality and the previously reported effects of the oostatic peptide Neb-TMOF are discussed in the context of our recent finding that Neb-TMOF represents a prime candidate for being the first known in vivo substrate for circulating insect ACE. Arch.


Exploring plant growth-promotion actinomycetes from vermicompost and rhizosphere soil for yield enhancement in chickpea.
The main objective of the present study was to isolate and characterize actinomycetes for their plant growth-promotion in chickpea. A total of 89 actinomycetes were screened for their antagonism against fungal pathogens of chickpea by dual culture and metabolite production assays. Four most promising actinomycetes were evaluated for their physiological and plant growth-promotion properties under in vitro and in vivo conditions. All the isolates exhibited good growth at temperatures from 20°C to 40°C, pH range of 7-11 and NaCl concentrations up to 8%. These were also found highly tolerant to Bavistin, slightly tolerant to Thiram and Captan (except VAI-7 and VAI-40) but susceptible to Benlate and Ridomil at field application levels and were found to produce siderophore, cellulase, lipase, protease, chitinase (except VAI-40), hydrocyanic acid (except VAI-7 and VAI-40), indole acetic acid and β-1,3-glucanase. When the four actinomycetes were evaluated for their plant growth-promotion properties under field conditions on chickpea, all exhibited increase in nodule number, shoot weight and yield. The actinomycetes treated plots enhanced total N, available P and organic C over the un-inoculated control. The scanning electron microscope studies exhibited extensive colonization by actinomycetes on the root surface of chickpea. The expression profiles for indole acetic acid, siderophore and β-1,3-glucanase genes exhibited up-regulation for all three traits and in all four isolates. The actinomycetes were identified as Streptomyces but different species in the 16S rDNA analysis. It was concluded that the selected actinomycetes have good plant growth-promotion and biocontrol potentials on chickpea.


After Mexico Implemented a Tax, Purchases of Sugar-Sweetened Beverages Decreased and Water Increased: Difference by Place of Residence, Household Composition, and Income Level.
Background: In January 2014, Mexico implemented a tax on sugar-sweetened beverage (SSB) purchases of 1 peso/L.Objective: We examined the heterogeneity of changes in nonalcoholic beverage (SSB and bottled water) purchases after the tax was implemented by household income, urban and rural strata, and household composition.Methods: We used 4 rounds of the National Income and Expenditure Surveys: 2008, 2010, 2012, and 2014. Changes in purchases in per capita liters per week were estimated with the use of 2-part models to adjust for nonpurchases. We compared absolute and relative differences between adjusted changes in observed purchases in 2014 with expected purchases in 2014 based on prior trends (2008-2012). The models were adjusted for sociodemographic characteristics of the households, place of residence, and lagged gross domestic product per capita.Results: We found a 6.3% reduction in the observed purchases of SSBs in 2014 compared with the expected purchases in that same year based on trends from 2008 to 2012. These reductions were higher among lower-income households, residents living in urban areas, and households with children. We also found a 16.2% increase in water purchases that was higher in low- and middle-income households, in urban areas, and among households with adults only.Conclusions: SSB purchases decreased and water purchases increased after an SSB tax was imposed in Mexico. The magnitude of these changes was greater in lower-income and urban households.


Of chickens and men: avian influenza in humans.
Highly pathogenic H5N1 avian influenza virus can infect humans and is currently the most deadly influenza virus that has crossed the species barrier. As of December 2007, the spread of H5N1 virus from human to human has been rare. Nobody can predict if H5N1 may cause a pandemic. However, the number of human cases is continuously increasing and changes in virulence and epidemiology have been detected. There are specific pathogenic features of H5N1 infection. In contrast to human-adapted influenza A strains, H5N1 preferentially infects cells of the lower respiratory tract and may spread to tissues outside the respiratory tract in humans. Moreover, H5N1 replication is prolonged in target organs and results in higher viral loads and increased tissue damage. These features will have to be considered for therapeutic protocols for H5N1 infection in humans. Rapid genetic and antigenic changes observed in H5N1 virus isolates represent a challenge for the development of vaccines. In the present review, current knowledge about epidemiology, virulence factors and pathology of H5N1 infections in humans are summarised and discussed. Moreover, the possible roles of anti-influenza drugs in the pandemic situation as well as the development of effective vaccines are subject of this overview.


Phase extraction from interferograms with unknown tilt phase shifts based on a regularized optical flow method.
A novel method is presented to extract phase distribution from phase-shifted interferograms with unknown tilt phase shifts. The proposed method can estimate the tilt phase shift between two temporal phase-shifted interferograms with high accuracy, by extending the regularized optical flow method with the spatial image processing and frequency estimation technology. With all the estimated tilt phase shifts, the phase component encoded in the interferograms can be extracted by the least-squares method. Both simulation and experimental results have fully proved the feasibility of the proposed method. Particularly, a flat-based diffractive optical element with quasi-continuous surface is tested by the proposed method with introduction of considerably large tilt phase shift amounts (i.e., the highest estimated tilt phase shift amount between two consecutive frame reaches 6.18λ). The phase extraction result is in good agreement with that of Zygo's MetroPro software under steady-state testing conditions, and the residual difference between them is discussed. In comparison with the previous methods, the proposed method not only has relatively little restrictions on the amounts or orientations of the tilt phase shifts, but also works well with interferograms including open and closed fringes in any combination.


Characterization of protein-protein interaction interfaces from a single species.
Most proteins attain their biological functions through specific interactions with other proteins. Thus, the study of protein-protein interactions and the interfaces that mediate these interactions is of prime importance for the understanding of biological function. In particular the precise determinants of binding specificity and their contributions to binding energy within protein interfaces are not well understood. In order to better understand these determinants an appropriate description of the interaction surface is needed. Available data from the yeast Saccharomyces cerevisiae allow us to focus on a single species and to use all the available structures, correcting for redundancy, instead of using structural representatives. This allows us to control for potentially confounding factors that may affect sequence propensities. We find a significant contribution of main-chain atoms to protein-protein interactions. These include interactions both with other main-chain and side-chain atoms on the interacting chain. We find that the type of interaction depends on both amino acid and secondary structure type involved in the contact. For example, residues in α-helices and large amino acids are the most likely to be involved in interactions through their side-chain atoms. We find an intriguing homogeneity when calculating the average solvation energy of different areas of the protein surface. Unexpectedly, homo- and hetero-complexes have quite similar results for all analyses. Our findings demonstrate that the manner in which protein-protein interactions are formed is determined by the residue type and the secondary structure found in the interface. However the homogeneity of the desolvation energy despite heterogeneity of interface properties suggests a complex relationship between interface composition and binding energy.


Impact of Diabetes, Insulin, and Metformin Use on the Outcome of Patients With Human Epidermal Growth Factor Receptor 2-Positive Primary Breast Cancer: Analysis From the ALTTO Phase III Randomized Trial.
Purpose Previous studies have suggested an association between metformin use and improved outcome in patients with diabetes and breast cancer. In the current study, we aimed to explore this association in human epidermal growth factor receptor 2 (HER2 ) -positive primary breast cancer in the context of a large, phase III adjuvant trial. Patients and Methods The ALTTO trial randomly assigned patients with HER2-positive breast cancer to receive 1 year of either trastuzumab alone, lapatinib alone, their sequence, or their combination. In this substudy, we evaluated whether patients with diabetes at study entry-with or without metformin treatment-were associated with different disease-free survival (DFS), distant disease-free survival (DDFS), and overall survival (OS) compared with patients without diabetes. Results A total of 8,381 patients were included in the current analysis: 7,935 patients (94.7%) had no history of diabetes at diagnosis, 186 patients (2.2%) had diabetes with no metformin treatment, and 260 patients (3.1%) were diabetic and had been treated with metformin. Median follow-up was 4.5 years (0.16 to 6.31 years), at which 1,205 (14.38%), 929 (11.08%), and 528 (6.3%) patients experienced DFS, DDFS, and OS events, respectively. Patients with diabetes who had not been treated with metformin experienced worse DFS (multivariable hazard ratio [HR], 1.40; 95% CI, 1.01 to 1.94; P = .043), DDFS (multivariable HR, 1.56; 95% CI, 1.10 to 2.22; P = .013), and OS (multivariable HR, 1.87; 95% CI, 1.23 to 2.85; P = .004). This effect was limited to hormone receptor-positive patients. Whereas insulin treatment was associated with a detrimental effect, metformin had a salutary effect in patients with diabetes who had HER2-positive and hormone receptor-positive breast cancer. Conclusion Metformin may improve the worse prognosis that is associated with diabetes and insulin treatment, mainly in patients with primary HER2-positive and hormone receptor-positive breast cancer.


Reconstructed Homo habilis type OH 7 suggests deep-rooted species diversity in early Homo.
Besides Homo erectus (sensu lato), the eastern African fossil record of early Homo has been interpreted as representing either a single variable species, Homo habilis, or two species. In the latter case, however, there is no consensus over the respective groupings, and which of the two includes OH 7, the 1.8-million-year-old H. habilis holotype. This partial skull and hand from Olduvai Gorge remains pivotal to evaluating the early evolution of the Homo lineage, and by priority names one or other of the two taxa. However, the distorted preservation of the diagnostically important OH 7 mandible has hindered attempts to compare this specimen with other fossils. Here we present a virtual reconstruction of the OH 7 mandible, and compare it to other early Homo fossils. The reconstructed mandible is remarkably primitive, with a long and narrow dental arcade more similar to Australopithecus afarensis than to the derived parabolic arcades of Homo sapiens or H. erectus. We find that this shape variability is not consistent with a single species of early Homo. Importantly, the jaw morphology of OH 7 is incompatible with fossils assigned to Homo rudolfensis and with the A.L. 666-1 Homo maxilla. The latter is morphologically more derived than OH 7 but 500,000 years older, suggesting that the H. habilis lineage originated before 2.3 million years ago, thus marking deep-rooted species diversity in the genus Homo. We also reconstructed the parietal bones of OH 7 and estimated its endocranial volume. At between 729 and 824 ml it is larger than any previously published value, and emphasizes the near-complete overlap in brain size among species of early Homo. Our results clarify the H. habilis hypodigm, but raise questions about its phylogenetic relationships. Differences between species of early Homo appear to be characterized more by gnathic diversity than by differences in brain size, which was highly variable within all taxa.


Hepatitis-C multimedia prevention program in poor Hispanic HIV-infected injecting drug users: six months after intervention.
In order to prevent the spread of the hepatitis C virus (HCV) amongst Hispanic injection drug users (IDUs), we developed, validated, and implemented a multimedia educational intervention program. A pre-post intervention study design was used to evaluate long-lasting knowledge and behavior changes in a group of 88 low-income Hispanic HIV-infected IDUs. Pre-intervention data was compared with data measured six months after the intervention. A significant increase in the awareness regarding HCV clinical manifestations, HCV risky behaviors, HCV prevention practices, and HIV/HCV co-infection synergisms was observed in the group six months post-intervention. Our study confirms the long-lasting benefits of multimedia based intervention programs for disseminating HCV prevention strategies in IDUs. Preventive educational approaches that use images, figures, and animations tools can be recommended to target and tailor interventions for vulnerable populations.


Identifying indicator species in ecological habitats using Deep Optimal Feature Learning.
Much of the current research on supervised modelling is focused on maximizing outcome prediction accuracy. However, in engineering disciplines, an arguably more important goal is that of feature extraction, the identification of relevant features associated with the various outcomes. For instance, in microbial communities, the identification of keystone species can often lead to improved prediction of future behavioral shifts. This paper proposes a novel feature extractor based on Deep Learning, which is largely agnostic to underlying assumptions regarding the training data. Starting from a collection of microbial species abundance counts, the Deep Learning model first trains itself to classify the selected distinct habitats. It then identifies indicator species associated with the habitats. The results are then compared and contrasted with those obtained by traditional statistical techniques. The indicator species are similar when compared at top taxonomic levels such as Domain and Phylum, despite visible differences in lower levels such as Class and Order. More importantly, when our estimated indicators are used to predict final habitat labels using simpler models (such as Support Vector Machines and traditional Artificial Neural Networks), the prediction accuracy is improved. Overall, this study serves as a preliminary step that bridges modern, black-box Machine Learning models with traditional, domain expertise-rich techniques.


Expression of p27BBP/eIF6 is highly modulated during Xenopus laevis embryogenesis.
Protein p27BBP/eIF6 is necessary for ribosomal function of all cells. Previous data showed that from mammals to yeast p27BBP/eIF6 is involved in the biogenesis of ribosomal subunit 60S and its association with the 60S prevents premature 80S formation regulated by PKC signaling, indicating that phosphorylation of p27BBP/eIF6 is needed for translation to occur. While in vitro p27BBP/eIF6 is constitutively expressed, and it has a high level of expression in cycling cells, in vivo its expression varies according to tissues and appears regulated by factors up to now unknown. p27BBP/eIF6 has never been investigated in developing organisms where its upregulation can be correlated with tissue growth and differentiation. In this study we have sequenced p27BBP/eIF6 cDNA and studied its expression during development of Xenopus laevis, as the first step for studying its regulation. The amino acid sequence is highly conserved with two putative PKC phosphorylation sites in serine, one site being typical of Xenopus. At the end of gastrulation, the p27BBP/eIF6 riboprobe localizes in the neural plate and in the paraxial mesoderm. In particular, from stage 24, a clear-cut localization occurs in the perspective head. In embryos exposed to teratogens, the localization of p27BBP/eIF6 riboprobe varies according to the change of head size caused by the treatment. p27BBP/eIF6 expression is particularly evident in differentiating olfactory pits, the lens, otic vesicles, and in branchial arches. Features of particular interest are p27BBP/eIF6 high level of expression in the eye field, and in the mid-hindbrain-boundary, two regions with high proliferative activity. Altogether, data indicate that a modulated expression of p27BBP/eIF6 occurs in developing anlagens in addition to a basal level of expression, and may suggest a correlation between p27BBP/eIF6 and proliferative activity. Moreover, the X. laevis cDNA isolation and characterization offer new hints for further studies in relation to potential p27BBP/eIF6 phosphorylation.


Genetics of obesity in adult adoptees and their biological siblings.
An adoption study of genetic effects on obesity in adulthood was carried out in which adoptees separated from their natural parents very early in life were compared with their biological full and half siblings reared by their natural parents. The adoptees represented four groups who by sampling from a larger population were categorised as either thin, medium weight, overweight, or obese. Weight and height were obtained for 115 full siblings of 57 adoptees and for 850 half siblings of 341 adoptees. In full siblings body mass index (kg/m2) significantly increased with weight of the adoptees. Body mass index of the half siblings showed a steady but weaker increase across the four weight groups of adoptees. There were no significant interactions with sex of the adoptees, sex of the siblings, or (for the half siblings) sex of the common parent. In contrast with the findings in half siblings and (previously) the natural parents there was a striking, significant increase in body mass index between full siblings of overweight and obese adoptees. The degree of fatness in adults living in the same environment appears to be influenced by genetic factors independent of sex, which may include polygenic as well as major gene effects on obesity.


Prediction of caregiver quality of life in amyotrophic lateral sclerosis using explainable machine learning.
Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative, fatal and currently incurable disease. People with ALS need support from informal caregivers due to the motor and cognitive decline caused by the disease. This study aims to identify caregivers whose quality of life (QoL) may be impacted as a result of caring for a person with ALS. In this study, we worked towards the identification of the predictors of a caregiver's QoL in addition to the development of a model for clinical use to alert clinicians when a caregiver is at risk of experiencing low QoL. The data were collected through the Irish ALS Registry and via interviews on several topics with 90 patient and caregiver pairs at three time-points. The McGill QoL questionnaire was used to assess caregiver QoL-the MQoL Single Item Score measures the overall QoL and was selected as the outcome of interest in this work. The caregiver's existential QoL and burden, as well as the patient's depression and employment before the onset of symptoms were the features that had the highest impact in predicting caregiver quality of life. A small subset of features that could be easy to collect was used to develop a second model to use it in a clinical setting. The most predictive features for that model were the weekly caregiving duties, age and health of the caregiver, as well as the patient's physical functioning and age of onset.


Genetic manipulation in Sulfolobus islandicus and functional analysis of DNA repair genes.
Recently, a novel gene-deletion method was developed for the crenarchaeal model Sulfolobus islandicus, which is a suitable tool for addressing gene essentiality in depth. Using this technique, we have investigated functions of putative DNA repair genes by constructing deletion mutants and studying their phenotype. We found that this archaeon may not encode a eukarya-type of NER (nucleotide excision repair) pathway because depleting each of the eukaryal NER homologues XPD, XPB and XPF did not impair the DNA repair capacity in their mutants. However, among seven homologous recombination proteins, including RadA, Hel308/Hjm, Rad50, Mre11, HerA, NurA and Hjc, only the Hjc nuclease is dispensable for cell viability. Sulfolobus encodes redundant BER (base excision repair) enzymes such as two uracil DNA glycosylases and two putative apurinic/apyrimidinic lyases, but inactivation of one of the redundant enzymes already impaired cell growth, highlighting their important roles in archaeal DNA repair. Systematically characterizing these mutants and generating mutants lacking two or more DNA repair genes will yield further insights into the genetic mechanisms of DNA repair in this model organism.


Immunohistochemical Classification of Ampullary Carcinomas: Critical Reappraisal Fails to Confirm Prognostic Relevance for Recently Proposed Panels, and Highlights MUC5AC as a Strong Prognosticator.
Recently, immunohistochemistry-based classifications of ampullary carcinomas have been proposed (Ang and colleagues [PMID: 24832159]; Chang and colleagues [PMID: 23439753]). In this study, the prognostic value of Ang/Chang panel markers (CK20, MUC1, MUC2, CDX2) as well as other markers (CK7, MUC5AC, and MUC6) were tested on full-faced sections of 136 ampullary carcinoma resections with substantial (>5 mm) invasion. Immunohistochemistry was correlated with both histologic classification (intestinal [INT], pancreatobiliary [PB], or nontubular based on ≥3/5 observer agreement) and clinical outcome. No prognostic correlation was found with MUC1, CDX2, MUC2 or CK20 despite testing with different quantitative cutoffs. CK7 and CK20 were nonspecific. Ang classification had reasonable correlation with histologic subclassification of tubular cases as INT versus PB with high specificity but low sensitivity and ambiguous category was large (29%) and included also some classical cases. Prognostically, Ang classification approached but did not reach statistical significance, even when their large "ambiguous" group was eliminated and only tubular cases were analyzed (Ang-INT vs. Ang-PB; P=0.08). The Chang panel, in which the definition of the INT subcategory is not clearly defined, only marginally reached prognostic significance when tested as MUC1+/CDX2- versus MUC1-/CDX2+ and only by Wilcoxon test (P=0.0485) but 31% of the cases were "unclassifiable." The only individual marker that was found to have direct and strong correlation with the clinical outcome was MUC5AC (not used in the Ang or Chang panels), with statistically significant survival differences found with various cutoffs tested (for 20% cutoff, 5-y survival, 68% vs. 31%; P=0.0002). In addition, MUC5AC significantly stratified the histologically PB and INT cases (P=0.01 and 0.03, respectively), as well as Ang's ambiguous and Chang's unclassified cases (P=0.006 and 0.007, respectively). In conclusion, the widely used putative lineage markers, MUC1/MUC2/CK7/CK20/CDX2, do not seem to have direct/significant prognostic correlation either individually or in combination of Ang and Chang panels. Ang panel is helpful as an adjunct in determining the cell lineage with a few caveats. MUC5AC proves to be a significant independent prognosticator and should be incorporated into evaluation of ampullary carcinomas.


Antigenic oscillations and shifting immunodominance in HIV-1 infections.
A typical protein antigen contains several epitopes that can be recognized by cytotoxic T lymphocytes (CTL), but in a characteristic antiviral immune response in vivo, CTL recognize only a small number of these potential epitopes, sometimes only one, this phenomenon is known as immunodominance. Antigenic variation within CTL epitopes has been demonstrated for the human immunodeficiency virus HIV-1 (ref. 11) and other viruses and such 'antigenic escape' may be responsible for viral persistence. Here we develop a new mathematical model that deals with the interaction between CTL and multiple epitopes of a genetically variable pathogen, and show that the nonlinear competition among CTL responses against different epitopes can explain immunodominance. This model suggests that an antigenically homogeneous pathogen population tends to induce a dominant response against a single epitope, whereas a heterogeneous pathogen population can stimulate complicated fluctuating responses against multiple epitopes. Antigenic variation in the immunodominant epitope can shift responses to weaker epitopes and thereby reduce immunological control of the pathogen population. These ideas are consistent with detailed longitudinal studies of CTL responses in HIV-1 infected patients. For vaccine design, the model suggests that the major response should be directed against conserved epitopes even if they are subdominant.


Relations Among Some Low-Rank Subspace Recovery Models.
Recovering intrinsic low-dimensional subspaces from data distributed on them is a key preprocessing step to many applications. In recent years, a lot of work has modeled subspace recovery as low-rank minimization problems. We find that some representative models, such as robust principal component analysis (R-PCA), robust low-rank representation (R-LRR), and robust latent low-rank representation (R-LatLRR), are actually deeply connected. More specifically, we discover that once a solution to one of the models is obtained, we can obtain the solutions to other models in closed-form formulations. Since R-PCA is the simplest, our discovery makes it the center of low-rank subspace recovery models. Our work has two important implications. First, R-PCA has a solid theoretical foundation. Under certain conditions, we could find globally optimal solutions to these low-rank models at an overwhelming probability, although these models are nonconvex. Second, we can obtain significantly faster algorithms for these models by solving R-PCA first. The computation cost can be further cut by applying low-complexity randomized algorithms, for example, our novel l2,1 filtering algorithm, to R-PCA. Although for the moment the formal proof of our l2,1 filtering algorithm is not yet available, experiments verify the advantages of our algorithm over other state-of-the-art methods based on the alternating direction method.


Mange mite infestation in small ruminants in Ethiopia: Systematic review and meta-analysis.
Mange mites are economically important ectoparasites of sheep and goats responsible for rejection or downgrading of skins in tanneries or leather industries in Ethiopia. The objective of this systematic review was to compute the pooled prevalence estimate and identify factors influencing mange mite prevalence in sheep and goats at national level based on the available research evidence. Articles on mange mite infestation of small ruminants in Ethiopia were searched in PubMed, Web of Science, Google scholar and African journals on-line. The review was based on 18 cross-sectional studies carried out between 2003 and 2015 in four administrative states of Ethiopia. Accordingly, the pooled prevalence estimate in a random effects meta-analysis was estimated to be 4.4% (95% CI 3.0, 6.3) although there were evidence of a substantial amount of between-study variance (I(2)=98.4%). In subgroup and multivariable meta-regression analyses, animal species, agro-ecology and administrative state were found to have significant effect on the prevalence estimate (P<0.05) and explained 32.87% of the explainable proportion of the heterogeneity noted between studies The prevalence was found to be higher in goats in lowland agro-ecology. Region wise the highest estimate was calculated for Amhara (6.4%) followed by Oromia (4.7%), Tigray (3.6%) and Southern Nations, Nationalities and People Region (SNNPR) (3.1%). Significant difference was noted between Amhara and SNNPR. The study further revealed that mites of the genus Sarcoptes, Demodex and Psoroptes are the most prevalent mites infesting small ruminants in Ethiopia. Valid studies were lacking from five regional states. As some of these regions are known for their large small ruminant population, further studies are warranted to produce better picture of the infestation at a national level. Meanwhile, the need for monitoring the ongoing control intervention is suggested.


Sensitive, Rapid, Robust, and Reproducible Workflow for Host Cell Protein Profiling in Biopharmaceutical Process Development.
There is a growing industry and regulatory need to detect host cell protein (HCP) impurities in the production of protein biopharmaceuticals, as certain HCPs can impact product stability, safety, and efficacy, even at low levels. In some cases, regulatory agencies require the identification and the quantification of HCPs in drug products (DPs) for risk assessment, and this is an active and growing topic of conversation in the industry and amongst regulators. In this study, we developed a sensitive, robust, and reproducible workflow for HCP detection and quantification in a significantly shorter turnaround time than that previously reported using an Evosep ONE LC system coupled to an Orbitrap Fusion Lumos mass spectrometer. Because of its fast turnaround time, this HCP workflow can be integrated into process development for the high-throughput (60 samples analyzed per day) identification of HCPs. The ability to rapidly measure HCPs and follow their clearance throughout the downstream process can be used to pinpoint sources of HCP contamination, which can be used to optimize biopharmaceutical production to minimize HCP levels. Analysis of the NIST monoclonal antibody reference material using the rapid HCP profiling workflow detected the largest number of HCPs reported to date, underscoring an improvement in performance along with an increased throughput. The HCP workflow can be readily implemented and adapted for different purposes to guide biopharmaceutical process development and enable better risk assessment of HCPs in drug substances and DPs.


Metabolic programming a lean phenotype by deregulation of RNA polymerase III.
As a master negative regulator of RNA polymerase (Pol) III, Maf1 modulates transcription in response to nutrients and stress to balance the production of highly abundant tRNAs, 5S rRNA, and other small noncoding RNAs with cell growth and maintenance. This regulation of Pol III transcription is important for energetic economy as mice lacking Maf1 are lean and resist weight gain on normal and high fat diets. The lean phenotype of Maf1 knockout (KO) mice is attributed in part to metabolic inefficiencies which increase the demand for cellular energy and elevate catabolic processes, including autophagy/lipophagy and lipolysis. A futile RNA cycle involving increased synthesis and turnover of Pol III transcripts has been proposed as an important driver of these changes. Here, using targeted metabolomics, we find changes in the liver of fed and fasted Maf1 KO mice consistent with the function of mammalian Maf1 as a chronic Pol III repressor. Differences in long-chain acylcarnitine levels suggest that energy demand is higher in the fed state of Maf1 KO mice versus the fasted state. Quantitative metabolite profiling supports increased activity in the TCA cycle, the pentose phosphate pathway, and the urea cycle and reveals changes in nucleotide levels and the creatine system. Metabolite profiling also confirms key predictions of the futile RNA cycle hypothesis by identifying changes in many metabolites involved in nucleotide synthesis and turnover. Thus, constitutively high levels of Pol III transcription in Maf1 KO mice reprogram central metabolic pathways and waste metabolic energy through a futile RNA cycle.


Study on post-harvest quantitative and qualitative losses of potato tubers from two different road access districts of Jimma zone, South West Ethiopia.
One of the main challenges facing Ethiopia today is to ensure food security for its highly increasing population growth. Although the country's production is much lower than the national demand, there are high post-harvest food losses. Among several causes at different levels, absence of infrastructure plays a critical role in quantitative and qualitative losses of mainly fruit, vegetable, root and tuber crops. This study was initiated to investigate impact of two different production areas having different road access on qualitative and quantitative loss of potato tuber in extended storage time. Load tracking approach was used by simulating existing postharvest handling and storage practice of potato tuber in the study area. Quantitative post-harvest loss assessment along the supply chains, qualitative loss and shelf life estimation at retail market were investigated using standard methods. Results showed that more mechanical damages and seepage loss were observed in case of poor road quality and transportation. During extended storage time (30 days) at ambient environment, potato tubers transported from Dedo to Jimma resulted in high average weight loss (17.9 ± 0.9%), significant loss in firmness (7.0 ± 0.6N in average) and average vitamin C content of 9.20 ± 0.05mg/100g as compared to tubers transported from Seka to Jimma market. This implies that, lack of good road access and transportation facility contributed for more external mechanical damages and internal injuries which will critically enhance rate of degradation of tubers in later stages of storage.


HPLC and mass spectrometry analysis of dolichol-phosphates at the cell culture scale.
Dolichols (Dol) are polyprenol lipids that are essential structural components of eukaryotic membranes. In addition, the phosphorylated derivatives of Dol function as lipid anchors of mono- and oligosaccharide precursors involved in protein glycosylation. The biological importance of Dol phosphates (Dol-P) is illustrated by the severe outcome of human disorders linked to Dol biosynthetic defects, such as Dol-kinase deficiency. For characterization of inherited human diseases and evaluation of therapeutic trials, cultured cells often serve as a sole possible source for experimentation. Limited amounts of cell culture material render the quantitative analysis of Dol a challenging task. Here, we present HPLC- and mass spectrometry-based approaches to analyze and quantitate Dol-P from cultured human cells. The composition of naturally occurring Dol-P and the saturation state of the alpha-isoprene units was identified by negative-ion electrospray ionization mass spectrometry. Furthermore, fluorescently labeled Dol-P were separated by HPLC and quantified by comparison to known amounts of the internal standard polyprenol-P. The effect of pravastatin, a 3-hydroxy-3-methyl-glutaryl coenzyme-A reductase inhibitor, on the formation of Dol-P in HeLa cells was investigated. As expected, this treatment led to a decrease of Dol-P down to 35% of normal levels.


Association of statin use and development and progression of hip osteoarthritis in elderly women.
. To examine the association between use of HMG CoA reductase inhibitor (statin) and development and progression of radiographic findings of hip osteoarthritis (OA) in elderly women. Baseline and followup anteroposterior pelvic radiographs were obtained, a mean of 8 years apart, in 5674 women (69% of the total survivors) age 65 years and older from the Study of Osteoporotic Fractures. Hips were scored for individual radiographic features (IRF) and assigned a summary grade of radiographic hip osteoarthritis (RHOA) based on the number and type of IRF present. Statin use was recorded from prescription labels provided by participants at Year 6 and Year 8 of followup. We estimated odds ratios for the development of RHOA in hips without disease at baseline and for progression of RHOA in hips with baseline disease, using generalized estimating equations to account for correlations of right and left hips. RHOA was present in 936 hips of 745 women at baseline and there were 9318 hips without baseline RHOA. Overall, 7% of women in this cohort were statin users. Statin use was associated with increased odds of developing incident summary grade > or = 3 RHOA (OR 1.92, 95% CI 1.03-4.43, p = 0.045), but was not significantly related to other measures of new RHOA. In hips with baseline RHOA there was a consistent but nonsignificant trend toward decreased progression of RHOA in statin users (all OR between 0.69 and 0.76, with 95% CI between 0.29 and 1.67). Statin use may be associated with an increased risk of developing incident RHOA but does not adversely influence progression of established disease.


Understanding how local authorities in England address obesity: A wider determinants of health perspective.
Local government organisations (LAs) have a major role in the prevention and treatment of obesity in England. This study aims to 1) understand what actions are being taken by LAs to address obesity, and 2) determine how actions counter the perceived causes of obesity when mapped against the Wider Determinants of Health (WDoH) model. Thirty-two LAs were invited to complete an Action Mapping Tool, 10 participated. The tool requires LAs to document actions being implemented locally to address obesity. This then enables LAs to map their actions against the perceived causes of obesity, using the WDoH model as an analytical lens. We collated data from the 10 LAs and used an adapted framework synthesis method for analysis. 280 actions were documented across the 10 LAs; almost 60% (n = 166) targeted Individual Lifestyle Factors (ILF), with 7.1% (n = 20), 16.8% (n = 47) and 16.4% (n = 46) targeting Social and Community Factors (SCF), Living and Working Conditions (LWC) and Wider Conditions (WC) respectively. Conversely, 60% of causes were spread across the LWC and WC, with 16.4% regarded as ILF. Physical activity-, weight management-, and health improvement- programmes were most frequently implemented by LAs. There is a stark mismatch between LA actions on obesity and its perceived causes. Given that LAs acknowledge the complex aetiology of obesity, an equally comprehensive approach should be implemented in the future.


An Epitope Platform for Safe and Effective HTLV-1-Immunization: Potential Applications for mRNA and Peptide-Based Vaccines.
Human T-cell lymphotropic virus type 1 (HTLV-1) infection affects millions of individuals worldwide and can lead to severe leukemia, myelopathy/tropical spastic paraparesis, and numerous other disorders. Pursuing a safe and effective immunotherapeutic approach, we compared the viral polyprotein and the human proteome with a sliding window approach in order to identify oligopeptide sequences unique to the virus. The immunological relevance of the viral unique oligopeptides was assessed by searching them in the immune epitope database (IEDB). We found that HTLV-1 has 15 peptide stretches each consisting of uniquely viral non-human pentapeptides which are ideal candidate for a safe and effective anti-HTLV-1 vaccine. Indeed, experimentally validated HTLV-1 epitopes, as retrieved from the IEDB, contain peptide sequences also present in a vast number of human proteins, thus potentially instituting the basis for cross-reactions. We found a potential for cross-reactivity between the virus and the human proteome and described an epitope platform to be used in order to avoid it, thus obtaining effective, specific, and safe immunization. Potential advantages for mRNA and peptide-based vaccine formulations are discussed.


Rituximab as therapy to induce remission after relapse in ANCA-associated vasculitis.
Evaluation of rituximab and glucocorticoids as therapy to induce remission after relapse in ANCA-associated vasculitis (AAV) in a prospective observational cohort of patients enrolled into the induction phase of the RITAZAREM trial. Patients relapsing with granulomatosis with polyangiitis or microscopic polyangiitis were prospectively enrolled and received remission-induction therapy with rituximab (4×375 mg/m2) and a higher or lower dose glucocorticoid regimen, depending on physician choice: reducing from either 1 mg/kg/day or 0.5 mg/kg/day to 10 mg/day by 4 months. Patients in this cohort achieving remission were subsequently randomised to receive one of two regimens to prevent relapse. 188 patients were studied: 95/188 (51%) men, median age 59 years (range 19-89), prior disease duration 5.0 years (range 0.4-34.5). 149/188 (79%) had previously received cyclophosphamide and 67/188 (36%) rituximab. 119/188 (63%) of relapses had at least one major disease activity item, and 54/188 (29%) received the higher dose glucocorticoid regimen. 171/188 (90%) patients achieved remission by 4 months. Only six patients (3.2% of the study population) did not achieve disease control at month 4. Four patients died in the induction phase due to pneumonia (2), cerebrovascular accident (1), and active vasculitis (1). 41 severe adverse events occurred in 27 patients, including 13 severe infections. This large prospective cohort of patients with relapsing AAV treated with rituximab in conjunction with glucocorticoids demonstrated a high level of efficacy for the reinduction of remission in patients with AAV who have relapsed, with a similar safety profile to previous studies.


Renal oxygenation during robot-assisted laparoscopic partial nephrectomy: characterization using laparoscopic digital light processing hyperspectral imaging.
Abstract Background and Purpose: Digital light processing-based hyperspectral imaging (DLP(®)-HSI) was adapted for use during laparoscopic surgery by coupling the spectral illumination source with a conventional laparoscopic light guide and incorporating a customized digital charge-coupled device camera for image acquisition. The system was used to characterize renal oxygenation during robot-assisted laparoscopic partial nephrectomy (RALPN) in humans. After Institutional Review Board approval, laparoscopic DLP-HSI was performed in consecutive patients undergoing RALPN at our institution. Time trends in relative tissue oxygen saturation (%HbO2) were descriptively analyzed. Associations between %HbO2 and patient age, comorbidities, and estimated glomerular filtration rate (eGFR) were investigated using the Kendall tau test. Laparoscopic DLP-HSI was performed in 18 patients between May 2011 and February 2012. Median (interquartile range; IQR) age was 55.9 (49-67.5) years. Of the patients, 10/18 (56%) were men and 12/18 (66.7%) had a history of hypertension, diabetes, and/or tobacco use. Median (IQR) %HbO2 before, during, and after ischemia was 60.8% (57.9-68.2%), 53.6% (46.8-55.1%), and 61.5% (54.9-67.6%), respectively. Baseline %HbO2 was inversely associated with preoperative eGFR (τ=-0.38; P=0.036), and eGFR at most recent follow-up (τ=-0.38; P=0.036). Baseline or ischemic %HbO2 did not correlate with hypertension, diabetes, and/or tobacco history. Younger patients (<56 years) had a lower median baseline %HbO2 (P=0.07) and a higher median preoperative eGFR (P=0.038), than their older counterparts. The laparoscopic HSI system successfully characterized dynamic changes in renal oxygenation during RALPN. Intraoperative laparoscopic HSI outcomes have the potential to predict postoperative individual kidney function.


Severe hypoglycemia and risks of vascular events and death.
Severe hypoglycemia may increase the risk of a poor outcome in patients with type 2 diabetes assigned to an intensive glucose-lowering intervention. We analyzed data from a large study of intensive glucose lowering to explore the relationship between severe hypoglycemia and adverse clinical outcomes. We examined the associations between severe hypoglycemia and the risks of macrovascular or microvascular events and death among 11,140 patients with type 2 diabetes, using Cox proportional-hazards models with adjustment for covariates measured at baseline and after randomization. During a median follow-up period of 5 years, 231 patients (2.1%) had at least one severe hypoglycemic episode; 150 had been assigned to intensive glucose control (2.7% of the 5571 patients in that group), and 81 had been assigned to standard glucose control (1.5% of the 5569 patients in that group). The median times from the onset of severe hypoglycemia to the first major macrovascular event, the first major microvascular event, and death were 1.56 years (interquartile range, 0.84 to 2.41), 0.99 years (interquartile range, 0.40 to 2.17), and 1.05 years (interquartile range, 0.34 to 2.41), respectively. During follow-up, severe hypoglycemia was associated with a significant increase in the adjusted risks of major macrovascular events (hazard ratio, 2.88; 95% confidence interval [CI], 2.01 to 4.12), major microvascular events (hazard ratio, 1.81; 95% CI, 1.19 to 2.74), death from a cardiovascular cause (hazard ratio, 2.68; 95% CI, 1.72 to 4.19), and death from any cause (hazard ratio, 2.69; 95% CI, 1.97 to 3.67) (P<0.001 for all comparisons). Similar associations were apparent for a range of nonvascular outcomes, including respiratory, digestive, and skin conditions (P<0.01 for all comparisons). No relationship was found between repeated episodes of severe hypoglycemia and vascular outcomes or death. Severe hypoglycemia was strongly associated with increased risks of a range of adverse clinical outcomes. It is possible that severe hypoglycemia contributes to adverse outcomes, but these analyses indicate that hypoglycemia is just as likely to be a marker of vulnerability to such events. (Funded by Servier and the National Health and Medical Research Council of Australia; ClinicalTrials.gov number, NCT00145925.).