A voltage-dependent gap junction in Drosophila melanogaster.
Steady-state and kinetic analyses of gap junctional conductance, gi, in salivary glands of Drosophila melanogaster third instar larvae reveal a strong and complex voltage dependence that can be elicited by two types of voltages. Voltages applied between the cells, i.e., transjunctional voltages, Vj, and those applied between the cytoplasm and the extracellular space, inside-outside voltages, Vi,o, markedly alter gj. Alteration of Vi-o while holding Vj = O,i.e., by equal displacement of the voltages in the cells, causes gj to increase to a maximum on hyperpolarization and to decrease to near zero on depolarization. These conductance changes associated with Vi-o are fit by a model in which there are two independent gates in series, one in each series, one in each membrane, where each gate is equally sensitive to Vi-o and exhibits first order kinetics. Vj's generated by applying voltage steps of either polarity to either cell, substantially reduce gj. These conductance changes exhibit complex kinetics that depend on Vi-o as well as Vj. At more positive Vi-o's, the changes in gj have two phases, an early phase consisting of of a decrease in gj for either polarity of Vj and a later phase consisting of an increase in gj on hyperpolarizing either cell and a decrease on depolarizing either cell. At negative Vi-o's in the plateau region of the gj-Vi-o relation, the later slow increase in gj is absent on hyperpolarizing either cell. Also, the early decrease in gj for either polarity of Vj is faster the more positive the Vi-o. The complex time course elicited by applying voltage steps to one cell can be explained as combined actions of Vi-o and Vj, with the early phase ascribable to Vj, but influenced by Vi-o, and the later phase to the changes in Vi-o associated with the generation of Vj. The substantially different kinetics and sensitivity of changes in gj by Vi-o and Vj suggests that the mechanisms of gating by these two voltages are different. Evidently, these gap-junction channels are capable of two distinct, but interactive forms of voltage dependence.


Three types of dural suturing for closure of CSF leak after endoscopic transsphenoidal surgery.
OBJECTIVE: Transsphenoidal surgery (TSS) is commonly used for anterior skull base surgery, especially in the sella turcica (sellar) region. However, because of its anatomical position, CSF leakage is a major complication of this approach. The authors introduced a new grading reconstruction strategy for anterior skull base surgery with continuous dural suturing in 2013. In this paper the authors report on their methods and results. METHODS: All patients with sellar or anterior skull base lesions that were removed with TSS or extended TSS by a single neurosurgeon between April 2013 and March 2017 at Nagoya University Hospital and several cooperating hospitals were retrospectively identified. Three methods of suturing dura were considered, depending on the dural defect. RESULTS: There were 176 TSS cases (141 conventional TSS cases and 35 extended endoscopic TSS cases) and 76 cases of Esposito's grade 2 or 3 intradural high-flow CSF leakage. In the high-flow CSF leak group, there were 3 cases of CSF leakage after the operation. The rates of CSF leakage after surgery corresponding to grades 2 and 3 were 2.9% (1/34) and 4.7% (2/42), respectively. CONCLUSIONS: Dural suturing is a basic and key method for reconstruction of the skull base, and continuous suturing is the most effective approach. Using this approach, the frequency of cases requiring a nasoseptal flap and lumbar drainage can be reduced.


Patterns of host genome-wide gene transcript abundance in the peripheral blood of patients with acute dengue hemorrhagic fever.
Responses by peripheral blood leukocytes may contribute to the pathogenesis of dengue hemorrhagic fever (DHF). We used DNA microarrays to reveal transcriptional patterns in the blood of 14 adults with DHF. Acute DHF was defined by an abundance of transcripts from cell cycle- and endoplasmic reticulum (ER)-related genes, suggesting a proliferative response accompanied by ER stress. Transcript-abundance levels for immunoresponse-associated genes, including cell surface markers, immunoglobulin, and innate response elements, were also elevated. Twenty-four genes were identified for which transcript abundance distinguished patients with dengue shock syndrome (DSS) from those without DSS. All the gene transcripts associated with DSS, many of which are induced by type I interferons, were less abundant in patients with DSS than in those without DSS. To our knowledge, these data provide the first snapshot of gene-expression patterns in peripheral blood during acute dengue and suggest that DSS is associated with attenuation of selected aspects of the innate host response.


Remedies sought and obtained in healthcare complaints.
In the wake of adverse events, injured patients and their families have a complex range of needs and wants. The tort system, even when operating at its best, will inevitably fall far short of addressing them. In Australia and New Zealand, government-run health complaints commissions take a more flexible and expansive approach to providing remedies for patients injured by or disgruntled with care. Unfortunately, survey research has shown that many patients in these systems are dissatisfied with their experience. We hypothesised that an important explanation for this dissatisfaction is an 'expectations gap'; discordance between what complainants want and what they eventually get out of the process. Analysing a sample of complaints relating to informed consent from the Commission in Victoria (Australia's second largest state, with 5.2 million residents), we found evidence of such a gap. One-third (59/189) of complainants who sought restoration received it; 1 in 5 complainants (17/101) who sought correction received assurances that changes had been or would be made to reduce the risk of others suffering a similar harm; and fewer than 1 in 10 (3/37) who sought sanctions saw steps taken to achieve this outcome initiated. We argue that bridging the expectations gap would go far toward improving patient satisfaction with complaints systems, and suggest several ways this might be done.


Superior immune reconstitution using Treg-expanded donor cells versus PTCy treatment in preclinical HSCT models.
Posttransplant cyclophosphamide (PTCy) has been found to be effective in ameliorating acute graft-versus-host disease (GVHD) in patients following allogeneic hematopoietic stem cell transplantation (aHSCT). Adoptive transfer of high numbers of donor Tregs in experimental aHSCT has shown promise as a therapeutic modality for GVHD regulation. We recently described a strategy for in vivo Treg expansion targeting two receptors: TNFRSF25 and CD25. To date, there have been no direct comparisons between the use of PTCy and Tregs regarding outcome and immune reconstitution within identical groups of transplanted mice. Here, we assessed these two strategies and found both decreased clinical GVHD and improved survival long term. However, recipients transplanted with Treg-expanded donor cells (TrED) exhibited less weight loss early after HSCT. Additionally, TrED recipients demonstrated less thymic damage, significantly more recent thymic emigrants, and more rapid lymphoid engraftment. Three months after HSCT, PTCy-treated and TrED recipients showed tolerance to F1 skin allografts and comparable immune function. Overall, TrED was found superior to PTCy with regard to weight loss early after transplant and initial lymphoid engraftment. Based on these findings, we speculate that morbidity and mortality after transplant could be diminished following TrED transplant into aHSCT recipients, and, therefore, that TrED could provide a promising clinical strategy for GVHD prophylaxis.


Optimal achieved blood pressure in acute intracerebral hemorrhage: INTERACT2.
To investigate the effects of intensive blood pressure (BP) lowering according to baseline BP levels and optimal achieved BP levels in patients with acute intracerebral hemorrhage (ICH). INTERACT2 was an open, blinded endpoint, randomized controlled trial in 2,839 patients with ICH within 6 hours of onset and elevated systolic BP (SBP) (150-220 mm Hg) who were allocated to receive intensive (target SBP <140 mm Hg within 1 hour, with lower limit of 130 mm Hg for treatment cessation) or guideline-recommended (target SBP <180 mm Hg) BP-lowering treatment. Outcome was physical function across all 7 levels of the modified Rankin Scale at 90 days. Analysis of the randomized comparisons showed that intensive BP lowering produced comparable benefits on physical function at 90 days in 5 subgroups defined by baseline SBP of <160, 160-169, 170-179, 180-189, and ≥190 mm Hg (p homogeneity = 0.790). Analyses of achieved BP showed linear increases in the risk of physical dysfunction for achieved SBP above 130 mm Hg for both hyperacute (1-24 hours) and acute (2-7 days) phases while modest increases were also observed for achieved SBP below 130 mm Hg. Intensive BP lowering appears beneficial across a wide range of baseline SBP levels, and target SBP level of 130-139 mm Hg is likely to provide maximum benefit in acute ICH. This study provides Class I evidence that the effect of intensive BP lowering on physical function is not influenced by baseline BP.


The cell-cycle state of stem cells determines cell fate propensity.
Self-renewal and differentiation of stem cells are fundamentally associated with cell-cycle progression to enable tissue specification, organ homeostasis, and potentially tumorigenesis. However, technical challenges have impaired the study of the molecular interactions coordinating cell fate choice and cell-cycle progression. Here, we bypass these limitations by using the FUCCI reporter system in human pluripotent stem cells and show that their capacity of differentiation varies during the progression of their cell cycle. These mechanisms are governed by the cell-cycle regulators cyclin D1-3 that control differentiation signals such as the TGF-β-Smad2/3 pathway. Conversely, cell-cycle manipulation using a small molecule directs differentiation of hPSCs and provides an approach to generate cell types with a clinical interest. Our results demonstrate that cell fate decisions are tightly associated with the cell-cycle machinery and reveal insights in the mechanisms synchronizing differentiation and proliferation in developing tissues.


The effect of cocaine-amphetamine regulated transcript (CART) on the activation of the pituitary-adrenal axis.
It has been reported that the cocaine and amphetamine regulated transcript (CART) is widely expressed in the hypothalamic nuclei involved in the mechanism of hormonal secretion. In order to evaluate the effect of CART on the activation of the pituitary-adrenal axis in the rat, CART (55-102) in a dose of 0.5 microg in 5 microl CSF was infused into the third ventricle (icv) with an automatic pump. At 10, 30, 60, 120 min after the infusion of CART or vehicle, the animals were decapitated and the trunk blood was collected. Serum ACTH and corticosterone concentrations were measured with RIA kits provided by Phoenix Pharmaceuticals and ICN Biomedicals USA, respectively. The stimulating effect of CART on ACTH concentration was observed only 30 min after icv injection. However, CART stimulated corticosterone release at 10, 30, 60 min after icv injection. CART (55-102) administered intracerebroventricularly (icv) stimulated ACTH and corticosterone release. The stimulating effect of CART on ACTH was short-lived.


Clinical characteristics of pruritus in chronic idiopathic urticaria.
Although pruritus is a predominant symptom of chronic idiopathic urticaria (CIU) its clinical characteristics have not been explored. To characterize the clinical pattern and sensory and affective dimensions of the itch experience, utilizing a comprehensive itch questionnaire. A structured questionnaire based on the McGill pain questionnaire was used in 100 patients suffering from CIU randomly recruited from a tertiary referral centre. All 100 patients recruited with CIU completed the questionnaire. In 68 patients pruritus appeared on a daily basis. Most patients experienced their pruritus at night and in the evening (n = 83), and 62 reported difficulty in falling asleep. Pruritus involved all body areas, but mostly the arms (n = 86), back (n = 78) and legs (n = 75). Accompanying symptoms were a sensation of heat in 45 patients and sweating in 15. Most patients (n = 98) were prescribed antihistamines (mainly sedating), of whom 34 experienced long-term relief. The sensation of itch was reported to be stinging (n = 27), tickling (n = 25) and burning (n = 23). Seventy-six patients found their pruritus bothersome, 66 annoying and 14 complained of depression. The itch intensity at its peak was more than double that felt after a mosquito bite. The worst itch scores of those who felt depressed were significantly higher than of those who did not (P = 0.018). There was a positive correlation between the sensory and affective scores during worst itch (P < 0.001). This study describes the itch experienced in CIU, highlighting sensory and affective dimensions. The itch questionnaire was found to be a valuable tool for evaluating pruritus in CIU and its unique features.


Plasma membrane calcium ATPases are important components of receptor-mediated signaling in plant immune responses and development.
Plasma membrane-resident receptor kinases (RKs) initiate signaling pathways important for plant immunity and development. In Arabidopsis (Arabidopsis thaliana), the receptor for the elicitor-active peptide epitope of bacterial flagellin, flg22, is encoded by FLAGELLIN SENSING2 (FLS2), which promotes plant immunity. Despite its relevance, the molecular components regulating FLS2-mediated signaling remain largely unknown. We show that plasma membrane ARABIDOPSIS-AUTOINHIBITED Ca(2+)-ATPase (ACA8) forms a complex with FLS2 in planta. ACA8 and its closest homolog ACA10 are required for limiting the growth of virulent bacteria. One of the earliest flg22 responses is the transient increase of cytosolic Ca(2+) ions, which is crucial for many of the well-described downstream responses (e.g. generation of reactive oxygen species and the transcriptional activation of defense-associated genes). Mutant aca8 aca10 plants show decreased flg22-induced Ca(2+) and reactive oxygen species bursts and exhibit altered transcriptional reprogramming. In particular, mitogen-activated protein kinase-dependent flg22-induced gene expression is elevated, whereas calcium-dependent protein kinase-dependent flg22-induced gene expression is reduced. These results demonstrate that the fine regulation of Ca(2+) fluxes across the plasma membrane is critical for the coordination of the downstream microbe-associated molecular pattern responses and suggest a mechanistic link between the FLS2 receptor complex and signaling kinases via the secondary messenger Ca(2+). ACA8 also interacts with other RKs such as BRI1 and CLV1 known to regulate plant development, and both aca8 and aca10 mutants show morphological phenotypes, suggesting additional roles for ACA8 and ACA10 in developmental processes. Thus, Ca(2+) ATPases appear to represent general regulatory components of RK-mediated signaling pathways.


Tissue factor expression on monocyte subpopulations during normal pregnancy.
Pregnancy is often referred to as a hypercoagulable state due to changes in the haemostatic system. Tissue factor (TF) is the initiator of blood clotting in vivo. The effect of pregnancy on monocyte TF expression was determined in a longitudinal case control study (89 pregnant, 39 non-pregnant). Using whole blood flow cytometry and CD14 as a monocyte marker, TF expression was measured on all CD14 positive, CD14Bright and CD14Dim cells. TF expression was significantly lower in pregnant women than in non-pregnant control subjects, on all CD14 positive cells at 20 and 35 weeks, on CD14Bright cells at 12 and 35 weeks and on CD14Dim cells at 20 weeks. Additionally, we report that a higher percentage of CD14Dim than CD14Bright cells express TF. These results suggest that, in order to maintain homeostasis in haemostasis in an otherwise hypercoagulable state, monocyte TF expression is reduced during normal pregnancy.


A meta-analysis of the effect of antibody therapy for the prevention of severe respiratory syncytial virus infection.
The primary objective of this meta-analytic study was to determine the impact of RSV-IGIV and palivizumab on risk of respiratory syncytial virus (RSV)-related hospitalization. Secondary objectives were to determine if antibody therapy decreases the risk of RSV infection, intensive care admission, mechanical ventilation, and mortality in high risk infant populations. We performed searches of electronic data bases from 1966 to April 2009. Inclusion and exclusion criteria were defined a priori. Inclusion criteria were as follows: 1) There was randomization between polyclonal or monoclonal antibodies and placebo or no therapy, and 2) Polyclonal or monoclonal antibodies were given as prophylaxis. Of the six included studies, three utilized RSV-IGIV (total of 533 randomized to treatment groups) and three utilized palivizumab (total of 1,663 randomized to treatment groups). The absolute risk of hospitalization in the control arms was 12% and overall RR for all 2,196 children who received one of the antibody products was 0.53 (95% CI 0.43, 0.66), P < 0.00001. When looking only at the children who received palivizumab, the RR for hospitalization was 0.50 (95% CI 0.38, 0.66), P < 0.00001. For the children receiving RSV-IGIV, the RR for hospitalization was 0.59 (95% CI 0.42, 0.83, P < 0.002). The use of palivizumab resulted in a significant decrease in admission to the ICU (RR 0.29 (95% CI 0.14, 0.59; P = 0.0007). There was no significant reduction in the risk of mechanical ventilation or mortality with the use of antibody prophylaxis. Infants born at less than 35 weeks gestational age, and those with chronic lung and congenital heart disease all had a significant reduction in the risk of RSV hospitalization with children born under 35 weeks gestational age showing a trend towards the greatest benefit. Both palivizumab and RSV-IGIV decrease the incidence of RSV hospitalization and ICU admission and their effect appears to be qualitatively similarly. There was neither a statistically significant reduction in the incidence of mechanical ventilation nor in all cause mortality. This meta-analysis separately quantifies the impact of RSV-IGIV and palivizumab on various measures of severe RSV disease and builds upon a previous study that was only able to examine the pooled effect of all antibody products together.


Cochlear implantation improves hearing and vertigo in patients after removal of vestibular schwannoma.
With rising quality of speech perception in cochlear implant users, the indication widens. Nowadays, cochlear implantation is reasonable even in vestibular schwannoma patients. Speech perception with a cochlear implant is in these patients as promising as in patients with sensorineural hearing loss. However, the impact of cochlear implantation on vertigo and tinnitus after removal of vestibular schwannoma has not been investigated yet. In a retrospective study, we analysed 12 patients treated with a cochlear implant after removal of vestibular schwannoma. In addition to a promising hearing perception - all patients reported improvement of vertigo. This improvement was also demonstrated by postural analysis. Improvement of tinnitus was achieved in 50% of the patients. Cochlear implantation seems a promising treatment for hearing loss, vertigo and even tinnitus in patients after removal of vestibular schwannoma. However, for successful cochlear implantation with adequate speech perception and improvement of vertigo and tinnitus, functional hearing nerve and intact inner ear anatomy is necessary.


Causal Contribution of Awake Post-encoding Processes to Episodic Memory Consolidation.
Stable representations of past experience are thought to depend on processes that unfold after events are initially encoded into memory. Post-encoding reactivation and hippocampal-cortical interactions are leading candidate mechanisms thought to support memory retention and stabilization across hippocampal-cortical networks. Although putative consolidation mechanisms have been observed during sleep and periods of awake rest, the direct causal contribution of awake consolidation mechanisms to later behavior is unclear, especially in humans. Moreover, it has been argued that observations of putative consolidation processes are epiphenomenal and not causally important, yet there are few tools to test the functional contribution of these mechanisms in humans. Here, we combined transcranial magnetic stimulation (TMS) and fMRI to test the role of awake consolidation processes by targeting hippocampal interactions with lateral occipital cortex (LOC). We applied theta-burst TMS to LOC (and a control site) to interfere with an extended window (approximately 30-50 min) after memory encoding. Behaviorally, post-encoding TMS to LOC selectively impaired associative memory retention compared to multiple control conditions. In the control TMS condition, we replicated prior reports of post-encoding reactivation and memory-related hippocampal-LOC interactions during periods of awake rest using fMRI. However, post-encoding LOC TMS reduced these processes, such that post-encoding reactivation in LOC and memory-related hippocampal-LOC functional connectivity were no longer present. By targeting and manipulating post-encoding neural processes, these findings highlight the direct contribution of awake time periods to episodic memory consolidation. This combined TMS-fMRI approach provides an opportunity for causal manipulations of human memory consolidation.


Pain and suicidality in children and adolescents: a longitudinal population-based study.
Suicidality is a common public health concern in young people. Previous research has highlighted pain as a key correlate of suicidality in young people. However, the long-term experience of pain may vary between individuals, and the relationship between distinct pain trajectories and suicidality is poorly understood. This study aims to describe the number and nature of distinct pain trajectories, their demographic and clinical correlates, including baseline suicidality, and whether identified pain trajectories may predict future suicidality. Secondary data analyses were performed, using longitudinal data from the British Child and Adolescent Mental Health Survey (N = 7977), collected at five timepoints between 2004 and 2007 on a population-based sample of UK youth (5-16 years). Data were collected from up to three respondents (parents, teachers, and 11 + year-olds). Latent Class Growth Analysis was used to identify distinct pain trajectories, explore predictors of these trajectories, and establish whether trajectories predicted future suicidality. We identified the following four pain trajectories: increasing (33.6%), decreasing (4.5%), persistent/recurrent probability of pain (15.7%), and no pain (46.2%). Pain trajectories were associated with unique demographic and clinical correlates. Only the persistent/recurrent (vs. no-pain) trajectory was predicted by baseline suicidality (aOR = 2.24; 95% bootstrap-CI = 1.59-3.26). Furthermore, the persistent/recurrent trajectory predicted future suicidality (aOR = 1.03, 95% bootstrap-CI = 1.01-1.06), after controlling for baseline suicidality, psychiatric disorder, age, and gender. Findings provide a better understanding of correlates associated with distinct pain trajectories and long-term risk of suicidality in young people, suggesting a bidirectional pain-suicidality association and emphasising the need of targeted support for young people with persistent/recurrent pain.


Open retropubic colposuspension for urinary incontinence in women.
Urinary incontinence is a common and potentially debilitating problem. Stress urinary, incontinence as the most common type of incontinence, imposes significant health and economic burdens on society and the women affected. Open retropubic colposuspension is a surgical treatment which involves lifting the tissues near the bladder neck and proximal urethra in the area behind the anterior pubic bones to correct deficient urethral closure to correct stress urinary incontinence. The review aimed to determine the effects of open retropubic colposuspension for the treatment of urinary incontinence in women. A secondary aim was to assess the safety of open retropubic colposuspension in terms of adverse events caused by the procedure. We searched the Cochrane Incontinence Group Specialised Register, which contains trials identified from the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, MEDLINE in process, ClinicalTrials.gov, WHO ICTRP and handsearching of journals and conference proceedings (searched 5 May 2015), and the reference lists of relevant articles. We contacted investigators to locate extra studies. Randomised or quasi-randomised controlled trials in women with symptoms or urodynamic diagnoses of stress or mixed urinary incontinence that included open retropubic colposuspension surgery in at least one trial group. Studies were evaluated for methodological quality or susceptibility to bias and appropriateness for inclusion and data extracted by two of the review authors. Trial data were analysed by intervention. Where appropriate, a summary statistic was calculated. This review included 55 trials involving a total of 5417 women.Overall cure rates were 68.9% to 88.0% for open retropubic colposuspension. Two small studies suggested lower incontinence rates after open retropubic colposuspension compared with conservative treatment. Similarly, one trial suggested lower incontinence rates after open retropubic colposuspension compared to anticholinergic treatment. Evidence from six trials showed a lower incontinence rate after open retropubic colposuspension than after anterior colporrhaphy. Such benefit was maintained over time (risk ratio (RR) for incontinence 0.46; 95% CI 0.30 to 0.72 before the first year, RR 0.37; 95% CI 0.27 to 0.51 at one to five years, RR 0.49; 95% CI 0.32 to 0.75 in periods beyond five years).Evidence from 22 trials in comparison with suburethral slings (traditional slings or trans-vaginal tape or transobturator tape) found no overall significant difference in incontinence rates in all time periods evaluated (as assessed subjectively RR 0.90; 95% CI 0.69 to 1.18, within one year of treatment, RR 1.18; 95%CI 1.01 to 1.39 between one and five years, RR 1.11; 95% CI 0.97 to 1.27 at five years and more, and as assessed objectively RR 1.24; 95% CI 0.93 to 1.67 within one year of treatment, RR 1.12; 95% CI 0.82 to 1.54 for one to five years follow up, RR 0.70; 95% CI 0.30 to 1.64 at more than five years). However, subgroup analysis of studies comparing traditional slings and open colposuspension showed better effectiveness with traditional slings in the medium and long term (RR 1.35; 95% CI 1.11 to 1.64 from one to five years follow up, RR 1.19; 95% CI 1.03 to 1.37).In comparison with needle suspension, there was a lower incontinence rate after colposuspension in the first year after surgery (RR 0.66; 95% CI 0.42 to 1.03), after the first year (RR 0.56; 95% CI 0.39 to 0.81), and beyond five years (RR 0.32; 95% CI 15 to 0.71).Patient-reported incontinence rates at short, medium and long-term follow-up showed no significant differences between open and laparoscopic retropubic colposuspension, but with wide confidence intervals. In two trials incontinence was less common after the Burch (RR 0.38; 95% CI 0.18 to 0.76) than after the Marshall Marchetti Krantz procedure at one to five year follow-up. There were few data at any other follow-up times.In general, the evidence available does not show a higher morbidity or complication rate with open retropubic colposuspension compared to the other open surgical techniques, although pelvic organ prolapse is more common than after anterior colporrhaphy and sling procedures. Voiding problems are also more common after sling procedures compared to open colposuspension. Open retropubic colposuspension is an effective treatment modality for stress urinary incontinence especially in the long term. Within the first year of treatment, the overall continence rate is approximately 85% to 90%. After five years, approximately 70% of women can expect to be dry. Newer minimal access sling procedures look promising in comparison with open colposuspension but their long-term performance is limited and closer monitoring of their adverse event profile must be carried out. Open colposuspension is associated with a higher risk of pelvic organ prolapse compared to sling operations and anterior colporrhaphy, but with a lower risk of voiding dysfunction compared to traditional sling surgery. Laparoscopic colposuspension should allow speedier recovery but its relative safety and long-term effectiveness is not yet known. A Brief Economic Commentary (BEC) identified five studies suggesting that tension-free vaginal tape (TVT) and laparoscopic colposuspension may be more cost-effective compared with open retropubic colposuspension.


The Effectiveness of Standardized Patient Simulation in Training Hospital Ethics Committees.
Clinical simulation using standardized patients has become standard in medical education--and is now being incorporated into some graduate programs in bioethics--for both formative and summative evaluation. In most hospitals, though, clinical ethics consultation is done by the ethics committee (or a subset of it). This study is the first, to our knowledge, to examine the effectiveness of standardized patient simulation in training hospital ethics committees to deal with ethically complex and emotionally fraught clinical situations. Following a substantial revision of the institution's nonbeneficial treatment policy, ethics committee members underwent a simulation to determine whether a specific requested treatment should be withheld on the basis of futility. Pre- and post-intervention surveys showed improvement in all domains, although the small sample size limited the power of the study, with only one measure showing a statistically significant difference. An interesting incidental finding was that one-quarter of committee members voted against a determination of futility, even though the case clearly met the definition set forth in the policy. This highlights the emotional challenges in implementing an ethically rigorous, unanimously accepted policy that ultimately determines the timing and manner of a patient's death.


CheB is required for behavioural responses to negative stimuli during chemotaxis in Bacillus subtilis.
The methyl-accepting chemotaxis protein, McpB, is the sole receptor mediating asparagine chemotaxis in Bacillus subtilis. In this study, we show that wild-type B. subtilis cells contain approximately 2,000 copies of McpB per cell, that these receptors are localized polarly, and that titration of only a few receptors is sufficient to generate a detectable behavioural response. In contrast to the wild type, a cheB mutant was incapable of tumbling in response to decreasing concentrations of asparagine, but the cheB mutant was able to accumulate to low concentrations of asparagine in the capillary assay, as observed previously in response to azetidine-2-carboxylate. Furthermore, net demethylation of McpB is logarithmically dependent on asparagine concentration, with half-maximal demethylation of McpB occurring when only 3% of the receptors are titrated. Because the corresponding methanol production is exponentially dependent on attractant concentration, net methylation changes and increased turnover of methyl groups must occur on McpB at high concentrations of asparagine. Together, the data support the hypothesis that methylation changes occur on asparagine-bound McpB to enhance the dynamic range of the receptor complex and to enable the cell to respond to a negative stimulus, such as removal of asparagine.


Literacy and health outcomes: a systematic review of the literature.
To review the relationship between literacy and health outcomes. We searched MEDLINE, Cumulative Index to Nursing and Allied Health (CINAHL), Educational Resources Information Center (ERIC), Public Affairs Information Service (PAIS), Industrial and Labor Relations Review (ILLR), PsychInfo, and Ageline from 1980 to 2003. We included observational studies that reported original data, measured literacy with any valid instrument, and measured one or more health outcomes. Two abstractors reviewed each study for inclusion and resolved disagreements by discussion. One reviewer abstracted data from each article into an evidence table; the second reviewer checked each entry. The whole study team reconciled disagreements about information in evidence tables. Both data extractors independently completed an 11-item quality scale for each article; scores were averaged to give a final measure of article quality. We reviewed 3,015 titles and abstracts and pulled 684 articles for full review; 73 articles met inclusion criteria and, of those, 44 addressed the questions of this report. Patients with low literacy had poorer health outcomes, including knowledge, intermediate disease markers, measures of morbidity, general health status, and use of health resources. Patients with low literacy were generally 1.5 to 3 times more likely to experience a given poor outcome. The average quality of the articles was fair to good. Most studies were cross-sectional in design; many failed to address adequately confounding and the use of multiple comparisons. Low literacy is associated with several adverse health outcomes. Future research, using more rigorous methods, will better define these relationships and guide developers of new interventions.


Diabetes UK evidence-based nutrition guidelines for the prevention and management of diabetes.
A summary of the latest evidence-based nutrition guidelines for the prevention and management of diabetes is presented. These guidelines are based on existing recommendations last published in 2011, and were formulated by an expert panel of specialist dietitians after a literature review of recent evidence. Recommendations have been made in terms of foods rather than nutrients wherever possible. Guidelines for education and care delivery, prevention of Type 2 diabetes, glycaemic control for Type 1 and Type 2 diabetes, cardiovascular disease risk management, management of diabetes-related complications, other considerations including comorbidities, nutrition support, pregnancy and lactation, eating disorders, micronutrients, food supplements, functional foods, commercial diabetic foods and nutritive and non-nutritive sweeteners are included. The sections on pregnancy and prevention of Type 2 diabetes have been enlarged and the weight management section modified to include considerations of remission of Type 2 diabetes. A section evaluating detailed considerations in ethnic minorities has been included as a new topic. The guidelines were graded using adapted 'GRADE' methodology and, where strong evidence was lacking, grading was not allocated. These 2018 guidelines emphasize a flexible, individualized approach to diabetes management and weight loss and highlight the emerging evidence for remission of Type 2 diabetes. The full guideline document is available at www.diabetes.org.uk/nutrition-guidelines.


A modified approach to supraclavicular subclavian vein catheter placement: the pocket approach.
Central venous access is often necessary for the administration of fluids, blood products, and medications. Several approaches to supraclavicular subclavian venous access have been described. This study examines the effectiveness of central venous catheter placement utilizing an alternative set of anatomic landmarks for supraclavicular subclavian vein access. This was a two phase study. The first portion involved subclavian vein cannulation using a supraclavicular approach in 28 cadavers. The specific set of anatomic landmarks for the supraclavicular approach, termed the "pocket approach," is described. Cadavers were subsequently dissected to verify appropriate line placement. The second portion was a chart review of Emergency Department (ED) patients who underwent attempted subclavian vein catheter placement utilizing the pocket approach. Charts were extracted following education of the ED faculty and resident staff to determine: 1) Success of subclavian line placement, 2) The incidence of pneumothorax, and 3) The use of supraclavicular subclavian access in the trauma setting, during cardiopulmonary resuscitation (CPR), and in patients who had cervical collars. In 28 cadavers, the success rate of the pocket approach was 100% (34/34; 95% CI 90% to 100%). Chart review of the 68 patients revealed a success rate of 90% (61/68; CI 80% to 96%). No pneumothoraces were recorded (0/68; CI 0% to 5%). The pocket approach was used successfully in 11 patients with cervical collars, (100%, CI 72% to 100%) and in 15 of 16 patients undergoing CPR (94%, CI 70% to 100%). In four fresh cadavers, the average distance from the posterior subclavian vein to the subclavian artery was 0.40cm, and the dome of the pleura was 1.75cm posterior to the vein. Our data suggest that the supraclavicular pocket approach to subclavian vein cannulation is a useful and safe method of adult central venous catheterization, with complication and success rates comparable to more common approaches. The anatomic advantage of a great vein that is closer to the skin and farther from the pleural dome makes this an approach worthy of further investigation.


Twelve-month-olds disambiguate new words using mutual-exclusivity inferences.
Representing objects in terms of their kinds enables inferences based on the long-term knowledge made available through kind concepts. For example, children readily use lexical knowledge linked to familiar kind concepts to disambiguate new words (e.g., "find the toma"): they exclude members of familiar kinds falling under familiar kind labels (e.g., a ball) as potential referents and link new labels to available unfamiliar objects (e.g., a funnel), a phenomenon dubbed as 'mutual exclusivity'. Younger infants' failure in mutual exclusivity tasks has been commonly interpreted as a limitation of early word-learning or inferential abilities. Here, we investigated an alternative explanation, according to which infants do not spontaneously represent familiar objects under kind concepts, hence lacking access to the information necessary for rejecting them as referents of novel labels. Building on findings about conceptual development and communication, we hypothesized that nonverbal communication could prompt infants to set up kind-based representations which, in turn, would promote mutual exclusivity inferences. This hypothesis was tested in a looking-while-listening task involving novel word disambiguation. Twelve-month-olds saw pairs of objects, one familiar and one unfamiliar, and heard familiar kind labels or novel words. Across two experiments providing a cross-lab replication in two different languages, infants successfully disambiguated novel words when the familiar object had been pointed at before labeling, but not when it had been highlighted in a non-communicative manner (Experiment 1) or not highlighted at all (Experiment 2). Nonverbal communication induced infants to recruit kind-based representations of familiar objects that they failed to recruit in its absence and that, once activated, supported mutual-exclusivity inferences. Developmental changes in children's appreciation of communicative contexts may modulate the expression of early inferential and word learning competences.


Plasticization-enhanced hydrogen purification using polymeric membranes.
Polymer membranes are attractive for molecular-scale separations such as hydrogen purification because of inherently low energy requirements. However, membrane materials with outstanding hydrogen separation performance in feed streams containing high-pressure carbon dioxide and impurities such as hydrogen sulfide and water are not available. We report highly permeable, reverse-selective membrane materials for hydrogen purification, as exemplified by molecularly engineered, highly branched, cross-linked poly(ethylene oxide). In contrast to the performance of conventional materials, we demonstrate that plasticization can be harnessed to improve separation performance.


Visual quality of life after macular hole surgery: outcome and predictive factors.
In the present study we evaluated the functional success after macular hole surgery in correlation to visual quality of life and looked for predictive factors determining surgical success. Fifty-nine patients that underwent pars plana vitrectomy for idiopathic macular hole were included. Follow-up visits were performed in regular intervals after surgery and included a clinical examination, optical coherence tomography (OCT) and measurement of visual acuity. To assess the visual quality of life patients filled out the National Eye Institute 25-item Visual Function Questionnaire (VFQ-25) before and three months and one year after surgery. Macular hole closure was achieved in 57 of 59 patients (97%). Mean visual acuity increased from 20/100 preoperatively to 20/34 one year after surgery (p = 0.02). Despite good visual acuity (20/27) in the fellow eye, visual quality of life (VFQ composite score) rose from 75.9 +/- 14.4 (SD) to 81.5 +/- 14.2 one year after surgery (p<0.001). Although there was no correlation between the increase in visual quality of life and visual acuity, the increase in VFQ-25 could be well predicted: low visual acuity and significant impairment on VFQ-25 testing preoperatively made patients most likely to benefit from macular hole surgery. A relatively high retinal thickness measurement at the hole border measured on OCT further increases the predictive value. Macular hole surgery is associated with an increase in visual quality of life despite good visual acuity of the fellow eye. Preoperative visual acuity, VFQ-25 value and partly OCT may help to predict the increase in patients' vision related quality of life after surgery.


Altered cell adhesion and cell viability in a p38alpha mitogen-activated protein kinase-deficient mouse embryonic stem cell line.
p38 mitogen-activated protein (MAP) kinase alpha (p38alpha) is a broadly expressed protein kinase that regulates growth and development. Most studies of p38alpha have been in somatic cells. Little is known about its function in embryonic stem (ES) cells. Using a ES cell line isolated from p38alpha knockout mouse embryos (p38alpha (-/-) ES cells), we investigated roles of p38alpha in the regulation of ES cell activities. p38alpha (-/-) ES cells displayed several altered features different from wild-type cells. The major findings are that p38alpha (-/-) ES cells have significantly increased cell adhesion to several extracelluar matrix proteins, correlating with elevated phosphorylation of focal adhesion kinase and paxillin. p38alpha (-/-) ES cells also showed increased cell viability, correlating with increased expression of survivin and activation of AKT (protein kinase B), two molecules that are known to improve cell viability. p38alpha (-/-) ES cells reach confluence faster than wild-type cells in routine cell culture. However, this is not due to a higher cell proliferation rate in p38alpha (-/-) ES cells, but rather is likely a result of improved cell adhesion and/or cell viability. Together our results indicated that p38alpha may negatively regulate mouse ES cell adhesion and viability.


Stabilization of lysozyme mass extracted from lotrafilcon silicone hydrogel contact lenses.
Lysozyme deposits extracted from lotrafilcon silicone hydrogel (SH) contact lens materials demonstrate a loss in total mass as a function of storage time when assessed by Western blotting. This loss represents a potential source of error when quantifying total lysozyme deposition on SH lenses. The purpose of this study was to devise a method whereby lysozyme mass would be preserved over time to allow for its accurate quantitation after its removal from SH lenses. Lysozyme deposits from 12 human worn lotrafilcon lenses were extracted using a 50:50 mixture of 0.2% trifluoroacetic acid and acetonitrile. Extracts were lyophilized to dryness, then resuspended in either reconstitution buffer (10 mM Tris-HCl, 1 mM EDTA) or modified reconstitution buffer (reconstitution buffer + 0.9% saline). BIOSTAB Biomolecule Storage Solution (Sigma-Aldrich) was added to one half of the samples from each buffer group. One microliter of each of the samples was immediately subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting, whereas the remaining volume was aliquoted and stored at -20 degrees C or -70 degrees C and subjected to the same procedures after 48 h of storage. Comparison of lysozyme band intensity in stored vs. fresh samples enabled calculation of percentage mass loss of lysozyme. Samples stored at -20 degrees C in reconstitution buffer with no BIOSTAB demonstrated a 33% loss in mass over 48 h of storage. Identical samples stored at -70 degrees C in modified reconstitution buffer with BIOSTAB added demonstrated <1% loss in mass. Statistical analysis indicated that buffer composition (p < 0.001), storage temperature (p = 0.04), and addition of BIOSTAB (p < 0.001) were all important in controlling loss of mass over time. We have optimized a procedure whereby the extracted mass of lysozyme deposits found on lotrafilcon SH lenses can be preserved, thus enabling accurate quantitation after extraction and resuspension.


Relationships between Fusarium population structure, soil nutrient status and disease incidence in field-grown asparagus.
Fusarium species cause important diseases in many crops. Lack of knowledge on how Fusarium species and strains interact with their environment hampers growth management strategies to control root diseases. A field experiment involving asparagus as host plant and three phosphorus fertilization levels was designed to examine the seasonal changes and ecological relationships between Fusarium populations and their soil and plant environments. Fusarium taxa were identified and assessed using PCR-denaturing gradient electrophoresis of the EF1-alpha gene. Resulting profiles were analyzed with respect to 17 ecological parameters measured during the three main asparagus phenological phases across a growing season. Multivariate statistical analysis showed that Fusarium population structure was strongly influenced by soil P level while seasonal variation was less important. A significant relationship between Fusarium population composition and Fusarium crown and root rot incidence was also found in September. Canonical analysis further revealed significant relationships between Fusarium population structure, and plant manganese and iron contents, soil dehydrogenase activity and soil calcium concentration. If higher Fusarium crown and root rot incidence is related to the Fusarium community structure, strategies to reduce the incidence in asparagus plantations may be found through manipulation of the soil fertility.


Biomarker Potential of Extracellular miRNAs in Duchenne Muscular Dystrophy.
miRNAs are small, noncoding RNAs that not only regulate gene expression within cells, but might also constitute promising extracellular biomarkers for a variety of pathologies, including the progressive muscle-wasting disorder Duchenne Muscular Dystrophy (DMD). A set of muscle-enriched miRNAs, the myomiRs (miR-1, miR-133, and miR-206) are highly elevated in the serum of patients with DMD and in dystrophin-deficient animal models. Furthermore, circulating myomiRs might be used as pharmacodynamic biomarkers, given that their levels can be restored towards wild-type levels following exon skipping therapy in dystrophic mice. The relationship between muscle pathology and extracellular myomiR release is complex, and incompletely understood. Here, we discuss current progress leading towards the clinical utility of extracellular miRNAs as putative DMD biomarkers, and their possible contribution to muscle physiology.


Medial frontal hyperactivity to sad faces in generalized social anxiety disorder and modulation by oxytocin.
Generalized social anxiety disorder (GSAD) is associated with heightened limbic and prefrontal activation to negative social cues conveying threat (e.g. fearful faces), but less is known about brain response to negative non-threatening social stimuli. The neuropeptide oxytocin (Oxt) has been shown to attenuate (and normalize) fear-related brain activation and reactivity to emotionally negative cues. Here, we examined the effects of intranasal Oxt on cortical activation to non-threatening sad faces in patients with GSAD and matched controls (Con). In a double-blind placebo-controlled within-subjects design, the cortical activation to sad and happy (vs. neutral) faces was examined using functional magnetic resonance imaging following acute intranasal administration of 24 IU Oxt and placebo. Relative to the Con group, GSAD patients exhibited heightened activity to sad faces in the medial prefrontal cortex (mPFC/BA 10) extending into anterior cingulate cortex (ACC/BA 32). Oxt significantly reduced this heightened activation in the mPFC/ACC regions to levels similar to that of controls. These findings suggest that GSAD is associated with cortical hyperactivity to non-threatening negative but not positive social cues and that Oxt attenuates this exaggerated cortical activity. The modulation of cortical activity by Oxt highlights a broader mechanistic role of this neuropeptide in modulating socially negative cues in GSAD.


Sinomenine Inhibits the Progression of Rheumatoid Arthritis by Regulating the Secretion of Inflammatory Cytokines and Monocyte/Macrophage Subsets.
Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory arthropathy associated with articular damage and attendant comorbidities. Even although RA treatment has advanced remarkably over the last decade, a significant proportion of patients still do not achieve sustained remission. The cause of RA is not yet known despite the many potential mechanisms proposed. It has been confirmed that RA is associated with dysregulated immune system and persistent inflammation. Therefore, management of inflammation is always the target of therapy. Sinomenine (SIN) is the prescription drug approved by the Chinese government for RA treatment. A previous study found that SIN was a robust anti-inflammation drug. In this study, we screened the different secretory cytokines using inflammation antibody arrays and qRT-PCR in both LPS-induced and SIN-treated RAW264.7 cells followed by evaluation of the ability of SIN to modulate cytokine secretion in a cell model, collagen-induced arthritis (CIA) mouse model, and RA patients. Several clinical indexes affecting the 28-joint disease activity score (DAS28) were determined before and after SIN treatment. Clinical indexes, inflammatory cytokine secretion, and DAS28 were compared among RA patients treated with either SIN or methotrexate (MTX). To explore the mechanism of SIN anti-inflammatory function, RA-associated monocyte/macrophage subsets were determined using flow cytometry in CIA mouse model and RA patients, both treated with SIN. The results demonstrated that SIN regulated IL-6, GM-CSF, IL-12 p40, IL-1α, TNF-α, IL-1β, KC (CXCL1), Eotaxin-2, IL-10, M-CSF, RANTES, and MCP-1 secretion in vivo and in vitro and reduced RA activity and DAS28 in a clinical setting. Furthermore, SIN attenuated CD11b+F4/80+CD64+ resident macrophages in the synovial tissue, CD11b+Ly6C+CD43+ macrophages in the spleen and draining lymph nodes of CIA mice. The percentage of CD14+CD16+ peripheral blood mononuclear cells was reduced by SIN in RA patients. These data indicated that SIN regulates the secretion of multiple inflammatory cytokines and monocyte/macrophage subsets, thereby suppressing RA progression. Therefore, along with MTX, SIN could be an alternative cost-effective anti-inflammatory agent for treating RA.


Effect of regular inhaled albuterol on allergen-induced late responses and sputum eosinophils in asthmatic subjects.
Treatment with inhaled beta(2)-agonists immediately before allergen inhalation inhibits allergen-induced early, but not late asthmatic responses (LAR). By contrast, 2 wk treatment with inhaled albuterol increases airway responses to inhaled allergen. We examined the effects of regular albuterol treatment on allergen-induced increases in inflammatory cells in blood and induced sputum. Ten mild, stable allergic asthmatics inhaled albuterol (800 micrograms/day) or placebo for 7 d in a controlled, randomized, double-blind, crossover study. Allergen inhalation was performed 12 h after the final dose. Methacholine airway responsiveness and blood samples were analyzed before and 24 h after, and induced sputum was obtained before, 7 h and 24 h after allergen. Allergen significantly reduced methacholine PC20, increased blood eosinophil numbers, and numbers of sputum neutrophils, EG2 positive and metachromatic cells (p < 0.05), without significant differences between treatments. Albuterol treatment significantly increased the LAR compared to placebo treatment (p = 0.003) and significantly enhanced the number of sputum eosinophils (p = 0.009) and sputum ECP (p = 0.04) at 7 h but not 24 h post-allergen (p > 0.05). We conclude that regular use of inhaled albuterol significantly increases the LAR to inhaled allergen, in association with an increase in the number of sputum eosinophils and the release of ECP, suggesting albuterol increases the late response by increasing eosinophil influx into the airways.


Effects of chlorthalidone, oxprenolol, and their combination in hypertensive blacks: a randomized double-blind crossover study.
One hundred twenty black patients with mild to moderate essential hypertension participated in a double-blind placebo-controlled crossover study of the efficacy and tolerability of slow release oxprenolol versus chlorthalidone singly and in combination. Oxprenolol as monotherapy produced no effect on blood pressure as compared with placebo even after doubling the dose. Chlorthalidone as monotherapy produced a significant decrease in blood pressure (p less than 0.01). Combining oxprenolol with chlorthalidone yielded hypotensive effects in excess of those of either of the components given singly. Oxprenolol produced a significant decrease in plasma renin activity (PRA) whereas chlorthalidone produced a significant increase in PRA. These results indicate that a beta-blocking agent alone is ineffective in lowering blood pressure in hypertensive blacks, even when the dose is high. Oxprenolol may increase the hypotensive effect of chlorthalidone by counteracting the hypokalemic effect of the diuretic and by attenuating the diuretic-induced increase in plasma renin activity.


Chronic administration of statins alters multiple gene expression patterns in mouse cerebral cortex.
Statins have been reported to lower the risk of developing Alzheimer's disease; however, the mechanism of this potentially important neuroprotective action is not understood. Lowering cholesterol levels does not appear to be the primary mechanism. Statins have pleiotropic effects in addition to lowering cholesterol, and statins may act on several different pathways involving distinct gene expression patterns that would be difficult to determine by focusing on a few genes or their products in a single study. In addition, gene expression patterns may be specific to a particular statin. To understand the molecular targets of statins in brain, DNA microarrays were used to identify gene expression patterns in the cerebral cortex of mice chronically treated with lovastatin, pravastatin, and simvastatin. Furthermore, brain statin levels were determined using liquid chromatography/tandem mass spectrometry. These studies revealed 15 genes involved in cell growth and signaling and trafficking that were similarly changed by all three statins. Overall, simvastatin had the greatest influence on expression as demonstrated by its ability to modify the expression of 23 genes in addition to those changed by all three drugs. Of particular interest was the expression of genes associated with apoptotic pathways that were altered by simvastatin. Reverse transcription-polymerase chain reaction experiments confirmed the microarray findings. All three drugs were detected in the cerebral cortex, and acute experiments revealed that statins are relatively rapidly removed from the brain. These results provide new insight into possible mechanisms for the potential efficacy of statins in reducing the risk of Alzheimer's disease and lay the foundation for future studies.


Repeated intravenous ketamine therapy in a patient with treatment-resistant major depression.
The intravenous administration of ketamine, an N-methyl-D-aspartate receptor antagonist, results in a great improvement of depression symptoms, but it is not clear for how long. This single-case trial was conducted to explore the duration of improvement and the effects of a second administration on the clinical outcome. In an open label trial, a 55-year-old male patient with treatment-resistant major depression and a co-occurring alcohol and benzodiazepine dependence received two intravenous infusions of 0.5 mg/kg ketamine over the course of 6 weeks. Depression severity was assessed by means of a weekly clinical interview, the 21-item Hamilton Depression Rating Scale (HDRS), and the 21-item Beck Depression Inventory (BDI). The first ketamine infusion lead to a pronounced improvement of symptoms, peaking on the second day post infusion (HDRS -56.6%, BDI -65.4%). Positive effects started fading by day 7, reaching baseline by day 35. The second infusion was less efficacious: HDRS and BDI were reduced by 43 and 35%, respectively, and returned to baseline by day 7. In this patient with a co-occurring substance use disorder, repeated administrations of ketamine produced positive results. Since the second application has been less efficacious, doses and schedule of administrations need to be further investigated.


Scientific mobilization of keystone actors for biosphere stewardship.
The biosphere crisis requires changes to existing business practices. We ask how corporations can become sustainability leaders, when constrained by multiple barriers to collaboration for biosphere stewardship. We describe how scientists motivated, inspired and engaged with ten of the world's largest seafood companies, in a collaborative process aimed to enable science-based and systemic transformations (2015-2021). CEOs faced multiple industry crises in 2015 that incentivized novel approaches. New scientific insights, an invitation to collaborate, and a bold vision of transformative change towards ocean stewardship, created new opportunities and direction. Co-creation of solutions resulted in new knowledge and trust, a joint agenda for action, new capacities, international recognition, formalization of an organization, increased policy influence, time-bound goals, and convergence of corporate change. Independently funded scientists helped remove barriers to cooperation, provided means for reflection, and guided corporate strategies and actions toward ocean stewardship. By 2021, multiple individuals exercised leadership and the initiative had transitioned from preliminary and uncomfortable conversations, to a dynamic, operational organization, with capacity to perform global leadership in the seafood industry. Mobilizing transformational agency through learning, collaboration, and innovation represents a cultural evolution with potential to redirect and accelerate corporate action, to the benefit of business, people and the planet.


Crosslinkers and motors organize dynamic microtubules to form stable bipolar arrays in fission yeast.
Microtubule (MT) nucleation not only occurs from centrosomes, but also in large part from dispersed nucleation sites. The subsequent sorting of short MTs into networks like the mitotic spindle requires molecular motors that laterally slide overlapping MTs and bundling proteins that statically connect MTs. How bundling proteins interfere with MT sliding is unclear. In bipolar MT bundles in fission yeast, we found that the bundler ase1p localized all along the length of antiparallel MTs, whereas the motor klp2p (kinesin-14) accumulated only at MT plus ends. Consequently, sliding forces could only overcome resistant bundling forces for short, newly nucleated MTs, which were transported to their correct position within bundles. Ase1p thus regulated sliding forces based on polarity and overlap length, and computer simulations showed these mechanisms to be sufficient to generate stable bipolar bundles. By combining motor and bundling proteins, cells can thus dynamically organize stable regions of overlap between cytoskeletal filaments.


Defining the phenotypical spectrum associated with variants in TUBB2A.
Variants in genes belonging to the tubulin superfamily account for a heterogeneous spectrum of brain malformations referred to as tubulinopathies. Variants in TUBB2A have been reported in 10 patients with a broad spectrum of brain imaging features, ranging from a normal cortex to polymicrogyria, while one patient has been reported with progressive atrophy of the cerebellar vermis. In order to further refine the phenotypical spectrum associated with TUBB2A, clinical and imaging features of 12 patients with pathogenic TUBB2A variants, recruited via the international network of the authors, were reviewed. We report 12 patients with eight novel and one recurrent variants spread throughout the TUBB2A gene but encoding for amino acids clustering at the protein surface. Eleven patients (91.7%) developed seizures in early life. All patients suffered from intellectual disability, and 11 patients had severe motor developmental delay, with 4 patients (36.4 %) being non-ambulatory. The cerebral cortex was normal in five individuals and showed dysgyria of variable severity in seven patients. Associated brain malformations were less frequent in TUBB2A patients compared with other tubulinopathies. None of the patients had progressive cerebellar atrophy. The imaging phenotype associated with pathogenic variants in TUBB2A is highly variable, ranging from a normal cortex to extensive dysgyria with associated brain malformations. For recurrent variants, no clear genotype-phenotype correlations could be established, suggesting the role of additional modifiers.


Reducing the Role of the Food, Tobacco, and Alcohol Industries in Noncommunicable Disease Risk in South Africa.
Noncommunicable diseases (NCDs) impose a growing burden on the health, economy, and development of South Africa. According to the World Health Organization, four risk factors, tobacco use, alcohol consumption, unhealthy diets, and physical inactivity, account for a significant proportion of major NCDs. We analyze the role of tobacco, alcohol, and food corporations in promoting NCD risk and unhealthy lifestyles in South Africa and in exacerbating inequities in NCD distribution among populations. Through their business practices such as product design, marketing, retail distribution, and pricing and their business practices such as lobbying, public relations, philanthropy, and sponsored research, national and transnational corporations in South Africa shape the social and physical environments that structure opportunities for NCD risk behavior. Since the election of a democratic government in 1994, the South African government and civil society groups have used regulation, public education, health services, and community mobilization to modify corporate practices that increase NCD risk. By expanding the practice of health education to include activities that seek to modify the practices of corporations as well as individuals, South Africa can reduce the growing burden of NCDs.


Hypoxia response and microRNAs: no longer two separate worlds.
MicroRNAs (miRs) are short non-coding transcripts involved in a wide variety of cellular processes. Several recent studies have established a link between hypoxia, a well-documented component of the tumour microenvironment, and specific miRs. One member of this class, miR-210, was identified as hypoxia inducible in all the cell types tested, and is overexpressed in most cancer types. Its hypoxic induction is dependent on a functional hypoxia-inducible factor (HIF), thus extending the transcriptional repertoire of the latter beyond 'classic' genes. From a clinical standpoint, miR-210 overexpression has been associated with adverse prognosis in breast tumours and been detected in serum of lymphoma patients and could serve as a tool to define hypoxic malignancies. We discuss the role of miR-210 and its emerging targets, as well as possible future directions for clinical applications in oncology and ischaemic disorders.


Density profile of blue-sensitive cones along the horizontal meridian of macaque retina.
Intravitreal injection of some fluorescent and nonfluorescent tissue-reactive dyes results in selective intracellular staining of a specific population of cones of macaque retina that have been identified tentatively as blue-sensitive cones. This paper describes quantitative density profiles of these cones as a function of retinal eccentricity. These profiles were measured from 0 deg to about 60 deg eccentricity along the nasal and temporal segments of the horizontal meridian of macaque retina. Stained cones were found to be absent from the very center of the fovea. These cones reach peak densities at 0.75-1.50 deg eccentricity, and decrease with greater eccentricity, more rapidly on the temporal than on the nasal segment of the horizontal meridian. Peak densities were found to be slightly closer to the foveal center of the retinas of adult male than of adult female macaques. Packing patterns of stained and unstained cones are discussed as is the mathematic expression of stained cone distribution. The spatial properties of the retinal distribution of stained cones agree very closely with those obtained in psychophysical human studies and other anatomic simian studies of blue-sensitive cones.


Genetic susceptibility to increased bacterial translocation influences the response to biological therapy in patients with Crohn's disease.
The aetiology of Crohn's disease (CD) has been related to nucleotide-binding oligomerisation domain containing 2 (NOD2) and ATG16L1 gene variants. The observation of bacterial DNA translocation in patients with CD led us to hypothesise that this process may be facilitated in patients with NOD2/ATG16L1-variant genotypes, affecting the efficacy of anti-tumour necrosis factor (TNF) therapies. 179 patients with Crohn's disease were included. CD-related NOD2 and ATG16L1 variants were genotyped. Phagocytic and bactericidal activities were evaluated in blood neutrophils. Bacterial DNA, TNFα, IFNγ, IL-12p40, free serum infliximab/adalimumab levels and antidrug antibodies were measured. Bacterial DNA was found in 44% of patients with active disease versus 23% of patients with remitting disease (p=0.01). A NOD2-variant or ATG16L1-variant genotype was associated with bacterial DNA presence (OR 4.8; 95% CI 1.1 to 13.2; p=0.001; and OR 2.4; 95% CI 1.4 to 4.7; p=0.01, respectively). This OR was 12.6 (95% CI 4.2 to 37.8; p=0.001) for patients with a double-variant genotype. Bacterial DNA was associated with disease activity (OR 2.6; 95% CI 1.3 to 5.4; p=0.005). Single and double-gene variants were not associated with disease activity (p=0.19). Patients with a NOD2-variant genotype showed decreased phagocytic and bactericidal activities in blood neutrophils, increased TNFα levels in response to bacterial DNA and decreased trough levels of free anti-TNFα. The proportion of patients on an intensified biological therapy was significantly higher in the NOD2-variant groups. Our results characterise a subgroup of patients with CD who may require a more aggressive therapy to reduce the extent of inflammation and the risk of relapse.


Adaptive evolution of the FADS gene cluster within Africa.
Long chain polyunsaturated fatty acids (LC-PUFAs) are essential for brain structure, development, and function, and adequate dietary quantities of LC-PUFAs are thought to have been necessary for both brain expansion and the increase in brain complexity observed during modern human evolution. Previous studies conducted in largely European populations suggest that humans have limited capacity to synthesize brain LC-PUFAs such as docosahexaenoic acid (DHA) from plant-based medium chain (MC) PUFAs due to limited desaturase activity. Population-based differences in LC-PUFA levels and their product-to-substrate ratios can, in part, be explained by polymorphisms in the fatty acid desaturase (FADS) gene cluster, which have been associated with increased conversion of MC-PUFAs to LC-PUFAs. Here, we show evidence that these high efficiency converter alleles in the FADS gene cluster were likely driven to near fixation in African populations by positive selection ∼85 kya. We hypothesize that selection at FADS variants, which increase LC-PUFA synthesis from plant-based MC-PUFAs, played an important role in allowing African populations obligatorily tethered to marine sources for LC-PUFAs in isolated geographic regions, to rapidly expand throughout the African continent 60-80 kya.


Predictive value of different glycemic control markers in total hip or knee arthroplasty: A prospective study.
The optimal glycemic control marker before total hip or knee arthroplasty remains inconclusive. Hemoglobin A1c (HbA1c) is widely used, while fructosamine may be valuable for predicting periprosthetic joint infection (PJI). Fructosamine levels can be affected by serum albumin levels; albumin-corrected fructosamine (AlbF) can be calculated to overcome this issue. The objective of this study was to evaluate the predictive value of different markers for complications after primary total hip or knee arthroplasty. This prospective cohort study included 304 patients (mean age: 65 years [range, 16-85), mean follow-up: 32 months (range, 12-49)] who underwent primary total hip or knee arthroplasty between 2018 and 2021. Of them, 156 patients had diabetes. Mean HbA1c was 6.5% (range, 4.8%-13%), fructosamine 244 µmol/L (range, 98-566 µmol/L), and AlbF 632 (range, 238-2308). Patients who did and did not have diabetes were matched 1 : 1. Hemoglobin A1c 7% and fructosamine 292 µmol/L were used as cutoff. Complications were documented. Glycemic markers were compared using logistic regression analyses, with a special focus on PJI. In the logistic regression analyses, HbA1c was strongly associated with total complications [adjusted odds ratio (OR): 3.61; 95% CI, 1.65-7.91, P = .001], while fructosamine was associated with PJI (adjusted OR: 13.68; 95% CI, 1.39-134.89, P = .025). Albumin-corrected fructosamine did not show any additional benefits. Preoperative assessment before total hip or knee arthroplasty must not focus on a single marker; HbA1c is a good predictor of total complications, while fructosamine is a better predictor of PJI. To the best of our knowledge, in its first orthopedic study, AlbF did not show any advantages. Level II, Prognostic Study.


Evaluating the impact of imputations for missing participant outcome data in a network meta-analysis.
In a meta-analysis of trials with missing outcome data, a parameter known as informative missing odds ratio (IMOR) can be used to quantify the relationship between informative missingness and a binary outcome. IMORs also account for the increased uncertainty due to missingness in the meta-analysis results. To extend the idea of IMOR into a network meta-analysis (NMA) setting in order to explore the impact of missing outcome data on the inferences about the relative effectiveness of several competing treatments in psychiatric trials. IMORs were estimated in two datasets comparing anti-manic treatments and antidepressants. The outcome was response to treatments. In the original meta-analyses, missing participants were assumed to have failed regardless the treatment they were allocated to. To evaluate the robustness of this assumption in each dataset, several imputations of the missing outcomes were studied by an IMOR parameter in the NMA model. By comparing the odds ratios for efficacy under the initial analysis and under several assumptions about the missingness, we assessed the consistency of the conclusions. The missing data mechanism was studied by comparing the prior with the posterior IMOR distribution. Models were fitted using Markov chain Monte Carlo (MCMC) in WinBUGS. In both datasets, the relative effectiveness of the treatments seems to be affected only by the two extreme imputation scenarios of worst- and best-case analyses. Moreover, heterogeneity increases in both datasets under these two extreme scenarios. Overall, there is a non-significant change on the ranking of the anti-manic and antidepressant treatments. The posterior and prior IMOR distributions are very similar showing that the data do not provide any information about the true outcome in missing participants. There is a very weak indication that missing participants tend to fail in placebo and paroxetine, while the opposite occurs for sertraline, fluoxetine, and fluvoxamine. Investigation of informative missingness was limited two classes of treatments and for dichotomous outcome measures. The proportion of missing outcomes was very low overall, and hence, the power of detecting any differences in effectiveness estimated under the various imputation methods is small. Sensitivity analysis to account for missing outcome data and their uncertainty in the NMA can be undertaken by extending the idea of IMOR. In two case examples, we found no differences between the various models due to low missing data rate. In line with previous observations, data carry little information about the reason of missingness.


A Newly Isolated Bacillus subtilis Strain Named WS-1 Inhibited Diarrhea and Death Caused by Pathogenic Escherichia coli in Newborn Piglets.
Bacillus subtilis is recognized as a safe and reliable human and animal probiotic and is associated with bioactivities such as production of vitamin and immune stimulation. Additionally, it has great potential to be used as an alternative to antimicrobial drugs, which is significant in the context of antibiotic abuse in food animal production. In this study, we isolated one strain of B. subtilis, named WS-1, from apparently healthy pigs growing with sick cohorts on one Escherichia coli endemic commercial pig farm in Guangdong, China. WS-1 can strongly inhibit the growth of pathogenic E. coli in vitro. The B. subtilis strain WS-1 showed typical Bacillus characteristics by endospore staining, biochemical test, enzyme activity analysis, and 16S rRNA sequence analysis. Genomic analysis showed that the B. subtilis strain WS-1 shares 100% genomic synteny with B. subtilis with a size of 4,088,167 bp. Importantly, inoculation of newborn piglets with 1.5 × 1010 CFU of B. subtilis strain WS-1 by oral feeding was able to clearly inhibit diarrhea (p < 0.05) and death (p < 0.05) caused by pathogenic E. coli in piglets. Furthermore, histopathological results showed that the WS-1 strain could protect small intestine from lesions caused by E. coli infection. Collectively, these findings suggest that the probiotic B. subtilis strain WS-1 acts as a potential biocontrol agent protecting pigs from pathogenic E. coli infection. Importance: In this work, one B. subtilis strain (WS-1) was successfully isolated from apparently healthy pigs growing with sick cohorts on one E. coli endemic commercial pig farm in Guangdong, China. The B. subtilis strain WS-1 was identified to inhibit the growth of pathogenic E. coli both in vitro and in vivo, indicating its potential application in protecting newborn piglets from diarrhea caused by E. coli infections. The isolation and characterization will help better understand this bacterium, and the strain WS-1 can be further explored as an alternative to antimicrobial drugs to protect human and animal health.


Reproductive mode and ovarian morphology regulation in chimeric planarians composed of asexual and sexual neoblasts.
Planarians are comprised of populations with different reproductive strategies: exclusively innately asexual (AS), exclusively innately sexual (InS), and seasonally switching. AS worms can be sexualized experimentally by feeding them with minced InS worms, and the resultant worms are characterized as acquired sexual (AqS). Differences between InS and AqS worms are expected to provide important clues to the poorly understood mechanism underlying the regulation of their reproductive mode. Morphological differences were found between InS and AqS worm ovaries, and we showed that the pluripotent stem cells (neoblasts) from InS worms, but not those of AqS worms, have the capacity to initiate the sexual state autonomously via neoblast fraction transplantation. To compare their reproductive mode and ovarian morphology regulation, InS donor neoblast fractions were transplanted into non-lethally X-ray-irradiated AS recipients. All transplants showed stable chimerism and reproduced sexually, suggesting that InS worm neoblasts can initiate sexual state autonomously, even when coexisting with AS worm neoblasts. The chimeras formed extraordinarily large and supernumerary ovaries equivalent to AqS worms, which were not seen in InS worms, suggesting that regulation of ovarian morphology in AS worm-derived cells in response to endogenous sexualizing stimulation distinctly differs from that of InS worms.


Spatial, Temporal, and Electrical Characteristics of Lightning in Reported Lightning-Initiated Wildfire Events.
Analysis was performed to determine if a lightning flash could be associated with every reported lightning-initiated wildfire that grew to at least 4 km2. In total, 905 lightning-initiated wildfires within CONUS between 2012 and 2015 were analyzed. Fixed and fire radius search methods showed that 81-88% of wildfires had a corresponding lightning flash within a 14 day period prior to the report date. The two methods showed that 52-60% of lightning-initiated wildfire were reported on the same day as the closest lightning flash. The fire radius method indicated the most promising spatial results, where the median distance between the closest lightning and the wildfire start location was 0.83 km, followed by a 75th percentile of 1.6 km, and a 95th percentile of 5.86 km. Ninety percent of the closest lightning flashes to wildfires were negative polarity. Maximum flash densities were less than 0.41 flashes km2 for the 24 hour period at the fire start location. The majority of lightning-initiated holdover events were observed in the Western CONUS, with a peak density in north-central Idaho. A twelve day holdover event from New Mexico was also discussed; outlining the opportunities and limitations of using lightning data to characterize wildfires.


Modeling the interactions between deformable capsules rolling on a compliant surface.
By integrating mesoscale models for hydrodynamics and micromechanics, we examine the fluid-driven motion of pairs of capsules on a compliant, adhesive substrate. The capsules, modeled as fluid filled elastic shells, represent cells or polymeric microcapsules. We show that both the relative and the average velocities of two closely spaced, rolling capsules depends on the elasticity of the capsules, the adhesive interaction between the capsules and the substrate, and the compliance of the substrate. We first focused on a stiff surface and found that pairs of rigid capsules always separate from each other, while for deformable capsules, the dynamic behavior depends critically on the strength of the adhesive interaction. For strong adhesion to the substrate, the capsules again roll away from each other, while for a relatively weak adhesion, the capsules actually approach each other. In the case of soft substrates, any significant deformations of the surface that are caused by the capsules give rise to a force that propels the particles to move rapidly apart. Thus, in the case of strong adhesion between the capsules and the soft substrates, both rigid and flexible capsules are driven to separate. On the other hand, for weak adhesion, the elastic particles approach each other, similar to the behavior on stiff surfaces. These findings reveal that the interactions between the capsules are mediated by the nature of the underlying layer. We can harness this information to design surfaces that actively control the relative separation between the capsules. This could be utilized to regulate the motion of biological cells, as well as polymeric microcapsules, and thus, could prove to be useful in various biological assays or tissue engineering studies.


Protein kinase C-alpha and -epsilon are enriched in growth cones of differentiating SH-SY5Y human neuroblastoma cells.
SH-SY5Y cells differentiate into neuronal-like cells and express marker proteins like growth-associated protein (GAP-43) and neuropeptide tyrosine when treated with a low concentration (16 nM) of the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) in the presence of growth factors or serum. Both control and differentiated cells expressed protein kinase C-alpha (PKC-alpha), PKC-epsilon, and PKC-zeta as revealed by Western blot analyses, but the subcellular distribution of the three isoforms was not uniform, indicating specific localized functions of the enzymes. In growth cones prepared from differentiating cells PKC-alpha and PKC-epsilon were enriched. In contrast, PKC-zeta was more evenly distributed within the differentiating cell. Cells treated with a high concentration of TPA (1.6 microM) differentiate poorly and continue to proliferate. In those cells, PKC-alpha and PKC-epsilon were found to be down-regulated while PKC-zeta remained present. Thus, down-regulation of PKC-alpha and PKC-epsilon appears to be incompatible with neuronal differentiation of SH-SY5Y cells. These cells also differentiate when treated with a combination of basic fibroblast growth factor and insulin-like growth factor I. Growth cones isolated from such cells are also enriched in PKC-alpha and PKC-epsilon, but not in PKC-zeta. Based on the subcellular distribution of PKC-alpha and epsilon, and that PKC substrates like GAP-43 and pp60c-src are enriched in SH-SY5Y growth cones, a role during neurite growth is suggested.


Endogenous low-level expression of the coxsackievirus and adenovirus receptor enables coxsackievirus B3 infection of RD cells.
Cells in which the appropriate viral receptor cannot be detected may paradoxically act as a host to the virus. For example, RD cells are often considered to be non-permissive for infection with coxsackievirus and adenovirus receptor (CAR)-dependent group B coxsackieviruses (CVB), insofar as inoculated cell monolayers show little or no cytopathic effect (CPE) and immunohistological assays for CAR have been consistently negative. Supernatants recovered from RD cells exposed to CVB, however, contained more virus than was added in the initial inoculum, indicating that productive virus replication occurred in the monolayer. When infected with a recombinant CVB type 3 (CVB3) chimeric strain expressing S-Tag within the viral polyprotein, 4-11 % of RD cells expressed S-Tag over 48 h. CAR mRNA was detected in RD cells by RT-PCR, and CAR protein was detected on Western blots of RD lysates; both were detected at much lower levels than in HeLa cells. Receptor blockade by an anti-CAR antibody confirmed that CVB3 infection of RD cells was mediated by CAR. These results show that some RD cells in the culture population express CAR and can thereby be infected by CVB, which explains the replication of CAR-dependent CVB in cell types that show little or no CPE and in which CAR has not previously been detected. Cells within cultures of cell types that have been considered non-permissive may express receptor transiently, leading to persistent replication of virus within the cultured population.


Multiple mechanisms of N-(phosphonoacetyl)-L-aspartate drug resistance in SV40-infected precrisis human fibroblasts.
Normal and SV40-infected human fibroblasts were grown in the presence of the drug N-(phosphonoacetyl)-L-aspartate (PALA) and examined for evidence of genetic instability. Both cell populations were precrisis and showed a normal, diploid karyotype at early passage. In contrast to the normal IMR-90 cells, which showed growth arrest and did not form colonies in PALA, the SV40-infected IMR-90 cells formed colonies at a very high frequency and continued to cycle in the drug. The drug-resistant colonies senesced after continued growth in culture, indicating that this change in ability to amplify preceded immortalization. This is the first observation of mortal human cells overcoming the drug-induced growth arrest. Although all previously isolated PALA-resistant colonies demonstrated CAD gene amplification as the mechanism of the drug-resistant phenotype, these SV40-infected human cells also showed alternative mechanisms, including increases in gene copy number by aneuploidy and formation of an isochromosome 2p.


Field-based physiological testing of wheelchair athletes.
The volume of literature on field-based physiological testing of wheelchair sports, such as basketball, rugby and tennis, is considerably smaller when compared with that available for individuals and team athletes in able-bodied (AB) sports. In analogy to the AB literature, it is recognized that performance in wheelchair sports not only relies on fitness, but also sport-specific skills, experience and technical proficiency. However, in contrast to AB sports, two major components contribute towards 'wheeled sports' performance, which are the athlete and the wheelchair. It is the interaction of these two that enable wheelchair propulsion and the sporting movements required within a given sport. Like any other athlete, participants of wheelchair sports are looking for efficient ways to train and/or analyse their technique and fitness to improve their performance. Consequently, laboratory and/or field-based physiological monitoring tools used at regular intervals at key time points throughout the year must be considered to help with training evaluation. The present review examines methods available in the literature to assess wheelchair sports fitness in a field-based environment, with special attention on outcome variables, validity and reliability issues, and non-physiological influences on performance. It also lays out the context of field-based testing by providing details about the Paralympic court sports and the impacts of a disability on sporting performance. Due to the limited availability of specialized equipment for testing wheelchair-dependent participants in the laboratory, the adoption of field-based testing has become the preferred option by team coaches of wheelchair athletes. An obvious advantage of field-based testing is that large groups of athletes can be tested in less time. Furthermore, athletes are tested in their natural environment (using their normal sports wheelchair set-up and floor surface), potentially making the results of such testing more relevant than laboratory testing. However, given that many tests, such as the multistage fitness test and the Yo-Yo intermittent test, have originally been developed for AB games players, the assumption that these can also be used for wheelchair athletes may be erroneous. With the array of AB aerobic and anaerobic field tests available, it is difficult to ascertain which ones may be best suited for wheelchair athletes. Therefore, new, wheelchair sport-specific tests have been proposed and validated. Careful selection of tests to enable coaches to distinguish between disability classifications, wheelchair proficiency and actual performance improvements is paramount as this will not only enhance the value of field-based testing, but also help with the development of meaningful normative data.


Bak compensated for Bax in p53-null cells to release cytochrome c for the initiation of mitochondrial signaling during Withanolide D-induced apoptosis.
The goal of cancer chemotherapy to induce multi-directional apoptosis as targeting a single pathway is unable to decrease all the downstream effect arises from crosstalk. Present study reports that Withanolide D (WithaD), a steroidal lactone isolated from Withania somnifera, induced cellular apoptosis in which mitochondria and p53 were intricately involved. In MOLT-3 and HCT116p53+/+ cells, WithaD induced crosstalk between intrinsic and extrinsic signaling through Bid, whereas in K562 and HCT116p53-/- cells, only intrinsic pathway was activated where Bid remain unaltered. WithaD showed pronounced activation of p53 in cancer cells. Moreover, lowered apoptogenic effect of HCT116p53-/- over HCT116p53+/+ established a strong correlation between WithaD-mediated apoptosis and p53. WithaD induced Bax and Bak upregulation in HCT116p53+/+, whereas increase only Bak expression in HCT116p53-/- cells, which was coordinated with augmented p53 expression. p53 inhibition substantially reduced Bax level and failed to inhibit Bak upregulation in HCT116p53+/+ cells confirming p53-dependent Bax and p53-independent Bak activation. Additionally, in HCT116p53+/+ cells, combined loss of Bax and Bak (HCT116Bax-Bak-) reduced WithaD-induced apoptosis and completely blocked cytochrome c release whereas single loss of Bax or Bak (HCT116Bax-Bak+/HCT116Bax+Bak-) was only marginally effective after WithaD treatment. In HCT116p53-/- cells, though Bax translocation to mitochondria was abrogated, Bak oligomerization helped the cells to release cytochrome c even before the disruption of mitochondrial membrane potential. WithaD also showed in vitro growth-inhibitory activity against an array of p53 wild type and null cancer cells and K562 xenograft in vivo. Taken together, WithaD elicited apoptosis in malignant cells through Bax/Bak dependent pathway in p53-wild type cells, whereas Bak compensated against loss of Bax in p53-null cells.


A Multiday Evaluation of Real-Time Intramuscular EMG Usability with ANN.
Recent developments in implantable technology, such as high-density recordings, wireless transmission of signals to a prosthetic hand, may pave the way for intramuscular electromyography (iEMG)-based myoelectric control in the future. This study aimed to investigate the real-time control performance of iEMG over time. A novel protocol was developed to quantify the robustness of the real-time performance parameters. Intramuscular wires were used to record EMG signals, which were kept inside the muscles for five consecutive days. Tests were performed on multiple days using Fitts' law. Throughput, completion rate, path efficiency and overshoot were evaluated as performance metrics using three train/test strategies. Each train/test scheme was categorized on the basis of data quantity and the time difference between training and testing data. An artificial neural network (ANN) classifier was trained and tested on (i) data from the same day (WDT), (ii) data collected from the previous day and tested on present-day (BDT) and (iii) trained on all previous days including the present day and tested on present-day (CDT). It was found that the completion rate (91.6 ± 3.6%) of CDT was significantly better (p < 0.01) than BDT (74.02 ± 5.8%) and WDT (88.16 ± 3.6%). For BDT, on average, the first session of each day was significantly better (p < 0.01) than the second and third sessions for completion rate (77.9 ± 14.0%) and path efficiency (88.9 ± 16.9%). Subjects demonstrated the ability to achieve targets successfully with wire electrodes. Results also suggest that time variations in the iEMG signal can be catered by concatenating the data over several days. This scheme can be helpful in attaining stable and robust performance.


In vitro evaluation of herpes simplex virus type 1 thymidine kinase reporter system in dynamic studies of transcriptional gene regulation.
The herpes simplex virus type 1 thymidine kinase (HSV1-TK) reporter system is being used to directly and indirectly monitor therapeutic gene expression, immune cell trafficking and protein-protein interactions in various living animals. However, the issues of HSV1-TK enzyme stability in living cells and whether this reporter system is optimal for dynamic studies of gene expression events in genetic imaging have not be addressed. The purpose of the present study was to evaluate the application of this reporter system in dynamic studies of transcriptional gene regulation. To achieve this purpose, we established two tetracycline-inducible murine sarcoma cell lines, tetracycline-turn-off HSV1-tk-expressing cell line (NG4TL4/tet-off-HSV1-tk) and tetracycline-turn-off Luc-expressing cell line (NG4TL4/tet-off-Luc), to create an artificially regulated gene expression model in vitro. The dynamic transcriptional events mediating a series of doxycycline (Dox) inductions were monitored by HSV1-TK or by the firefly luciferase reporter gene using HSV1-TK enzyme activity assay and luciferase assay, respectively. The results of dynamic gene expression studies showed that the luciferase gene is an optimal reporter gene for monitoring short-timescale, dynamic transcriptional events mediating a series of Dox inductions, whereas the HSV1-tk is not optimal to achieve this purpose. Furthermore, the enzyme half-life of HSV1-TK in NG4TL4 cells is about 35 h after cycloheximide-induced protein inhibition. On the other hand, the results of an efflux assay of [(131)I] FIAU and [(3)H] GCV revealed that the molecular probe phosphorylated by HSV1-TK can be trapped long term within HSV1-TK stably transformed cells. Therefore, a long half-life radionuclide is not suitable for dynamic gene expression studies. Based on these results, we suggest that the HSV1-TK reporter system is not optimal for monitoring short-timescale dynamic processes such as kinetic gene expression controlled by inducible promoters or a less stable protein with a more rapid turnover due to the limitations of the half-life of the HSV1-TK enzyme and the cellular retention time of their phosphorylated molecular probes.


Search for dark matter annihilations in the sun with the 79-string IceCube detector.
We have performed a search for muon neutrinos from dark matter annihilation in the center of the Sun with the 79-string configuration of the IceCube neutrino telescope. For the first time, the DeepCore subarray is included in the analysis, lowering the energy threshold and extending the search to the austral summer. The 317 days of data collected between June 2010 and May 2011 are consistent with the expected background from atmospheric muons and neutrinos. Upper limits are set on the dark matter annihilation rate, with conversions to limits on spin-dependent and spin-independent scattering cross sections of weakly interacting massive particles (WIMPs) on protons, for WIMP masses in the range 20-5000 GeV/c2. These are the most stringent spin-dependent WIMP-proton cross section limits to date above 35 GeV/c2 for most WIMP models.


Prevalence of HIV, HBV and HCV among street and labour children in Tehran, Iran.
The existence of street and working children in Iran is undeniable. The precarious conditions of these children (including disrupted family, poverty, high prevalence of crime among relatives, family members and peers) cause social harm and high-risk behaviours, including drug addiction, selling sex or having sex with adolescents or peers. Here we explore the HIV, hepatitis B and hepatitis C status of street and working children in Tehran. One thousand street and labour children, aged 10-18 years, were recruited by using the time-location sampling method, and semistructured questionnaires were used to find demographic information and information on HIV/AIDS-related high-risk sexual behaviours. Blood samples were collected from children, with use of the dried blood sampling method. 4.5% of children were HIV infected, 1.7% were infected with hepatitis B virus and 2.6% were infected with hepatitis C virus (HCV). Having parents who used drug, infected with HCV and having experience in trading sex significantly increased the likelihood of getting HIV among the street children of Tehran. HIV prevalence among street children is much higher than general population (<0.1%), and in fact ,the rate of positivity comes close to that among female sex workers in Iran. These findings must be an alarm for HIV policymakers to consider immediate and special interventions for this at-risk group.


Anxiety in first year medical students taking gross anatomy.
To study anxiety levels in first-year medical students taking gross anatomy. Thirty medical students per year, for 2 years, completed the Beck Anxiety Inventory (BAI) 10 times during a 13-week gross anatomy course. In addition, behavioral observations were made by a psychiatrist during gross anatomy for demonstrations of assertive, destructive, neutral, or passive behavior. Additional qualitative outcome measures were group exit interviews with the faculty and students. The mean BAI for all 60 students per year, for 2 years, was 2.19 ± 3.76, 93% of the scores indicated minimal anxiety, and 89% of BAI values were less than five which confirmed a minimal level of anxiety. The low level of reported BAI contrasted sharply with verbal reports by the same students and face-to-face exit interviews with the psychiatrist. Symptoms of stress and anxiety emerged as a result of these conversations. The high levels of subjective stress and anxiety revealed by the interviews were unknown to the gross anatomy faculty. The low scores of students on the BAI's stand in sharp contrast to the BAI's reported for medical students in other published reports. Although it is possible that our students were truthfully devoid of anxiety, it is more likely that our students were denying even minimal anxiety levels. There have been reports that medical students feel that admitting stress, depression, or anxiety put their competitiveness for a residency at risk. We conclude that students may be in frank denial of experiencing anxiety and, if so, this behavior is not conducive to good mental health.