The Fanconi anemia gene product FANCF is a flexible adaptor protein.
The Fanconi anemia (FA) protein FANCF is an essential component of a nuclear core complex that protects the genome against chromosomal instability, but the specific function of FANCF is still poorly understood. Based upon the homology between human and Xenopus laevis FANCF, we carried out an extensive mutagenesis study to examine which domains are functionally important and to gain more insight into the function of FANCF. In contrast to previous suggestions, we show that FANCF does not have a ROM-like function. We found that the C terminus of FANCF interacts directly with FANCG and allows the assembly of other FA proteins into a stable complex. The N terminus appears to stabilize the interaction with FANCA and FANCG and is essential for the binding of the FANCC/FANCE subcomplex. We identified several important amino acids in this N-terminal region but, surprisingly, many amino acid changes failed to affect the function of the FANCF protein. Our data demonstrate that FANCF acts as a flexible adaptor protein that plays a key role in the proper assembly of the FA core complex.


Extreme body messages: themes from Facebook posts in extreme fitness and nutrition online support groups.
Extreme fitness and nutrition programs, that are said to "jump start" a healthy lifestyle, are becoming more pervasive. While some research has explored the harmful impact of fitspiration, thinspiration, and pro-anorexia/bulimia websites, no studies have examined the content associated with these extreme programs that advertised as promoting a healthy lifestyle. The objective of this study was to analyze posts found on extreme fitness and nutrition program support pages on Facebook. As these programs have become more popular, along with Facebook platforms for their participants, examining online user-generated posts related to these programs can provide information about the types of messages being promoted, and whether these can contribute to disordered eating and other harmful health behaviors. User data were selected from one month from public posts on two popular Facebook fitness groups. Both groups were short-term programs that set extreme guidelines for fitness and nutrition. Researchers examined the data for harmful health messages using an individual coding strategy with inter-rater reliability (Kappa =0.92). Majority (88.6%) of messages analyzed promoted harmful health messages. Common categories of messages included losing weight/fat, promoting dieting/restraint, and harmful body messages, which promoted unsafe ways to attain certain body types. The data analyzed in this study represent overwhelmingly negative commentary relating to harmful health and body messages. Messages in this study normalized dysfunctional behaviors and promoted fixating on certain body parts and objectifying bodies. These Facebook groups, though intended to be a sort of online support forum, provide an open space for body negativity and promotion of extreme behaviors for the sake of thinness. Most concerning, participants of these programs, which are advertised as promoting health, are expressing unhealthy thoughts and behaviors surrounding food, exercise, and body image. Given the ease of accessibility to this content using online platforms, these harmful messages can reach large groups of people and continue adding to a culture that values physical appearance to the detriment of health.


Molecular anatomy of breast cancer stroma and its prognostic value in estrogen receptor-positive and -negative cancers.
The purpose of this study was to identify genes enriched in breast cancer stroma, assess the stromal gene expression differences between estrogen receptor (ER) -positive and -negative cancers, and separately determine their prognostic value in these two subtypes of breast cancers. We compared gene expression profiles of pairs of fine-needle (stroma-poor) and core-needle (stroma-rich) biopsies from 37 cancers to identify stroma-associated genes. We defined stromal metagenes and tested their prognostic values in 684 node-negative patients who received no systemic adjuvant therapy and 259 tamoxifen-treated patients. We identified 293 probe sets overexpressed in core biopsies; these included five highly coexpressed gene clusters (metagenes) corresponding to immune functions and extracellular matrix components. These genes showed quantitative and qualitative differences between ER-positive and ER-negative cancers. A B-cell/plasma cell metagene showed strong prognostic value in ER-positive highly proliferative cancers, a lesser prognostic value in ER-negative cancers, and no prognostic value in ER-positive cancers with low proliferation. The hazard ratio for distant relapse in the lowest compared with the highest tertile of the pooled prognostic data set was 4.29 (95% CI, 2.04 to 9.01; P = .001) in ER-positive cancers and 3.34 (95% CI, 1.60 to 6.97; P = .001) in ER-negative cancers. This remained significant in multivariate analysis including routine variables and other genomic prognostic scores. As a result of quantitative differences in this metagene between ER-positive and ER-negative cancers, different thresholds apply in the two subgroups. Other stromal metagenes had inconsistent prognostic value. Among ER-negative and ER-positive highly proliferative cancers, a subset of tumors with high expression of a B-cell/plasma cell metagene carries a favorable prognosis.


Rapid and Culture Free Identification of Francisella in Hare Carcasses by High-Resolution Tandem Mass Spectrometry Proteotyping.
Zoonotic pathogens that can be transmitted via food to humans have a high potential for large-scale emergencies, comprising severe effects on public health, critical infrastructures, and the economy. In this context, the development of laboratory methods to rapidly detect zoonotic bacteria in the food supply chain, including high-resolution mass spectrometry proteotyping are needed. In this work, an optimized sample preparation method for liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteome profiling was established for Francisella isolates, and a cluster analysis, as well as a phylogenetic tree, was generated to shed light on evolutionary relationships. Furthermore, this method was applied to tissues of infected hare carcasses from Germany. Even though the non-informative data outnumbered by a manifold the information of the zoonotic pathogen in the resulting proteome profiles, the standardized evaluation of MS data within an established automated analysis pipeline identified Francisella (F.) tularensis and, thus, could be, in principle, an applicable method to monitor food supply chains.


Chemical Generation of Hydroxyl Radical for Oxidative 'Footprinting'.
For almost four decades, hydroxyl radical chemically generated by Fenton chemistry has been a mainstay for the oxidative 'footprinting' of macromolecules. In this article, we start by reviewing the application of chemical generation of hydroxyl radical to the development of oxidative footprinting of DNA and RNA and the subsequent application of the method to oxidative footprinting of proteins. We next discuss a novel strategy for generating hydroxyl radicals by Fenton chemistry that immobilizes catalytic iron on a solid surface (Pyrite Shrink Wrap laminate) for the application of nucleic acid and protein footprinting. Pyrite Shrink-Wrap Laminate is fabricated by depositing pyrite (Fe-S2, aka 'fool's gold') nanocrystals onto thermolabile plastic (Shrinky Dink). The laminate can be thermoformed into a microtiter plate format into which samples are deposited for oxidation. We demonstrate the utility of the Pyrite Shrink-Wrap Laminate for the chemical generation of hydroxyl radicals by mapping the surface of the T-cell co-stimulatory protein Programmed Death - 1 (PD-1) and the interface of the complex with its ligand PD-L1. We have developed and validated an affordable and reliable benchtop method of hydroxyl radical generation that will broaden the application of protein oxidative footprinting. Due to the minimal equipment required to implement this method, it should be easily adaptable by many laboratories with access to mass spectrometry.


Interaction of the p53 DNA-binding domain with its n-terminal extension modulates the stability of the p53 tetramer.
The tetrameric tumor suppressor p53 plays a pivotal role in the control of the cell cycle and provides a paradigm for an emerging class of oligomeric, multidomain proteins with structured and intrinsically disordered regions. Many of its biophysical and functional properties have been extrapolated from truncated variants, yet the exact structural and functional role of certain segments of the protein is unclear. We found from NMR and X-ray crystallography that the DNA-binding domain (DBD) of human p53, usually defined as residues 94-292, extends beyond these domain boundaries. Trp91, in the hinge region between the disordered proline-rich N-terminal domain and the DBD, folds back onto the latter and has a cation-π interaction with Arg174. These additional interactions increase the melting temperature of the DBD by up to 2 °C and inhibit aggregation of the p53 tetramer. They also modulate the dissociation of the p53 tetramer. The absence of the Trp91/Arg174 packing presumably allows nonnative DBD-DBD interactions that both nucleate aggregation and stabilize the interface. These data have important implications for studies of multidomain proteins in general, highlighting the fact that weak ordered-disordered domain interactions can modulate the properties of proteins of complex structure.


Influence of Exogenous β-Hydroxybutyrate on Walking Economy and Rating of Perceived Exertion.
This study investigates the effect of a supplementary ketone, β-hydroxybutyrate (BHB), on walking economy and ratings of perceived exertion in apparently healthy individuals. In a repeated-measures, crossover design, ten non-aerobically trained participants (three males; seven females) performed two stages of a duration-modified Bruce treadmill protocol. Participants blindly consumed either 1 ounce of an exogenous BHB solution (KETO) or a noncaloric placebo (CON) 30 minutes prior to exercise testing. Blood ketone and glucose concentrations were measured prior to supplementation (baseline), immediately before exercise, and after exercise. Oxygen consumption (VO2), respiratory exchange ratio (RER), energy expenditure (EE), and rating of perceived exertion (RPE) were recorded during the last two minutes of each stage. Blood BHB concentrations were significantly elevated at the pre-exercise and postexercise time points as compared to the CON condition (p < .001), and blood glucose was significantly elevated postexercise in both conditions as compared to baseline levels (p < .001). No significant between-trial differences (p > .05) were found for VO2, RER, EE, or RPE. The intervention of this study did not produce evidence of an ergogenic benefit from BHB supplementation in a healthy subject pool.


The role of zinc in calprotectin expression in human myeloid cells.
Elevated levels of calprotectin and other inflammatory mediators have been observed in inflammatory diseases paralleling serum hypozincemia. While a role of zinc in the regulation of tumor necrosis factor α, interleukin (IL)-1β and IL-6 expression has been established, the direct interrelation of zinc and calprotectin (S100A8/S100A9 heterodimer) expression is so far missing. In the present study, we analyzed mRNA and protein levels of S100A8 and S100A9 in monocytic Mono Mac (MM)1 and early myeloid THP-1 and U937 cells to elucidate the effect of zinc deficiency on their expression. We could depict that zinc deficiency alone enhances mRNA and protein expression of calprotectin in myeloid cells, independently from maturity stage. Moreover, pre-existing zinc deficiency augmented lipopolysaccharide (LPS)-induced calprotectin expression in CD14+ MM1, but not in CD14- U937 or CD14- THP-1 cells. Zinc deficiency and LPS seem therefore to activate different intracellular pathways. Our findings suggest that zinc does not only regulate the activity of calprotectin but also its expression by human myeloid cells.


The refractory epilepsy screening tool for Lennox-Gastaut syndrome (REST-LGS).
The complex clinical presentation and progression of Lennox-Gastaut syndrome (LGS) can complicate the accurate diagnosis of this severe, lifelong, childhood-onset epilepsy, often resulting in suboptimal treatment. The Refractory Epilepsy Screening Tool for LGS (REST-LGS) was developed to improve the identification of patients with LGS. Using the Modified Delphi Consensus, a group of experts developed and tested the REST-LGS Case Report Form (CRF) comprising 8 criteria (4 major, 4 minor) considered potentially indicative of LGS. Diagnosis-blinded specialist and nonspecialist raters at 2 epilepsy centers applied the CRF to deidentified patient records, including 1:1 records of patients with drug-resistant epilepsy or confirmed LGS. Interrater reliability was measured by Cohen's κ. Diagnosis was then unblinded to reveal common criteria for LGS or drug-resistant epilepsy. Cronbach's α was used to measure internal consistency between raters for all criteria combined. Of 200 patients, 81% to 85% met 1 to 3 major criteria. At both sites, moderate (κ, 0.41-0.60) to good (κ, 0.61-0.80) agreement on most criteria was reached between expert and nonexpert raters. Unblinding revealed that most patients with LGS met 3 major and 2 to 3 minor criteria, while patients with drug-resistant epilepsy met ≤1 major and only 1 to 2 minor criteria. Cronbach's α of raters at both sites was 0.64. The combined number of major/minor criteria on the CRF may be particularly indicative of LGS. Therefore, the REST-LGS may be a valuable clinical tool in identifying patients requiring further diagnostic evaluation for LGS.


Epithelioid hemangioma (angiolymphoid hyperplasia with eosinophilia) of the orbit: a case report.
Angiolymphoid hyperplasia with eosinophilia (ALHE) and Kimura's Disease (KD) share many clinical and histopathological features. Although they were once considered different stages of the same disease, they are now known to represent separate entities. Recently, ALHE is being called epithelioid hemangioma (EH), a term that better describes the possible neoplastic nature of the entity. An eighteen year-old Asian female presented with a three-month history of fluctuating swelling and ptosis of the left upper eyelid. Computed tomography disclosed a distinct homogeneous lesion in the left superior orbit, molding to the globe and other orbital structures. At histopathological evaluation the lesion was composed of numerous blood vessels lined by plump endothelial cells with oval nuclei protruding into the lumen. Surrounding the vessels, there was a chronic inflammatory infiltrate with a large proportion of eosinophils. Based on clinical and histopathological findings, the diagnosis of EH was made. Although exams like blood count, urinalysis and whole body scans can assist in the differential diagnosis, EH can be diagnosed and differentiated from KD on histopathological grounds. The presence of vascular hyperplasia with plump endothelial cells protruding into the lumen is the most important discriminator in establishing the diagnosis of EH. Such distinction is crucial for the patient because EH is not associated with any of the systemic manifestations present in KD.


Quantitative analysis of venation patterns of Arabidopsis leaves by supervised image analysis.
The study of transgenic Arabidopsis lines with altered vascular patterns has revealed key players in the venation process, but details of the vascularization process are still unclear, partly because most lines have only been assessed qualitatively. Therefore, quantitative analyses are required to identify subtle perturbations in the pattern and to test dynamic modeling hypotheses using biological measurements. We developed an online framework, designated Leaf Image Analysis Interface (LIMANI), in which venation patterns are automatically segmented and measured on dark-field images. Image segmentation may be manually corrected through use of an interactive interface, allowing supervision and rectification steps in the automated image analysis pipeline and ensuring high-fidelity analysis. This online approach is advantageous for the user in terms of installation, software updates, computer load and data storage. The framework was used to study vascular differentiation during leaf development and to analyze the venation pattern in transgenic lines with contrasting cellular and leaf size traits. The results show the evolution of vascular traits during leaf development, suggest a self-organizing mechanism for leaf venation patterning, and reveal a tight balance between the number of end-points and branching points within the leaf vascular network that does not depend on the leaf developmental stage and cellular content, but on the leaf position on the rosette. These findings indicate that development of LIMANI improves understanding of the interaction between vascular patterning and leaf growth.


Visualization of fluid drainage pathways in lymphatic vessels and lymph nodes using a mouse model to test a lymphatic drug delivery system.
Curing/preventing micrometastasis to lymph nodes (LNs) located outside the surgically resected area is essential for improving the morbidity and mortality associated with breast cancer and head and neck cancer. However, no lymphatic therapy system exists that can deliver drugs to LNs located outside the dissection area. Here, we demonstrate proof of concept for a drug delivery system using MXH10/Mo-lpr/lpr mice that exhibit systemic lymphadenopathy, with some peripheral LNs being as large as 10 mm in diameter. We report that a fluorescent solution injected into the subiliac LN (defined as the upstream LN within the dissection area) was delivered successfully to the proper axillary LN (defined as the downstream LN outside the dissection area) through the lymphatic vessels. Our results suggest that this approach could be used before surgical resection to deliver drugs to downstream LNs outside the dissection area. We anticipate that our methodology could be applied clinically, before surgical resection, to cure/prevent micrometastasis in LNs outside the dissection area, using techniques such as ultrasound-guided internal jugular vein catheterization.


Substrate-Competitive Activity-Based Profiling of Ester Prodrug Activating Enzymes.
Understanding the mechanistic basis of prodrug delivery and activation is critical for establishing species-specific prodrug sensitivities necessary for evaluating preclinical animal models and potential drug-drug interactions. Despite significant adoption of prodrug methodologies for enhanced pharmacokinetics, functional annotation of prodrug activating enzymes is laborious and often unaddressed. Activity-based protein profiling (ABPP) describes an emerging chemoproteomic approach to assay active site occupancy within a mechanistically similar enzyme class in native proteomes. The serine hydrolase enzyme family is broadly reactive with reporter-linked fluorophosphonates, which have shown to provide a mechanism-based covalent labeling strategy to assay the activation state and active site occupancy of cellular serine amidases, esterases, and thioesterases. Here we describe a modified ABPP approach using direct substrate competition to identify activating enzymes for an ethyl ester prodrug, the influenza neuraminidase inhibitor oseltamivir. Substrate-competitive ABPP analysis identified carboxylesterase 1 (CES1) as an oseltamivir-activating enzyme in intestinal cell homogenates. Saturating concentrations of oseltamivir lead to a four-fold reduction in the observed rate constant for CES1 inactivation by fluorophosphonates. WWL50, a reported carbamate inhibitor of mouse CES1, blocked oseltamivir hydrolysis activity in human cell homogenates, confirming CES1 is the primary prodrug activating enzyme for oseltamivir in human liver and intestinal cell lines. The related carbamate inhibitor WWL79 inhibited mouse but not human CES1, providing a series of probes for analyzing prodrug activation mechanisms in different preclinical models. Overall, we present a substrate-competitive activity-based profiling approach for broadly surveying candidate prodrug hydrolyzing enzymes and outline the kinetic parameters for activating enzyme discovery, ester prodrug design, and preclinical development of ester prodrugs.


Surveillance of recent HIV infections among newly diagnosed HIV cases in Germany between 2008 and 2014.
The HIV surveillance system in Germany is based on mandatory, anonymous notification of newly diagnosed HIV cases by laboratories. Because the time between HIV infection and the diagnosis of HIV varies widely between persons, it is difficult to determine the number of cases of recent HIV infection among newly diagnosed cases of HIV. In Germany, the BED-capture-enzyme immunoassay (BED-CEIA) has been used to distinguish between recent and long-standing HIV infection. The aim of this analysis is to report the proportion of cases of recent HIV infection among newly diagnosed cases in Germany between 2008 and 2014 and to identify factors associated with recent infections. A sample of voluntary laboratories among all HIV diagnostic laboratories was recruited. Residual blood from HIV diagnostic tests was spotted on filter paper as dried serum or dried plasma spots and was sent along with the notification form of the HIV cases. The BED-CEIA test was performed. A case was defined as recent HIV infection with a BED-CEIA test result of less than 0.8 normalized optical density, with the exclusion of CDC stage C. The proportion of recent newly diagnosed HIV infections among different groups (such as transmission groups, gender or age groups) was calculated. We used logistic regression to identify factors associated with recent HIV infection and to identify subpopulations with high proportions of recent HIV infections. Approximately 10,257 newly diagnosed cases were tested for recency using the BED-CEIA. In total, 3084 (30.4%) of those were recently infected with HIV. The highest proportion of recent HIV infections was found among men who had sex with men (MSM) (35%) and persons between 18 and 25 years of age (43.0%). Logistic regression revealed that female German intravenous drug users with a recent HIV infection had a higher chance of being detected than German MSM (OR 2.27). Surveillance of recent HIV infection is a useful additional tool to monitor the HIV epidemic in Germany. We could observe ongoing HIV transmission in Germany in general and in different subgroups, and we could identify factors associated with recent HIV infection in Germany.


Prediction of cerebral ischemia by ophthalmoscopy after carotid occlusion in gerbils.
The Mongolian gerbil provides a unique model of unilateral focal cerebral ischemia because of the lack of posterior communicating arteries in all gerbils as well as an absence of an anterior communicating artery in approximately 20% of the gerbil population. It is unclear how to identify unequivocably the subpopulation of animals that would suffer a severe focal cerebral ischemia after unilateral carotid occlusion. Ninety-three male gerbils were exposed to unilateral occlusion of the right common carotid artery. The severity of neuronal loss was evaluated histologically in gerbils selected as having significant focal ischemia based on either behavioral criteria (i.e., the demonstration of stereotypical behavior within 1 hour after occlusion) or ophthalmoscopic criteria (i.e., interruption of the retinal arterial perfusion within 10 minutes of carotid ligation as assessed with an ophthalmoscope). After 3 hours of unilateral carotid occlusion, cerebral blood flow was reinstated for 24 hours before fixation for histological analysis. The viability of the CA1 region of the hippocampus, lateral cortex, and medial cortex was scored on a scale of 0-4 based on the percentage of apparent neuronal loss (e.g., 0, no damage; 4, > 75% damage (the Viability Index). Twenty-eight percent of the gerbils met the behavioral selection criteria, and 17% met the ophthalmoscopic criteria. In the specimens selected by behavioral criteria (n = 7), 30% demonstrated no evidence of postischemic neuronal loss; the mean +/- SEM Viability Index scores for CA1, lateral cortex, and medial cortex were 1.6 +/- 0.6, 1.0 +/- 0.3, and 0.3 +/- 0.2, respectively. Of the animals selected by ophthalmoscopic criteria (n = 12), 100% had severe ischemic tissue damage to the ipsilateral hemisphere; the Viability Index scores for CA1, lateral cortex, and medial cortex were 3.5 +/- 0.1, 3.1 +/- 0.2, and 1.2 +/- 0.2, respectively; all scores were significantly larger than those observed in the behaviorally selected group. Selection of animals by ophthalmoscopic criteria provides a reliable, consistent method to predict animals with severe focal cerebral ischemia.


Simple sequence repeats as advantageous mutators in evolution.
Simple sequence repeats (SSRs) often serve to modify genes with which they are associated. The influence of SSRs on gene regulation, transcription and protein function typically depends on the number of repeats, while mutations that add or subtract repeat units are both frequent and reversible. SSRs thus provide a prolific source of quantitative and qualitative variation. Over the past decade, researchers have found that this spontaneous variation has been tapped by natural and artificial selection to adjust almost every aspect of gene function. These studies support the hypothesis that SSRs, by virtue of their special mutational and functional qualities, have a major role in generating the genetic variation underlying adaptive evolution.


Brain signature of working memory for sentence structure: enriched encoding and facilitated maintenance.
Sentences are easier to memorize than ungrammatical word strings, a phenomenon known as the sentence superiority effect. Yet, it is unclear how higher-order linguistic information facilitates verbal working memory and how this is implemented in the neural system. The goal of the current fMRI study was to specify the brain mechanisms underlying the sentence superiority effect during encoding and during maintenance in working memory by manipulating syntactic structure and working memory load. The encoding of sentence material, as compared with the encoding of ungrammatical word strings, recruited not only inferior frontal (BA 47) and anterior temporal language-related areas but also the medial-temporal lobe, which is not classically reported for language tasks. During maintenance, it was sentence structure as contrasted with ungrammatical word strings that led to activation decrease in Broca's area, SMA, and parietal regions. Furthermore, in Broca's area, an interaction effect revealed a load effect for ungrammatical word strings but not for sentences. The sentence superiority effect, thus, is neurally reflected in a twofold pattern, consisting of increased activation in classical language as well as memory areas during the encoding phase and decreased maintenance-related activation. This pattern reflects how chunking, based on sentential syntactic and semantic information, alleviates rehearsal demands and thus leads to improved working memory performance.


Switching to preservative-free latanoprost: impact on tolerability and patient satisfaction.
Patient satisfaction with glaucoma treatment has been poorly studied to date. Because glaucoma is a chronic condition in which the therapeutic response is dependent on adherence to treatment, patient acceptability is an important factor in achieving satisfactory outcomes. This multicenter, international (Belgium, the Netherlands, and Spain), epidemiological convenience sample survey among patients commencing treatment with preservative-free latanoprost collected data on patient satisfaction with particular regard to tolerability. A total of 1,541 patients were recruited who were predominantly elderly (74% were over 60 years of age) and female (61%). Most of the patients had previously received preserved topical glaucoma medication (69%), 6.7% had previously received preservative-free medication, whereas 24% had not previously been treated for glaucoma. The great majority of patients (>95%) were satisfied with the preservative-free latanoprost treatment. Among the patients who had previously received preserved medication, 73% of patients found preservative-free latanoprost to be better tolerated and 89% found it at least as easy to use as their prior treatment. Patient satisfaction (determined by a 0-100 mm visual analog scale) was improved by 47% on a switch from preserved treatment to preservative-free latanoprost. Intraocular pressure was similar in patients who had previously received preserved (18.3 mmHg), preservative-free (17.8 mmHg) glaucoma medication or who were naïve to treatment (20.3 mmHg). Preservative-free latanoprost provided effective reduction of intraocular pressure with better tolerability and patient satisfaction than preserved glaucoma medication. This tolerability profile can be expected to improve adherence to treatment in glaucoma patients.


Interaction of mitochondrial initiation factor 2 with mitochondrial fMet-tRNA.
The mammalian mitochondrial genome contains a single tRNA(Met) gene that gives rise to the initiator and elongator tRNA(Met). It is generally believed that mitochondrial protein synthesis begins with formylmethionyl-tRNA, which indicates that the formylation of mitochondrial Met-tRNA specifies its participation in initiation through its interaction with initiation factor 2 (IF-2). However, recent studies in yeast mitochondria, suggest that formylation is not required for protein synthesis. In addition, bovine IF-2(mt) could replace yeast IF-2(mt) in strains that lack fMet-tRNA which suggests that this paradigm may extend to mammalian mitochondria. Here, the importance of the formylation of mitochondrial Met-tRNA for the interaction with IF-2(mt) was investigated by measuring the ability of bovine IF-2(mt) to bind mitochondrial fMet-tRNA. In direct binding experiments, bovine IF-2(mt) has a 25-fold greater affinity for mitochondrial fMet-tRNA than Met-tRNA, using either the native mitochondrial tRNA(Met) or an in vitro transcript of bovine mitochondrial tRNA(Met). In addition, IF-2(mt) will not effectively stimulate mitochondrial Met-tRNA binding to mitochondrial ribosomes, exhibiting a 50-fold preference for fMet-tRNA over Met-tRNA in this assay. Finally, the region of IF-2(mt) responsible for the interaction with fMet-tRNA was mapped to the C2 sub-domain of domain VI of this factor.


Proposed diagnostic criteria for the case definition of amniotic fluid embolism in research studies.
Amniotic fluid embolism is a leading cause of maternal mortality in developed countries. Our understanding of risk factors, diagnosis, treatment, and prognosis is hampered by a lack of uniform clinical case definition; neither histologic nor laboratory findings have been identified unique to this condition. Amniotic fluid embolism is often overdiagnosed in critically ill peripartum women, particularly when an element of coagulopathy is involved. Previously proposed case definitions for amniotic fluid embolism are nonspecific, and when viewed through the eyes of individuals with experience in critical care obstetrics, would include women with a number of medical conditions much more common than amniotic fluid embolism. We convened a working group under the auspices of a committee of the Society for Maternal-Fetal Medicine and the Amniotic Fluid Embolism Foundation whose task was to develop uniform diagnostic criteria for the research reporting of amniotic fluid embolism. These criteria rely on the presence of the classic triad of hemodynamic and respiratory compromise accompanied by strictly defined disseminated intravascular coagulopathy. It is anticipated that limiting research reports involving amniotic fluid embolism to women who meet these criteria will enhance the validity of published data and assist in the identification of risk factors, effective treatments, and possibly useful biomarkers for this condition. A registry has been established in conjunction with the Perinatal Research Branch of the Eunice Kennedy Shriver National Institute of Child Health and Human Development to collect both clinical information and laboratory specimens of women with suspected amniotic fluid embolism in the hopes of identifying unique biomarkers of this condition.


Native macrophyte density and richness affect the invasiveness of a tropical poaceae species.
The role of the native species richness and density in ecosystem invasibility is a matter of concern for both ecologists and managers. We tested the hypothesis that the invasiveness of Urochloa arrecta (non-native in the Neotropics) is negatively affected by the species richness and abundance of native aquatic macrophytes in freshwater ecosystems. We first created four levels of macrophyte richness in a greenhouse (richness experiment), and we then manipulated the densities of the same native species in a second experiment (density experiment). When the native macrophytes were adults, fragments of U. arrecta were added, and their growth was assessed. Our results from the richness experiment corroborated the hypothesis of a negative relationship between the native species richness and the growth of U. arrecta, as measured by sprout length and root biomass. However, the resistance to invasion was not attributed to the presence of a particular native species with a greater competitive ability. In the density experiment, U. arrecta growth decreased significantly with an increased density of all five of the native species. Density strongly affected the performance of the Poaceae in a negative manner, suggesting that patches that are densely colonized by native macrophytes and less subject to disturbances will be more resistant to invasion than those that are poorly colonized and more commonly subjected to disturbances. Our density experiment also showed that some species exhibit a higher competitive ability than others (sampling effect). Although native richness and abundance clearly limit the colonization and establishment of U. arrecta, these factors cannot completely prevent the invasion of aquatic ecosystems by this Poaceae species.


T cell receptor V-segment frequencies in peripheral blood T cells correlate with human leukocyte antigen type.
We compared T cell receptor (TCR) V-segment frequencies in human leukocyte antigen (HLA) identical siblings to sibling pairs who differ at one or both HLA haplotypes using four V beta-specific and one V alpha-specific monoclonal antibody. In every one of nine families HLA-identical sibs had the most similar patterns of V-segment frequencies in their peripheral blood, whereas totally mismatched sibs were, in general, the most dissimilar; HLA haploidentical sibs tended to be intermediate between the two groups. The degree of similarity among HLA-identical sibs was comparable to that observed among three pairs of identical twins suggesting that HLA is the major genetic component influencing TCR V-segment frequency. Consistent with this observation, it was found that the frequency of T cells expressing particular V beta segments was skewed towards either CD4+ or CD8+ cells indicating that T cells expressing some V beta genes may be positively selected primarily by class I or class II major histocompatibility complex proteins. Finally, it was observed that individuals who express the HLA class I specificity, B38, tend to express high levels of V alpha 2.3+ cells among their CD8+ T cells. These observations represent definitive proof that human V-segment frequencies are profoundly influenced by the HLA complex.


SU-E-I-79: Correction of Dark Current and Image Lag in Multi-Source Carbon Nanotube Imaging Systems.
To correct for dark current and detector image lag in multi-source carbon nanotube (CNT) based imaging systems. CNTs can be used as a novel electron source for X-ray generation. Here a small (∼2 kV) gate voltage induces a large electric field at the CNT tip that causes field emission as opposed to thermionic emission. This enables faster switching times and allows miniaturization which aids in the development of stationary multi-source systems. These systems may encounter two key problems: dark current in non-active sources and detector lag. In damaged and perhaps end-of-life devices, sources may develop significant electron output without gate voltage, interfering with images from non-active sources. Although the output is small, it is constant, and continuous integration of detectors compared with small X-ray exposure time magnifies its contribution to the signal. A method based on binary multiplexing is developed to mitigate the dark current contributions possibly extending device lifetime. Having discrete sources, these multi-source systems have coarser angular sampling than rotating systems. This coarser sampling means that the detector signal changes more in adjacent projections than rotating systems exacerbating the effect of detector lag. Since flat panel detectors have a capacitive behavior, readout only removes some charge from pixels. As a result, a device dependent amount (6.6% in our device) of the signal from the current image contributes to the subsequent image. A fast, stable inverse filter separating detector lag has been developed. Projection data were simulated based on parameters measured from a multi-source CNT system. Incorporating dark current and image lag phenomena into reconstruction improves contrast to noise ratio. These correction methods can be easily implemented on clinical multi-source systems to improve image quality and extend the device's usable lifetime. Supported in part by Siemens Medical.


Drug-loaded nanoparticles and supramolecular nanotubes formed from a volatile microemulsion with bile salt derivatives.
The main objective of this study was to form nanoparticles of a model hydrophobic drug, celecoxib, from a volatile microemulsion stabilized by a bile salt derivative. Nanoparticles were obtained by conversion of the microemulsion nanodroplets with the dissolved drug into solid nanometric particles. The use of bile salt derivatives as the surfactants for the formation of a microemulsion enabled significantly higher loading of the drug in both the microemulsion and nanoparticles, compared with the native bile salt. In addition, superior stability of the particles was achieved with the bile salt derivatives, and drug crystallization was inhibited. Interestingly, differences in particle stability and crystallization inhibition were observed between two bile salt derivatives differing only by one hydroxyl group on the bile salt backbone, indicating the delicate balance of interactions in the system. For one of the derivatives, upon dispersion of the nanoparticles in water, they spontaneously arranged into well-defined elongated nanometric tubules as detected and attested by cryo-TEM. It was found that the drug present in nanoparticles induces formation of the nanotubes.


Hydrogen-rich saline attenuates eosinophil activation in a guinea pig model of allergic rhinitis via reducing oxidative stress.
It is well considered that reactive oxygen species (ROS) plays a prominent causative role in the development of allergic rhinitis (AR), and eosinophils cells as important allergic inflammatory cells contribute to elevating oxidative stress. Hydrogen, emerging as a novel antioxidant, has been proven effective in selectively reducing ROS in animals models of oxidative damage. We herein aim to verify protective effects of hydrogen on eosinophils cells in guinea pigs models of AR. Thirty two guinea pigs were random divided into four groups, and AR model was established through ovalbumin sensitization. The guinea pigs were injected with hydrogen-rich saline (Normal-HRS and AR-HRS group) or normal saline (control and AR group). The frequencies of sneezing and scratching were recorded. The IgE level, blood eosinophil count and eosinophil cationic protein (ECP) level in serum were measured. The serum malondialdehyde (MDA) and superoxide dismutase (SOD) assays were also measured to evaluate oxidative stress. The expression levels of eotaxin mRNA and protein in the nasal mucosa were also determined by real-time RT-PCR, Western blot and immunofluorescence. HRS reduced the ROS and MDA levels and increased SOD level in guinea pigs of AR-HRS group accompanied with decreased frequency of sneezing and scratches. Meanwhile, there was a decline of the number of eosinophils cells in blood and of thelevel of ECP in serum in the AR-HRS group. HRS also significantly decreased the expression of eotaxin in nasal mucosa. HRS may play a protective role in attenuating allergic inflammation, and suppressing the increase and activation of eosinophils in AR possibly through antioxidation effect of hydrogen.


Metabolic engineering of tomato fruit organic acid content guided by biochemical analysis of an introgression line.
Organic acid content is regarded as one of the most important quality traits of fresh tomato (Solanum lycopersicum). However, the complexity of carboxylic acid metabolism and storage means that it is difficult to predict the best way to engineer altered carboxylic acid levels. Here, we used a biochemical analysis of a tomato introgression line with increased levels of fruit citrate and malate at breaker stage to identify a metabolic engineering target that was subsequently tested in transgenic plants. Increased carboxylic acid levels in introgression line 2-5 were not accompanied by changes in the pattern of carbohydrate oxidation by pericarp discs or the catalytic capacity of tricarboxylic acid cycle enzymes measured in isolated mitochondria. However, there was a significant decrease in the maximum catalytic activity of aconitase in total tissue extracts, suggesting that a cytosolic isoform of aconitase was affected. To test the role of cytosolic aconitase in controlling fruit citrate levels, we analyzed fruit of transgenic lines expressing an antisense construct against SlAco3b, one of the two tomato genes encoding aconitase. A green fluorescent protein fusion of SlAco3b was dual targeted to cytosol and mitochondria, while the other aconitase, SlAco3a, was exclusively mitochondrial when transiently expressed in tobacco (Nicotiana tabacum) leaves. Both aconitase transcripts were decreased in fruit from transgenic lines, and aconitase activity was reduced by about 30% in the transgenic lines. Other measured enzymes of carboxylic acid metabolism were not significantly altered. Both citrate and malate levels were increased in ripe fruit of the transgenic plants, and as a consequence, total carboxylic acid content was increased by 50% at maturity.


The notch 3 intracellular domain represses notch 1-mediated activation through Hairy/Enhancer of split (HES) promoters.
The Notch signaling pathway is important for cellular differentiation. The current view is that the Notch receptor is cleaved intracellularly upon ligand activation. The intracellular Notch domain then translocates to the nucleus, binds to Suppressor of Hairless (RBP-Jk in mammals), and acts as a transactivator of Enhancer of Split (HES in mammals) gene expression. In this report we show that the Notch 3 intracellular domain (IC), in contrast to all other analysed Notch ICs, is a poor activator, and in fact acts as a repressor by blocking the ability of the Notch 1 IC to activate expression through the HES-1 and HES-5 promoters. We present a model in which Notch 3 IC interferes with Notch 1 IC-mediated activation at two levels. First, Notch 3 IC competes with Notch 1 IC for access to RBP-Jk and does not activate transcription when positioned close to a promoter. Second, Notch 3 IC appears to compete with Notch 1 IC for a common coactivator present in limiting amounts. In conclusion, this is the first example of a Notch IC that functions as a repressor in Enhancer of Split/HES upregulation, and shows that mammalian Notch receptors have acquired distinct functions during evolution.


Lack of beta 1 integrin gene in embryonic stem cells affects morphology, adhesion, and migration but not integration into the inner cell mass of blastocysts.
A gene trap-type targeting vector was designed to inactivate the beta 1 integrin gene in embryonic stem (ES) cells. Using this vector more than 50% of the ES cell clones acquired a disruption in the beta 1 integrin gene and a single clone was mutated in both alleles. The homozygous mutant did not produce beta 1 integrin mRNA or protein, while alpha 3, alpha 5, and alpha 6 integrin subunits were transcribed but not detectable on the cell surface. Heterozygous mutants showed reduced beta 1 expression and surface localization of alpha/beta 1 heterodimers. The alpha V subunit expression was not impaired on any of the mutants. Homozygous ES cell mutants lacked adhesiveness for laminin and fibronectin but not for vitronectin and showed a reduced association with a fibroblast feeder layer. Furthermore, they did not migrate towards chemoattractants in fibroblast medium. None of these functions were impaired in heterozygous mutants. Scanning electron microscopy revealed that homozygous cells showed fewer cell-cell junctions and had many microvilli not usually found on wild type and heterozygous cells. This profound change in cell shape is not associated with gross alterations in the expression and distribution of cytoskeletal components. Unexpectedly, microinjection into blastocysts demonstrated full integration of homozygous and heterozygous mutants into the inner cell mass. This will allow studies of the consequences of beta 1 integrin deficiency in several in vivo situations.


Amines and a peptide as neurohormones in lobsters: actions on neuromuscular preparations and preliminary behavioural studies.
In this communication we report that four substances, thought to function as neurohormones in Crustacea, all produce long-term changes in the physiological properties of lobster opener muscle preparations. The substances are the amines, octopamine, serotonin and dopamine, and the peptide, proctolin. The actions of these substances are superimposed on the normal synaptic apparatus that utilizes the amino acids GABA and glutamate (probably) as the inhibitory and excitatory neurotransmitter compounds. Serotonin acts on excitatory and inhibitory nerve endings to facilitate transmitter release and directly on muscle fibres to produce a contracture and to induce the appearance of Ca2+ action potentials. The latter two actions of serotonin are shared by proctolin and octopamine as well. Dopamine, on the other hand, relaxes muscle baseline tension. The mechanism of action of these substances at their target site (or sites) has been explored with electrophysiological and biochemical techniques and the results will be presented. In addition preliminary behavioural experiments have been carried out with serotonin and octopamine. These substances produce opposite postures when injected into lobsters. The amines act on central ganglia to produce these effects where they cause a programmed readout of firing of neurones that will produce either a flexed posture (serotonin) or an extended posture (octopamine).


Algae consumption and risk of type 2 diabetes: Korean National Health and Nutrition Examination Survey in 2005.
The purpose of this study is to examine the relationship between dietary algae (seaweed) consumption and the risk of Type 2 diabetes mellitus in the Korean population. We analyzed data from the Korean National Health and Nutrition Examination Survey in 2005, a nationally representative survey. The study participants included 3,405 males and females aged 20-65 y. Participants were classified into four groups according to the quartiles of total algae consumption frequencies. Proportional odds models were used to assess the relationship between algae consumption and the risk of having diabetes or prediabetes, after adjustment for age, family history of diabetes, education, smoking, alcohol consumption, physical activity, body mass index, waist circumference, triglycerides, total energy intake and food group intakes. The frequency of algae consumption was positively correlated to the consumption of legumes, fruits, fish, and dairy products in both genders (p<0.001). The odds ratios (95% confidence interval) for diabetes vs. prediabetes vs. normoglycemia was 0.66 (0.43-0.99) for males and 0.80 (0.51-1.24) for females in the highest quartile of algae consumption compared to the lowest quartile. Our results suggest that dietary algae consumption may decrease the risk of diabetes mellitus in Korean men. A well-designed prospective study is needed to confirm this association.


Imaging metals in biology: balancing sensitivity, selectivity and spatial resolution.
Metal biochemistry drives a diverse range of cellular processes associated with development, health and disease. Determining metal distribution, concentration and flux defines our understanding of these fundamental processes. A comprehensive analysis of biological systems requires a balance of analytical techniques that inform on metal quantity (sensitivity), chemical state (selectivity) and location (spatial resolution) with a high degree of certainty. A number of approaches are available for imaging metals from whole tissues down to subcellular organelles, as well as mapping metal turnover, protein association and redox state within these structures. Technological advances in micro- and nano-scale imaging are striving to achieve multi-dimensional and in vivo measures of metals while maintaining the native biochemical environment and physiological state. This Tutorial Review discusses state-of-the-art imaging technology as a guide to obtaining novel insight into the biology of metals, with sensitivity, selectivity and spatial resolution in focus.


Multimodal optimization using a bi-objective evolutionary algorithm.
In a multimodal optimization task, the main purpose is to find multiple optimal solutions (global and local), so that the user can have better knowledge about different optimal solutions in the search space and as and when needed, the current solution may be switched to another suitable optimum solution. To this end, evolutionary optimization algorithms (EA) stand as viable methodologies mainly due to their ability to find and capture multiple solutions within a population in a single simulation run. With the preselection method suggested in 1970, there has been a steady suggestion of new algorithms. Most of these methodologies employed a niching scheme in an existing single-objective evolutionary algorithm framework so that similar solutions in a population are deemphasized in order to focus and maintain multiple distant yet near-optimal solutions. In this paper, we use a completely different strategy in which the single-objective multimodal optimization problem is converted into a suitable bi-objective optimization problem so that all optimal solutions become members of the resulting weak Pareto-optimal set. With the modified definitions of domination and different formulations of an artificially created additional objective function, we present successful results on problems with as large as 500 optima. Most past multimodal EA studies considered problems having only a few variables. In this paper, we have solved up to 16-variable test problems having as many as 48 optimal solutions and for the first time suggested multimodal constrained test problems which are scalable in terms of number of optima, constraints, and variables. The concept of using bi-objective optimization for solving single-objective multimodal optimization problems seems novel and interesting, and more importantly opens up further avenues for research and application.


Flexible method based on four-beam interference lithography for fabrication of large areas of perfectly periodic plasmonic arrays.
A novel nanofabrication technique based on 4-beam interference lithography is presented that enables the preparation of large macroscopic areas (>50 mm2) of perfectly periodic and defect-free two-dimensional plasmonic arrays of nanoparticles as small as 100 nm. The technique is based on a special interferometer, composed of two mirrors and a sample with photoresist that together form a right-angled corner reflector. In such an interferometer, the incoming expanded laser beam is split into four interfering beams that yield an interference pattern with rectangular symmetry. The interferometer allows setting the periods of the array from about 220 nm to 1500 nm in both directions independently through the rotation of the corner-reflector assembly around horizontal and vertical axes perpendicular to the direction of the incident beam. Using a theoretical model, the implementation of the four-beam interference lithography is discussed in terms of the optimum contrast as well as attainable periods of the array. Several examples of plasmonic arrays (on either glass or polymer substrate layers) fabricated by this technique are presented.


Renal unit practitioners' knowledge, attitudes and practice regarding the safety of unfractionated heparin for chronic haemodialysis.
Chronic haemodialysis for adult patients with end-stage kidney failure requires a patent extracorporeal circuit, maintained by anticoagulants such as unfractionated heparin (UFH). Incorrect administration of UFH has safety implications for patients. Firstly, to describe renal practitioners' self-reported knowledge, attitudes and practice (KAP) regarding the safe use of UFH and its effects; secondly, to determine an association between KAP and selected independent variables. A cross-sectional descriptive survey by self-administered questionnaire and non-probability convenience sampling was conducted in two tertiary hospital dialysis units and five private dialysis units in 2013. The mean age of 74/77 respondents (96.1%), was 41.1 years. Most (41/77, 53.2%) had 0-5 years of renal experience. The odds of enrolled nurses having poorer knowledge of UFH than registered nurses were 18.7 times higher at a 95% Confidence Interval (CI) (1.9-187.4) and statistically significant (P = 0.013). The odds of delivering poor practice having ≤ five years of experience and no in-service education were 4.6 times higher at a 95% CI (1.4-15.6), than for respondents who had ≥ six years of experience (P = 0.014) and 4.3 times higher (95% CI 1.1-16.5) than for respondents who received in-service education (P = 0.032), the difference reaching statistical significance in both cases. Results suggest that the category of the professional influences knowledge and, thus, safe use of UFH, and that there is a direct relationship between years of experience and quality of haemodialysis practice and between having in-service education and quality of practice.


Targeted multiplex next-generation sequencing: advances in techniques of mitochondrial and nuclear DNA sequencing for population genomics.
Next-generation sequencing (NGS) is emerging as an efficient and cost-effective tool in population genomic analyses of nonmodel organisms, allowing simultaneous resequencing of many regions of multi-genomic DNA from multiplexed samples. Here, we detail our synthesis of protocols for targeted resequencing of mitochondrial and nuclear loci by generating indexed genomic libraries for multiplexing up to 100 individuals in a single sequencing pool, and then enriching the pooled library using custom DNA capture arrays. Our use of DNA sequence from one species to capture and enrich the sequencing libraries of another species (i.e. cross-species DNA capture) indicates that efficient enrichment occurs when sequences are up to about 12% divergent, allowing us to take advantage of genomic information in one species to sequence orthologous regions in related species. In addition to a complete mitochondrial genome on each array, we have included between 43 and 118 nuclear loci for low-coverage sequencing of between 18 kb and 87 kb of DNA sequence per individual for single nucleotide polymorphisms discovery from 50 to 100 individuals in a single sequencing lane. Using this method, we have generated a total of over 500 whole mitochondrial genomes from seven cetacean species and green sea turtles. The greater variation detected in mitogenomes relative to short mtDNA sequences is helping to resolve genetic structure ranging from geographic to species-level differences. These NGS and analysis techniques have allowed for simultaneous population genomic studies of mtDNA and nDNA with greater genomic coverage and phylogeographic resolution than has previously been possible in marine mammals and turtles.


Arl13b Regulates Breast Cancer Cell Migration and Invasion by Controlling Integrin-Mediated Signaling.
Breast cancer is the first cause of cancer-related mortality among women worldwide, according to the most recent estimates. This mortality is mainly caused by the tumors' ability to form metastases. Cancer cell migration and invasion are essential for metastasis and rely on the interplay between actin cytoskeleton remodeling and cell adhesion. Therefore, understanding the mechanisms by which cancer cell invasion is controlled may provide new strategies to impair cancer progression. We investigated the role of the ADP-ribosylation factor (Arf)-like (Arl) protein Arl13b in breast cancer cell migration and invasion in vitro, using breast cancer cell lines and in vivo, using mouse orthotopic models. We show that Arl13b silencing inhibits breast cancer cell migration and invasion in vitro, as well as cancer progression in vivo. We also observed that Arl13b is upregulated in breast cancer cell lines and patient tissue samples. Moreover, we found that Arl13b localizes to focal adhesions (FAs) and interacts with β3-integrin. Upon Arl13b silencing, β3-integrin cell surface levels and FA size are increased and integrin-mediated signaling is inhibited. Therefore, we uncover a role for Arl13b in breast cancer cell migration and invasion and provide a new mechanism for how ARL13B can function as an oncogene, through the modulation of integrin-mediated signaling.


Electronic cigarette aerosols suppress cellular antioxidant defenses and induce significant oxidative DNA damage.
Electronic cigarette (EC) aerosols contain unique compounds in addition to toxicants and carcinogens traditionally found in tobacco smoke. Studies are warranted to understand the public health risks of ECs. The aim of this study was to determine the genotoxicity and the mechanisms induced by EC aerosol extracts on human oral and lung epithelial cells. Cells were exposed to EC aerosol or mainstream smoke extracts and DNA damage was measured using the primer anchored DNA damage detection assay (q-PADDA) and 8-oxo-dG ELISA assay. Cell viability, reactive oxygen species (ROS) and total antioxidant capacity (TAC) were measured using standard methods. mRNA and protein expression were evaluated by RT-PCR and western blot, respectively. EC aerosol extracts induced DNA damage in a dose-dependent manner, but independently of nicotine concentration. Overall, EC aerosol extracts induced significantly less DNA damage than mainstream smoke extracts, as measured by q-PADDA. However, the levels of oxidative DNA damage, as indicated by the presence of 8-oxo-dG, a highly mutagenic DNA lesion, were similar or slightly higher after exposure to EC aerosol compared to mainstream smoke extracts. Mechanistically, while exposure to EC extracts significantly increased ROS, it decreased TAC as well as the expression of 8-oxoguanine DNA glycosylase (OGG1), an enzyme essential for the removal of oxidative DNA damage. Exposure to EC aerosol extracts suppressed the cellular antioxidant defenses and led to significant DNA damage. These findings emphasize the urgent need to investigate the potential long-term cancer risk of exposure to EC aerosol for vapers and the general public.


The role of vocal tract filtering in identity cueing in rhesus monkey (Macaca mulatta) vocalizations.
The importance of individual identity and kinship has been demonstrated in the social behavior of many nonhuman primates, with some evidence suggesting that individually distinctive acoustic features are present in their vocalizations as well. In order to systematically test whether acoustic cues to identity are reliably present across the vocal repertoire of rhesus monkeys (Macaca mulatta), we examined coos, grunts, and noisy screams produced by adult females of two free-ranging groups. First, acoustic analyses were used to characterize spectral patterning, the fundamental frequency, and temporal characteristics of these three distinct call types. Vocalizations were then classified by caller identity, based on discriminant function analyses. Results showed that coos (rich, harmonically structured sounds) were markedly more distinctive by caller than were either grunts or noisy screams, and that spectral-patterning measures related to vocal tract filtering effects were the most reliable markers of individual identity. Grunts (pulsed, noisy calls) were classified at lower, but above-chance rates and spectral patterning cues were again critical in this sorting. Noisy screams (continuous, broadband noise bursts that could include a high-frequency, periodic component) could not be reliably sorted by caller. Playback experiments conducted with the screams showed no response differences when listening animals heard vocalizations produced by kin or nonkin individuals. This result was strikingly different from the corresponding outcome of a previous test with coo calls, but consistent with the acoustic analysis. Implications of these findings for vocal production mechanisms in nonhuman primates and previous studies of rhesus monkey vocalizations are discussed.


[Independent Components Analysis (ICA) in the study of electroencephalographic signals].
Independent Component Analysis (ICA) is a mathematical tool able to separate complex signals in statistically independent components. It solves the blind source separation problem (BSS). The EEG satisfies most of the assumptions of ICA, so it may be an adequate signal for ICA for its use. In this paper we review the method and the applications of ICA in EEG. The studied applications are: a) artifacts removal; b) source estimation of spikes; c) analysis of seizures. Several studies have demonstrated that ICA is useful to remove artifacts from contaminated EEG records without distorting cerebral activity. It is able to decompose epileptic discharges and seizures in independent spatio-temporal components. In combination with techniques of source localisation, as dipole modelling, ICA improves the localization of the epileptic focus. Finally, we discuss the future role of ICA in the study of epileptic patients.


Amyloid-β oligomers induce synaptic damage via Tau-dependent microtubule severing by TTLL6 and spastin.
Mislocalization and aggregation of Aβ and Tau combined with loss of synapses and microtubules (MTs) are hallmarks of Alzheimer disease. We exposed mature primary neurons to Aβ oligomers and analysed changes in the Tau/MT system. MT breakdown occurs in dendrites invaded by Tau (Tau missorting) and is mediated by spastin, an MT-severing enzyme. Spastin is recruited by MT polyglutamylation, induced by Tau missorting triggered translocalization of TTLL6 (Tubulin-Tyrosine-Ligase-Like-6) into dendrites. Consequences are spine loss and mitochondria and neurofilament mislocalization. Missorted Tau is not axonally derived, as shown by axonal retention of photoconvertible Dendra2-Tau, but newly synthesized. Recovery from Aβ insult occurs after Aβ oligomers lose their toxicity and requires the kinase MARK (Microtubule-Affinity-Regulating-Kinase). In neurons derived from Tau-knockout mice, MTs and synapses are resistant to Aβ toxicity because TTLL6 mislocalization and MT polyglutamylation are prevented; hence no spastin recruitment and no MT breakdown occur, enabling faster recovery. Reintroduction of Tau re-establishes Aβ-induced toxicity in TauKO neurons, which requires phosphorylation of Tau's KXGS motifs. Transgenic mice overexpressing Tau show TTLL6 translocalization into dendrites and decreased MT stability. The results provide a rationale for MT stabilization as a therapeutic approach.


Origin of activation energy in a superionic conductor.
The characteristics of cation diffusion with many-body effects are discussed using Ag β-alumina as an example of a superionic conductor. Polarized Raman spectra of Ag β-alumina have been measured at room temperature. The interatomic potentials were determined by a non-linear least square fitting between the phonon eigenvalues from the Raman observations and a dynamical matrix calculation based on a rigid-ion model. The obtained potential parameters for the model crystal of Ag β-alumina successfully reproduce the macroscopic properties with respect to the heat capacity, isothermal compressibility and self-diffusion constant. A molecular dynamics (MD) calculation has been carried out using the model crystal of Ag β-alumina to understand the many-body effects for the fast ionic diffusion. It was found that the Ag-Ag repulsion by excess Ag defects significantly reduced the cost of the energy difference of the occupancy between the stable and metastable sites. It is possible for the system to take various configurations of the mobile ions through defects easily, and then the fast ionic diffusion will appear. On the other hand, the Ag-Ag repulsion changes the dynamics of the Ag ions from a random hopping to a cooperative motion. In the cooperative motion, the ionic transport becomes difficult due to the additional energy required for the structural relaxation of the surrounding Ag ions. We propose a new insight into the superionic conduction, that is, the activation energy for the ionic transport is composed of two kinds of elements: a 'static' activation energy and a 'dynamic' one. The static activation energy is the cost of the averaged energy difference in the various structural configurations in the equilibrium state. The dynamic activation energy is the additional energy required for the structural relaxation induced by the jump process.


Decrease of theta response in euthymic bipolar patients during an oddball paradigm.
Theta oscillations are related to cognitive functions and reflect functional integration of frontal and medial temporal structures into coherent neurocognitive networks. This study assessed event-related theta oscillations in medication-free, euthymic patients with bipolar disorder upon auditory oddball paradigm. Twenty-two DSM-IV euthymic bipolar I (n = 19) and II (n = 3) patients and twenty-two healthy subjects were included. Patients were euthymic for at least 6 months, and psychotropic-free for at least 2 weeks. EEG was recorded at 30 electrode sites. Auditory oddball paradigm and sensory stimuli were used. Event-related Oscillations were analyzed using adaptive filtering in two different theta frequency bands (4-6 Hz, 6-8 Hz). In healthy subjects, slow theta (4-6 Hz) responses were significantly higher than those of euthymic patients upon target, non-target and sensory stimuli (p < 0.05). Fast theta (6-8 Hz) responses of healthy subjects were significantly higher than those of euthymic patients upon target-only stimuli (p < 0.05). Reduced theta oscillations during auditory processing provide strong quantitative evidence of activation deficits in related networks in bipolar disorder. Fast theta responses are related to cognitive functions, whereas slow theta responses are related to sensory processes more than cognitive processes.


Risk of Major Adverse Cardiovascular Events and All-Cause Mortality in Patients With Hidradenitis Suppurativa.
Hidradenitis suppurativa (HS) is a common inflammatory skin disease. The disease has been associated with cardiovascular (CV) risk factors, but the risk of CV disease in patients with HS is unknown. To investigate CV risk in patients with HS. A population-based cohort study was conducted from January 1, 1997, to December 31, 2011, using individual-level linkage of nationwide administrative registers. In a study population of 35 368 Danish individuals, 5964 patients aged 18 years or older with a hospital-based diagnosis of HS (cases) were matched 1:5 on age, sex, and calendar time with 29 404 individuals serving as controls. Outcomes were myocardial infarction (MI), stroke, CV-associated death, major adverse CV events (MACEs), and all-cause mortality. Incidence rate ratios (IRRs) were estimated by Poisson regression. The 5964 patients with HS had a mean (SD) age of 37.7 (11.7) years; 4346 (72.9%) were women. In this sample, a total of 62 (42 749.0 person-years) MIs, 74 (42 647.8 person-years) ischemic strokes, 63 (42 941.7 person-years) CV-associated deaths, 169 (42 463.5 person-years) MACEs, and 231 (42 941.7 person-years) all-cause deaths occurred during follow-up. Adjusted (age, sex, socioeconomic status, smoking, comorbidity, and medication) IRRs (95% CIs) were 1.57 (1.14-2.17) for MI, 1.33 (1.01-1.76) for ischemic stroke, 1.95 (1.42-2.67) for CV-associated death, 1.53 (1.27-1.86) for MACEs, and 1.35 (1.15-1.59) for all-cause mortality. When patients with severe psoriasis were used as controls, the adjusted IRRs in patients with HS were 1.00 (0.74-1.35) for MI, 0.93 (0.71-1.22) for ischemic stroke, 1.58 (1.17-2.12) for CV-associated death, 1.08 (0.90-1.29) for MACEs, and 1.09 (0.94-1.28) for all-cause mortality. Hidradenitis suppurativa was associated with a significantly increased risk of adverse CV outcomes and all-cause mortality independent of measured confounders. The risk of CV-associated death was higher in patients with HS compared with the risk in those with severe psoriasis. The results call for increased awareness of this association and for studies of its clinical consequences.


Expression of the DF3-P epitope in human ovarian carcinomas.
Recent studies have described the generation of a mAb, designated DF3-P, which reacts with underglycosylated precursors of the DF3/MUC1 mucin-like glycoprotein. The present work demonstrates that the epitope recognized by mAb DF3-P is expressed by cell lines derived from human epithelial ovarian carcinomas and not a teratocarcinoma. Indirect immunofluorescence assays of single-cell suspensions support expression of the DF3-P epitope on the surface of ovarian carcinomas. Immunofluorescence studies on chamber slides further demonstrate that the mAb DF3-P-reactive cells are present in clusters. We also demonstrate that 125I-labeled mAb DF3-P selectively localizes to human ovarian carcinoma xenografts in athymic mice. The percentage of injected 125I dose/g tissue ranged between 10 and 17% for implanted CAOV-3 and OVCAR-3 tumors. Finally, the results of immunoperoxidase staining studies demonstrate that the DF3-P epitope is detectable in formalin-fixed sections of ovarian tumors and that mAb DF3-P exhibits little if any reactivity with normal surrounding tissues. Selective expression of the DF3-P epitope may be useful as a target for radioimaging or immunotherapeutic approaches to ovarian cancer.


Direct cone beam SPECT reconstruction with camera tilt.
A filtered backprojection (FBP) algorithm is derived to perform cone beam (CB) single-photon emission computed tomography (SPECT) reconstruction with camera tilt using circular orbits. This algorithm reconstructs the tilted angle CB projection data directly by incorporating the tilt angle into it. When the tilt angle becomes zero, this algorithm reduces to that of Feldkamp. Experimentally acquired phantom studies using both a two-point source and the three-dimensional Hoffman brain phantom have been performed. The transaxial tilted cone beam brain images and profiles obtained using the new algorithm are compared with those without camera tilt. For those slices which have approximately the same distance from the detector in both tilt and non-tilt set-ups, the two transaxial reconstructions have similar profiles. The two-point source images reconstructed from this new algorithm and the tilted cone beam brain images are also compared with those reconstructed from the existing tilted cone beam algorithm which requires rebinning the projection data. These comparisons demonstrate that the new algorithm, compared with the existing tilted cone beam algorithm, can provide better image contrast and improved image resolution.


Standalone performance of artificial intelligence for upper GI neoplasia: a meta-analysis.
Artificial intelligence (AI) may reduce underdiagnosed or overlooked upper GI (UGI) neoplastic and preneoplastic conditions, due to subtle appearance and low disease prevalence. Only disease-specific AI performances have been reported, generating uncertainty on its clinical value. We searched PubMed, Embase and Scopus until July 2020, for studies on the diagnostic performance of AI in detection and characterisation of UGI lesions. Primary outcomes were pooled diagnostic accuracy, sensitivity and specificity of AI. Secondary outcomes were pooled positive (PPV) and negative (NPV) predictive values. We calculated pooled proportion rates (%), designed summary receiving operating characteristic curves with respective area under the curves (AUCs) and performed metaregression and sensitivity analysis. Overall, 19 studies on detection of oesophageal squamous cell neoplasia (ESCN) or Barrett's esophagus-related neoplasia (BERN) or gastric adenocarcinoma (GCA) were included with 218, 445, 453 patients and 7976, 2340, 13 562 images, respectively. AI-sensitivity/specificity/PPV/NPV/positive likelihood ratio/negative likelihood ratio for UGI neoplasia detection were 90% (CI 85% to 94%)/89% (CI 85% to 92%)/87% (CI 83% to 91%)/91% (CI 87% to 94%)/8.2 (CI 5.7 to 11.7)/0.111 (CI 0.071 to 0.175), respectively, with an overall AUC of 0.95 (CI 0.93 to 0.97). No difference in AI performance across ESCN, BERN and GCA was found, AUC being 0.94 (CI 0.52 to 0.99), 0.96 (CI 0.95 to 0.98), 0.93 (CI 0.83 to 0.99), respectively. Overall, study quality was low, with high risk of selection bias. No significant publication bias was found. We found a high overall AI accuracy for the diagnosis of any neoplastic lesion of the UGI tract that was independent of the underlying condition. This may be expected to substantially reduce the miss rate of precancerous lesions and early cancer when implemented in clinical practice.


Reflection-mode optical-resolution photoacoustic microscopy based on a reflective objective.
We developed a new reflection-mode optical-resolution photoacoustic microscopy (OR-PAM) based on the cooperation of a reflective objective and an ultrasonic transducer. The reflective objective is used to produce nearly diffraction-limited optical focusing, and the excited ultrasound waves are then directly detected by an ultrasonic transducer that was placed in the central cone of the objective. This new design avoids the coupling between optical focusing and ultrasound transmission in the reflection mode. Moreover, the proposed system is able to provide lateral resolution of 1.2 μm at 580 nm, penetration depth of 0.9 mm in biological tissues, and a work distance of 6.0 mm. We present in vivo imaging of the microvasculature in mouse ears and in vitro imaging of red blood cells (RBCs), which demonstrate the capability of the system to study microcirculation.


Prevalence of human immunodeficiency virus and hepatitis C virus among French prison inmates in 2010: a challenge for public health policy.
We evaluated prevalence of human immunodeficiency virus (HIV) and hepatitis C virus (HCV) among prison inmates in France in 2010, in a cross-sectional single-day study based on a two-stage design. Sampling favoured larger establishments and included all types of prisons. Establishments were stratified by geographical region. Estimates were adjusted by post-stratification of the total population of inmates in France. From 60,975 inmates in all 188 prisons on the sampling day, 2,154 were selected from 27 prisons, and 1,876 questionnaires completed. HIV prevalence was estimated at 2.0% (95% confidence interval (CI): 0.9–4.2), 2.6% (95% CI: 0.7–8.8) in women and 2.0% (95% CI: 0.9–4.3) in men; 75% of inmates were receiving treatment for HIV. HCV prevalence was estimated at 4.8% (95% CI: 3.5–6.5) and was higher for women (11.8%; 95% CI: 8.5–16.1) than men (4.5%; 95% CI: 3.3–6.3). Almost half of HCV-infected inmates had chronic hepatitis C and 44% were receiving or had received treatment. HIV and HCV prevalence was six times higher than in the general population, and 2.5% of inmates had viraemic hepatitis C. The moment of incarceration provides an ideal opportunity for testing and treating, limiting spread of HCV and improving patients' prognosis.


Hotspot SF3B1 mutations induce metabolic reprogramming and vulnerability to serine deprivation.
Cancer-associated mutations in the spliceosome gene SF3B1 create a neomorphic protein that produces aberrant mRNA splicing in hundreds of genes, but the ensuing biologic and therapeutic consequences of this missplicing are not well understood. Here we have provided evidence that aberrant splicing by mutant SF3B1 altered the transcriptome, proteome, and metabolome of human cells, leading to missplicing-associated downregulation of metabolic genes, decreased mitochondrial respiration, and suppression of the serine synthesis pathway. We also found that mutant SF3B1 induces vulnerability to deprivation of the nonessential amino acid serine, which was mediated by missplicing-associated downregulation of the serine synthesis pathway enzyme PHGDH. This vulnerability was manifest both in vitro and in vivo, as dietary restriction of serine and glycine in mice was able to inhibit the growth of SF3B1MUT xenografts. These findings describe a role for SF3B1 mutations in altered energy metabolism, and they offer a new therapeutic strategy against SF3B1MUT cancers.


Purine transport and metabolism in man: the effect of exercise on concentrations of purine bases, nucleosides and nucleotides in plasma, urine, leucocytes and erythrocytes.
1. After decreasing muscle ATP by a 2 min period of intense exercise, we have studied purine metabolism by using high-pressure liquid chromatography. 2. A major increase in hypoxanthine concentration in plasma and urine was found with increases in xanthine and, in plasma, inosine. Erythrocyte hypoxanthine rose with the level in plasma, but there was no corresponding rise in IMP, the first intracellular metabolite of hypoxanthine. No rises in uridine or urate were found in plasma. 3. Plasma adenosine did not rise and fall significantly after exercise, but a small rise and fall in adenine nucleotide concentrations in plasma was found. 4. Running, swimming and games, which tended to be at the weekend, were associated with a rise in hypoxanthine and xanthine excretion; exercise was probably the cause of the higher excretion during the day than at night. Such activities do not produce changes in concentrations of ATP in muscle, although turnover must rise. 5. The results are consistent with widespread purine exchange between tissues and a 'circulating hypoxanthine pool'.


Increasing comorbidity and health services utilization in older adults with prior stroke.
To characterize comorbid chronic conditions, describe health services use, and estimate health care costs among community-dwelling older adults with prior stroke. This is a retrospective cohort study using administrative data from Ontario, Canada. We identified all community-dwelling individuals aged 66 and over on April 1, 2008 (baseline), who had experienced a stroke at least 6 months prior. We estimated the prevalence of 14 comorbid conditions at baseline; we captured all physician visits, emergency department visits, hospital admissions, home care contacts, and associated costs over 5 years stratifying by number of comorbid conditions. Where possible, we distinguished between health services use for stroke- and non-stroke-related reasons. A total of 29,673 individuals met our criteria. Only 1% had no comorbid conditions, while 74.9% had 3 or more. The most common conditions were hypertension (89.8%) and arthritis (65.8%); 5 other conditions had a prevalence of 20% or more (ischemic heart disease, diabetes, chronic obstructive pulmonary disease, inflammatory bowel disease, and dementia). Use of all health services doubled with increasing comorbidity and was largely attributed to non-stroke-related reasons. Total and per-patient costs increased with comorbidity. Main cost drivers shifted from physician and home care visits to hospital admissions with greater comorbidity. Our findings demonstrate the importance of community-based patient-centered care strategies for stroke survivors that address their range of health needs and prevent more costly acute care use.


The Influence of Preoperative Interventions on Postoperative Surgical Wound Healing in Patients Without Risk Factors: A Systematic Review.
Poor wound healing and scar formation remain critical problems in daily surgical practice. Generally, most attention is paid to intra- and postoperative interventions to improve wound healing after surgery, while preoperative interventions remain unsatisfactorily explored. In this systematic review, the available literature on the beneficial effects of preoperative interventions on wound healing and scar formation have been summarized and compared. A comprehensive and systematic search has been conducted in MEDLINE, Pubmed, Embase, Web of Science, and Cochrane, supplemented by reference and citation tracking. All preoperative interventions and all clinically relevant outcome parameters have been considered for inclusion, due to the expected limited availability of literature. A total of 13 studies were included, which were all randomized trials. No cohort studies or retrospective studies have been identified. All studies described different preoperative interventions and outcome parameters and could hence not be pooled and compared. Eight studies showed significantly better wound healing after a preoperative intervention. The individual studies have been summarized in this review. This systemic review shows that preoperative interventions can be beneficial in improving wound healing and scar formation. In selected cases, wound healing was found to benefit from a higher preoperative body temperature, topical vitamin E application, and low patient stress levels.


Updates on clinical studies of selenium supplementation in radiotherapy.
To establish guidelines for the selenium supplementation in radiotherapy we assessed the benefits and risks of selenium supplementation in radiotherapy. Clinical studies on the use of selenium in radiotherapy were searched in the PubMed electronic database in January 2013. Sixteen clinical studies were identified among the 167 articles selected in the initial search. Ten articles were observational studies, and the other 6 articles reported studies on the effects of selenium supplementation in patients with cancer who underwent radiotherapy. The studies were conducted worldwide including European, American and Asian countries between 1987 and 2012. Plasma, serum or whole blood selenium levels were common parameters used to assess the effects of radiotherapy and the selenium supplementation status. Selenium supplementation improved the general conditions of the patients, improved their quality of life and reduced the side effects of radiotherapy. At the dose of selenium used in these studies (200-500 μg/day), selenium supplementation did not reduce the effectiveness of radiotherapy, and no toxicities were reported. Selenium supplementation may offer specific benefits for several types of cancer patients who undergo radiotherapy. Because high-dose selenium and long-term supplementation may be unsafe due to selenium toxicity, more evidence-based information and additional research are needed to ensure the therapeutic benefits of selenium supplementation.


Measuring the accuracy of self-reported height and weight in a community-based sample of young people.
Self-reported anthropometric data are commonly used to estimate prevalence of obesity in population and community-based studies. We aim to: 1) Determine whether survey participants are able and willing to self-report height and weight; 2) Assess the accuracy of self-reported compared to measured anthropometric data in a community-based sample of young people. Participants (16-29 years) of a behaviour survey, recruited at a Melbourne music festival (January 2011), were asked to self-report height and weight; researchers independently weighed and measured a sub-sample. Body Mass Index was calculated and overweight/obesity classified as ≥25 kg/m². Differences between measured and self-reported values were assessed using paired t-test/Wilcoxon signed ranks test. Accurate report of height and weight were defined as <2 cm and <2 kg difference between self-report and measured values, respectively. Agreement between classification of overweight/obesity by self-report and measured values was assessed using McNemar's test. Of 1405 survey participants, 82% of males and 72% of females self-reported their height and weight. Among 67 participants who were also independently measured, self-reported height and weight were significantly less than measured height (p=0.01) and weight (p<0.01) among females, but no differences were detected among males. Overall, 52% accurately self-reported height, 30% under-reported, and 18% over-reported; 34% accurately self-reported weight, 52% under-reported and 13% over-reported. More females (70%) than males (35%) under-reported weight (p=0.01). Prevalence of overweight/obesity was 33% based on self-report data and 39% based on measured data (p=0.16). Self-reported measurements may underestimate weight but accurately identified overweight/obesity in the majority of this sample of young people.


Final height and growth hormone secretion after completion of growth hormone therapy in patients with idiopathic growth hormone deficiency and with abnormalities of the hypothalamic-pituitary region.
The aim of the study was an evaluation of final height and growth hormone (GH) secretion after completion of GH therapy (retesting) in patients with GH deficiency (GHD). The analysis comprised 53 patients (43 boys, 10 girls) with childhood-onset GHD, who completed GH therapy and reached final height. Magnetic resonance imaging (MRI), performed in all the patients, led to the following groups: pituitary stalk interruption syndrome (PSIS), pituitary hypoplasia (HP), craniopharyngioma (CP) -- patients after tumour excision, patients with normal hypothalamic-pituitary region (NP). In 51 patients, final height was normal. The height gain was significantly (p<0.05) greater in PSIS than in that other groups. In retesting, GH secretion was significantly (p<0.005) lower in PSIS and CP than in HP and in NP and also (p<0.05) in HP than in NP. Permanent severe GHD was confirmed in all the patients with PSIS and CP and in some patients with HP (37.5%), while it was excluded in all the patients with normal pituitary in MRI. It seems that in patients with PSIS and CP, the confirmation of persistent character of GHD needs no retesting, while in patients with normal MRI results, GHD diagnosis should be established with special attention.


Recognition without awareness in humans and its implications for animal models of episodic memory.
Animal models of episodic memory-the ability to recall events from the past along with relevant spatial, temporal and other contextual detail-have been criticized on the grounds that it is difficult to determine if animals engage in the "mental time travel" that accompanies episodic recall in humans. It is possible that animals rely on mechanisms different from those operative in humans. We have added a new dimension to these concerns by showing that a test widely considered to measure episodic memory in humans could be constructed such that it measures nonconscious (implicit) memory under certain test parameters. The same parameters are common in tests frequently used to study recognition in nonhuman animals. In particular, recognition was driven by implicit memory when (1) recognition decisions could not be guided by semantic information, and (2) decisions were made with relatively high automaticity. These findings suggest that nonhuman animals may rely on implicit memory processing in many episodic-memory testing circumstances, as semantic content is minimal and automaticity is likely after the extended practice usually required. Identifying factors that promote and discourage recognition based on implicit memory is therefore necessary for creating accurate cognitive and neurobiological models of memory processing in humans and other animals.


Sepsis-induced liver dysfunction was ameliorated by propofol via suppressing hepatic lipid peroxidation, inflammation, and drug interactions.
Our previous study showed that propofol can protect against sepsis-induced insults through suppressing liver nitrosation and inflammation. This study further evaluated the mechanisms of propofol-caused protection from sepsis-induced liver dysfunction. Male Wistar rats were subjected to cecal ligation and puncture (CLP) and then exposed to propofol. Levels of hepatic oxidative stress and lipid peroxidation were consecutively measured. Expressions of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-4 messenger (m)RNA or proteins were quantified. Effects of propofol on microsomal pentoxyresorufin O-dealkelase (PROD) and ethoxycoumarin O-deethylase (ECOD) activities were determined. Administration of propofol to CLP-treated rats significantly attenuated sepsis-induced insults. CLP caused augmented serum aspartate aminotransferase and alanine aminotransferase activities and concurrently triggered liver damage. In contrast, treatment with propofol protected against CLP-induced liver dysfunction. As to the mechanisms, the CLP-induced increases in oxidative stress and lipid peroxidation levels and TNF-α and IL-1β mRNA and protein expressions were subsequently attenuated by propofol. Furthermore, administration of CLP-treated rats with propofol augmented levels of IL-4 in the liver. Phenobarbital treatment of liver microsomes in CLP-treated rats produced less amplification of PROD and ECOD activities, and a smaller amount of 4-hydroxypropofol was metabolized from propofol by liver microsomes. In contrast, more drug interactions occurred with propofol, which decreased PROD and ECOD activities in liver microsomes of CLP-treated rats. Taken together, the present study showed that propofol can protect against sepsis-induced liver dysfunction through suppressing hepatic oxidative stress, lipid peroxidation, inflammation, and drug biotransformation and interactions in the liver.


The genetic architecture of biofilm formation in a clinical isolate of Saccharomyces cerevisiae.
Biofilms are microbial communities that form on surfaces. They are the primary form of microbial growth in nature and can have detrimental impacts on human health. Some strains of the budding yeast Saccharomyces cerevisiae form colony biofilms, and there is substantial variation in colony architecture between biofilm-forming strains. To identify the genetic basis of biofilm variation, we developed a novel version of quantitative trait locus mapping, which leverages cryptic variation in a clinical isolate of S. cerevisiae. We mapped 13 loci linked to heterogeneity in biofilm architecture and identified the gene most closely associated with each locus. Of these candidate genes, six are members of the cyclic AMP-protein kinase A pathway, an evolutionarily conserved cell signaling network. Principal among these is CYR1, which encodes the enzyme that catalyzes production of cAMP. Through a combination of gene expression measurements, cell signaling assays, and gene overexpression, we determined the functional effects of allelic variation at CYR1. We found that increased pathway activity resulting from protein coding and expression variation of CYR1 enhances the formation of colony biofilms. Four other candidate genes encode kinases and transcription factors that are targets of this pathway. The protein products of several of these genes together regulate expression of the sixth candidate, FLO11, which encodes a cell adhesion protein. Our results indicate that epistatic interactions between alleles with both positive and negative effects on cyclic AMP-protein kinase A signaling underlie much of the architectural variation we observe in colony biofilms. They are also among the first to demonstrate genetic variation acting at multiple levels of an integrated signaling and regulatory network. Based on these results, we propose a mechanistic model that relates genetic variation to gene network function and phenotypic outcomes.


Scary and nasty beasts: Self-reported fear and disgust of common phobic animals.
Animal phobias are one of the most prevalent mental disorders. We analysed how fear and disgust, two emotions involved in their onset and maintenance, are elicited by common phobic animals. In an online survey, the subjects rated 25 animal images according to elicited fear and disgust. Additionally, they completed four psychometrics, the Fear Survey Schedule II (FSS), Disgust Scale - Revised (DS-R), Snake Questionnaire (SNAQ), and Spider Questionnaire (SPQ). Based on a redundancy analysis, fear and disgust image ratings could be described by two axes, one reflecting a general negative perception of animals associated with higher FSS and DS-R scores and the second one describing a specific aversion to snakes and spiders associated with higher SNAQ and SPQ scores. The animals can be separated into five distinct clusters: (1) non-slimy invertebrates; (2) snakes; (3) mice, rats, and bats; (4) human endo- and exoparasites (intestinal helminths and louse); and (5) farm/pet animals. However, only snakes, spiders, and parasites evoke intense fear and disgust in the non-clinical population. In conclusion, rating animal images according to fear and disgust can be an alternative and reliable method to standard scales. Moreover, tendencies to overgeneralize irrational fears onto other harmless species from the same category can be used for quick animal phobia detection.


Opportunistic pathogens relative to physicochemical factors in water storage tanks.
Household water in Oman, as well as in other countries in the region, is stored in tanks placed on house roofs that can be subjected to physicochemical factors which can promote microbial growth, including pathogens and opportunistic pathogens which pose health risks. Water samples were collected from 30 houses in a heavily populated suburb of Muscat. The tanks used were either glass reinforced plastic (GRP), polyethylene or galvanised iron (GI). Heterotrophic bacteria, coliforms, faecal coliforms and iron sulphur bacteria varied significantly in the three tanks. Yeast and mould count showed significant variations. Isolation of Aeromonas spp., fluorogenic and pathogenic Pseudomonas, Pasteurella, Salmonella, Serratia and Tatumella, and Yersinia and Legionella in biofilms varied in the three tanks. The fungi isolates in the three tanks were Penicillium, Cladosporium and Aspergillus. Nephelometric turbidity unit, threshold odour number and free chlorine varied significantly in the three tanks. True colour unit values did not show a significant difference; however, GRP tanks had algae, autotrophic and pigmented microorganisms. In addition, GI tanks had sediments and corrosion. The results of this investigation are important to evaluate the status of the present household water tanks in countries with high annual temperatures, which may affect public health.


Prolonged-acting, multi-targeting gallium nanoparticles potently inhibit growth of both HIV and mycobacteria in co-infected human macrophages.
Human immunodeficiency virus (HIV) infection and Mycobacterium tuberculosis (TB) are responsible for two of the major global human infectious diseases that result in significant morbidity, mortality and socioeconomic impact. Furthermore, severity and disease prevention of both infections is enhanced by co-infection. Parallel limitations also exist in access to effective drug therapy and the emergence of resistance. Furthermore, drug-drug interactions have proven problematic during treatment of co-incident HIV and TB infections. Thus, improvements in drug access and simplified treatment regimens are needed immediately. One of the key host cells infected by both HIV and TB is the mononuclear phagocyte (MP; monocyte, macrophage and dendritic cell). Therefore, we hypothesized that one way this can be achieved is through drug-targeting by a nanoformulated drug that ideally would be active against both HIV and TB. Accordingly, we validated macrophage targeted long acting (sustained drug release) gallium (Ga) nanoformulation against HIV-mycobacterium co-infection. The multi-targeted Ga nanoparticle agent inhibited growth of both HIV and TB in the macrophage. The Ga nanoparticles reduced the growth of mycobacterium and HIV for up to 15 days following single drug loading. These results provide a potential new approach to treat HIV-TB co-infection that could eventually lead to improved clinical outcomes.