Feminisation of the health workforce and wage conditions of health professions: an exploratory analysis.
The feminisation of the global health workforce presents a unique challenge for human resource policy and health sector reform which requires an explicit gender focus. Relatively little is known about changes in the gender composition of the health workforce and its impact on drivers of global health workforce dynamics such as wage conditions. In this article, we use a gender analysis to explore if the feminisation of the global health workforce leads to a deterioration of wage conditions in health. We performed an exploratory, time series analysis of gender disaggregated WageIndicator data. We explored global gender trends, wage gaps and wage conditions over time in selected health occupations. We analysed a sample of 25 countries over 9 years between 2006 and 2014, containing data from 970,894 individuals, with 79,633 participants working in health occupations (48,282 of which reported wage data). We reported by year, country income level and health occupation grouping. The health workforce is feminising, particularly in lower- and upper-middle-income countries. This was associated with a wage gap for women of 26 to 36% less than men, which increased over time. In lower- and upper-middle-income countries, an increasing proportion of women in the health workforce was associated with an increasing gender wage gap and decreasing wage conditions. The gender wage gap was pronounced in both clinical and allied health professions and over lower-middle-, upper-middle- and high-income countries, although the largest gender wage gaps were seen in allied healthcare occupations in lower-middle-income countries. These results, if a true reflection of the global health workforce, have significant implications for health policy and planning and highlight tensions between current, purely economic, framing of health workforce dynamics and the need for more extensive gender analysis. They also highlight the value of a more nuanced approach to health workforce planning that is gender sensitive, specific to countries' levels of development, and considers specific health occupations.


Phase-II study of recombinant alpha 2-interferon in advanced malignant melanoma.
Fifteen patients with metastatic malignant melanoma, including 10 who had not previously received systemic therapy, were treated with recombinant alpha2-interferon (IFN-alpha 2) in a dose of 20 million IU/m2 by 30-min i.v. infusion daily for 5 days each 14 days. Evaluable metastatic sites included lung, subcutaneous tissue, liver, nodes, adrenals, and bone. Subjective toxicity was generally mild to moderate, with fever (38.2-40.2 degrees C), occasional rigors, fatigue, myalgia, headache, and nausea. Objective toxicity included transient neutropenia and elevation of hepatic enzymes, particularly gamma-glutamyl transpeptidase. In 1 of the 10 patients receiving more than one cycle, IFN dosage was reduced because of toxicity, but later reescalated. All patients were evaluated for response. No overall partial or complete responses were observed, but two site responses (lung and subcutaneous tissue) were seen. Median survival from start of IFN treatment was 19 weeks. High doses of IFN were reasonably well tolerated in this study, but the results suggest little activity against malignant melanoma.


Virologic models of chronic relapsing demyelinating disease.
The present report, compares two murine models of virus induced chronic relapsing demyelination. MHV-induced demyelination in the BALB/c mouse results from the direct virus mediated cytolysis of oligodendrocytes. Extensive remyelination by oligodendrocytes is noted. Recurrent demyelination occurs in small areas. Infectious virus persists and viral antigens are localized within oligodendrocytes and their processes. TMEV-induced demyelination in SJL/J mice is associated with perivascular inflammatory infiltrates and is diminished by immunosuppressive measures. Remyelination by oligodendrocytes is delayed and incomplete. Chronic demyelination is widespread and associated with perivascular inflammatory infiltrates. The virus persists and viral antigen is localized within oligodendrocytes. The findings indicate virus persistence in oligodendrocytes in both models. Demyelination follows the disintegration of infected oligodendrocytes. Virus replication in oligodendrocytes is responsible for cell lysis in the MHV model whereas immune mediated injury of infected oligodendrocytes is considered to play a role in the pathogenesis of demyelination in the TMEV model.


Clinical and histopathologic features of recurrent vestibular schwannoma (acoustic neuroma) after stereotactic radiosurgery.
Stereotactic radiosurgery for vestibular schwannoma entails uncertain long-term risk of tumor recurrence and delayed cranial neuropathies. In addition, the underlying histopathologic changes to the tumor bed are not fully characterized. We seek to understand the clinical and histologic features of recurrent vestibular schwannoma after stereotactic radiation therapy. Retrospective review. Tertiary referral center. Four patients who underwent microsurgical resection of vestibular schwannoma after primary stereotactic radiation therapy. Patients were treated primarily with gamma knife radiosurgery or fractionated stereotactic radiotherapy followed by salvage microsurgery. Retrosigmoid craniotomy was used in all cases. Histopathologic review. Preoperative and postoperative facial nerve function was assessed with the House-Brackmann scale. We observed highly inconsistent radiation changes in the cerebellopontine angle and internal auditory canal. Fibrosis outside and within the tumor bed varied markedly, complicating microsurgical dissection. Light microscopy confirmed the presence of viable tumor in all cases. Histopathologic features were typical of vestibular schwannoma, and there was no significant scarring that could be attributed to radiation effect. The variable fibrosis in the cerebellopontine angle and lack of radiation changes seen histopathologically in irradiated vestibular schwannoma suggest that a uniform treatment effect was not achieved in these cases. Although all four patients with preoperative cranial neuropathies were found intraoperatively to have fibrosis in the cerebellopontine angle, excellent preservation of facial nerve anatomy and function was possible with salvage microsurgical resection. Additional analyses are needed to clarify the histopathologic and molecular characteristics associated with vestibular schwannoma growth after stereotactic radiation.


Potential syntrophic relationship between coral-associated Prosthecochloris and its companion sulfate-reducing bacterium unveiled by genomic analysis.
Endolithic microbial symbionts in the coral skeleton may play a pivotal role in maintaining coral health. However, compared to aerobic micro-organisms, research on the roles of endolithic anaerobic micro-organisms and microbe-microbe interactions in the coral skeleton are still in their infancy. In our previous study, we showed that a group of coral-associated Prosthecochloris (CAP), a genus of anaerobic green sulphur bacteria, was dominant in the skeleton of the coral Isopora palifera. Though CAP is diverse, the 16S rRNA phylogeny presents it as a distinct clade separate from other free-living Prosthecochloris. In this study, we build on previous research and further characterize the genomic and metabolic traits of CAP by recovering two new high-quality CAP genomes - Candidatus Prosthecochloris isoporae and Candidatus Prosthecochloris sp. N1 - from the coral I. palifera endolithic cultures. Genomic analysis revealed that these two CAP genomes have high genomic similarities compared with other Prosthecochloris and harbour several CAP-unique genes. Interestingly, different CAP species harbour various pigment synthesis and sulphur metabolism genes, indicating that individual CAPs can adapt to a diversity of coral microenvironments. A novel high-quality genome of sulfate-reducing bacterium (SRB)- Candidatus Halodesulfovibrio lyudaonia - was also recovered from the same culture. The fact that CAP and various SRB co-exist in coral endolithic cultures and coral skeleton highlights the importance of SRB in the coral endolithic community. Based on functional genomic analysis of Ca. P. sp. N1, Ca. P. isoporae and Ca. H. lyudaonia, we also propose a syntrophic relationship between the SRB and CAP in the coral skeleton.


Altered squat jumping mechanics after specific exercise.
This study examined squat jumping (SJ) mechanics following length restricted strength training regimens of the knee extensors and flexors. SJ from a 110 degrees knee joint angle starting position and isometric moment-knee angle relations of knee extensors and flexors were analysed in 16 athletes before and directly after 8 weeks of strength training regimens that were restricted to knee joint angles corresponding to long muscle-tendon unit (MTU) length for the knee extensors and flexors. SJ mechanics were studied using a two-dimensional kinematic model with three rigid bodies (upper leg, lower leg, foot) in combination with force plates measurements of ground reaction force in the right and left lower extremity. Centre of mass jumping height significantly (p<0.05) increased post training, but this was not explained by enhanced absolute power generation in the knee joint. However, post training small but significant (p<0.05) shifts to smaller knee joint angles occurred in the normalized [% Max.] knee joint angle dependent power generation in the right and left knee joint during SJ. The isometric moment-knee angle relation of the knee extensors was also significantly (p<0.05) shifted to longer MTU lengths of knee extensors. Length restricted strength training may alter the mechanical situation during both isometric contractions and dynamic athletic movements.


Role for B-cell adapter for PI3K (BCAP) as a signaling adapter linking Toll-like receptors (TLRs) to serine/threonine kinases PI3K/Akt.
Toll like receptors (TLRs) use Toll-IL-1 receptor (TIR) domain-containing adapters, such as myeloid differentiation primary response gene 88 (MyD88) and TIR domain-containing adapter inducing IFN-β (TRIF), to induce activation of transcription factors, including NF-κB, MAP kinases, and IFN regulatory factors. TLR signaling also leads to activation of PI3K, but the molecular mechanism is not understood. Here we have discovered a unique role for B-cell adapter for PI3K (BCAP) in the TLR-signaling pathway. We find that BCAP has a functional N-terminal TIR homology domain and links TLR signaling to activation of PI3K. In addition, BCAP negatively regulates proinflammatory cytokine secretion upon TLR stimulation. In vivo, the absence of BCAP leads to exaggerated recruitment of inflammatory myeloid cells following infections and enhanced susceptibility to dextran sulfate sodium-induced colitis. Our results demonstrate that BCAP is a unique TIR domain-containing TLR signaling adapter crucial for linking TLRs to PI3K activation and regulating the inflammatory response.


Pharmacological interventions for alcoholic liver disease (alcohol-related liver disease): an attempted network meta-analysis.
Alcohol-related liver disease is due to excessive alcohol consumption. It includes a spectrum of liver diseases such as alcohol-related fatty liver, alcoholic hepatitis, and alcoholic cirrhosis. Mortality associated with alcoholic hepatitis is high. The optimal pharmacological treatment of alcoholic hepatitis and other alcohol-related liver disease remains controversial. To assess the comparative benefits and harms of different pharmacological interventions in the management of alcohol-related liver disease through a network meta-analysis and to generate rankings of the available pharmacological interventions according to their safety and efficacy in order to identify potential treatments. However, even in the subgroup of participants when the potential effect modifiers appeared reasonably similar across comparisons, there was evidence of inconsistency by one or more methods of assessment of inconsistency. Therefore, we did not report the results of the network meta-analysis and reported the comparative benefits and harms of different interventions using standard Cochrane methodology. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, Embase, Science Citation Index Expanded, World Health Organization International Clinical Trials Registry Platform and randomised controlled trials registers until February 2017 to identify randomised clinical trials on pharmacological treatments for alcohol-related liver diseases. Randomised clinical trials (irrespective of language, blinding, or publication status) including participants with alcohol-related liver disease. We excluded trials that included participants who had previously undergone liver transplantation and those with co-existing chronic viral diseases. We considered any of the various pharmacological interventions compared with each other or with placebo or no intervention. Two review authors independently identified trials and independently extracted data. We calculated the odds ratio (OR) and rate ratio with 95% confidence intervals (CIs) using both fixed-effect and random-effects models based on available-participant analysis with Review Manager. We assessed risk of bias according to Cochrane, controlled risk of random errors with Trial Sequential Analysis, and assessed the quality of the evidence using GRADE. We identified a total of 81 randomised clinical trials. All the trials were at high risk of bias, and the overall quality of the evidence was low or very low for all outcomes. Alcoholic hepatitisFifty randomised clinical trials included 4484 participants with alcoholic hepatitis. The period of follow-up ranged from one to 12 months. Because of concerns about transitivity assumption, we did not perform the network meta-analysis. None of the active interventions showed any improvement in any of the clinical outcomes reported in the trials, which includes mortality (at various time points), cirrhosis, decompensated cirrhosis, liver transplantation. None of the trials reported health-related quality of life or incidence of hepatocellular carcinoma. Severe alcoholic hepatitisOf the trials on alcoholic hepatitis, 19 trials (2545 participants) included exclusively participants with severe alcoholic hepatitis (Maddrey Discriminat Function > 32). The period of follow-up ranged from one to 12 months. There was no alteration in the conclusions when only people with severe alcoholic hepatitis were included in the analysis. Eleven trials were funded by parties with vested interest in the results. Sixteen trials were funded by parties without vested interest in the results. The source of funding was not reported in 23 trials. Other alcohol-related liver diseasesThirty-one randomised clinical trials included 3695 participants with other alcohol-related liver diseases (with a wide spectrum of alcohol-related liver diseases). The period of follow-up ranged from one to 48 months. The mortality at maximal follow-up was lower in the propylthiouracil group versus the no intervention group (OR 0.45, 95% CI 0.26 to 0.78; 423 participants; 2 trials; low-quality evidence). However, this result is based on two small trials at high risk of bias and further confirmation in larger trials of low risk of bias is necessary to recommend propylthiouracil routinely in people with other alcohol-related liver diseases. The mortality at maximal follow-up was higher in the ursodeoxycholic acid group versus the no intervention group (OR 2.09, 95% CI 1.12 to 3.90; 226 participants; 1 trial; low-quality evidence). Twelve trials were funded by parties with vested interest in the results. Three trials were funded by parties without vested interest in the results. The source of funding was not reported in 16 trials. Because of very low-quality evidence, there is uncertainty in the effectiveness of any pharmacological intervention versus no intervention in people with alcoholic hepatitis or severe alcoholic hepatitis. Based on low-quality evidence, propylthiouracil may decrease mortality in people with other alcohol-related liver diseases. However, these results must be confirmed by adequately powered trials with low risk of bias before propylthiouracil can be considered effective.Future randomised clinical trials should be conducted with approximately 200 participants in each group and follow-up of one to two years in order to compare the benefits and harms of different treatments in people with alcoholic hepatitis. Randomised clinical trials should include health-related quality of life and report serious adverse events separately from adverse events. Future randomised clinical trials should have a low risk of bias and low risk of random errors.


SECIS-binding protein 2 interacts with the SMN complex and the methylosome for selenoprotein mRNP assembly and translation.
Selenoprotein synthesis requires the co-translational recoding of a UGASec codon. This process involves an RNA structural element, called Selenocysteine Insertion Sequence (SECIS) and the SECIS binding protein 2 (SBP2). Several selenoprotein mRNAs undergo unusual cap hypermethylation by the trimethylguanosine synthase 1 (Tgs1), which is recruited by the ubiquitous Survival of MotoNeurons (SMN) protein. SMN, the protein involved in spinal muscular atrophy, is part of a chaperone complex that collaborates with the methylosome for RNP assembly. Here, we analyze the role of individual SMN and methylosome components in selenoprotein mRNP assembly and translation. We show that SBP2 interacts directly with four proteins of the SMN complex and the methylosome core proteins. Nevertheless, SBP2 is not a methylation substrate of the methylosome. We found that both SMN and methylosome complexes are required for efficient translation of the selenoprotein GPx1 in vivo. We establish that the steady-state level of several selenoprotein mRNAs, major regulators of oxidative stress damage in neurons, is specifically reduced in the spinal cord of SMN-deficient mice and that cap hypermethylation of GPx1 mRNA is affected. Altogether we identified a new function of the SMN complex and the methylosome in selenoprotein mRNP assembly and expression.


Multiple components of lateral posterior parietal activation associated with cognitive set shifting.
Posterior parietal activation has commonly been observed in previous neuroimaging studies in association with flexible shifting of cognitive set. However, it is not clear whether the parietal activation reflects cognitive processes intrinsic to the shifting itself or other confounding factors such as spatial attention. To address this issue, the Wisconsin Card Sorting Task (WCST) was modified such that spatial components were eliminated from the sensory and motor aspects of the task. Moreover, a visual instruction of a next dimension was introduced to eliminate cognitive processes related to trial and error identification of a next rule, and a control null-instruction was also introduced to eliminate perceptual/oddball effects of the instruction cue. Localizer scans using a visually guided saccade task were also conducted to identify eye movement/spatial attention-related areas. Activity related to set shifting with trial and error was revealed in the lateral parts of the intraparietal regions, while activity related to eye movements/spatial attention was revealed in the medial parts of the intraparietal regions, confirming little spatial contribution to the modified WCST as indexed by the double dissociation. The lateral intraparietal activity was bilateral, but when the instructed shifting was contrasted with the null-instructed shifting to purify the shift-related activity further, the left intraparietal activation was significantly greater than that in the right hemisphere. These results reveal the left hemisphere dominance of purified shifting-related activity in the lateral posterior parietal cortex that may cooperate with the lateral prefrontal cortex whose left hemisphere dominance has already been reported.


A systematic review of the reliability and validity of discrete choice experiments in valuing non-market environmental goods.
While discrete choice experiments (DCEs) are increasingly used in the field of environmental valuation, they remain controversial because of their hypothetical nature and the contested reliability and validity of their results. We systematically reviewed evidence on the validity and reliability of environmental DCEs from the past thirteen years (Jan 2003-February 2016). 107 articles met our inclusion criteria. These studies provide limited and mixed evidence of the reliability and validity of DCE. Valuation results were susceptible to small changes in survey design in 45% of outcomes reporting reliability measures. DCE results were generally consistent with those of other stated preference techniques (convergent validity), but hypothetical bias was common. Evidence supporting theoretical validity (consistency with assumptions of rational choice theory) was limited. In content validity tests, 2-90% of respondents protested against a feature of the survey, and a considerable proportion found DCEs to be incomprehensible or inconsequential (17-40% and 10-62% respectively). DCE remains useful for non-market valuation, but its results should be used with caution. Given the sparse and inconclusive evidence base, we recommend that tests of reliability and validity are more routinely integrated into DCE studies and suggest how this might be achieved.


Clinical impact and a prognostic marker of early rituximab treatment after rituximab reimbursement in Korean pemphigus patients.
Pemphigus is an autoimmune mucocutaneous blistering disease caused by autoantibodies against desmogleins. Rituximab effectively treats pemphigus by inducing remission and rapidly reducing corticosteroid dosage. In Korea, the high cost of rituximab had been a burden until the National Health Insurance began to cover 90% of rituximab costs via reimbursement for severe pemphigus patients. We analyzed 214 patients with pemphigus who were treated with their first round of rituximab. The time to initiate rituximab and the time to partial remission under minimal therapy (PRMT) were both significantly shorter after the rituximab reimbursement policy. The total steroid intake for PRMT and complete remission (CR) was less in patients who were diagnosed after the reimbursement. The interrupted time series (ITS) model, a novel analysis method to evaluate the effects of an intervention, showed a decrease in total systemic corticosteroid intake until PRMT after reimbursement began. In peripheral blood mononuclear cells from patients with pemphigus vulgaris, the relative frequencies of desmoglein 3-specific CD11c+CD27-IgD- atypical memory B cells positively correlated with the periods from disease onset to rituximab treatment and to PRMT and the total systemic corticosteroid intake until PRMT. We found that early rituximab therapy, induced by the reimbursement policy, shortened the disease course and reduced the total corticosteroid use by pemphigus patients. The decreased frequency of circulating desmoglein-specific atypical memory B cells can be used as a surrogate marker for a good prognosis after rituximab.


[Oxidative-reductive activity of the mycelium as a criterion for assessing inoculate quality in gentamycin biosynthesis].
The development of the seed mycelium at various stages and temperatures was studied with the method of methylene blue decolorization characterizing the respiration level of the culture and its oxidation-reduction (dehydrogenase) activity. Relationship between the time of methylene blue decolorization by the seed mycelium, the quantity of the inoculum transferred from one stage to the other and the temperature of cultivation was shown. The seed mycelium decolorizing methylene blue within 2 minutes was most productive at the biosynthetic stage irrespective of the growth period, biomass volume, cultivation temperature and volume of the inoculum transferred from one stage to the other.


Stressors, Repetitive Negative Thinking, and Insomnia Symptoms in Adolescents Beginning High School.
Although the first year of high school may represent a particularly stressful time for adolescents, no research addresses how stressors are related to insomnia symptoms during this time. Thus, we examined how stress relates to concurrent and prospective insomnia symptoms in adolescents beginning high school (Aim 1). Additionally, we assessed repetitive negative thinking (RNT) as a mediator (Aim 2). We also evaluated whether the pattern of associations differed for boys and girls (Aim 3). Adolescents (N = 502; M age = 14.22 years; 58.2% girls; 91.2% Hispanic/Latinx) completed questionnaires about stressors related to beginning high school (e.g., school performance, peer pressure), family support, RNT, and insomnia symptoms at the beginning and end of their first year of high school. Multiple group structural equation models assessed relationships between these variables and evaluated differences between boys and girls. School/leisure conflict and low family support were directly associated with insomnia symptoms at both times, and RNT mediated these relationships in both boys and girls. In girls, peer pressure and low family support were indirectly associated with Time 1 and Time 2 insomnia symptoms via RNT. In boys, school performance was indirectly associated with Time 1 and Time 2 insomnia symptoms via RNT. Stressful experiences at the beginning of high school negatively affect sleep in adolescents both in the short and long term. Pediatric psychologists should educate adolescents and their parents about the risk of sleep problems during this time period and provide strategies for stress management and for proper sleep hygiene.


Functional lacrimal gland regeneration by transplantation of a bioengineered organ germ.
The lacrimal gland has a multifaceted role in maintaining a homeostatic microenvironment for a healthy ocular surface via tear secretion. Dry-eye disease, which is caused by lacrimal gland dysfunction, is one of the most prevalent eye diseases that cause corneal epithelial damage and results in significant loss of vision and a reduction in the quality of life. Here we demonstrate orthotopic transplantation of bioengineered lacrimal gland germs into adult mice with an extra-orbital lacrimal gland defect, a mouse model that mimics the corneal epithelial damage caused by lacrimal gland dysfunction. The bioengineered lacrimal gland germs and harderian gland germs both develop in vivo and achieve sufficient physiological functionality, including tear production in response to nervous stimulation and ocular surface protection. This study demonstrates the potential for bioengineered organ replacement to functionally restore the lacrimal gland.


Topology-regulated nanocatalysts for ferroptosis-mediated cancer phototherapy.
Ferroptosis-mediated tumor treatment is constrained by the absence of single-component, activatable multifunctional inducers. Given this, a topological synthesis strategy is employed to develop an efficient bismuth-based semiconductor nano-photocatalyst (Bi2O3:S) for tumor ferroptosis therapy. Photo-excited electrons can participate in the reduction reaction to produce harmful reactive oxygen species (ROS) when exposed to near-infrared light. Meanwhile, photo-excited holes can contribute to the oxidation reaction to utilize extra glutathione (GSH) in tumors. In the acidic tumor microenvironment, bismuth ions generated from Bi2O3:S may further cooperate with GSH to amplify oxidative stress damage and achieve biodegradation. Both promote ferroptosis by downregulating glutathione peroxidase 4 (GPX4) expression. Besides, sulfur doping optimizes its near-infrared light-induced photothermal conversion efficiency, benefiting its therapeutic effect. Thus, bismuth ions and holes synergistically drive photo-activable ferroptosis in this nanoplatform, opening up new avenues for tumor therapy.


The influence of posture on parasternal intercostal muscle activity in healthy young adults.
Parasternal intercostal muscle electromyography (EMGpara) has been used as an index of respiratory load in health and disease. While reference values are available, such data have been obtained with subjects in the seated position only. The objective of the current study was to determine the influence of posture on measurements of EMGpara. Fifty-one healthy adult participants underwent measurement of EMGpara, respiratory flow and volume in the seated, reclined at 45°, and supine positions. Resting peak EMGpara activity per breath was determined and expressed both as the raw signal and normalised to that obtained during a maximum inspiratory effort (EMGpara%max). Neural respiratory drive index (NRDI, the product of EMGpara%max and respiratory rate) and neuroventilatory efficiency (NVE, tidal volume divided by EMGpara) were also calculated. No significant differences were observed in raw EMGpara, EMGpara%max, NRDI, NVE or tidal volume from the seated to reclined or supine positions. Respiratory rate and minute ventilation were significantly lower in the supine position compared to seated (p = 0.0043 and 0.0266 respectively). Poor agreement was observed between raw EMGpara and EMGpara%max, likely due to submaximal efforts or cross-talk from adjacent musculature during the maximal manoeuvres. Agreement was notably poorer in the supine posture. Posture does not have a significant effect on EMGpara activity, suggesting that measurements can be made in the reclined or supine position if required or requested by the participant. Normalising the EMGpara signal to a maximal respiratory effort may give unreliable estimates of respiratory load.


Infant Walker-Related Injuries in the United States.
To investigate the epidemiologic characteristics of infant walker-related injuries among children <15 months old who were treated in US emergency departments and to evaluate the effect of the 2010 federal mandatory safety standard on these injuries. National Electronic Injury Surveillance System data from 1990 to 2014 were analyzed. An estimated 230 676 children <15 months old were treated for infant walker-related injuries in US emergency departments from 1990 to 2014. Most of the children sustained head or neck injuries (90.6%) and 74.1% were injured by falling down the stairs in an infant walker. Among patients who were admitted to the hospital (4.5%), 37.8% had a skull fracture. From 1990 to 2003, overall infant walker-related injuries and injuries related to falling down the stairs decreased by 84.5% and 91.0%, respectively. The average annual number of injuries decreased by 22.7% (P = .019) during the 4-year period after the implementation of the federal mandatory safety standard compared with the 4-year period before the standard. Infant walker-related injuries decreased after the implementation of the federal mandatory safety standard in 2010. This decrease may, in part, be attributable to the standard as well as other factors, such as decreased infant walker use and fewer older infant walkers in homes. Despite the decline in injuries, infant walkers remain an important and preventable source of injury among young children, which supports the American Academy of Pediatrics' call for a ban on their manufacture and sale in the United States.


Continuing or adding IL-2 in patients treated with antiretroviral therapy (ACTG Protocol A5051, a rollover trial of ACTG Protocol A328).
Effective antiretroviral therapy reduces HIV-1 RNA levels, improves CD4 T-cell counts, and lowers the risk of opportunistic infections and malignancies. Interleukin-2 (IL-2) has been shown to increase CD4 T-cell numbers mainly by expanding CD4 cells and by prolonging their half-lives. HIV-infected patients previously enrolled into A328 had been randomized to antiretroviral therapy (ART) alone or ART followed by IL-2. In A5051, 53 patients from A328 who had previously received IL-2 were allowed to continue IL-2 for an additional 80 weeks; 27 patients who had received ART alone received IL-2 for 80 weeks. The patients previously receiving IL-2 continued to have elevated CD4 levels with extended use of IL-2. The prior ART-alone recipients had increases in CD4 levels to comparable levels as the prior IL-2 recipients (median 804 versus 847 cells/mm3 at week 72; 60% versus 9% had >50% increase in A5051 to week 72, p < 0.001). Those who had previously received IL-2 required fewer IL-2 cycles to maintain their CD4 T-cell counts compared to those newly initiating IL-2. The treatments were well tolerated with no significant differences in toxicity or discontinuations between those newly versus previously receiving IL-2. There were few clinical events observed. Although sustained CD4 T-cell count increases were seen with IL-2 administration as in other studies, the absence of clinical benefit in two recent randomized trials has demonstrated no apparent role for IL-2 as a therapy in HIV disease. A5051 ClinicalTrials.gov Identifier: NCT00000923.


Cushing's syndrome secondary to ectopic ACTH secretion from metastatic breast carcinoma.
Breast carcinoma is a rare cause of ectopic ACTH syndrome. There are only two previously reported cases in which ACTH secretion is documented. We describe the case of a 56-year-old woman who presented with clinical and biochemical features of ectopic ACTH syndrome in the setting of metastatic breast carcinoma. Despite aggressive management of her ectopic ACTH syndrome, her course was complicated by opportunistic infection, respiratory failure and death. Immunostaining of the breast metastases for ACTH was positive and in situ hybridization revealed proopiomelanocortin gene expression. This is the first reported case of ectopic ACTH syndrome associated with metastatic breast cancer in which the technique of in situ hybridization has been used to confirm the breast cancer metastases as the source of ectopic ACTH secretion.


Ligand-dependent interactions of coactivators steroid receptor coactivator-1 and peroxisome proliferator-activated receptor binding protein with nuclear hormone receptors can be imaged in live cells and are required for transcription.
Members of the nuclear receptor superfamily are thought to activate transcription by recruitment of one or more recently identified coactivator complexes. Here we demonstrate that both peroxisome proliferator-activated receptor binding protein (PBP) and steroid receptor coactivator-1 (SRC-1) are required for ligand-dependent transcription of transiently transfected and chromosomally integrated reporter genes by the estrogen receptor (ER) and retinoic acid receptor (RAR). To examine ligand-dependent interactions between nuclear receptors and specific coactivators in living cells, these proteins were tagged with cyan (CFP) and yellow (YFP) mutants of the green fluorescent protein. Fluorescence resonance energy transfer (FRET) from the CFP to the YFP indicated interaction between the receptor and coactivator. CFP fusions to RAR or its ligand-binding domain exhibited rapid ligand-dependent FRET to YFP-tagged nuclear receptor interaction domains of the coactivators SRC-1 and PBP. The ER-ligand-binding domain, unlike RAR, also exhibited some basal interaction with coactivators in unstimulated cells that was abolished by the receptor antagonists tamoxifen or ICI182,780. Inhibition of FRET by tamoxifen but not ICI182,780 could be reversed by estradiol, whereas estradiol-enhanced FRET could not be inhibited by either antagonist, indicating that ligand effects can show varying degrees of hysteresis. These findings suggest that ligand-dependent transcriptional activities of the RAR and ER require concurrent or sequential recruitment of SRC-1 and PBP-containing coactivator complexes.


Factors associated with the spatial heterogeneity of the first wave of COVID-19 in France: a nationwide geo-epidemiological study.
The objective of this study was to better understand the factors associated with the heterogeneity of in-hospital COVID-19 morbidity and mortality across France, one of the countries most affected by COVID-19 in the early months of the pandemic. This geo-epidemiological analysis was based on data publicly available on government and administration websites for the 96 administrative departments of metropolitan France between March 19 and May 11, 2020, including Public Health France, the Regional Health Agencies, the French national statistics institute, and the Ministry of Health. Using hierarchical ascendant classification on principal component analysis of multidimensional variables, and multivariate analyses with generalised additive models, we assessed the associations between several factors (spatiotemporal spread of the epidemic between Feb 7 and March 17, 2020, the national lockdown, demographic population structure, baseline intensive care capacities, baseline population health and health-care services, new chloroquine and hydroxychloroquine dispensations, economic indicators, degree of urbanisation, and climate profile) and in-hospital COVID-19 incidence, mortality, and case fatality rates. Incidence rate was defined as the cumulative number of in-hospital COVID-19 cases per 100 000 inhabitants, mortality rate as the cumulative number of in-hospital COVID-19 deaths per 100 000, and case fatality rate as the cumulative number of in-hospital COVID-19 deaths per cumulative number of in-hospital COVID-19 cases. From March 19 to May 11, 2020, hospitals in metropolitan France notified a total of 100 988 COVID-19 cases, including 16 597 people who were admitted to intensive care and 17 062 deaths. There was an overall cumulative in-hospital incidence rate of 155·6 cases per 100 000 inhabitants (range 19·4-489·5), in-hospital mortality rate of 26·3 deaths per 100 000 (1·1-119·2), and in-hospital case fatality rate of 16·9% (4·8-26·2). We found clear spatial heterogeneity of in-hospital COVID-19 incidence and mortality rates, following the spread of the epidemic. After multivariate adjustment, the delay between the first COVID-19-associated death and the onset of the national lockdown was positively associated with in-hospital incidence (adjusted standardised incidence ratio 1·02, 95% CI 1·01-1·04), mortality (adjusted standardised mortality ratio 1·04, 1·02-1·06), and case fatality rates (adjusted standardised fatality ratio 1·01, 1·01-1·02). Mortality and case fatality rates were higher in departments with older populations (adjusted standardised ratio for populations with a high proportion older than aged >85 years 2·17 [95% CI 1·20-3·90] for mortality and 1·43 [1·08-1·88] for case fatality rate). Mortality rate was also associated with incidence rate (1·0004, 1·0002-1·001), but mortality and case fatality rates did not appear to be associated with baseline intensive care capacities. We found no association between climate and in-hospital COVID-19 incidence, or between economic indicators and in-hospital COVID-19 incidence or mortality rates. This ecological study highlights the impact of the epidemic spread, national lockdown, and reactive adaptation of intensive care capacities on the spatial distribution of COVID-19 morbidity and mortality. It provides information for future geo-epidemiological analyses and has implications for preparedness and response policies to current and future epidemic waves in France and elsewhere. None.


CD3ζ-chain expression of human T lymphocytes is regulated by TNF via Src-like adaptor protein-dependent proteasomal degradation.
Decreased expression of the TCR ζ-chain has been reported in several autoimmune, inflammatory, and malignant diseases, suggesting that ζ-chain downregulation is common at sites of chronic inflammation. Although ζ-chain is critically important in T lymphocyte activation, the mechanism of the decreased ζ-chain expression is less clear. Src-like adaptor protein (SLAP) is a master regulator of T cell activation; previous data have reported that SLAP regulates immunoreceptor signaling. We have examined the mechanism and the functional consequences of CD3 ζ-chain downregulation. TNF treatment of human T lymphocytes (15-40 ng/ml) selectively downregulates CD3 ζ-chain expression in a dose-dependent manner (p < 0.05) and decreases activation-induced IL-2 expression (p < 0.01). Although blocking of the lysosomal compartment fails to restore TNF-induced CD3 ζ-chain downregulation, inhibition of the proteasome prevented the effect of TNF. Both SLAP expression and the colocalization of SLAP with CD3 ζ-chain was enhanced by TNF treatment (p < 0.05 and p < 0.01, respectively), whereas TNF-induced ζ-chain downregulation was inhibited by gene silencing of SLAP with small interfering RNA. SLAP levels of the CD4(+) T lymphocytes isolated from patients with rheumatoid arthritis were more than 2-fold higher than that of the healthy donors' (p < 0.05); moreover, TNF treatment did not alter the SLAP expression of the CD4(+) cells of anti-TNF therapy-treated patients. Our present data suggest that TNF modulates T cell activation during inflammatory processes by regulating the amount of CD3 ζ-chain expression via a SLAP-dependent mechanism. These data provide evidence for SLAP-dependent regulation of CD3 ζ-chain in the fine control of TCR signaling.


Case report: Emergence of dolutegravir resistance in a patient on second-line antiretroviral therapy.
The integrase strand transfer inhibitor dolutegravir (DTG) has a high genetic barrier to resistance. Only rare cases of resistance to DTG have been reported when it is used as a component of antiretroviral therapy regimens in treatment-experienced patients unless there was prior use of a first-generation integrase inhibitor. A 38-year-old woman diagnosed with tuberculosis was switched to a second-line antiretroviral regimen of zidovudine, lamivudine and dolutegravir 50 mg 12-hourly together with rifampicin-based TB treatment. Based on treatment history and a previous resistance test there was resistance to lamivudine but full susceptibility to zidovudine. The patient did not suppress her viral load on this regimen and later admitted to only taking dolutegravir 50 mg in the morning because of insomnia. A second resistance test was performed which showed intermediate level of resistance to dolutegravir. Her regimen was changed to tenofovir, emtricitabine and ritonavir-boosted atazanavir with rifabutin replacing rifampicin for the remainder of her TB treatment. She achieved viral suppression on this regimen. To our knowledge this is the first case report from South Africa of emergent dolutegravir resistance in a treatment-experienced, integrase inhibitor-naïve patient. Factors that may have contributed to resistance emergence in this patient were that there was only one fully active nucleoside reverse transcriptase inhibitor in the regimen and lower exposure to dolutegravir because of the reduced dosing frequency while on rifampicin.


The controllable growth of ultrathin MnO2 on polydopamine nanospheres as a single nanoplatform for the MRI-guided synergistic therapy of tumors.
The integration of two-dimensional (2D) nanosheets with biocompatible photothermal nanoparticles may produce effective multifunctional nanotheranostic agents. However, such explorations are hindered due to the lack of suitable methods for the incorporation of both materials. In this work, a simple redox strategy was designed for the controllable growth of ultrathin manganese dioxide nanosheets on polydopamine nanospheres. After loading with methylene blue, the obtained polydopamine@ultrathin manganese dioxide/methylene blue nanoflowers (termed PDA@ut-MnO2/MB NFs) show excellent multiple capabilities for tumor synergistic therapy, for instance: (1) they efficiently modulate the tumor microenvironment (TME) via the generation of O2 and the depletion of glutathione (GSH); (2) they demonstrate favorable photothermal effects during PTT to speed up intratumoral blood flow and chemical reactions, enhancing oxygen distribution and synergistically increasing PDT efficacy; and (3) they support ultrasensitive reduction-responsive magnetic resonance imaging (MRI). Using mice bearing HCT116 colorectal tumors as a model system, PDA@MnO2/MB NFs were shown to achieve near-complete suppression of tumor growth following MRI-guided PDT/PTT synergistic therapy.


3-benzazecine-based cyclic allene derivatives as highly potent P-glycoprotein inhibitors overcoming doxorubicin multidrug resistance.
Aim: Enamino 3-benzazecine compounds, incorporating the C6-C8 allene system, were synthesized and evaluated in vitro as inhibitors of P-glycoprotein (P-gp) and/or multidrug resistance-associated protein 1 (MRP1), two efflux pumps mainly connected with multidrug resistance (MDR) in cancer cells. Results & methodology: Most of the synthesized compounds were selective P-gp inhibitors in Calcein-AM uptake assay. Structure-activity relationships (SARs) pointed out that CO2Me derivatives are more potent than acetyl derivatives, and 10,11-dimethoxy compounds are five to tenfold more potent inhibitors than the respective unsubstituted compounds, and that the P-gp inhibition potency is mainly related to volume parameters. Conclusion: Nanomolar P-gp inhibitors, such as 23 (IC50 = 4.2 nM), restored the antiproliferative activity of doxorubicin in multidrug-resistant cells. The observed activities showed that 3-benzazecine-based compounds may be promising MDR reversers.


COVID-19 mortality rates in the European Union, Switzerland, and the UK: effect of timeliness, lockdown rigidity, and population density.
To date, the European experience with COVID-19 mortality has been different to that observed in China and Asia. We aimed to forecast mortality trends in the 27 countries of the European Union (EU), plus Switzerland and the UK, where lockdown dates and confinement interventions have been heterogeneous, and to explore its determinants. We have adapted our predictive model of COVID-19-related mortality, which rested on the observed mortality within the first weeks of the outbreak and the date of the respective lockdown in each country. It was applied in a training set of three countries (Italy, Germany and Spain), and then applied to the EU plus the UK and Switzerland. In addition, we explored the effects of timeliness and rigidity of the lockdown (on a five-step scale) and population density in our forecasts. We report r2, and percent variation of expected versus observed deaths, all following TRIPOD guidance. We identified a homogeneous distribution of deaths, and found a median of 24 days after lockdown adoption to reach the maximum daily deaths. Strikingly, cumulative deaths up to April 25th, 2020 observed in Europe separated countries in three waves, according to the time lockdown measures were adopted following the onset of the outbreak: after a week, within a week, or even prior to the outbreak (r2=0.876). In contrast, no correlation neither with lockdown rigidity nor population density were observed. The European experience confirms that early, effective interventions of lockdown are fundamental to minimizing the COVID-19 death toll.


7 to 12 Hz activity in rat gustatory cortex reflects disengagement from a fluid self-administration task.
The 7 to 12 Hz rhythm is a high-voltage oscillatory phenomenon recorded in many rat neocortical regions, largely analogous to the rodent and human somatosensory mu rhythm. Central to any interpretation of the functional significance of this pattern is the analysis of the behavioral context associated with it. Much of the debate on the function of mu, variously believed to represent either an environment-oriented or -isolated state, has relied primarily on its association with quiet immobility. In this report, we describe the relationship between the 7 to 12 Hz rhythm and a more complex behavioral setting, in which we were able to dissociated task orientation from disengagement. We trained head-restrained, water-restricted rats to perform a simple variant of a timed fluid self-administration task, while recording local field potentials from gustatory cortex (GC). Rats progressed through two behavioral states that were clearly distinguishable on the basis of lever-pressing regimes: a task-oriented state and a second state that reflected disengagement from the task. Concurrent GC neural recordings revealed bilaterally coherent oscillations in the 7 to 12 Hz range associated solely with the latter state. Consistent with published recordings of mu rhythm from somatosensory cortex, these rhythmic episodes were endogenously quenched when the rats prepared to lever-press; this inhibition of rhythmic episodes lasted through fluid delivery and consumption, making it clear that GC rhythms are not related to gustatory processing itself. By showing a direct relationship between the 7 to 12 Hz rhythm and disengagement from a task, these data provide strong and novel evidence that this gustatory rhythm in rats is associated with withdrawal from experimental contingencies.


Population fluctuations of the fungiid coral Cycloseris curvata, Galápagos Islands, Ecuador.
Fungiid corals (Cnidaria: Anthozoa: Scleractinia) occur at isolated locations scattered throughout the eastern tropical Pacific. They can be reef-associated but are often found on sand and rubble substrata distant from reef coral habitat. Cycloseris curvata is known in this region from the southern Gulf of California, through Mexico, Costa Rica, and Panamá, and with the southern-most populations occurring in the Galápagos Islands, Ecuador. During Archipelago-wide surveys (1988-2019), living individuals of Cycloseris curvata were observed at only two locations, Devil's Crown (near Floreana Island) and Xarifa Island (near Española Island). The Devil's Crown population was observed from 1988 to 2017, whereas living individuals in the Xarifa population were observed from 2005 to 2009. In 2012 a death assemblage (dead skeletons) was discovered at Darwin Island, at the northern-most extent of the Archipelago. At Devil's Crown, visual surveys were performed annually or biennially from 1990 to 2012, with two more surveys in 2017 and 2019. The living Cycloseris curvata population consisted of 15 individuals in 1990 that gradually increased to 78 individuals by 1995. Over 200 individuals were observed in 1996, and high numbers persisted through 1998 with 335 individuals. Live tissue surface area per polyp ranged from 0.5 to 95.0cm2. The population decreased to 112 individuals in 1999 (following warming associated with the 1997-98 El Niño), with further declines to 20 in 2009 (following cooling associated with the 2007 La Niña) and a rebound to 91 in 2012. After a 5y break in data collection, only one individual (28.3cm2) was observed in 2017, and in 2019 none were observed. Although undetected living Cycloseris curvata populations may exist, and renewed recruitment provides some hope for population reestablishment, it is possible that this fungiid coral species is now extirpated from the Galápagos Archipelago.


New transport peptides broaden the horizon of applications for peptidic pharmaceuticals.
Protein transduction domains (PTDs) have proven to be an invaluable tool to transduce a wide variety of cargo's including peptides across the plasma membrane and into intact tissue. The PTDs are able to deliver biologically active molecules both in vitro and in vivo. This study describes many new polybasic PTDs of which some are just as potent as the PTDs derived from extracellular RNAses or other published PTDs. Large differences in potency became apparent when the PTDs are coupled to particular cargoes. Therefore, the unique characteristic of a PTD may only become apparent when it is selected for a particular application. Rules for optimization of PTDs for particular applications are now emerging and open the way for a new generation of drug delivery agents. Because fixation artifacts and irreversible membrane binding may cause misinterpretation of the amount of internalization of polybasic peptides, we have developed an enzyme transduction assay based on the intracellular loading of a cell permeable substrate. In this assay, a fluorescent signal is generated by internalized enzyme in intact cells and not by membrane-bound or extracellular enzyme.


Effects of metabolic acidosis, alkalosis, and dietary hydrogen ion intake on phosphate transport in the proximal convoluted tubule.
Early proximal convoluted tubules were perfused in vivo with equilibrium Ringer buffered at pH 7.65 and 6.5 to characterize phosphate absorption due to changes in extracellular and intracellular hydrogen ion concentration. Phosphate absorption is normally greater from intraluminal pH 7.65 compared with pH 6.5 in thyroparathyroidectomized rats with fractional phosphate excretion of 0.5%. Metabolic alkalosis produced by bicarbonate infusion in rats ingesting normal amounts of phosphate (45 mg/day) resulted in an increase in overall renal phosphate reabsorption (fractional phosphate excretion 0.3%). The Jmax and Km values were: pH 7.65, 33.67 +/- 13.49 pmol X min-1 X mm-1, and 1.74 +/- 0.30 mM; pH 6.5, 24.87 +/- 6.22 and 0.50 +/- 0.25, respectively. By contrast, rats on a high dietary phosphate intake (180 mg/day) demonstrated a large increase in urinary phosphate excretion (18%) following bicarbonate infusion, which was due to a decrease in tubular phosphate absorption from both alkaline and acidic luminal pH values. Acute metabolic acidosis did not significantly alter tubular transport at either intraluminal pH value. In contrast, rats maintained on an elevated dietary acid intake for 5 days had a phosphaturia (fractional excretion 7.1%) and diminished reabsorptive capacity. Dietary acidosis also decreased tubular phosphate transport in rats previously maintained on phosphate-restricted diets. These data suggest that acid-base balance may modulate tubular phosphate transport independent of intraluminal pH and phosphate concentration. Further, these changes depend on the chronicity of exposure and act independent but integral to the effects of parathyroid hormone and the intrinsic adaptation to dietary phosphate availability.


Spectrophotometric monitoring of O2 delivery to the exposed rat kidney.
Optical spectrophotometry was used to measure changes in the oxidation-reduction state of cytochrome c oxidase in the in situ rat kidney. Alterations in the redox state of cytochrome c oxidase were monitored during variations in the amount of oxygen being delivered to the animal. Spectral analyses were performed on whole kidney, cortical tubule suspensions, and blood (the latter flowing freely through surgically implanted femoral arteriovenous shunts). Reaction spectra identified the location of the absorption maxima for reduced cytochrome c oxidase to be at approx. 605 nm. Additional spectral analysis indicated that hemoglobin oxygenation-deoxygenation changes had minimal artifactual interference on the cytochrome redox signals. The present results indicate that mitochondrial cytochrome c oxidase is not maximally oxidized in the in situ kidney of anesthetized rats at arterial oxygen in the in situ kidney of anesthetized rats at arterial oxygen tensions within the normal physiological range. The redox state of the enzyme in vivo is altered by changes in the level of inspired oxygen over a wide range. The current data are consistent with the existence of nonuniform regions of aerobic respiration occurring in the in situ kidney.


Diversity and abundance of Crenarchaeota in terrestrial habitats studied by 16S RNA surveys and real time PCR.
Novel phylogenetic lineages of as yet uncultivated crenarchaeota have been frequently detected in low to moderate-temperature, marine and terrestrial environments. In order to gain a more comprehensive view on the distribution and diversity of Crenarchaeota in moderate habitats, we have studied 18 different terrestrial and freshwater samples by 16S rDNA-based phylogenetic surveys. In seven different soil samples of diverse geographic areas in Europe (forest, grassland, ruderal) and Asia (permafrost, ruderal) as well as in two microbial mats, we have consistently found one particular lineage of crenarchaeota. The diversity of Crenarchaeota in freshwater sediments was considerably higher with respresentative 16S rDNA sequences distributed over four different groups within the moderate crenarchaeota. Systematic analysis of a 16S rDNA universal library from a sandy ecosystem containing 800 clones exclusively revealed the presence of the soil-specific crenarchaeotal cluster. With primers specific for non-thermophilic crenarchaeota we established a rapid method to quantify archaeal 16S rDNA in real time PCR. The relative abundance of crenarchaeotal rDNA was 0.5-3% in the bulk soil sample and only 0.16% in the rhizosphere of the sandy ecosystem. A nearby agricultural setting yielded a relative abundance of 0.17% crenarchaeotal rDNA. In total our data suggest that soil crenarchaeota represent a stable and specific component of the microbiota in terrestrial habitats.


Risk of serious bacterial infection in children with fever without a source in the post-Haemophilus influenzae era when antibiotics are reserved for culture-proven bacteremia.
To determine the rate of serious bacterial infection in children aged 2 to 36 months with fever without a source in the post-Haemophilus influenzae era, when antibiotic therapy is reserved until blood culture results turn positive. Retrospective review of emergency department, urgent care center, and hospital medical records from an urban children's hospital. Eligible participants were identified from hospital medical record and microbiology laboratory databases. Immunocompetent individuals aged 2 to 36 months with fever without a source were eligible for enrollment. Exclusion criteria were temperature less than 39.0 degrees C, identifiable focus of infection, current or recent antibiotic use, and hospital admission. Enrolled participants were assigned to group 1 (blood culture obtained) or group 2 (no blood culture) and did not receive empiric antibiotic treatment in the emergency department, in the urgent care center, or for home use. Demographic and outcome data were collected on all enrolled patients. During the study, 9241 febrile children were identified; 2641 (29%) met the enrollment criteria. Blood cultures (group 1) were performed on 1202 patients (46%), and 37 (3%) had culture-proven occult bacteremia (95% confidence interval, 2.2%-4.2%). Streptococcus pneumoniae was the most prevalent organism (84%). The mean +/- SD time for reporting a positive blood culture finding was 17.5 +/- 8.5 hours. Two patients (0.08%; 95% confidence interval, 0.009%-0.27%) developed serious bacterial infection, and both recovered completely. Reserving antibiotic therapy for culture-proven occult bacteremia was not associated with increased risk of developing serious bacterial infection compared with previously published data.


Caring for the patient with multiple somatic complaints.
Patients with multiple somatic complaints but little disease are commonly seen in most medical practices. Their behavior can be understood by considering the contribution of psychologic, perceptual, and social factors in the etiology and maintenance of the illness complaint. Because these patients cling to their symptoms, care rather than cure is the cornerstone of management. Recommendations for patient care are based on a multifactorial model of symptom etiology. The doctor-patient relationship is central to therapy.


Impact evaluation of India's 'Yeshasvini' community-based health insurance programme.
Using propensity score matching techniques, the study evaluates the impact of India's Yeshasvini community-based health insurance programme on health-care utilisation, financial protection, treatment outcomes and economic well-being. The programme offers free out-patient diagnosis and lab tests at discounted rates when ill, but, more importantly, it covers highly catastrophic and less discretionary in-patient surgical procedures. For its impact evaluation, 4109 randomly selected households in villages in rural Karnataka, an Indian state, were interviewed using a structured questionnaire. A comprehensive set of indicators was developed and the quality of matching was tested. Generally, the programme is found to have increased utilisation of health-care services, reduced out-of-pocket spending, and ensured better health and economic outcomes. More specifically, however, these effects vary across socio-economic groups and medical episodes. The programme operates by bringing the direct price of health-care down but the extent to which this effectively occurs across medical episodes is an empirical issue. Further, the effects are more pronounced for the better-off households. The article demonstrates that community insurance presents a workable model for providing high-end services in resource-poor settings through an emphasis on accountability and local management.


Problematic Drinking Among Postgraduate Students: Binge Drinking, Prepartying, and Mixing Alcohol With Energy Drinks.
Although problematic alcohol use has been studied extensively in undergraduate students, little is known about problematic drinking among postgraduate students. This study examined binge drinking, prepartying, and mixing alcohol with energy drinks to determine: (1) the extent to which postgraduate students engage in these drinking behaviors, (2) how postgraduate students differ from undergraduate students in these behaviors, and (3) the demographic risk factors for these behaviors in postgraduate (and undergraduate) students. This study utilized data from n = 695 students (n = 298 postgraduate; n = 397 undergraduate) who participated in the Healthy Minds Study at a large, public university in the Midwestern US. Past-two-week binge drinking, past-year and past-30-day prepartying, and past-30-day mixing alcohol with energy drinks were reported by 26.2%, 28.6%, 14.9%, and 8.1% of postgraduate students, respectively. Multivariate analyses indicated that postgraduate status was a significant negative predictor of binge drinking and prepartying, and that status interacted with age in predicting prepartying such that the effect of age on prepartying was negative for postgraduate students and nonsignificant for undergraduates. Age was a significant negative predictor of mixing alcohol with energy drinks for all students. This study makes a unique contribution to the literature by providing information on problematic drinking in postgraduate students. Although there was evidence of "maturing out," a substantial number of postgraduate students were found to engage in binge drinking and prepartying, and a not insubstantial number of them were found to mix alcohol with energy drinks.


Stochasticity of TOL plasmid catabolic promoters sets a bimodal expression regime in Pseudomonas putida mt-2 exposed to m-xylene.
The expression dynamics of the catabolic promoters of the TOL plasmid pWW0 has been examined in single cells of Pseudomonas putida mt-2 exposed to m-xylene. To this end, we employed an á la carte bi-cistronic gfp-lacZ reporter system for generating monocopy transcriptional fusions to each of the four promoters (Pr, Ps, Pu and Pm) of the regulatory network. Whereas expression of xylR (the master regulatory gene of the TOL system) behaved in a unimodal fashion, the activation of Pu and Pm displayed a high degree of stochasticity. This event resulted in a time-dependent shift in the bacterial population between two induction states without intermediate expression levels. Bimodality of the Pm promoter for the lower TOL pathway was largely due to the propagation of the Ps output throughout the network, which caused stochastic overproduction of the second regulator of the system, XylS. Surprisingly, activation of Pm by the form of XylS that was bound to the pathway intermediate 3-methylbenzoate was relatively minor. These stochastic phenomena in response to m-xylene disappeared in the stationary phase. Taken together, the data suggests that genetically homogeneous populations of P. putida mt-2 employ a strategy of phenotypic variation (metabolic bet-hedging) when confronted with mixtures of nutrients.


Interactions Between Intensive Care and Palliative Care Are Influenced by Training, Professionals' Perceptions and Institutional Barriers.
Background: There is growing interest in the use of a Palliative care approach in Intensive care. However, it tends to remain inconsistent, infrequent or non-existent, as does its acceptance by intensive care physicians. This study sought to explore the perceptions, level of knowledge, perceived barriers, and practices of physicians regarding palliative care practices (PC) in Intensive Care Units (ICU). Methods: Descriptive-correlational study. Participating physicians working in ICU in Colombia (n = 101) completed an ad hoc questionnaire that included subscales of perceptions, knowledge, perceived barriers, and PC practices in ICU. A Structural Equation Model (PLS-SEM) was used to examine the reciprocal relationships between the measured variables and those that could predict interaction practices between the 2 specialties. Results: First, results from the measurement model to examine the validity and reliability of the latent variables found (PC training, favorable perceptions about PC, institutional barriers, and ICU-PC interaction practices) and their indicators were obtained. Second, the structural model found that, a greater number of hours of PC training, a favorable perception of PC and a lower perception of institutional barriers are related to greater interaction between PC and ICU, particularly when emotional or family problems are detected. Conclusions: PC-ICU interactions are influenced by training, a positive perception of PC and less perceived institutional barriers. An integrated ICU-PC model that strengthens the PC training of those who work in ICU and provides clearer guidelines for interaction practices, may help overcome perceived barriers and improve the perception of the potential impact of PC.


Imaging of Subsurface Corrosion Using Gradient-Field Pulsed Eddy Current Probes with Uniform Field Excitation.
A corrosive environment leaves in-service conductive structures prone to subsurface corrosion which poses a severe threat to the structural integrity. It is indispensable to detect and quantitatively evaluate subsurface corrosion via non-destructive evaluation techniques. Although the gradient-field pulsed eddy current technique (GPEC) has been found to be superior in the evaluation of corrosion in conductors, it suffers from a technical drawback resulting from the non-uniform field excited by the conventional pancake coil. In light of this, a new GPEC probe with uniform field excitation for the imaging of subsurface corrosion is proposed in this paper. The excited uniform field makes the GPEC signal correspond only to the field perturbation due to the presence of subsurface corrosion, which benefits the corrosion profiling and sizing. A 3D analytical model of GPEC is established to analyze the characteristics of the uniform field induced within a conductor. Following this, experiments regarding the imaging of subsurface corrosion via GPEC have been carried out. It has been found from the results that the proposed GPEC probe with uniform field excitation not only applies to the imaging of subsurface corrosion in conductive structures, but provides high-sensitivity imaging results regarding the corrosion profile and opening size.


Multicentric warfarin-induced skin necrosis complicating heparin-induced thrombocytopenia.
Two patients developed catastrophic multicentric skin necrosis while receiving warfarin to treat venous thromboembolism complicated by immune-mediated heparin-induced thrombocytopenia (HIT). Patient 1 developed skin necrosis involving the breasts, thighs, and face, as well as venous limb gangrene and bilateral hemorrhagic necrosis of the adrenal glands, resulting in death. The second patient developed bilateral mammary necrosis necessitating mastectomies, as well as skin necrosis involving the thigh. Neither patient had an identifiable hypercoagulable syndrome, other than HIT. HIT may represent a risk factor for the development of multicentric warfarin-induced skin necrosis (WISN).


Molecular and point-of-care diagnostics for Ebola and new threats: National POCT policy and guidelines will stop epidemics.
US hospitals that admitted Ebola virus disease (EVD) patients mitigated risk by using point-or-care testing (POCT) for critical support in isolation units. Success proved unequivocally the need for POCT. Additionally, molecular diagnostics have been used to help stop new outbreaks, and even handheld diagnostic solutions are emerging. Areas covered: This update of 'Molecular detection and point-of-care testing in Ebola virus disease and other threats' [Expert Reviews 2015;15(10):1249-1255], assesses the impact of EVD epidemics, documents insights from recent reviews, summarizes evolving POC molecular technologies, presents General Accountability Office (GAO) recommendations, identifies the role of POC Coordinators, and casts a vision for national POCT policies and guidelines. Factual updating comprised summarizing EVD outbreaks including 2017-2018, analyzing reviews and evidence-based publications since the 2014-2016 epidemic, and tabulating published technical and molecular diagnostics. New graphics illustrate POC error mitigation/risk reduction, a framework for national POCT policy and guidelines, modular adaptations for country-specific solutions, and a logic diagram for future progress embedding artificial intelligence. Expert commentary: The USA is still not prepared for highly infectious diseases. Key is lack of community rapid response and resilience, which must be enhanced not via mechanisms distant, but instead by molecular diagnostics directly at critical points of need.


Role of microsatellite instability, immunohistochemistry and mismatch repair germline aberrations in immunosuppressed transplant patients: a phenocopy dilemma in Muir-Torre syndrome.
Sebaceous tumours and keratoacanthomas are uncommon neoplasms that constitute important clinical criteria for Muir-Torre syndrome (MTS) diagnosis. In MTS patients, the increased risk of developing synchronous or metachronous visceral malignancies is characterised by autosomal dominant inheritance. However, there are further conditions, other than MTS, that increase the risk of sebaceous neoplasms, e.g. iatrogenic immunosuppression. In this latter scenario, the sebaceous tumours can present microsatellite instability (MSI) and loss of mismatch repair (MMR) proteins, characteristic of hereditary syndromes, even in the absence of MMR germline mutations. In this article, we examine transplant probands in which the immunosuppressive therapies unmask the MTS cutaneous phenotypes, showing MSI and loss of MMR protein expression, as demonstrated by immunohistochemistry (IHC). Furthermore, MMR genes sequencing analysis identified the presence of germline mutations in MTS-suspected individuals, in the absence of a visceral MTS phenotype. It is well known that immunosuppression plays a central role in the development of sebaceous tumours in both MTS and in non-syndromic settings. Sebaceous skin tumours' MSI status and IHC profiles can be influenced by epigenetic or iatrogenic factors; however, they constitute valuable tools and a cost-effective approach to screen individuals who otherways should undergo MMR genes direct sequencing in the context of immunosuppression. In this complex setting, the choice of the immunosuppressive drug becomes a critical decision for the management of both MTS and sporadic transplant patients, which may benefit from the administration of immunosuppressive drugs, resulting in a low impact on skin cancerogenesis.


Blastogenesis as an in vitro correlate of delayed hypersensitivity in guinea pigs infected with Listeria monocytogenes.
Randomly bred guinea pigs of both sexes were injected intracardially with one-half a 50% lethal dose of Listeria monocytogenes. When these animals were skin tested with 30 mug of a water-soluble extract of sonically disrupted Listeria, animals had uniformly detectable levels of delayed-type hypersensitivity (DTH) 6 days after infection. Histological examination of skin test reaction sites, after fixation in Helly fixative and Giemsa staining, revealed a classical tuberculin-type infiltrate consisting primarily of mononuclear cells with few polymorphonuclear cells. Many of the small vessels showed perivascular cuffing. When purified peritoneal exudate lymphocytes from these animals were cultured in vitro in the presence of various concentrations of Listeria antigen, it was fount that the optimal antigenic dose for specific antigen-induced incorporation of [3H]thymidine varied for individual animals. In contrast to the early onset of uniformly detectable levels of in vivo DTH, in vitro lymphocyte blastogenesis was not uniformly demonstrable until 14 days postinfection and remained highly significant on days 21, 28, and 84 postinfection. At 7 days postinfection, lymphocytes from 7 of 17 animals were capable of undergoing sifnificant blastogenesis. The Listeria antigen preparation was not mitogenic for peritoneal exudate lymphocytes from normal animals. It was found that no direct correlation exists between the in vivo levels of DTH and in vitro blastogenesis. Cell donors showing significant in vitro blastogenesis nevertheless were also skin test positive for most animals tested. Humoral antibody was found to play no significant role in the immune response of guinea pigs to a primary infection with Listeria monocytogenes.


Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers.
Komagataella phaffii (syn. Pichia pastoris) is one of the most commonly used host systems for recombinant protein expression. Achieving targeted genetic modifications had been hindered by low frequencies of homologous recombination (HR). Recently, a CRISPR/Cas9 genome editing system has been implemented for P. pastoris enabling gene knockouts based on indels (insertion, deletions) via non-homologous end joining (NHEJ) at near 100% efficiency. However, specifically integrating homologous donor cassettes via HR for replacement studies had proven difficult resulting at most in ∼20% correct integration using CRISPR/Cas9. Here, we demonstrate the CRISPR/Cas9 mediated integration of markerless donor cassettes at efficiencies approaching 100% using a ku70 deletion strain. The Ku70p is involved in NHEJ repair and lack of the protein appears to favor repair via HR near exclusively. While the absolute number of transformants in the Δku70 strain is reduced, virtually all surviving transformants showed correct integration. In the wildtype strain, markerless donor cassette integration was also improved up to 25-fold by placing an autonomously replicating sequence (ARS) on the donor cassette. Alternative strategies for improving donor cassette integration using a Cas9 nickase variant or reducing off targeting associated toxicity using a high fidelity Cas9 variant were so far not successful in our hands in P. pastoris. Furthermore we provide Cas9/gRNA expression plasmids with a Geneticin resistance marker which proved to be versatile tools for marker recycling. The reported CRSIPR-Cas9 tools can be applied for modifying existing production strains and also pave the way for markerless whole genome modification studies in P. pastoris.


Outcomes after two years of providing antiretroviral treatment in Khayelitsha, South Africa.
A community-based antiretroviral therapy (ART) programme was established in 2001 in a South African township to explore the operational issues involved in providing ART in the public sector in resource-limited settings and demonstrate the feasibility of such a service. Data was analysed on a cohort of patients with symptomatic HIV disease and a CD4 lymphocyte count < 200 x 10 cells/l. The programme used standardized protocols (using generic medicines whenever possible), a team-approach to clinical care and a patient-centred approach to promote adherence. Two-hundred and eighty-seven adults naive to prior ART were followed for a median duration of 13.9 months. The median CD4 lymphocyte count was 43 x 10 cells/l at initiation of treatment, and the mean log10 HIV RNA was 5.18 copies/ml. The HIV RNA level was undetectable (< 400 copies/ml) in 88.1, 89.2, 84.2, 75.0 and 69.7% of patients at 3, 6, 12, 18 and 24 months respectively. The cumulative probability of remaining alive was 86.3% at 24 months on treatment for all patients, 91.4% for those with a baseline CD4 lymphocyte count > or =50 x 10 cells/l, and 81.8% for those with a baseline CD4 lymphocyte count < 50 x 10 cells/l. The cumulative probability of changing a single antiretroviral drug by 24 months was 15.1% due to adverse events or contraindications, and 8.4% due to adverse events alone. ART can be provided in resource-limited settings with good patient retention and clinical outcomes. With responsible implementation, ART is a key component of a comprehensive response to the epidemic in those communities most affected by HIV.


Biased to learn language.
Some recent publications that explore the foundations of early language development are reviewed in this article. The review adopts the pivotal idea that infants' advancements are helped by the existence of different types of biases. The infant's discovery of the phonological properties of the language of the environment, as well as their learning of words, is assisted by different types of processing biases, ranging from perceptual to social factors.


Fiber Bragg grating ring resonators under rotation for angular velocity sensing.
In this paper we investigate the possibility of using hybrid resonators based on fiber Bragg grating ring resonators (FBGRRs) and π-shifted FBGRRs (i.e., defective FBGRRs) as rotation sensitive elements for gyroscope applications. In particular, we model the conventional fiber Bragg grating (FBG) with the coupled mode theory by taking into account how the Sagnac effect, induced by the rotation, modifies the eigenvalues, the photonic band gap, and the spectral response of the FBG. Then, on the basis of the FBG model under rotation conditions, the spectral responses of the FBGRR and π-FBGRR have been evaluated, confirming that the Sagnac effect manifests itself with a spectral shift of the eigensolutions. This physical investigation can be exploited for opening new ways in the optical gyroscope platforms.


Management considerations for the pregnant or nursing emergency patient.
It is important to realize the risk-benefit ratio in all therapeutic modalities in any patient and that these variables vary from patient to patient as do circumstances. The emergency patient who is pregnant or nursing should not be feared or be denied appropriate treatment. It is always best to err on the side of safety and remain conservative. The dental problem can usually be treated with the use of adequate local anaesthesia and supplemental nitrous oxide in the second or third trimester. Nevertheless, situations arise that may warrant consultation with the patient's physician or specialist. This brief overview should refamiliarize the dental practitioner with the medications available to treat the gravid patient. It is important to have an understanding of our common therapeutics as well as the inherent treatment risks and benefits as they relate to the changing maternal-fetal physiology.


Sperm from beta 1,4-galactosyltransferase-null mice are refractory to ZP3-induced acrosome reactions and penetrate the zona pellucida poorly.
A variety of sperm surface components have been suggested to mediate gamete recognition by binding to glycoside ligands on the egg coat glycoprotein ZP3. The function of each of these candidate receptors is based upon varying degrees of circumstantial and direct evidence; however, the effects on fertilization of targeted mutations in any of these candidate receptors have not yet been reported. In this paper, we describe the effects of targeted mutations in beta1,4-galactosyltransferase, the best studied of the candidate receptors for ZP3. Surprisingly, galactosyltransferase-null (gt[-/-]) males are fertile; however, sperm from gt(-/-) males bind less radiolabeled ZP3 than wild-type sperm, and are unable to undergo the acrosome reaction in response to either ZP3 or anti-galactosyltransferase antibodies, as do wild-type sperm. In contrast, gt(-/-) sperm undergo the acrosome reaction normally in response to calcium ionophore, which bypasses the requirement for ZP3 binding. The inability of gt(-/-) sperm to undergo a ZP3-induced acrosome reaction renders them physiologically inferior to wild-type sperm, as assayed by their relative inability to penetrate the egg coat and fertilize the oocyte in vitro. Thus, although ZP3 binding and subsequent induction of the acrosome reaction are dispensable for fertilization, they impart a physiological advantage to the fertilizing sperm. A second strain of mice was created that is characterized by a loss of of the long galactosyltransferase isoform responsible for ZP3-dependent signal transduction, but which maintains normal levels of Golgi galactosylation. Sperm from these mice show that the defective sperm-egg interactions in gt(-/-) mice are due directly to a loss of the long galactosyltransferase isoform from the sperm surface and are independent of the state of intracellular galactosylation during spermatogenesis.


Metoidioplasty With Urethral Lengthening: A Stepwise Approach.
Metoidioplasty is a gender-affirming surgical option for individuals who desire masculine genitalia while preserving erogenous sensation and avoiding the morbidity of phalloplasty. Concurrent urethral lengthening offers patients the potential to stand to urinate. To demonstrate an adaptation of the Belgrade technique of gender-affirming metoidioplasty and describe outcomes. We identified 33 patients of which 12 underwent simple metoidioplasty and 21 underwent metoidioplasty with urethral lengthening between 2016 and 2020. Prior to surgery, all patients underwent at least 1 year of testosterone therapy to maximize clitoral growth. The clitoris is degloved and the superficial suspensory ligament divided to maximize phallic length. Labia minora flaps are developed and the urethral plate is divided to allow for maximal ventral extension. The resultant urethral defect is bridged with a buccal mucosa graft. To construct the ventral aspect of the urethra, an anterior vaginal wall flap and labia minora flap are sutured to the urethral plate and tubularized over a 16 Fr catheter. The medial aspect of the contralateral labia minora is de-epithelialized and overlaps the urethra, serving as ventral skin coverage for the phallic shaft. Bilateral labia majora flaps are then rotated anteriorly and superiorly to create a neoscrotum using the Ghent technique. For metoidioplasty with urethral lengthening, the median operative time was 408 minutes, estimated blood loss 400 mL, and length of stay 3 days. Of the 21 patients, 10 (47%) elected to undergo second stage scrotoplasty, 7 (33%) underwent testicular implant placement, and 2 (9%) required revision urethroplasty. Of the 10 patients (48%) who experienced postoperative complications, 7 were Clavien-Dindo grade I-II. There were no fistulae at a median follow-up time of 5.5 months (range 1-27.2). We provide a stepwise approach to metoidioplasty with urethral lengthening using a modified Belgrade technique, which was associated with a low rate of urethral complications.


Soluble expression and partial purification of recombinant human erythropoietin from E. coli.
Human erythropoietin (hEpo) is an essential regulator of erythrocyte production that induces the division and differentiation of erythroid progenitor cells in the bone marrow into mature erythrocytes. It is widely used for the treatment of anemia resulting from chronic kidney disease, chemotherapy, and cancer-related therapies. Active hEpo, and hEpo analogs, have been purified primarily from mammalian cells, which has several disadvantages, including low yields and high production costs. Although an Escherichia coli (E. coli) expression system may provide economic production of therapeutic proteins, it has not been used for the production of recombinant hEpo (rhEpo) because it aggregates in inclusion bodies in the E. coli cytoplasm and is not modified post-translationally. We investigated the soluble overexpression of active rhEpo with various protein tags in E. coli, and found out that several tags increased the solubility of rhEpo. Among them maltose binding protein (MBP)-tagged rhEpo was purified using affinity and gel filtration columns. Non-denaturing electrophoresis and MALDI-TOF MS analysis demonstrated that the purified rhEpo had two intra-disulfide bonds identical to those of the native hEpo. An in vitro proliferation assay showed that rhEpo purified from E. coli had similar biological activity as rhEpo derived from CHO cells. Therefore, we report for the first time that active rhEpo was overexpressed as a soluble form in the cytoplasm of E. coli and purified it in simple purification steps. We hope that our results offer opportunities for progress in rhEpo therapeutics.


The Neural Correlates of Similarity- and Rule-based Generalization.
The idea that there are multiple learning systems has become increasingly influential in recent years, with many studies providing evidence that there is both a quick, similarity-based or feature-based system and a more effortful rule-based system. A smaller number of imaging studies have also examined whether neurally dissociable learning systems are detectable. We further investigate this by employing for the first time in an imaging study a combined positive and negative patterning procedure originally developed by Shanks and Darby [Shanks, D. R., & Darby, R. J. Feature- and rule-based generalization in human associative learning. Journal of Experimental Psychology: Animal Behavior Processes, 24, 405-415, 1998]. Unlike previous related studies employing other procedures, rule generalization in the Shanks-Darby task is beyond any simple non-rule-based (e.g., associative) account. We found that rule- and similarity-based generalization evoked common activation in diverse regions including the pFC and the bilateral parietal and occipital lobes indicating that both strategies likely share a range of common processes. No differences between strategies were identified in whole-brain comparisons, but exploratory analyses indicated that rule-based generalization led to greater activation in the right middle frontal cortex than similarity-based generalization. Conversely, the similarity group activated the anterior medial frontal lobe and right inferior parietal lobes more than the rule group did. The implications of these results are discussed.


Long-term outcome of renal transplant recipients with chronic hepatitis B infection-impact of antiviral treatments.
Antiviral treatment has improved the short-term outcome of kidney transplant recipients with chronic hepatitis B infection, but its long-term impact, especially in patients who have developed drug resistance, remains uncertain. Sixty-three hepatitis B surface antigen positive (HBsAg+) and 63 HBsAg- patients who have undergone kidney transplantation from 1985 to 2008 were retrospectively reviewed and their clinical outcomes were compared. With lamivudine as initial treatment, 62% of patients developed drug resistance after 4 years. Lamivudine resistance was associated with a higher incidence of chronic hepatitis but had no significant impact on liver stiffness score or patient survival during follow-up. Salvage treatment with adefovir or entecavir was well tolerated, and resulted in a three-log decrease in hepatitis B deoxynucleic acid after 6 months and normalization of alanine aminotransferase in 75% of patients. The survival rate of HBsAg+ patients transplanted in the recent era of antiviral treatment was 81% at 10 years. Treatment of hepatitis B with nucleoside/nucleotide analogues resulted in significantly improved patient survival (83% vs. 34% at 20 years, P=0.006). Although antiviral treatment was associated with reduced mortality because of liver complications (P=0.036), liver-related deaths still accounted for 40% of mortalities in HBsAg+ patients in the era of antiviral therapies and 22.2% of all deaths that occurred in patients who had received antiviral treatment. Treatment of HBsAg+ renal transplant recipients with nucleoside/nucleotide analogues confers long-term survival benefit, and that rescue therapy with adefovir or entecavir is effective and well tolerated in patients who had developed resistance to lamivudine.


The effect of inhaled budesonide on symptoms, lung function, and cold air and methacholine responsiveness in 2- to 5-year-old asthmatic children.
We hypothesized that measurement of lung function (LF) and bronchial hyperresponsiveness (BHR) could serve as supplemental tools in evaluating the efficacy of treatment with inhaled corticosteroids in asthmatic children aged 2 to 5 yr. We studied 38 children (mean age: 53 mo; range: 35 to 71 mo) with moderately severe asthma in a single-center, randomized, double-blind, parallel-group, placebo-controlled study involving 8 wk of treatment. Budesonide (BUD) 400 microgram twice daily was administered via a pressurized metered-dose inhaler and metal spacer device. Symptom scores (SSc) and use of short-acting beta(2)-agonist were monitored with diary cards. LF in awake children was measured as the specific airway resistance (sRaw), using whole-body plethysmography; as resistance by the interrupter technique (Rint); and as resistance and reactance at 5 Hz (Rrs5, Xrs5) by the impulse oscillation technique. Cold air challenge (CACh) and methacholine challenge (MCh) were used to assess BHR. Children in the BUD group experienced significantly fewer night- and daytime symptoms (p < 0.05) and more symptom-free days (p < 0.05), but not nights (p = 0.07), than children in the placebo group. Daytime (p < 0.05) but not nighttime (p = 0.09) use of rescue medication and asthma exacerbation rates (3.7 versus 9.3 exacerbations/yr) (p = 0.006) were both in favor of BUD. LF measured with the Rint technique, Rrs5, and Xrs5 were significantly improved by BUD. BHR as measured by CACh improved significantly with BUD, whereas no improvement was found on MCh. In conclusion, inhaled BUD at a total dose of 800 microgram daily significantly improved SSc, asthma exacerbation rates, lung function, and BHR as assessed by CACh in asthmatic children aged 2 to 5 yr.


18O labeling method for identification and quantification of succinimide in proteins.
We have developed a new method for identification and quantification of succinimide in proteins. The method utilizes 18O water to monitor succinimide hydrolysis. 18O-labeled isoaspartic acid and aspartic acid peptides were produced by hydrolysis of a succinimide-containing protein in 18O water (H218O) followed by tryptic digestion in regular water (H216O). The peptides that had 18O incorporated were 2 Da heavier than their 16O native counterparts. The mass difference was detected and quantified by electrospray time-of-flight mass spectrometry. The amount of 18O incorporation into the isoaspartic acid- and aspartic acid-containing peptides was used to quantify the amount of succinimide present in the native sample. The method was applied to analyze a degraded recombinant monoclonal antibody, which exhibited the accumulation of succinimide after storage in mildly acidic buffers at elevated temperatures for a few weeks. We unambiguously identified amino acid residue 30 located in the antibody light chain as the site of aspartic acid isomerization. At this site, there were 20% isoaspartic acid and 80% aspartic acid detected by peptide mapping in the degraded sample (8 weeks, 45 degrees C, pH 5.0). Hydrolysis in 18O water showed that 80% of the isoaspartic acid and 6% of the aspartic acid had 18O incorporated. The only explanation of 18O incorporation was the presence of succinimide in the sample. Together, a total of 21% (0.8x20% isoaspartic acid+0.06x80% aspartic acid) of aspartic acid residue 30 was found to be present in the form of succinimide in this degraded sample. As a control, the same sample, analyzed using regular 16O water did not show any incorporation of 18O water. By monitoring the amount of 18O-labeled isoaspartic acid and aspartic acid over time under both denaturing and native conditions at pH 8.2, we found that, at denaturing conditions, succinimide at light chain residue 30 hydrolyzed very rapidly (in less than 5 s), but slower (succinimide half-life of approximately 6 h) under native conditions. We also found that, under denaturing conditions, succinimide hydrolyzed at an isoaspartic acid/aspartic acid ratio of 3.5:1, but hydrolyzed almost exclusively to aspartic acid under native conditions. This finding indicates that protein structure plays an important role in the kinetics of succinimide hydrolysis as well as in the generation of the hydrolysis products isoaspartic acid and aspartic acid.


A 'Total Unique Variation Analysis' for Brain-Machine Interfaces.
When designing a fully implantable brain-machine interface (BMI), the primary aim is to detect as much neural information as possible with as few channels as possible. In this paper, we present a total unique variance analysis (TUVA) for evaluating the signal unique to each channel that cannot be predicted by linear combination of signals on other channels. TUVA is a statistical method for determining the total unique variance in multidimensional data, ordering channels from most to least informative, to aid in the design of maximally-efficacious BMIs. We demonstrate how this method can be applied to the design of BMIs by comparing TUVA values computed for simulated lead-field maps for high-channel-count electrocorticography (ECoG) with values computed for recordings in the interictal period in the context of surgery planning for epileptic resection.Clinical Relevance- This paper introduces a new statistical method for comparison of neural interface designs, focused on quantifying recording efficiency by minimizing channel crosstalk, which may help improve the risk-benefit profile of invasive neural recording.


Multidimensional knapsack problem: a fitness landscape analysis.
Fitness landscape analysis techniques are used to better understand the influence of genetic representations and associated variation operators when solving a combinatorial optimization problem. Five representations are investigated for the multidimensional knapsack problem. Common mutation operators, such as bit-flip mutation, are employed to generate fitness landscapes. Measures such as fitness distance correlation and autocorrelation are applied to examine the landscapes associated with the tested genetic encodings. Furthermore, additional experiments are made to observe the effects of adding heuristics and local optimization to the representations. Encodings with a strong heuristic bias are more efficient, and the addition of local optimization techniques further enhances their performance.


Analyses of cancer incidence and other morbidities in gamma irradiated B6CF1 mice.
With increasing medical radiation exposures, it is important to understand how different modes of delivery of ionizing radiation as well as total doses of exposure impact health outcomes. Our lab studied the risks associated with ionizing radiation by analyzing the Northwestern University Radiation Archive for animals (NURA). NURA contains detailed data from a series of 10 individual neutron and gamma irradiation experiments conducted on over 50,000 mice. Rigorous statistical testing on control mice from all Janus experiments enabled us to select studies that could be compared to one another and uncover unexpected differences among the controls as well as experimental animals. For controls, mice sham irradiated with 300 fractions died significantly earlier than those with fewer sham fractions and were excluded from the pooled dataset. Using the integrated dataset of gamma irradiated and control mice, we found that fractionation significantly decreased the death hazard for animals dying of lymphomas, tumors, non-tumors, and unknown causes. Gender differences in frequencies of causes of death were identified irrespective of irradiation and dose fractionation, with female mice being at a greater risk for all causes of death, except for lung tumors. Irradiated and control male mice were at a significantly greater risk for lung tumors, the opposite from observations noted in humans. Additionally, we discovered that lymphoma deaths can occur quickly after exposures to high doses of gamma rays. This study systematically cross-compared outcomes of different modes of fractionation evaluated across different Janus experiments and across a wide span of total doses. It demonstrates that protraction modulated survival and disease status differently based on the total dose, cause of death, and sex of an animal. This novel method for analyzing the Janus datasets will lead to insightful new mechanistic hypotheses and research in the fields of radiation biology and protection.