PKCθ and HIV-1 Transcriptional Regulator Tat Co-exist at the LTR Promoter in CD4(+) T Cells.
PKCθ is essential for the activation of CD4(+) T cells. Upon TCR/CD28 stimulation, PKCθ is phosphorylated and migrates to the immunological synapse, inducing the activation of cellular transcription factors such as NF-κB and kinases as ERK that are critical for HIV-1 replication. We previously demonstrated that PKCθ is also necessary for HIV-1 replication but the precise mechanism is unknown. Efficient HIV-1 transcription and elongation are absolutely dependent on the synergy between NF-κB and the viral regulator Tat. Tat exerts its function by binding a RNA stem-loop structure proximal to the viral mRNA cap site termed TAR. Besides, due to its effect on cellular metabolic pathways, Tat causes profound changes in infected CD4(+) T cells such as the activation of NF-κB and ERK. We hypothesized that the aberrant upregulation of Tat-mediated activation of NF-κB and ERK occurred through PKCθ signaling. In fact, Jurkat TetOff cells with stable and doxycycline-repressible expression of Tat (Jurkat-Tat) expressed high levels of mRNA for PKCθ. In these cells, PKCθ located at the plasma membrane was phosphorylated at T(538) residue in undivided cells, in the absence of stimulation. Treatment with doxycycline inhibited PKCθ phosphorylation in Jurkat-Tat, suggesting that Tat expression was directly related to the activation of PKCθ. Both NF-κB and Ras/Raf/MEK/ERK signaling pathway were significantly activated in Jurkat-Tat cells, and this correlated with high transactivation of HIV-1 LTR promoter. RNA interference for PKCθ inhibited NF-κB and ERK activity, as well as LTR-mediated transactivation even in the presence of Tat. In addition to Tat-mediated activation of PKCθ in the cytosol, we demonstrated by sequential ChIP that Tat and PKCθ coexisted in the same complex bound at the HIV-1 LTR promoter, specifically at the region containing TAR loop. In conclusion, PKCθ-Tat interaction seemed to be essential for HIV-1 replication in CD4(+) T cells and could be used as a therapeutic target.


Neuroprosthetic technologies to augment the impact of neurorehabilitation after spinal cord injury.
Spinal cord injury leads to a range of disabilities, including limitations in locomotor activity, that seriously diminish the patients' autonomy and quality of life. Electrochemical neuromodulation therapies, robot-assisted rehabilitation and willpower-based training paradigms restored supraspinal control of locomotion in rodent models of severe spinal cord injury. This treatment promoted extensive and ubiquitous remodeling of spared circuits and residual neural pathways. In four chronic paraplegic individuals, electrical neuromodulation of the spinal cord resulted in the immediate recovery of voluntary leg movements, suggesting that the therapeutic concepts developed in rodent models may also apply to humans. Here, we briefly review previous work, summarize current developments, and highlight impediments to translate these interventions into medical practice to improve functional recovery of spinal-cord-injured individuals.


GABA(A) receptor alpha1 subunit deletion prevents developmental changes of inhibitory synaptic currents in cerebellar neurons.
Developmental changes in miniature IPSC (mIPSC) kinetics have been demonstrated previously in cerebellar neurons in rodents. We report that these kinetic changes in mice are determined primarily by developmental changes in GABA(A) receptor subunit expression. mIPSCs were studied by whole-cell recordings in cerebellar slices, prepared from postnatal day 11 (P11) and P35 mice. Similar to reports in granule neurons, wild-type cerebellar stellate neuron mIPSCs at P11 had slow decay kinetics, whereas P35 mIPSCs decayed five times faster. When mIPSCs in cerebellar stellate neurons were compared between wild-type (+/+) and GABA(A) receptor alpha1 subunit-deficient (-/-) littermates at P35, we observed dramatically slower mIPSC decay rates in -/- animals. We took advantage of the greater potency of imidazopyridines for GABA current potentiation with alpha1 subunit-containing receptors to characterize the relative contribution of alpha1 subunits in native receptors on inhibitory synapses of cerebellar granule neurons. Zolpidem-induced prolongation of mIPSC decay was variable among distinct cells, but it increased during development in wild-type mice. Similarly, Zolpidem prolongation of mIPSC decay rate was significantly greater in adult +/+ mice than in knock-outs. We propose that an increased alpha1 subunit assembly in postsynaptic receptors of cerebellar inhibitory synapses is responsible for the fast inhibitory synaptic currents that are normally observed during postnatal development.


UMD (Universal mutation database): a generic software to build and analyze locus-specific databases.
The human genome is thought to contain about 80,000 genes and presently only 3,000 are known to be implicated in genetic diseases. In the near future, the entire sequence of the human genome will be available and the development of new methods for point mutation detection will lead to a huge increase in the identification of genes and their mutations associated with genetic diseases as well as cancers, which is growing in frequency in industrial states. The collection of these mutations will be critical for researchers and clinicians to establish genotype/phenotype correlations. Other fields such as molecular epidemiology will also be developed using these new data. Consequently, the future lies not in simple repositories of locus-specific mutations but in dynamic databases linked to various computerized tools for their analysis and that can be directly queried on-line. To meet this goal, we devised a generic software called UMD (Universal Mutation Database). It was developed as a generic software to create locus-specific databases (LSDBs) with the 4(th) Dimension(R) package from ACI. This software includes an optimized structure to assist and secure data entry and to allow the input of various clinical data. Thanks to the flexible structure of the UMD software, it has been successfully adapted to nine genes either involved in cancer (APC, P53, RB1, MEN1, SUR1, VHL, and WT1) or in genetic diseases (FBN1 and LDLR). Four new LSDBs are under construction (VLCAD, MCAD, KIR6, and COL4A5). Finally, the data can be transferred to core databases.


High prevalence of equine-like G3P[8] rotavirus in children and adults with acute gastroenteritis in Thailand.
Group A rotavirus (RVA) is a major cause of acute gastroenteritis in infants and young children worldwide. This study aims to clarify the distribution of G/P types and genetic characteristics of RVAs circulating in Thailand. Between January 2014 and September 2016, 1867 stool specimens were collected from children and adults with acute gastroenteritis in six provinces in Thailand. RVAs were detected in 514/1867 (27.5%) stool specimens. G1P[8] (44.7%) was the most predominant genotype, followed by G3P[8] (33.7%), G2P[4] (11.5%), G8P[8] (7.0%), and G9P[8] (1.3%). Unusual G3P[9] (0.8%), G3P[10] (0.4%), G4P[6] (0.4%), and G10P[14] (0.2%) were also detected at low frequencies. The predominant genotype, G1P[8] (64.4%), in 2014 decreased to 6.1% in 2016. In contrast, the frequency of G3P[8] markedly increased from 5.5% in 2014 to 65.3% in 2015 and 89.8% in 2016. On polyacrylamide gel electrophoresis, most (135/140; 96.4%) of the G3P[8] strains exhibited a short RNA profile. Successful determination of the nucleotide sequences of the VP7 genes of 98 G3P[8] strains with a short RNA profile showed that they are all equine-like G3P[8] strains. On phylogenetic analysis of genome segments of two representative Thai equine-like G3P[8] strains, it was noteworthy that they possessed distinct NSP4 genes, one bovine-like and the other human-like. Thus, we found that characteristic equine-like G3P[8] strains with a short RNA electropherotype are becoming highly prevalent in children and adults in Thailand.


Application of X-ray microtomography for the characterisation of hollow polymer-stabilised spray dried amorphous dispersion particles.
The aim of this study was to investigate the capability of X-ray microtomography to obtain information relating to powder characteristics such as wall thickness and solid volume fraction for hollow, polymer-stabilised spray dried dispersion (SDD) particles. SDDs of varying particle properties, with respect to shell wall thickness and degree of particle collapse, were utilised to assess the capability of the approach. The results demonstrate that the approach can provide insight into the morphological characteristics of these hollow particles, and thereby a means to understand/predict the processability and performance characteristics of the bulk material. Quantitative assessments of particle wall thickness, particle/void volume and thereby solid volume fraction were also demonstrated to be achievable. The analysis was also shown to be able to qualitatively assess the impact of the drying rate on the morphological nature of the particle surfaces, thus providing further insight into the final particle shape. The approach demonstrated a practical means to access potentially important particle characteristics for SDD materials which, in addition to the standard bulk powder measurements such as particle size and bulk density, may enable a better understanding of such materials, and their impact on downstream processability and dosage form performance.


Stress and parenting during the global COVID-19 pandemic.
Stress and compromised parenting often place children at risk of abuse and neglect. Child maltreatment has generally been viewed as a highly individualistic problem by focusing on stressors and parenting behaviors that impact individual families. However, because of the global coronavirus disease 2019 (COVID-19), families across the world are experiencing a new range of stressors that threaten their health, safety, and economic well-being. This study examined the impacts of the COVID-19 pandemic in relation to parental perceived stress and child abuse potential. Participants included parents (N = 183) with a child under the age of 18 years in the western United States. Tests of group differences and hierarchical multiple regression analyses were employed to assess the relationships among demographic characteristics, COVID-19 risk factors, mental health risk factors, protective factors, parental perceived stress, and child abuse potential. Greater COVID-19 related stressors and high anxiety and depressive symptoms are associated with higher parental perceived stress. Receipt of financial assistance and high anxiety and depressive symptoms are associated with higher child abuse potential. Conversely, greater parental support and perceived control during the pandemic are associated with lower perceived stress and child abuse potential. Results also indicate racial and ethnic differences in COVID-19 related stressors, but not in mental health risk, protective factors, perceived stress, or child abuse potential. Findings suggest that although families experience elevated stressors from COVID-19, providing parental support and increasing perceived control may be promising intervention targets.


Palaeoproteomic evidence identifies archaic hominins associated with the Châtelperronian at the Grotte du Renne.
In Western Europe, the Middle to Upper Paleolithic transition is associated with the disappearance of Neandertals and the spread of anatomically modern humans (AMHs). Current chronological, behavioral, and biological models of this transitional period hinge on the Châtelperronian technocomplex. At the site of the Grotte du Renne, Arcy-sur-Cure, morphological Neandertal specimens are not directly dated but are contextually associated with the Châtelperronian, which contains bone points and beads. The association between Neandertals and this "transitional" assemblage has been controversial because of the lack either of a direct hominin radiocarbon date or of molecular confirmation of the Neandertal affiliation. Here we provide further evidence for a Neandertal-Châtelperronian association at the Grotte du Renne through biomolecular and chronological analysis. We identified 28 additional hominin specimens through zooarchaeology by mass spectrometry (ZooMS) screening of morphologically uninformative bone specimens from Châtelperronian layers at the Grotte du Renne. Next, we obtain an ancient hominin bone proteome through liquid chromatography-MS/MS analysis and error-tolerant amino acid sequence analysis. Analysis of this palaeoproteome allows us to provide phylogenetic and physiological information on these ancient hominin specimens. We distinguish Late Pleistocene clades within the genus Homo based on ancient protein evidence through the identification of an archaic-derived amino acid sequence for the collagen type X, alpha-1 (COL10α1) protein. We support this by obtaining ancient mtDNA sequences, which indicate a Neandertal ancestry for these specimens. Direct accelerator mass spectometry radiocarbon dating and Bayesian modeling confirm that the hominin specimens date to the Châtelperronian at the Grotte du Renne.


Phase II Intergroup Trial of Alisertib in Relapsed and Refractory Peripheral T-Cell Lymphoma and Transformed Mycosis Fungoides: SWOG 1108.
Aurora A kinase (AAK) is upregulated in highly proliferative lymphomas, suggesting its potential as a therapeutic target. Alisertib is a novel oral AAK inhibitor without adverse safety signals in early-phase studies that demonstrated preliminary activity in T-cell lymphoma. This phase II study was conducted to further investigate the efficacy of alisertib in relapsed or refractory peripheral T-cell non-Hodgkin lymphoma (PTCL). Eligible patients with histologically confirmed relapsed/refractory PTCL or transformed Mycosis fungoides (tMF) received alisertib 50 mg twice a day for 7 days on 21-day cycles. Of 37 eligible patients, the histologic subtypes enrolled included PTCL not otherwise specified (n = 13), angioimmunoblastic T-cell lymphoma (n = 9), tMF (n = 7), adult T-cell lymphoma/leukemia (n = 4), anaplastic large-cell lymphoma (n = 2), and extranodal natural killer/T-cell lymphoma (n = 2). Grade 3 and 4 adverse events in ≥ 5% of patients included neutropenia (32%), anemia (30%), thrombocytopenia (24%), febrile neutropenia (14%), mucositis (11%), and rash (5%). Treatment was discontinued most commonly for disease progression. Among the PTCL subtypes, the overall response rate was 30%, whereas no responses were observed in tMF. Aurora B kinase was more commonly overexpressed than AAK in tumor specimens. Analysis of AAK, Aurora B kinase, MYC, BCL-2, phosphatidylinositol 3-kinase γ, and Notch1 expression revealed no association with response. Alisertib has antitumor activity in PTCL, including heavily pretreated patients. These promising results are being further investigated in an ongoing international, randomized phase III trial comparing alisertib with investigator's choice in PTCL.


Genotypic Prediction of Co-receptor Tropism of HIV-1 Subtypes A and C.
Antiretroviral treatment of Human Immunodeficiency Virus type-1 (HIV-1) infections with CCR5-antagonists requires the co-receptor usage prediction of viral strains. Currently available tools are mostly designed based on subtype B strains and thus are in general not applicable to non-B subtypes. However, HIV-1 infections caused by subtype B only account for approximately 11% of infections worldwide. We evaluated the performance of several sequence-based algorithms for co-receptor usage prediction employed on subtype A V3 sequences including circulating recombinant forms (CRFs) and subtype C strains. We further analysed sequence profiles of gp120 regions of subtype A, B and C to explore functional relationships to entry phenotypes. Our analyses clearly demonstrate that state-of-the-art algorithms are not useful for predicting co-receptor tropism of subtype A and its CRFs. Sequence profile analysis of gp120 revealed molecular variability in subtype A viruses. Especially, the V2 loop region could be associated with co-receptor tropism, which might indicate a unique pattern that determines co-receptor tropism in subtype A strains compared to subtype B and C strains. Thus, our study demonstrates that there is a need for the development of novel algorithms facilitating tropism prediction of HIV-1 subtype A to improve effective antiretroviral treatment in patients.


Dual Population Coding in the Neocortex: A Model of Interaction between Representation and Attention in the Visual Cortex.
The text describes a model that extends the population coding principles to any multidimensional attribute. The model distinguishes between the distribution of cell activity and the overall activity of a population. The distribution of cell activity is assumed to encode attribute information, while overall activity is assumed to reflect the significance or pertinence of the encoded attribute in the cerebral cortex, according to the dual coding principle. Three basic mechanisms of interaction between the representation of attribute and pertinence are defined and are applied to the motion (MT-MST) cortical pathway in the visual cortex. This framework determines three sources of pertinence that model cognitive processing, including preattentive processing, spatial-selective attention, and object-selective attention. The model accommodates most of the published psychophysical, neurophysiological, and neuroanatomical data and makes several testable predictions about the representations of attribute and enhanced effects in these areas.


Using mimicry of body movements by a virtual agent to increase synchronization behavior and rapport in individuals with schizophrenia.
Synchronization of behavior such as gestures or postures is assumed to serve crucial functions in social interaction but has been poorly studied to date in schizophrenia. Using a virtual collaborative environment (VCS), we tested 1) whether synchronization of behavior, i.e., the spontaneous initiation of gestures that are congruent with those of an interaction partner, was impaired in individuals with schizophrenia compared with healthy participants; 2) whether mimicry of the patients' body movements by the virtual interaction partner was associated with increased behavioral synchronization and rapport. 19 patients and 19 matched controls interacted with a virtual agent who either mimicked their head and torso movements with a delay varying randomly between 0.5 s and 4 s or did not mimic, and rated feelings of rapport toward the virtual agent after each condition. Both groups exhibited a higher and similar synchronization behavior of the virtual agent forearm movements when they were in the Mimicry condition rather than in the No-mimicry condition. In addition, both groups felt more comfortable with a mimicking virtual agent rather than a virtual agent not mimicking them suggesting that mimicry is able to increase rapport in individuals with schizophrenia. Our results suggest that schizophrenia cannot be considered anymore as a disorder of imitation, particularly as regards behavioral synchronization processes in social interaction contexts.


Amino Acid Uptake and Metabolism by Larvae of the Marine Worm Urechis caupo (Echiura), a New Species in Axenic Culture.
Axenic (bacteria-free) larval cultures of the marine echiuran worm, Urechis caupo, were reliably obtained by aseptically removing gametes directly from the gamete storage organs. Trochophore larvae only removed neutral amino acids from seawater as measured by high-performance liquid chromatography (HPLC). There was no detectable uptake, as measured by HPLC, of acidic or basic amino acids. Kinetic analysis showed that the transport system for alanine in 4-day-old larvae had a Kt of 4-6 µM and a Jmax of 9-10 pmol larva-1 h-1. Following a 50-min exposure, the majority of the radioactivity (95%) from 14C-alanine was found in the trichloroacetic acid-soluble fraction. Very little label appeared as acid-insoluble material, and there was no detectable lipid biosynthesis from 14C-alanine. Approximately 12% of the total alanine transported was released in the form of 14CO2. Thin-layer chromatography of intracellular free amino acid pools demonstrated that aspartic acid and glutamic acid were radiolabeled from the alanine precursor. A comparison of the energy acquired from the transport of alanine, with the metabolic rate of 4-day-old larvae, revealed that 51% of the metabolic demand could be provided by the transport and complete catabolism of this single amino acid at a concentration of 595 nM in seawater.


Characterization of forebrain neurons derived from late-onset Huntington's disease human embryonic stem cell lines.
Huntington's disease (HD) is an incurable neurodegenerative disorder caused by a CAG repeat expansion in exon 1 of the Huntingtin (HTT) gene. Recently, induced pluripotent stem cell (iPSC) lines carrying atypical and aggressive (CAG60+) HD variants have been generated and exhibit disparate molecular pathologies. Here we investigate two human embryonic stem cell (hESC) lines carrying CAG37 and CAG51 typical late-onset repeat expansions in comparison to wildtype control lines during undifferentiated states and throughout forebrain neuronal differentiation. Pluripotent HD lines demonstrate growth, viability, pluripotent gene expression, mitochondrial activity and forebrain specification that is indistinguishable from control lines. Expression profiles of crucial genes known to be dysregulated in HD remain unperturbed in the presence of mutant protein and throughout differentiation; however, elevated glutamate-evoked responses were observed in HD CAG51 neurons. These findings suggest typical late-onset HD mutations do not alter pluripotent parameters or the capacity to generate forebrain neurons, but that such progeny may recapitulate hallmarks observed in established HD model systems. Such HD models will help further our understanding of the cascade of pathological events leading to disease onset and progression, while simultaneously facilitating the identification of candidate HD therapeutics.


Coordination of transcription factors, NF-Y and C/EBP beta, in the regulation of the mdr1b promoter.
The expression of mdr genes that encode P-glycoprotein, an integral membrane drug transporter, has been associated with the emergence of the multidrug resistance phenotype during treatment with cancer chemotherapeutic drugs. To understand the regulation of the mdr genes, the murine mdr1b promoter has been isolated and characterized in our laboratory. Three nuclear protein binding sites that interact with nuclear proteins present in both drug-sensitive and -resistant murine macrophage-like 1774.2 cells have been localized to the promoter. In this report, transcription factor NF-Y has been identified as binding to the Y-box sequence in site 1 and as a major factor in the regulation of the murine mdr1b promoter in the mouse adrenal cell line, Y-1, that endogenously expresses the mdr1b gene. The expression of CCAAT/enhancer binding protein beta (C/EBP beta) in Y-1 cells augmented mdr1b promoter activity and resulted in an increased level of mdr1b mRNA. The effect of C/EBP beta expression on mdr1b promoter activity was sensitive to mutations in the Y-box, suggesting that coordination of NF-Y with C/EBP beta is required for further activation of the mdr1b promoter. Our studies have indicated that NF-Y is a critical factor for the mdr1b promoter, and its coordination with other factors, such as C/EBP beta, could be an important mechanism involved in mdr1b gene expression.


Modeling of in-use stability for tablets and powders in bottles.
A model is presented for determining the time when an active pharmaceutical ingredient in tablets/powders will remain within its specification limits during an in-use period; that is, when a heat-induction sealed bottle is opened for fixed time periods and where tablets are removed at fixed time points. This model combines the Accelerated Stability Assessment Program to determine the impact on degradation rates of relative humidity (RH) with calculations of the RH as a function of time for the dosage forms under in-use conditions. These calculations, in a conservative approach, assume that the air inside bottles with broached heat-induction seals completely exchanges with the external environment during periods when the bottle remains open. The solid dosages are assumed to sorb water at estimable rates during these openings. When bottles are capped, the moisture vapor transmission rate can be estimated to determine the changing RH inside the bottles between opening events. The impact of silica gel desiccants can also be included in the modeling.


Acid and non-acid reflux in patients refractory to proton pump inhibitor therapy: is gastroparesis a factor?
To determine whether an increased number and duration of non-acid reflux events as measured using the multichannel intraluminal impedance pH (MII-pH) is linked to gastroparesis (GP). A case control study was conducted in which 42 patients undergoing clinical evaluation for continued symptoms of gastroesophageal reflux disease (both typical and atypical symptoms) despite acid suppression therapy. MII-pH technology was used over 24 h to detect reflux episodes and record patients' symptoms. Parameters evaluated in patients with documented GP and controls without GP by scintigraphy included total, upright, and supine number of acid and non-acid reflux episodes (pH < 4 and pH > 4, respectively), the duration of acid and non-acid reflux in a 24-h period, and the number of reflux episodes lasting longer than 5 min. No statistical difference was seen between the patients with GP and controls with respect to the total number or duration of acid reflux events, total number and duration of non-acid reflux events or the duration of longest reflux episodes. The number of non-acid reflux episodes with a pH > 7 was higher in subjects with GP than in controls. In addition, acid reflux episodes were more prolonged (lasting longer than 5 min) in the GP patients than in controls; however, these values did not reach statistical significance. Thirty-five patients had recorded symptoms during the 24 h study and of the 35 subjects, only 9% (n = 3) had a positive symptom association probability (SAP) for acid/non-acid reflux and 91% had a negative SAP. The evaluation of patients with a documented history of GP did not show an association between GP and more frequent episodes of non-acid reflux based on MII-pH testing.


Humanization of Immu31, an alpha-fetoprotein-specific antibody.
Immu31 is a murine monoclonal antibody (Ab) specific for alpha-fetoprotein (AFP), a tumor-associated marker. The excellent tumor targeting ability of Immu31 has led to the development of a Immu31-based radioimmunodiagnostic agent, AFP-Scan, for hepatocellular carcinoma and other AFP-producing tumors. To enhance the capability of Immu31-based immunoconjugates being used in diagnostic and therapeutic procedures in humans, a humanized version of Immu31 (hImmu31) was constructed by grafting the complementarity determining regions (CDRs) of murine variable domains for the heavy (VH) and kappa (Vkappa) chain to the respective human VH and Vkappa framework regions (FRs). The cDNA encoding the VH and Vkappa of Immu31 was cloned by reverse transcription-PCR from hybridoma cells, and a chimeric Immu31 (cImmu31) composed of murine V and human C domains was constructed. Competitive ELISA assays showed identical AFP binding activity between the chimeric and murine Abs, confirming the authenticity of the cloned V genes. Based on sequence homology, the EU FR1, FR2, and FR3 and the NEWM FR4 were selected as the scaffold for grafting VH CDRs and REI FRs for Vkappa CDRs of Immu31. The amino acid residues in murine FRs that are considered to be in contact with the CDRs of the Ab were maintained in the humanized version. hImmu31, thus constructed and expressed, showed comparable immunoreactivity in a competitive binding ELISA assay to that of murine Immu31 and cImmu31. High-level production was achieved by expressing hImmu31 in a dhfr-based amplifiable system, and the productivity has exceeded 100 mg/liter in terminal cultures.


Committing to my mission: Faculty experiences with student veterans in baccalaureate nursing education.
Veterans transition to baccalaureate nursing education with significant experiences from their military service. Faculty play a role in facilitating this transition, in and out of the classroom. Approaches to enhance nursing education programs to better meet student veterans' needs have been proposed. However, less is known about the experiences of faculty teaching student veterans. A qualitative descriptive design was used to examine nursing faculty experiences teaching veterans, and in facilitating veterans' transfer of learning and knowledge from their military experience to baccalaureate nursing education. Semi-structured interviews with twelve faculty from the United States were conducted and data analyzed using content and thematic analysis. Three major themes included: committing to my mission, facilitating a shared understanding between military service and nursing, and appreciating a new duty. Results support faculty purposefully engaging in activities to increase their cultural sensitivity, while continuously developing humility and competence, to be successful teaching student veterans. Nursing faculty are positioned to leverage education as a means to improve the social status of veterans, ultimately improving their health and well-being, as well as academic success. It is recommended schools of nursing encourage and support faculty in attending professional development activities aimed at improving individual cultural sensitivity.


Funding and Service Organization to Achieve Universal Health Coverage for Medicines: An Economic Evaluation of the Best Investment and Service Organization for the Brazilian Scenario.
There are many health benefits since 31 years after the foundation of the National Health Service (NHS) in Brazil, especially the increase in life expectancy. However, family-income inequalities, insufficient funding, and suboptimal private sector-public sector collaboration are still areas for improvement. The efforts of Brazil to achieve universal health coverage (UHC) for medicines have resulted in increased public financing of medicines and their availability, reducing avoidable hospitalization and mortality. However, lack of access to medicines still remains. Due to historical reasons, pharmaceutical service organization in developing countries may have important differences from high-income countries. In some cases, developing countries finance and promote medicine access by using the public infrastructure of health care/medical units as dispensing sites and cover all costs of medicines dispensed. In contrast, many high-income countries use private community pharmacies and cover the costs of medicines dispensed plus a fee, which includes all logistic costs. In this study, we will undertake an economic evaluation to understand the funding needs of the Brazilian NHS to reduce inequalities in access to medicines through adopting a pharmaceutical service organization similar to that seen in many high-income countries with hiring/accrediting private pharmacies. We performed an economic evaluation of a model to provide access to medicines within public funds based on a decision tree model with two alternative scenarios public pharmacies (NHS, state-owned facilities) versus private pharmacies (NHS, agreements). The analysis assumed the perspective of the NHS. We identified the types of resources consumed, the amount, and costs in both scenarios. We also performed a budget impact forecast to estimate the incremental funding required to reduce inequalities in access to essential medicines in Brazil. The model without rebates for medicines estimated an incremental cost of US$3.1 billion in purchasing power parity (PPP) but with an increase in the average availability of medicines from 65% to 90% for citizens across the country irrespective of family income. This amount places the NHS in a very good position to negotiate extensive rebates without the need for external reference pricing for government purchases. Forecast scenarios above 35% rebates place the alternative of hiring private pharmacies as dominant. Higher rebate rates are feasible and may lead to savings of more than US$1.3 billion per year (30%). The impact of incremental funding is related to medicine access improvement of 25% in the second year when paying by dispensing fee. The estimate of the incremental budget in five years would be US$4.8 billion PPP. We have yet to explore the potential reduction in hospital and outpatient costs, as well as in lawsuits, with increased availability with the yearly expenses for these at US$9 billion and US$1.4 billion PPP respectively in 2017. The results of the economic evaluation demonstrate potential savings for the NHS and society. Achieving UHC for medicines reduces household expenses with health costs, health litigation, outpatient care, hospitalization, and mortality. An optimal private sector-public sector collaboration model with private community pharmacy accreditation is economically dominant with a feasible medicine price negotiation. The results show the potential to improve access to medicines by 25% for all income classes. This is most beneficial to the poorest families, whose medicines account for 76% of their total health expenses, with potential savings of lives and public resources.


Young HIV-infected children and their adult caregivers prefer tablets to syrup antiretroviral medications in Africa.
Provision of anti-retroviral therapy (ART) for HIV-infected children is complicated using syrup formulations, which are costlier than tablets, harder to transport and store and difficult for health-workers to prescribe and caregivers to administer. Dispersible/crushable tablets may be more appropriate. We studied the acceptability of syrups and scored tablets among young children who used both in the AntiRetroviral Research fOr Watoto (ARROW) trial. ARROW is an ongoing randomized trial of paediatric ART monitoring and treatment strategies in 1206 children in Uganda and Zimbabwe. 405 children initially received syrups of combination ART including Nevirapine, Zidovudine, Abacavir and Lamivudine before changing, when reaching the 12-<15 kg weightband, to scored adult-dose tablets prescribed according to WHO weightband tables. Caregiver expectations and experiences were collected in questionnaires at their last visit on syrups and after 8 and 24 weeks on tablets. Questionnaires were completed by caregivers of 267 children (median age 2.9 years (IQR 2.5, 3.4)). At last visit on syrups, 79% caregivers reported problems with syrups, mostly related to number, weight, transportation and conspicuousness of bottles. Difficulties taking tablets were expected by 127(48%) caregivers; however, after 8 and 24 weeks, only 26% and 18% reported their children had problems with tablets and no problems were reported with transportation/conspicuousness. Taste, swallowing or vomiting were reported as problems 'sometimes/often' for 14%, 9%, 22% children on syrups and 16%, 9%, 8% on tablets. At last visit on syrups, 74% caregivers expected to prefer tablets but only 27% thought their child would. After 8/24 weeks, 94%/97% caregivers preferred tablets and 57%/59% reported their child did. Most children at about 3 years can take tablets; caregivers and children themselves generally prefer tablets to liquid formulations of HIV medications above this age. Preferences of caregivers and children should be considered when designing and licensing paediatric drug formulations.


Precisely measured protein lifetimes in the mouse brain reveal differences across tissues and subcellular fractions.
The turnover of brain proteins is critical for organism survival, and its perturbations are linked to pathology. Nevertheless, protein lifetimes have been difficult to obtain in vivo. They are readily measured in vitro by feeding cells with isotopically labeled amino acids, followed by mass spectrometry analyses. In vivo proteins are generated from at least two sources: labeled amino acids from the diet, and non-labeled amino acids from the degradation of pre-existing proteins. This renders measurements difficult. Here we solved this problem rigorously with a workflow that combines mouse in vivo isotopic labeling, mass spectrometry, and mathematical modeling. We also established several independent approaches to test and validate the results. This enabled us to measure the accurate lifetimes of ~3500 brain proteins. The high precision of our data provided a large set of biologically significant observations, including pathway-, organelle-, organ-, or cell-specific effects, along with a comprehensive catalog of extremely long-lived proteins (ELLPs).


Glucose and lactate are equally effective in energizing activity-dependent synaptic vesicle turnover in purified cortical neurons.
This study examines the role of glucose and lactate as energy substrates to sustain synaptic vesicle cycling. Synaptic vesicle turnover was assessed in a quantitative manner by fluorescence microscopy in primary cultures of mouse cortical neurons. An electrode-equipped perfusion chamber was used to stimulate cells both by electrical field and potassium depolarization during image acquisition. An image analysis procedure was elaborated to select in an unbiased manner synaptic boutons loaded with the fluorescent dye N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl)pyridinium dibromide (FM1-43). Whereas a minority of the sites fully released their dye content following electrical stimulation, others needed subsequent K(+) depolarization to achieve full release. This functional heterogeneity was not significantly altered by the nature of metabolic substrates. Repetitive stimulation sequences of FM1-43 uptake and release were then performed in the absence of any metabolic substrate and showed that the number of active sites dramatically decreased after the first cycle of loading/unloading. The presence of 1 mM glucose or lactate was sufficient to sustain synaptic vesicle cycling under these conditions. Moreover, both substrates were equivalent for recovery of function after a phase of decreased metabolic substrate availability. Thus, lactate appears to be equivalent to glucose for sustaining synaptic vesicle turnover in cultured cortical neurons during activity.


GEOFFREY HARRIS PRIZE LECTURE 2018: Novel pathways regulating neuroendocrine function, energy homeostasis and metabolism in humans.
The discovery of leptin, an adipocyte-secreted hormone, set the stage for unraveling the mechanisms dictating energy homeostasis, revealing adipose tissue as an endocrine system that regulates appetite and body weight. Fluctuating leptin levels provide molecular signals to the brain regarding available energy reserves modulating energy homeostasis and neuroendocrine response in states of leptin deficiency and to a lesser extent in hyperleptinemic states. While leptin replacement therapy fails to provide substantial benefit in common obesity, it is an effective treatment for congenital leptin deficiency and states of acquired leptin deficiency such as lipodystrophy. Current evidence suggests that regulation of eating behavior in humans is not limited to homeostatic mechanisms and that the reward, attention, memory and emotion systems are involved, participating in a complex central nervous system network. It is critical to study these systems for the treatment of typical obesity. Although progress has been made, further studies are required to unravel the physiology, pathophysiology and neurobehavioral mechanisms underlying potential treatments for weight-related problems in humans.


Diagnostic and Management Pathways for Pulmonary Carcinoid Tumours in the United Kingdom: Results from the National Lung Neuroendocrine Tumour Pathway Project.
There is inconsistency among published guidelines for the optimal diagnostic and management pathways for patients with typical (TC) or atypical (AC) pulmonary carcinoid tumours. We conducted a UK-wide clinician survey to assess current practice for the diagnosis, management, and follow-up of patients with TC/AC and descriptively compared management between European Neuroendocrine Tumor Society (ENETS) accredited centres of excellence (CoE) and nonaccredited centres (non-CoE). Twenty-seven clinicians (10 CoE; 17 non-CoE) participated. Computed tomography of thorax, abdomen, and pelvis was the most commonly reported diagnostic tool (96% of respondents), and bone scans and gallium somatostatin receptor scintigraphy positron emission tomography (SRS PET) were the least commonly reported (30% and 37% of respondents, respectively). Adjuvant therapy is considered for resected TC/AC by <5% of respondents for patients with stage N0 M0 AC or TC, up to 48% of respondents for patients with AC with R1 disease. Somatostatin analogues were the most commonly reported first-line treatment (63% of respondents), and chemotherapy was the most commonly reported second-line therapy and third-line therapy (33% and 41%, respectively) for unresectable and metastatic disease. Reported frequency of initial follow-up after primary surgery ranged from every 2 months to annual, and total follow-up duration ranged from 2 years to indefinite depending on disease type (TC/AC) and stage. For most diagnostic investigations, the highest reported frequency of use was in CoE, most notably gallium SRS PET (70% CoE vs. 18% non-CoE respondents). 93% of respondents (100% CoE; 88% non-CoE) reported having neuroendocrine tumour- (NET-) specialist multidisciplinary team meetings at their centre; 59% (90% CoE; 41% non-CoE) had a NET Clinical Nurse Specialist (CNS) and 48% (80% CoE; 29% non-CoE) had a lung NET patient database. The survey results suggest variability between UK centres in diagnostic pathways and management of patients with TC/AC and suggest that CoE may be able to offer an improved service to patients.


Antidepressants for bipolar depression: a systematic review of randomized, controlled trials.
This study reviewed the evidence from randomized, controlled trials on the efficacy and safety of antidepressants in the short-term treatment of bipolar depression. The authors performed a systematic review and meta-analysis of randomized, controlled trials. They searched the Cochrane Collaboration Depression, Anxiety, and Neurosis Controlled Trials Register, incorporating results of searches of MEDLINE, EMBASE, CINAHL, PsycLIT, PSYNDEX, and LILACS. The main outcome measures were the proportion of patients who clinically responded to treatment and the rate of switching to mania. Twelve randomized trials were included, with a total of 1,088 randomly assigned patients. Five trials compared one or more antidepressants with placebo: 75% of these patients were receiving a concurrent mood stabilizer or an atypical antipsychotic. Antidepressants were more effective than placebo. Antidepressants did not induce more switching to mania (the event rate for antidepressants was 3.8% and for placebo, it was 4.7%). Six trials allowed comparison between two antidepressants. The rate of switching for tricyclic antidepressants was 10%, and for all other antidepressants combined, it was 3.2%. Antidepressants are effective in the short-term treatment of bipolar depression. The trial data do not suggest that switching is a common early complication of treatment with antidepressants. It may be prudent to use a selective serotonin reuptake inhibitor or a monoamine oxidase inhibitor rather than a tricyclic antidepressant as first-line treatment. Given the limited evidence, there is a compelling need for further studies with longer follow-up periods and careful definition and follow-up of emerging mania and partial remission.


Microbial diversity in an intensively managed landscape is structured by landscape connectivity.
Intensively managed land increases the rate of nutrient and particle transport within a basin, but the impact of these changes on microbial community assembly patterns at the basin scale is not yet understood. The objective of this study was to investigate how landscape connectivity and dispersal impacts microbial diversity in an agricultural-dominated watershed. We characterized soil, sediment and water microbial communities along the Upper Sangamon River basin in Illinois-a 3600 km2 watershed strongly influenced by human activity, especially landscape modification and extensive fertilization for agriculture. We employed statistical and network analyses to reveal the microbial community structure and interactions in the critical zone (water, soil and sediment media). Using a Bayesian source tracking approach, we predicted microbial community connectivity within and between the environments. We identified strong connectivity within environments (up to 85.4 ± 13.3% of sequences in downstream water samples sourced from upstream samples, and 44.7 ± 26.6% in soil and sediment samples), but negligible connectivity across environments, which indicates that microbial dispersal was successful within but not between environments. Species sorting based on sample media type and environmental parameters was the dominant driver of community dissimilarity. Finally, we constructed operational taxonomic unit association networks for each environment and identified a number of co-occurrence relationships that were shared between habitats, suggesting that these are likely to be ecologically significant.


Serum insulin-like growth factor-I is positively associated with serum prostate-specific antigen in middle-aged men without evidence of prostate cancer.
We have examined the relationship between serum insulin-like growth factor-I (IGF-I) and prostate-specific antigen in 367 healthy men without evidence of prostate cancer and found a positive association (P = 0.05). In men without prostate cancer, serum prostate-specific antigen is closely related to prostate size, and our findings, therefore, suggest that IGF-I may induce prostatic epithelial proliferation. Higher circulating levels of IGF-I have been associated with increased risk of both prostate cancer and possibly benign prostatic hyperplasia. Greater rates of cell proliferation induced by IGF-I may be a key biological pathway underlying these disorders.


The interface between the national tuberculosis control programme and district hospitals in Cameroon: missed opportunities for strengthening the local health system -a multiple case study.
Tuberculosis remains a major public health problem in sub-Saharan Africa. District hospitals (DHs) play a central role in district-based health systems, and their relation with vertical programmes is very important. Studies on the impact of vertical programmes on DHs are rare. This study aims to fill this gap. Its purpose is to analyse the interaction between the National Tuberculosis Control Programme (NTCP) and DHs in Cameroon, especially its effects on the human resources, routine health information system (HIS) and technical capacity at the hospital level. We used a multiple case study methodology. From the Adamaoua Region, we selected two DHs, one public and one faith-based. We collected qualitative and quantitative data through document reviews, semi-structured interviews with district and regional staff, and observations in the two DHs. The NTCP trained and supervised staff, designed and provided tuberculosis data collection and reporting tools, and provided anti-tuberculosis drugs, reagents and microscopes to DHs. However, these interventions were limited to the hospital units designated as Tuberculosis Diagnostic and Treatment Centres and to staff dedicated to tuberculosis control activities. The NTCP installed a parallel HIS that bypassed the District Health Services. The DH that performs well in terms of general hospital care and that is well managed was successful in tuberculosis control. Based on the available resources, the two hospitals adapt the organisation of tuberculosis control to their settings. The management teams in charge of the District Health Services are not involved in tuberculosis control. In our study, we identified several opportunities to strengthen the local health system that have been missed by the NTCP and the health system managers. Well-managed DHs perform better in terms of tuberculosis control than DHs that are not well managed. The analysis of the effects of the NTCP on the human resources, HIS and technical capacity of DHs indicates that the NTCP supports, rather than strengthens, the local health system. Moreover, there is potential for this support to be enhanced. Positive synergies between the NTCP and district health systems can be achieved if opportunities to strengthen the district health system are seized. The question remains, however, of why managers do not take advantage of the opportunities to strengthen the health system.


Antinociceptive effect of PnTx4(5-5), a peptide from Phoneutria nigriventer spider venom, in rat models and the involvement of glutamatergic system.
The venom of Phoneutria nigriventer spider is a source of numerous bioactive substances, including some toxins active in insects. An example is PnTx4(5-5) that shows a high insecticidal activity and no apparent toxicity to mice, although it inhibited NMDA-evoked currents in rat hippocampal neurons. In this work the analgesic activity of PnTx4(5-5) (renamed Γ-ctenitoxin-Pn1a) was investigated. The antinociceptive activity was evaluated using the paw pressure test in rats, after hyperalgesia induction with intraplantar injection of carrageenan or prostaglandin E2 (PGE2). PnTx4(5-5), subcutaneously injected, was able to reduce the hyperalgesia induced by PGE2 in rat paw, demonstrating a systemic effect. PnTx4(5-5) administered in the plantar surface of the paw caused a peripheral and dose-dependent antinociceptive effect on hyperalgesia induced by carrageenan or PGE2. The hyperalgesic effect observed in these two pain models was completely reversed with 5 µg of PnTx4(5-5). Intraplantar administration of L-glutamate induced hyperalgesic effect that was significantly reverted by 5 μg of PnTx4(5-5) injection in rat paw. The antinociceptive effect for PnTx4(5-5) was demonstrated against different rat pain models, i.e. induced by PGE2, carrageenan or glutamate. We suggest that the antinociceptive effect of PnTx4(5-5) may be related to an inhibitory activity on the glutamatergic system.


Dental insurance, dental care utilization, and perceived unmet dental needs in women living with HIV: Results from the Women's Interagency HIV Study.
Dental care is the most commonly cited unmet health-care service due to cost. Previous research has highlighted the unmet dental needs of people living with HIV (PLWH). Understanding associations among dental insurance availability, dental care utilization, and the presence of unmet dental needs among PLWH is a public health priority. Oral health surveys were collected cross-sectionally (April-October 2016) among 1,442 women living with HIV (WLWH) in the Women's Interagency HIV Study. Logistic regression models were used to analyze the association between having versus not having dental insurance by type (Ryan White, private, Medicaid/Medicare) and two primary outcomes: a) typical frequency of dental visits (at least annually, less than annually) and b) reporting an unmet dental need in the past 6 months. All dental insurance types were associated with higher odds of receiving annual dental care and, for those with either Medicare/Medicaid or private insurance, lower odds of having an unmet dental need. When WLWH were asked to describe their oral health, poor self-reported condition was associated with both an unmet dental need (odds ratio [OR]: 4.52, 95 percent Confidence Interval [CI] [3.29-6.20]) and lower odds of annual dental care utilization (OR: 0.44, 95 percent CI [0.34-0.57]). Self-reported depressive symptom burden was also linked to having an unmet dental need (OR: 2.10, 95 percent CI [1.46-3.01]). Dental insurance coverage increases dental care utilization and is associated with better oral health among WLWH. In the era of health-care reform, dental insurance coverage may be instrumental for enhancing treatment outcomes.


Cortical silent period following transcranial magnetic stimulation in epileptic patients.
Cortical silent period (SP) following transcranial magnetic stimulation is mainly due to cortical inhibitory mechanisms. SP may have a value for detecting inhibitory mechanisms in epileptic patients with or without treatment. The aim of this study was to evaluate the effect of both the epilepsy and the antiepileptic medication on these inhibitory mechanisms. The subgroups studied consisted of (a) normal subjects, (b) unmedicated epileptic patients, (c) epileptic patients with uncontrolled seizures under medication, (d) epileptic patients with controlled seizures under medication. SP following transcranial magnetic stimulation was measured in all subjects. The SP values from shortest to the longest were in the following order: 1) normal subjects; 2) epileptic patients with controlled seizures under medication; 3) unmedicated epileptic patients; 4) epileptic patients with uncontrolled seizures under medication. Our findings probably indicate the enhanced interictal inhibitory mechanisms in epilepsy which is resistant to antiepileptic medication.


Correlation of Virtual Reality Simulation and Dry Lab Robotic Technical Skills.
To examine whether a set of virtual reality (VR) surgical simulation drills have correlative validity when compared with the validated Robotic Objective Structured Assessment of Technical Skills (R-OSATS) dry lab drills. A prospective methods comparison study (Canadian Task Force classification II-2). A teaching hospital. Thirty current residents, fellows, and faculty from the Departments of Obstetrics and Gynecology, Urology, and General Surgery. Participants completed 5 VR drills on the da Vinci Skills Simulator and 5 dry lab drills. Participants were randomized to the order of completion. VR drills were scored automatically by the simulator. Dry lab drills were recorded, reviewed by 3 blinded experts, and scored using the R-OSATS assessment tool. Spearman correlation coefficients were calculated comparing simulator scores and R-OSATS scores for the same surgeon. The correlation for overall summary scores between VR and dry lab drills was strong (r = 0.83; p < .01). Each of the 5 VR drills was also found to have a statistically significant correlation to its corresponding dry lab drill, with correlation coefficients ranging from r = 0.49 to 0.73 (p < .01 for all). The performance on VR drills also confirmed construct validity. Faculty and fellows had consistently higher overall scores than residents (median VR scores: 458 for faculty, 425 for fellows, 339 for residents; p < .01). We selected a core set of VR drills that reliably correlate with validated dry lab R-OSATS drills. Because dry lab drills require significant time and effort on the part of the trainees and the evaluators, this set of VR drills could serve as an ancillary method of determining trainee competence.


[Gender inequalities in the impacts of informal elderly caregiving in Gipuzkoa: CUIDAR-SE study].
The provision of informal care determines the existence of health inequalities, as well as gender inequalities, because of women being the most frequent caregivers. The objectives of this study were to characterize the informal caregivers of elderly dependents in Gipuzkoa, to know the impact of caring on health and health related quality of life (HRQoL), and to explain the relationship between sex and consequences attributable to care. Cross-sectional study analyzing health (self-perceived health and GHQ-12) and HRQoL (EQ-5D-5L) data of 123 women and 103 men who took care of a dependent elderly and participated in the first wave of the CUIDAR-SE study in Gipuzkoa. A descriptive study was performed, and a logistic regression analysis was carried out to examine the association between sex and the consequences attributable to care. Women and men were over 60 years old, married, with primary education or less, and took care of their parents mainly. Women presented worse perceived health (46.3% vs 32.0%) and mental health (23.3% vs 13.9%), and more problems in the dimensions of anxiety/depression (35.0% vs 21.4%) and pain/discomfort (52.8% vs 31.1%) of EQ-5D-5L.The regression model showed greater risk of fatigue (OR = 2.83; 95% CI:1.53-5.24) and burden (OR = 1.87; 95% CI: 1.06-3.29) among women than among men. There are gender inequalities in the provision of informal care to dependent elderly people in Gipuzkoa, with women having a greater impact on their health and HRQoL than men. Considering the organization of care in this province, it will be necessary to design improvement actions more suited to the needs of caregivers.


Microbial Safety and Quality Evaluation of UV-Treated, Cold-Pressed Colored and Turbid Juices and Beverages.
The growing demand for fruit and vegetable juice blends, with improved nutritional and sensory attributes, has prompted the industrial adoption of nonthermal processing technologies, including UV light. Limited studies have explored conditions to overcome the well-known limitations of UV when treating liquid foods with a high content of particles that absorb or scatter UV light. This study addressed the effectiveness of the application of UV light, using a commercial processing unit, to inactivate pathogenic Escherichia coli O157:H7, Salmonella enterica (hereafter Salmonella), and Listeria monocytogenes, as well as spoilage microorganisms, in colored and turbid juices and beverages. The inactivation of cocktails of five strains (or serotypes) of E. coli O157:H7, Salmonella, and L. monocytogenes isolated from fruit- and vegetable-derived products linked to outbreaks was determined in seven colored and turbid cold-pressed juices and beverages. Juices and beverages were UV treated at a constant flow rate of 150 L/h through multiple consecutive passes. The inactivation of aerobic mesophilic bacteria, molds and yeasts, and lactic acid bacteria was also assessed at the cumulative dose that guaranteed a 5-log reduction of the most UV-tolerant pathogen for each product. A 5-log reduction of the three pathogens was achieved in all juices and beverages at a maximum cumulative UV dose of 12.0 ± 0.6 mJ/cm2. The dose required to ensure the targeted reduction varied depending on the tested product and the inoculated pathogen. The reduction of aerobic mesophiles, molds and yeasts, and lactic acid bacteria varied from 0.5 to 3.6, from 0.2 to 2.0, and from 0.5 to 3.6 log CFU/mL, respectively. Thus, the proposed treatment represents a suitable processing alternative to ensure the safety and extend the shelf life of colored and turbid cold-pressed juices and beverages.


Target size of 5'-nucleotidase in smooth muscle.
Target size of the 5'-nucleotidase in six different smooth muscles was determined by radiation inactivation. The enzyme in the soluble fraction of rat myometrium and vas deferens gave a target size of approximately 80,000 daltons. The plasma membrane bound 5'-nucleotidase however, gave target size of 80,000 to 110,000 daltons in rat gastric fundus and vas deferens and dog stomach and ileum, 135,000 daltons in rat mesenteric artery and 210,000 daltons in rat myometrium.


Sensory-motor index is useful parameter in electroneurographical diagnosis of carpal tunnel syndrome.
It was performed electroneurographic (ENG) studies with surface electrodes and examined nervus medianus (NM) in 60 patients (38 females), average age of 50,28 years (X+/-SD=50,28+/-11), with clinical diagnosis of carpal tunnel syndrome (CTS) and at least one border or discrete abnormal value of conventional electrophysiological tests. It was also examined 57 healthy individuals (33 females) as control group, average age of 45,65 years (X+/-SD=45,65+/-9,68). The sensitivity and specificity of sensory-motor index (SMI), terminal latency index (TLI) and residual latency (RL) were calculated and compared. SMI is determinate by using following formula: distal distance (DD) (in cm)/distal motor latency (DML) (in ms) + sensory conduction velocity (SCV) (in m/s)/motor conduction velocity (MCV) (in m/s) of NM. SCV of NM was measured by antidromic technique in segment wrist-index finger and MCV of NM in forearm segment above wrist. SMI mean value of control group was 3,45 (X+/-SD=3,45+/-0,45) with lower limit of normal value 2,82 and in patients with CTS 2,13 (X+/-SD=2,13 +/-0,37). The sensitivity of SMI in patients with CTS was 98,51%. SMI is useful parameter in electroneurographical diagnosis of CTS and it's determination is easy and fast and specially important in cases with border or discrete abnormal values of other NM electrophysiological parameters, when SMI values can indicate incipient phase of CTS evolution. In rare cases (about 1%) of CTS with selective NM motor axons affection, SMI may have normal value (false negative result), but DML is always prolonged in this cases. SMI is not dependent on age and DD values in patients with CTS and control subjects.


Medical students' attitudes to the physician's oath.
Students at the Ben-Gurion University medical school take the physician's oath at the beginning of their studies. Student attitudes towards the content, timing and relevance of the oath were examined before the ceremony, 3 months later and in the fourth and sixth years. Eight-seven percent of the students were positive about taking the oath, most commonly because the oath represented being part of a medical team that is bound by behavioural norms. Forty-three percent supported giving the oath legal force. There was a progressive decline between the first and final years in positive attitudes towards the oath. The timing most favoured for the oath was the beginning of clinical studies. Three months after the oath ceremony only 18% of the students were able to cite three obligations from the oath. Three percent of the students felt that the oath would affect their behaviour. In students, eyes the oath seems to be an emotionally important ritual, whose value probably transcends its actual content.


A Delphi yarn: applying Indigenous knowledges to enhance the cultural utility of SMART Recovery Australia.
Mutual support groups are a popular treatment for substance use and other addictive behaviours. However, little is known about the cultural utility of these programmes for Indigenous peoples. A three-round Delphi study, utilising Indigenous research yarning methods was conducted to: (1) Obtain expert opinion regarding the cultural utility of an Indigenous SMART Recovery handbook; (2) Gain consensus on areas within the SMART Recovery programme that require cultural modification and; (3) Seek advice on how modifications could be implemented in future programme design and delivery. The panellists were 11 culturally, geographically, and professionally diverse Indigenous Australian health and wellbeing experts. A group consensus level of 80% was set prior to each survey round. There was 100% participant retention across all three Delphi rounds. The panel reached consensus on five key programme modifications (composition of a separate facilitator and group member handbook; culturally appropriate language, terminology, and literacy level; culturally meaningful programme activities; supplementary storytelling resources; and customisation for diverse community contexts). The panel also developed a series of practical implementation strategies to guide SMART Recovery through a modification process. The findings highlight the importance of involving Indigenous peoples in the design, delivery and validation of mainstream mutual support programmes. Indigenous-led programme modifications could help improve accessibility and usefulness of mutual support groups for Indigenous peoples worldwide. This study is an example of how Indigenous research methods can be used alongside the Delphi technique. This approach demonstrated a way that Indigenous peoples from culturally and geographically diverse locations can participate in research anonymously, autonomously and without added burden on personal, community or professional obligations.


Early mammalian development: from basic research to the clinic.
Preimplantation embryonic development lays the foundations for the future individual. Fertilization, cleavage, differentiation of the first embryonic cell lineages and implantation of the embryo into the maternal uterus are absolutely critical for proper embryogenesis. Solving unanswered questions as well as creating new ideas and theories constitute the main axis of the basic research, which is driven by the curiosity of scientists and their desire to explore the unknown. We researchers have been exploring the development of mammalian embryos for decades, searching for the answer to the most fundamental question in the whole area of biology: how a complex organism derives from a single totipotent cell, a zygote. Due to obvious ethical concerns, animals, such as mice and, currently more and more often, cattle, pigs and rabbits, have become useful models for studying human embryonic development. Unprecedented advancement in cell and molecular biology techniques witnessed in the last years allows us to deepen our understanding of mammalian embryonic development.


The Burden of Depressive and Bipolar Disorders in Celiac Disease.
to measure the association between Celiac Disease (CD) and affective disorders, particularly Bipolar Disorder (BD), since it has not been studied yet, and to measure how much the quality of life (QoL) of a person with CD is affected by comorbidity with these disorders. Case-control study. 60 consecutive patients with CD. 240 subjects without CD, randomly selected after sex- and age-matching from a database of an epidemiological study. Psychiatric diagnoses according to DSM-IV carried out by physicians using structured interview tools (ANTAS-SCID). QoL was measured by means of SF-12. The lifetime prevalence of Major Depressive Disorder (MDD) was higher in CD than in controls (30.0% vs 8.3%, P<0.0001) as well as Panic Disorder (PD) (18.3% vs 5.4%, P<0.001) and BD (4.3% vs 0.4%, P<0.005). Patients with CD show a lower mean score than controls on SF12 (35.8±5.7 vs. 38.2±6.4; p=0.010), but those without comorbidity with MDD, PD and BD do not. The attributable burden of CD in worsening QoL - when comorbid with these disorders - was found comparable to that of serious chronic diseases like Wilson's Disease, and lower than Multiple Sclerosis only. MDD, PD and BD are strictly associated with CD. The comorbidity with these disorders is the key determinant of impaired quality of life in CD. Thus a preventive action on mood and anxiety disorders in patients suffering from CD is required. Moreover a screening for CD in people with affective disorders and showing key symptoms or family history of CD is recommended.


Monoclonal antibodies to adhesive cell coat glycoproteins secreted by zoospores of the green alga enteromorpha
Zoospores of Enteromorpha compressa (L.) Grev. secrete an adhesive cell coat which is involved in their attachment to various substrata. Two monoclonal antibodies (mAbs), designated Ent 1 and Ent 6, were raised against settled zoospores displaying secreted adhesive. Both antibodies labelled specifically the anterior region of the cell containing putative adhesive vesicles. During settlement the antigens recognised by both mAbs were secreted but whereas Ent 6 recognised a fibrillar material released within a few minutes of settlement, Ent 1 recognised components which were associated predominantly with the developing cell wall at later time points. Both mAbs also labelled a Golgi-rich region of settled spores, suggesting that these antigens are also synthesised after settlement. Both mAbs labelled the cell walls of vegetative tissue. Competitive enzyme-linked immunosorbent assay indicated that the two antibodies recognise separate, but overlapping epitopes. In spore settlement assays the Ent 6 immunoglobulin strongly reduced initial adhesion at low concentration whereas the inhibitory effects of Ent 1 occurred at later time points. On analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, (SDS-PAGE) both MAbs recognised a major buffer- and SDS-soluble, polydisperse 110-kDa antigen. The 110-kDa component was present in extracts of zoospores and sporulating tissue, but absent, in soluble form, from vegetative tissue. Deglycosylation of zoospore extract with anhydrous HF and peptide N-glycosidase digestion, showed that the major 110-kDa antigen is an N-linked glycan, and that the epitope is borne by the protein component. Time-course experiments showed that the Ent 6 antigen became progressively insoluble after zoospore attachment. Taken together, the data indicate that the two antibodies recognise separate but closely related antigens which have distinctive roles in adhesion and cell wall development.


Coarse-grain simulations of the R-SNARE fusion protein in its membrane environment detect long-lived conformational sub-states.
Coarse-grain molecular dynamics are used to look at conformational and dynamic aspects of an R-SNARE peptide inserted in a lipid bilayer. This approach allows carrying out microsecond-scale simulations which bring to light long-lived conformational sub-states potentially interesting in the context of the membrane fusion mechanism mediated by the SNARE proteins. We show that these coarse-grain models are in agreement with most experimental data on the SNARE system, but differ in some details that may have a functional interest, most notably in the orientation of the soluble part of R-SNARE that does not appear to be spontaneously accessible for SNARE complex formation. We also compare rat and yeast sequences of R-SNARE and find some minor differences in their behavior.


Identification of DNA sequences recognized by VirF, the transcriptional activator of the Yersinia yop regulon.
Pathogenic bacteria of the genus Yersinia harbor a 70-kb plasmid required for virulence. The plasmid-encoded virulence proteins of yersiniae are positively regulated at the transcriptional level by the product of the virF gene, the key activator of the system. virF encodes a DNA-binding protein related to the AraC family of transcriptional activators. The VirF protein from Yersinia enterocolitica is a 30-kDa protein that forms dimers in vitro and that specifically binds to the promoter region of VirF-regulated genes. In this work, we determined the sequences of eight VirF-binding sites from four different genes, by DNase I or hydroxyl radical footprinting. The protected regions, about 40 bases long, were aligned, and a number of conserved residues were identified. A 13-bp sequence resembling TTTTaGYcTtTat (in which nucleotides conserved in > or = 60% of the sequences are in uppercase letters and y indicates C or T) appeared, either isolated or as an inverted repeat in each of the eight sites.


The effects of breed and level of nutrition on whole-body and muscle protein metabolism in pure-bred Aberdeen angus and Charolais beef steers.
Eighteen pure-bred steers (live weight 350 kg) from each of two breeds, Aberdeen Angus (AA) and Charolais (CH), were split into three equal groups (six animals each) and offered three planes of nutrition during a 20-week period. The same ration formulation was offered to all animals with amounts adjusted at 3-week intervals to give predicted average weight gains of either 1.0 kg/d (M/M group) or 1.4 kg/d (H/H group). The remaining group (M/H) were offered the same amount of ration as the M/M group until 10 weeks before slaughter when the ration was increased to H. Data on animal performance, carcass characteristics and fibre-type composition in skeletal muscle are presented elsewhere (Maltin et al. 2000; Sinclair et al. 2000). On three occasions (17, 10 and 2 weeks before slaughter) the animals were transferred to metabolism stalls for 1 week, during which total urine collection for quantification of Ntau-methylhistidine (Ntau-MeH) elimination was performed for 4 d. On the last day, animals were infused for 11 h with [2H5]phenylalanine with frequent blood sampling (to allow determination of whole-body phenylalanine flux) followed by biopsies from m. longissimus lumborum and m. vastus lateralis to determine the fractional synthesis rate of mixed muscle protein. For both breeds, the absolute amount of Ntau-MeH eliminated increased with animal age or weight (P < 0.001) and was significantly greater for CH steers, at all intake comparisons, than for AA (P < 0.001). Estimates of fractional muscle breakdown rate (FBR; calculated from Ntau-MeH elimination and based on skeletal muscle as a fixed fraction of live weight) showed an age (or weight) decline for M/M and H/H groups of both breeds (P < 0.001). FBR was greater for the H/H group (P = 0.044). The M/H group also showed a lower FBR for the first two measurement periods (both at M intake) but increased when intake was raised to H. When allowance was made for differences in lean content (calculated from fat scores and eye muscle area in carcasses at the end of period 3), there were significant differences in muscle FBR with intake (P = 0.012) but not between breed. Whole-body protein flux (WBPF; g/d) based on plasma phenylalanine kinetics increased with age or weight (P < 0.001) and was similar between breeds. The WBPF was lower for M/M compared with H/H (P < 0.001) based on either total or per kg live weight0.75. Muscle protein fractional synthesis rate (FSR) declined with age for both breeds and tended to be higher at H/H compared with M intakes (intake x period effects, P < 0.05). Changing intake from M to H caused a significant increase (P < 0.001) in FSR. The FSR values for AA were significantly greater than for CH at comparable ages (P = 0. 044). Although FSR and FBR responded to nutrition, these changes in protein metabolism were not reflected in differences in meat eating quality (Sinclair et al. 2000).


Readmission after lung cancer resection is associated with a 6-fold increase in 90-day postoperative mortality.
Postoperative readmission affects patient care and healthcare costs. There is a paucity of nationwide data describing the clinical significance of readmission after thoracic operations. The purpose of this study was to evaluate the relationship between postoperative readmission and mortality after lung cancer resection. Data were extracted for patients undergoing lung cancer resection from the linked Surveillance Epidemiology and End Results-Medicare registry (2006-2011), including demographics, comorbidities, socioeconomic factors, readmission within 30 days from discharge, and 90-day mortality. Readmitting facility and diagnoses were identified. A hierarchical regression model clustered at the hospital level identified predictors of readmission. We identified 11,432 patients undergoing lung cancer resection discharged alive from 677 hospitals. The median age was 74.5 years, and 52% of patients received an open lobectomy. Thirty-day readmission rate was 12.8%, and 28.3% of readmissions were to facilities that did not perform the original operation. Readmission was associated with a 6-fold increase in 90-day mortality (14.4% vs 2.5%, P<.001). The most common readmitting diagnoses were respiratory insufficiency, pneumonia, pneumothorax, and cardiac complications. Patient factors associated with readmission included resection type; age; prior induction chemoradiation; preoperative comorbidities, including congestive heart failure and chronic obstructive pulmonary disease; and low regional population density. Factors associated with early readmission after lung cancer resection include patient comorbidities, type of operation, and socioeconomic factors. Metrics that only report readmissions to the operative provider miss one-fourth of all cases. Readmitted patients have an increased risk of death and demand maximum attention and optimal care.


Rare variants in SOX17 are associated with pulmonary arterial hypertension with congenital heart disease.
Pulmonary arterial hypertension (PAH) is a rare disease characterized by distinctive changes in pulmonary arterioles that lead to progressive pulmonary arterial pressures, right-sided heart failure, and a high mortality rate. Up to 30% of adult and 75% of pediatric PAH cases are associated with congenital heart disease (PAH-CHD), and the underlying etiology is largely unknown. There are no known major risk genes for PAH-CHD. To identify novel genetic causes of PAH-CHD, we performed whole exome sequencing in 256 PAH-CHD patients. We performed a case-control gene-based association test of rare deleterious variants using 7509 gnomAD whole genome sequencing population controls. We then screened a separate cohort of 413 idiopathic and familial PAH patients without CHD for rare deleterious variants in the top association gene. We identified SOX17 as a novel candidate risk gene (p = 5.5e-7). SOX17 is highly constrained and encodes a transcription factor involved in Wnt/β-catenin and Notch signaling during development. We estimate that rare deleterious variants contribute to approximately 3.2% of PAH-CHD cases. The coding variants identified include likely gene-disrupting (LGD) and deleterious missense, with most of the missense variants occurring in a highly conserved HMG-box protein domain. We further observed an enrichment of rare deleterious variants in putative targets of SOX17, many of which are highly expressed in developing heart and pulmonary vasculature. In the cohort of PAH without CHD, rare deleterious variants of SOX17 were observed in 0.7% of cases. These data strongly implicate SOX17 as a new risk gene contributing to PAH-CHD as well as idiopathic/familial PAH. Replication in other PAH cohorts and further characterization of the clinical phenotype will be important to confirm the precise role of SOX17 and better estimate the contribution of genes regulated by SOX17.


Canine helper-dependent vectors production: implications of Cre activity and co-infection on adenovirus propagation.
The importance of Cre recombinase to minimize helper vector (HV) contamination during helper-dependent adenovirus vectors (HDVs) production is well documented. However, Cre recombinase, by inducing DNA double-strand breaks (DSBs), can cause a reduced proliferation and genotoxic effects in cultured cells. In this work, Cre-expressing cell stability, co-infection and their relation to adenovirus amplification/HV contamination were evaluated to develop a production protocol for HD canine adenovirus type 2 (CAV-2) vectors. Long-term Cre expression reduced the capacity of MDCK-E1-Cre cells to produce CAV-2 by 7-fold, although cell growth was maintained. High HDV/HV MOI ratio (5:0.1) led to low HV contamination without compromising HDV yields. Indeed, such MOI ratio was sufficient to reduce HV levels, as these were similar either in MDCK-E1 or MDCK-E1-Cre cells. This raises the possibility of producing HDVs without Cre-expressing cells, which would circumvent the negative effects that this recombinase holds to the production system. Here, we show how Cre and MOI ratio impact adenovirus vectors yields and infectivity, providing key-information to design an improved manufacturing of HDV. Potential mechanisms to explain how Cre is specifically impacting cell productivity without critically compromising its growth are presented.


Direct Evidence for Prediction Signals in Frontal Cortex Independent of Prediction Error.
Predictive coding (PC) has been suggested as one of the main mechanisms used by brains to interact with complex environments. PC theories posit top-down prediction signals, which are compared with actual outcomes, yielding in turn prediction error (PE) signals, which are used, bottom-up, to modify the ensuing predictions. However, disentangling prediction from PE signals has been challenging. Critically, while many studies found indirect evidence for PC in the form of PE signals, direct evidence for the prediction signal is mostly lacking. Here, we provide clear evidence, obtained from intracranial cortical recordings in human surgical patients, that the human lateral prefrontal cortex evinces prediction signals while anticipating an event. Patients listened to task-irrelevant sequences of repetitive tones including infrequent predictable or unpredictable pitch deviants. The broadband high-frequency amplitude (HFA) was decreased prior to the onset of expected relative to unexpected deviants in the frontal cortex only, and its amplitude was sensitive to the increasing likelihood of deviants following longer trains of standards in the unpredictable condition. Single-trial HFA predicted deviations and correlated with poststimulus response to deviations. These results provide direct evidence for frontal cortex prediction signals independent of PE signals.


Mucoid degeneration of the anterior cruciate ligament: Management and outcome.
Mucoid degeneration (MD) is a rare pathological affection of the anterior cruciate ligament (ACL). Mucinous material within the substance of ACL produces pain and limited motion in the knee. This series describes the clinicoradiological presentation of patients with mucoid ACL, partial arthroscopic debridement of ACL and outcomes. During a period of 3 years, 11 patients were included based upon the clinical suspicion, magnetic resonance imaging (MRI) findings, arthroscopic features and histopathologic confirmation of MD of ACL. Six patients were male and five were female with median age of 40 years (range 21-59 years). All patients complained of knee pain with median duration of 5 months (range 1-24 months). All patients had painful deep flexion with 63.6% (N = 7) reporting trivial trauma before the onset of symptoms. MRI revealed MD of ACL in all with associated cyst in three patients. Partial debridement of ACL was done in ten and complete in one patient. None of them required notchplasty. Histopathology confirmed the diagnosis in all of them. At the mean followup of 13.81 months (range 6-28 months), all patients regained complete flexion and none complained of instability. Prior knowledge of condition with high index of suspicion and careful interpretation of MRI can establish the diagnosis preoperatively. It responds well to partial debridement of ACL and mucinous material without development of instability.


Evaluation of Nigerian traditional medicine: effects of Gakani, a herbal anti-asthmatic drug.
The anti-asthmatic potential of Gakani, a popular herbal drug in Nigeria was investigated. The LD50 values of the freeze-dried aqueous extract in mice and rats were 20.9 +/- 2.4 mg/kg and 18.6 +/- 4 mg/kg, respectively. The extract unsurmountably blocked the effects of histamine and isoprenaline on the guinea pig tracheaL chain. It produced initial dose-related contractions of the isolated guinea pig ileum and rat stomach strip, which was followed by persistent autoinhibition and inhibition of histamine- and 5-hydroxytryptamine (5-HT)-induced responses of the two preparations, respectively. The extract had good anti-inflammatory effect in rats, causing a dose-related inhibition of the increase in the paw circumference (acute inflammation) induced by subplantar injection of fresh egg albumin. These results highlight the anti-asthmatic and toxic potential of this preparation and the need for a systemic approach in the study of traditional medicines.


Connective tissue proteins and phagocytic cell function. Laminin enhances complement and Fc-mediated phagocytosis by cultured human macrophages.
Brief exposure of culture-derived human macrophages to laminin, a glycoprotein component of all mammalian basement membranes that has a molecular weight of 1,000,000, led to enhancement of subsequent macrophage phagocytosis of EAC4b, EAC3bi, and EAIgG (sheep erythrocytes sensitized with IgG anti-Forssman antibody). This effect on macrophage phagocytosis occurred with both substrate-adherent and fluid phase laminin. Preincubation of macrophages, but not of EAC4b, with laminin led to augmentation of phagocytosis, suggesting that interaction with the phagocytic cell, but not with the opsonized particle, was required for laminin's effect. Laminin-stimulated phagocytosis of EAC4b was blocked entirely by a monoclonal antibody to CR1. Direct comparison of the phagocytic ability of macrophages adherent to laminin- and fibronectin-coated glass slides showed that fibronectin had a somewhat greater enhancing effect on phagocytosis. Nonetheless, the phagocytosis-enhancing effect of laminin was not due to contamination of the purified laminin preparation by fibronectin, since the laminin preparation was free of fibronectin, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and enzyme-linked immunosorbent assay; in addition, laminin-enhanced phagocytosis was decreased in the presence of laminin-specific antibodies. Laminin inhibited macrophage adherence and spreading, but selection of a laminin-binding macrophage subpopulation could not account for the laminin-induced increases in phagocytosis. We hypothesize that interaction with extracellular matrix proteins may represent an important activation stimulus both to the macrophages normally present in the extravascular compartment and to the phagocytic cells that have emigrated from the blood-stream into areas of inflammation.


Diversity and oceanic distribution of prasinophytes clade VII, the dominant group of green algae in oceanic waters.
Prasinophytes clade VII is a group of pico/nano-planktonic green algae (division Chlorophyta) for which numerous ribosomal RNA (rRNA) sequences have been retrieved from the marine environment in the last 15 years. A large number of strains have also been isolated but have not yet received a formal taxonomic description. A phylogenetic analysis of available strains using both the nuclear 18S and plastidial 16S rRNA genes demonstrates that this group composes at least 10 different clades: A1-A7 and B1-B3. Analysis of sequences from the variable V9 region of the 18S rRNA gene collected during the Tara Oceans expedition and in the frame of the Ocean Sampling Day consortium reveal that clade VII is the dominant Chlorophyta group in oceanic waters, replacing Mamiellophyceae, which have this role in coastal waters. At some location, prasinophytes clade VII can even be the dominant photosynthetic eukaryote representing up to 80% of photosynthetic metabarcodes overall. B1 and A4 are the overall dominant clades and different clades seem to occupy distinct niches, for example, A6 is dominant in surface Mediterranean Sea waters, whereas A4 extend to high temperate latitudes. Our work demonstrates that prasinophytes clade VII constitute a highly diversified group, which is a key component of phytoplankton in open oceanic waters but has been neglected in the conceptualization of marine microbial diversity and carbon cycle.


Decrease of aquaporin-4 and excitatory amino acid transporter-2 indicate astrocyte dysfunction for pathogenesis of cortical degeneration in HIV-associated neurocognitive disorders.
Human immunodeficiency virus (HIV) encephalitis and degeneration of cerebral cortex are established histopathologies of HIV-associated neurocognitive disorders (HAND). We previously reported decreased excitatory amino acid transporter-2 (EAAT-2) and astrocytic apoptosis in cortical degeneration using SIVmac239 and simian-human immunodeficiency virus (SHIV)-infected macaques and human AIDS autopsy cases. In the present study, we added highly pathogenic SIVsm543-3-infected macaques. These animals showed similar degenerative changes in the frontal cortex. Using 11 SIV-infected macaques, three SIVsm543-3, five SIVmac239 and three SHIV, we compared brain pathology caused by three different viruses and further analyzed the pathogenic process of HAND. We noticed vacuolar changes in perivascular processes of astrocytes by electron microscopy, and examined expression of astrocyte-specific protein aquaporin-4 (AQP4) by immunohistochemistry. APQ4 was diffusely positive in the neuropil and perivascular area in control brains. There was patchy or diffuse decrease of AQP4 staining in the neuropil of SIV-infected macaques, which was associated with EAAT-2 staining by double immunostaining. A quantitative analysis demonstrated significant positive correlation between areas of AQP4 and EAAT-2. Some astrocytes express EAAT-2 but not AQP4, and decrease of EAAT-2 expression tended to be less than the decrease of AQP4. Active-caspase-3 immunostaining demonstrated apoptosis of neurons and astrocytes in the area of AQP4/EAAT-2 reduction. These results suggest that AQP4 is damaged first and decrease of EAAT-2 may follow in pathogenesis of cortical degeneration. This is the first demonstration of decrease of AQP4 and its association with EAAT-2 decrease in AIDS brain, suggesting a role in the pathogenesis of HAND.


Diffuse retinal nerve fiber layer defects identification and quantification in thickness maps.
To report retinal nerve fiber layer (RNFL) defect identification and quantification in RNFL thickness maps according to the structural RNFL loss, and to evaluate diffuse RNFL defects. A total of 170 patients with glaucoma and 186 normal subjects were consecutively enrolled. We defined RNFL defects in an RNFL thickness map by the degree of RNFL loss. The reference level for RNFL defect determination was set as a 20% to 70% degree of RNFL loss with a 1% interval. To identify RNFL defects, each individual RNFL thickness map was compared to the normative database map by using MATLAB software, and the region below the reference level was detected. The area, volume, location, and angular width of each RNFL defect were measured. Diffuse RNFL defects were defined as having an angular width > 30°. The optimal reference level for glaucomatous RNFL defects identification was 42% loss of RNFL. Retinal nerve fiber layer defects were identified in all (100%) of the 170 glaucoma patients and false-positive RNFL defects were detected in 16 (8.16%) cases among the 186 normal subjects. In all, 64.1% of glaucoma patients had diffuse RNFL defects, and 47.7% of diffuse RNFL defects were associated with mild glaucoma patients. The volume of diffuse RNFL defects was significantly associated with the severity of glaucomatous damage (P = 0.009). Diffuse RNFL defects were located closer to the center of the optic disc than localized RNFL defects (P < 0.001). Retinal nerve fiber layer thickness map analysis is an effective method for analyzing RNFL defects. Quantitative measurements (area, volume, location, and width) were useful to understanding diffuse RNFL defects.


Next generation probiotics in disease amelioration.
Studies on the role of gut commensal bacteria in health development have rapidly attracted much more attention beyond the classical pathogens over the last decade. Many important reports have highlighted the changes in the gut microbiota (dysbiosis) are closely related to development of intra- and extra-intestinal, chronic inflammation related diseases such as colitis, obesity/metabolic syndromes, diabetes mellitus, liver diseases, cardiovascular diseases and also cancer and neurodegenerative diseases. To circumvent these difficulties, the strategy of modulating the structure of the gut microbiota has been under intensive study and shed more light on amelioration of these inflammation related diseases. While traditional probiotics generally show marginal ameliorative effects, emerging next generation probiotics start to reveal as new preventive and therapeutic tools. Recent studies have unraveled many potential next generation probiotics (NGP). These include Prevotella copri and Christensenella minuta that control insulin resistance, Parabacteroides goldsteinii, Akkermansia muciniphila and Bacteroides thetaiotaomicron that reverse obesity and insulin resistance, Faecalibacterium prausnitzii that protects mice against intestinal diseases, and Bacteroides fragilis that reduces inflammation and shows anticancer effect. New agents will soon be revealed for targeted therapy on specific inflammation related diseases. The important roles of next generation probiotics and gut microbiota normobiosis on the maintenance of intestinal integrity and homeostasis are emphasized.


Observing the nonequilibrium dynamics of the quantum transverse-field Ising chain in circuit QED.
We show how a quantum Ising spin chain in a time-dependent transverse magnetic field can be simulated and experimentally probed in the framework of circuit QED with current technology. The proposed setup provides a new platform for observing the nonequilibrium dynamics of interacting many-body systems. We calculate its spectrum to offer a guideline for its initial experimental characterization. We demonstrate that quench dynamics and the propagation of localized excitations can be observed with the proposed setup and discuss further possible applications and modifications of this circuit QED quantum simulator.


Isolation, Development, and Genomic Analysis of Bacillus megaterium SR7 for Growth and Metabolite Production Under Supercritical Carbon Dioxide.
Supercritical carbon dioxide (scCO2) is an attractive substitute for conventional organic solvents due to its unique transport and thermodynamic properties, its renewability and labile nature, and its high solubility for compounds such as alcohols, ketones, and aldehydes. However, biological systems that use scCO2 are mainly limited to in vitro processes due to its strong inhibition of cell viability and growth. To solve this problem, we used a bioprospecting approach to isolate a microbial strain with the natural ability to grow while exposed to scCO2. Enrichment culture and serial passaging of deep subsurface fluids from the McElmo Dome scCO2 reservoir in aqueous media under scCO2 headspace enabled the isolation of spore-forming strain Bacillus megaterium SR7. Sequencing and analysis of the complete 5.51 Mbp genome and physiological characterization revealed the capacity for facultative anaerobic metabolism, including fermentative growth on a diverse range of organic substrates. Supplementation of growth medium with L-alanine for chemical induction of spore germination significantly improved growth frequencies and biomass accumulation under scCO2 headspace. Detection of endogenous fermentative compounds in cultures grown under scCO2 represents the first observation of bioproduct generation and accumulation under this condition. Culturing development and metabolic characterization of B. megaterium SR7 represent initial advancements in the effort toward enabling exploitation of scCO2 as a sustainable solvent for in vivo bioprocessing.


Prevalence and correlates of transactional sex among an urban emergency department sample: Exploring substance use and HIV risk.
Men and women involved in transactional sex (TS) report increased rates of HIV risk behaviors and substance use problems as compared with the general population. When people engaged in TS seek health care, they may be more likely to utilize the emergency department (ED) rather than primary care services. Our goal was to examine the prevalence and correlates of TS involvement among an ED sample of men and women. Adults ages 18-60 were recruited from an urban ED, as part of a larger randomized control trial. Participants (n = 4,575; 3,045 women, 1,530 men) self-administered a screening survey that assessed past 3-month substance use (including alcohol, marijuana, illicit drugs, and prescription drugs) and HIV risk behaviors, including TS (i.e., being paid in exchange of a sexual behavior), inconsistent condom use, multiple partners, and anal sex. Of the sample, 13.3% (n = 610) reported TS within the past 3 months (64.4% were female). Bivariate analysis showed TS was significantly positively associated with alcohol use severity, marijuana use, and both illicit and prescription drug use, and multiple HIV risk behaviors. These variables (except marijuana) remained significantly positively associated with TS in a binary logistic regression analysis. The prevalence of recent TS involvement among both male and female ED patients is substantial. These individuals were more likely to report higher levels of alcohol/drug use and HIV risk behaviors. The ED may be a prime location to engage both men and women who are involved in TS in behavioral interventions for substance use and sexual risk reduction.


Polyamines inhibit myosin phosphatase and increase LC20 phosphorylation and force in smooth muscle.
The increase in Ca(2+)-activated force caused by polyamines in beta-escin-permeabilized guinda pig ileum is shown to be associated with increased myosin 20-kDa light chain (LC20) phosphorylation and shortening velocity. Myosin LC20 dephosphorylation with arrested kinase activity was slower in the presence of 1 mM spermine. Smooth muscle phosphatases (SMP-I, -II, -III, and -IV) isolated from turkey gizzard are all active against phosphorylated LC20, but only SMP-III and -IV dephosphorylate heavy meromyosin (HMM). Spermine inhibited SMP-III activity toward LC20 but stimulated HMM dephosphorylation, whereas SMP-IV was inhibited with both substrates. In contrast, SMP-I and -II were stimulated by spermine. The relative effects of different polyamines correlated with an increasing number of positive charges. Spermine did not affect binding of SMP-IV to myosin and did not dissociate any of the subunits of the enzyme. Incubation of permeabilized strips with SMP-IV resulted in attenuated responses to Ca2+, an effect that was opposed by spermine and abolished by microcystin-LR. We conclude that spermine selectively inhibits myosin phosphatase activity and suggest that polyamines function as endogenous myosin phosphatase inhibitors.