Recognition of influenza virus hemagglutinin by subtype-specific and cross-reactive proliferative T cells: contribution of HA1 and HA2 polypeptide chains.
The recognition of influenza virus hemagglutinin (HA) by T lymphocytes was examined by assaying the T cell proliferative response of influenza virus-primed T cells to purified HA of different influenza A subtypes or to isolated heavy (HA1) or light (HA2) polypeptide chains of the HA molecule. The proliferative response to HA was dependent on the activation of an Ly-1+2- subset of T cells and required the presence of nylon wool-adherent, radiation-resistant accessory cells. T cells from mice primed by infection with one strain of type A influenza virus cross-reacted with other purified HA not only of the same subtype as the priming virus but also of serologically distinct subtypes of influenza A (but not B) virus. The response of virus-primed T cells to the homologous HA or to HA of the same subtype was shown to involve recognition of determinants on both the HA1 and the HA2 chains. The recognition of HA of different subtype by cross-reactive T cells appeared to be directed predominantly to determinants on HA2. Because the antibody response to influenza virus HA is not cross-reactive between subtypes and is directed predominantly to determinants on HA1, the present results indicate that at least some of the determinants on HA recognized by T cells are different from those recognized by B cells and that the HA2 chain may be involved primarily in stimulation of T cell rather than B cell immunity.


Intraoperative Ultrasound-Guided Excision of Cardiophrenic Lymph Nodes in an Advanced Ovarian Cancer Patient.
Ovarian cancer is commonly diagnosed at an advanced stage. Complete macroscopic eradication of the disease is associated with improved prognosis. In this setting, the surgical resection of enlarged cardiophrenic lymph nodes (CPLNs) can help to achieve cytoreduction to no gross residual disease. During surgery, CPLN removal is usually performed either via video-assisted thoracic surgery or through a large transdiaphragmatic or subxiphoid incision. In the present case, we propose the use of an intraoperative transdiaphragmatic ultrasound to confirm preoperative imaging and to obtain a precise localization of the suspicious CPLNs. A 50-year-old woman without peritoneal carcinomatosis was diagnosed with bilateral ovarian cancer and enlarged inguinal, pelvic, aortic, and cardiophrenic nodes. She underwent primary debulking surgery, including radical hysterectomy, bilateral salpingo-oophorectomy, omentectomy, peritoneal biopsies, and bulky nodes resection, at the iliac, inguinal, and lumboaortic regions. After obtaining complete abdominal cytoreduction, an intraoperative ultrasound scan was performed. Two enlarged CPLNs were ultrasonographically visualized using a convex contact probe through a transhepatic window, and their exact location was identified. After complete mobilization of the right liver, the right diaphragm was incised, proximal to the site of the lymphadenopathies. The 2 lymph nodes were identified, grasped, and removed by transdiaphragmatic approach. Absence of other residual disease was confirmed by thoracic inspection, palpation, and by a subsequent intraoperative ultrasound control. At final histology, CPLNs were positive for infiltration of high-grade serous ovarian carcinoma. Intraoperative transdiaphragmatic ultrasound represents a possible approach to localize suspicious CPLNs and to guide their surgical eradication.


Predictors of children's sleep onset and maintenance problems after road traffic accidents.
Sleep onset and maintenance problems are a frequent complaint after traumatic events in children. However, the association of traumatic experiences and disturbed sleep remains to be explained. To examine the incidence of sleep onset and maintenance problems in children after road traffic accidents and identify potential predictors of sleep onset and maintenance problems, including putative psychopathological mechanisms as well as stressors affecting the family system. In 33 children treated for injuries after road traffic accidents, sleep and measures of psychopathology were assessed 10 days, 2 months, and 6 months after hospital admission. The predictive value of four clusters of predictor variables for children's sleep onset and maintenance problems was prospectively tested by multiple regression analyses. These clusters included socio-demographic, injury- and accident-related, and psychopathological variable clusters as well as factors reflecting stressors concerning mothers and family. Children suffering from posttraumatic stress reported a prolonged subjective sleep latency. The severity of sleep onset and maintenance problems was predicted by female sex and the child's as well as mothers' posttraumatic stress disorder (PTSD) severity. Sleep onset and maintenance problems in children after trauma appear to result from a complex interaction of multiple factors. Our findings support the transactional model of sleep-wake regulation that bears implications for the development of adequate intervention strategies.


Sodium Pumps Mediate Activity-Dependent Changes in Mammalian Motor Networks.
Ubiquitously expressed sodium pumps are best known for maintaining the ionic gradients and resting membrane potential required for generating action potentials. However, activity- and state-dependent changes in pump activity can also influence neuronal firing and regulate rhythmic network output. Here we demonstrate that changes in sodium pump activity regulate locomotor networks in the spinal cord of neonatal mice. The sodium pump inhibitor, ouabain, increased the frequency and decreased the amplitude of drug-induced locomotor bursting, effects that were dependent on the presence of the neuromodulator dopamine. Conversely, activating the pump with the sodium ionophore monensin decreased burst frequency. When more "natural" locomotor output was evoked using dorsal-root stimulation, ouabain increased burst frequency and extended locomotor episode duration, whereas monensin slowed and shortened episodes. Decreasing the time between dorsal-root stimulation, and therefore interepisode interval, also shortened and slowed activity, suggesting that pump activity encodes information about past network output and contributes to feedforward control of subsequent locomotor bouts. Using whole-cell patch-clamp recordings from spinal motoneurons and interneurons, we describe a long-duration (∼60 s), activity-dependent, TTX- and ouabain-sensitive, hyperpolarization (∼5 mV), which is mediated by spike-dependent increases in pump activity. The duration of this dynamic pump potential is enhanced by dopamine. Our results therefore reveal sodium pumps as dynamic regulators of mammalian spinal motor networks that can also be affected by neuromodulatory systems. Given the involvement of sodium pumps in movement disorders, such as amyotrophic lateral sclerosis and rapid-onset dystonia parkinsonism, knowledge of their contribution to motor network regulation also has considerable clinical importance. The sodium pump is ubiquitously expressed and responsible for at least half of total brain energy consumption. The pumps maintain ionic gradients and the resting membrane potential of neurons, but increasing evidence suggests that activity- and state-dependent changes in pump activity also influence neuronal firing. Here we demonstrate that changes in sodium pump activity regulate locomotor output in the spinal cord of neonatal mice. We describe a sodium pump-mediated afterhyperpolarization in spinal neurons, mediated by spike-dependent increases in pump activity, which is affected by dopamine. Understanding how sodium pumps contribute to network regulation and are targeted by neuromodulators, including dopamine, has clinical relevance due to the role of the sodium pump in diseases, including amyotrophic lateral sclerosis, parkinsonism, epilepsy, and hemiplegic migraine.


Differential resource allocation in deer mice exposed to sin nombre virus.
The resource allocation hypothesis predicts that reproductive activity suppresses immunocompetence; however, this has never been tested in an endemic disease system with free-ranging mammals. We tested the resource allocation hypothesis in wild deer mice (Peromyscus maniculatus) with natural exposure to Sin Nombre Virus (SNV). Immunocompetence was estimated from the extent of swelling elicited after deer mice were injected with phytohemagglutinin (PHA); swelling is positively correlated with immunocompetence. After livetrapping deer mice, we determined their reproductive state and SNV infection status. Males were more likely to be seropositive for SNV than females (37% vs. 25%) and exhibited 10% less swelling after PHA injection. The swelling response of females differed with both infection status and reproductive condition. There was also a significant infection status by reproductive condition interaction: non-reproductive, seropositive females experienced the least amount of swelling, whereas females in all other categories experienced significantly greater swelling. The swelling response of males differed with both SNV infection status and reproductive condition, but there was no significant infection status by reproductive condition interaction. Seronegative males elicited greater swelling than seropositive males regardless of reproductive status. In contrast to the resource allocation hypothesis, these results do not indicate that reproductive activity suppresses immunocompetence of deer mice but rather suggest that chronic SNV infection reduces immunocompetence. Sex-based differences in swelling indicate that SNV modulates the immune system of female deer mice differently than it does that of males, particularly during reproduction. We propose that differences in resource allocation between males and females could result from inherent sex-based differences in parental investment.


Non-Invasive Acoustical sensing of Drug-Induced Effects on the Contractile Machinery of Human Cardiomyocyte Clusters.
There is an urgent need for improved models for cardiotoxicity testing. Here we propose acoustic sensing applied to beating human cardiomyocyte clusters for non-invasive, surrogate measuring of the QT interval and other characteristics of the contractile machinery. In experiments with the acoustic method quartz crystal microbalance with dissipation monitoring (QCM-D), the shape of the recorded signals was very similar to the extracellular field potential detected in electrochemical experiments, and the expected changes of the QT interval in response to addition of conventional drugs (E-4031 or nifedipine) were observed. Additionally, changes in the dissipation signal upon addition of cytochalasin D were in good agreement with the known, corresponding shortening of the contraction-relaxation time. These findings suggest that QCM-D has great potential as a tool for cardiotoxicological screening, where effects of compounds on the cardiomyocyte contractile machinery can be detected independently of whether the extracellular field potential is altered or not.


Human skeletal muscle protein: effect of malnutrition, elective surgery and total parenteral nutrition.
1. The concentration of alkali-soluble protein, DNA and RNA in percutaneous muscle biopsy specimens was analysed. Tissue alkali-soluble protein/DNA ratio is a measure of muscle protein concentration, while tissue RNA/DNA ratio may reflect the capacity for protein synthesis. 2. Patients with weight loss due to cancer (n = 6) were compared with metabolically healthy patients before elective surgery (n = 7). Alkali-soluble protein/DNA and RNA/DNA ratios in the weight loss group were 248 (14) g/g and 1.3 (0.1) g/g respectively as compared with 404 (13) g/g and 2.1 (0.1) g/g in otherwise healthy patients. All of the alkali-soluble protein/DNA ratios and 5/6 of the RNA/DNA ratios in the weight loss group were below the 95% confidence interval for the healthy control subjects. 3. Patients undergoing elective open cholecystectomy (n = 7) were studied preoperatively and on days 3, 10, 20 and 30 post-operatively. The alkali-soluble protein/DNA ratio remained unchanged on post-operative day 3 but decreased by 8.7% (P < 0.01), 9.6% (P < 0.05) and 20.4% (P < 0.01) on days 10, 20 and 30 respectively in patients eating at will after the operation. No significant post-operative changes in alkali-soluble protein/DNA ratio were seen in patients given post-operative total parenteral nutrition with (n = 9) or without (n = 7) glycyl-glutamine supplementation for 3 days after surgery. 4. In conclusion, patients with weight loss due to malignant disease have a low muscle protein concentration. Elective surgery of medium magnitude results in a decrease in muscle protein lasting for more than 30 days.(ABSTRACT TRUNCATED AT 250 WORDS)


Functional association between viral and cellular transcription during influenza virus infection.
Influenza viruses replicate and transcribe their segmented negative-sense single-stranded RNA genome in the nucleus of the infected host cell. All RNA synthesising activities associated with influenza virus are performed by the virally encoded RNA-dependent RNA polymerase (RdRp) that consists of three subunits, PA, PB1 and PB2. However, viral transcription is critically dependent on on-going cellular transcription, in particular, on activities associated with the cellular DNA-dependent RNA polymerase II (Pol II). Thus, the viral RdRp uses short 5' capped RNA fragments, derived from cellular Pol II transcripts, as primers for viral mRNA synthesis. These capped RNA primers are generated by cleavage of host Pol II transcripts by an endonuclease activity associated with the viral RdRp. Moreover, some viral transcripts require splicing and since influenza virus does not encode splicing machinery, it is dependent on host splicing, an activity also related to Pol II transcription. Despite these functional links between viral and host Pol II transcription, there has been no evidence that a physical association existed between the two transcriptional machineries. However, recently it was reported that there is a physical interaction between the trimeric viral RdRp and cellular Pol II. The viral RdRp was found to interact with the C-terminal domain (CTD) of initiating Pol II, at a stage in the transcription cycle when capping takes place. It was therefore proposed that this interaction may be required for the viral RNA (vRNA) polymerase to gain access to capped RNA substrates for endonucleolytic cleavage. The virus not only relies on cellular factors to support its own RNA synthesis, but also subverts cellular pathways in order to generate an environment optimised for viral multiplication. In this respect, the interaction of the viral NS1 protein with factors involved in cellular pre-mRNA processing is of particular relevance. The virus also alters the distribution of Pol II on cellular genes, leading to a reduction in elongating Pol II thereby contributing to the phenomenon known as host shut-off.


Experiences of primary care physicians and staff following lean workflow redesign.
In response to growing pressures on primary care, leaders have introduced a wide range of workforce and practice innovations, including team redesigns that delegate some physician tasks to nonphysicians. One important question is how such innovations affect care team members, particularly in view of growing dissatisfaction and burnout among healthcare professionals. We examine the work experiences of primary care physicians and staff after implementing Lean-based workflow redesigns. This included co-locating physician and medical assistant dyads, delegating significant responsibilities to nonphysician staff, and mandating greater coordination and communication among all care team members. The redesigns were implemented and scaled in three phases across 46 primary care departments in a large ambulatory care delivery system. We fielded 1164 baseline and 1333 follow-up surveys to physicians and other nonphysician staff (average 73% response rate) to assess workforce engagement (e.g., job satisfaction, motivation), perceptions of the work environment, and job-related burnout. We conducted multivariate regressions to detect changes in experiences after the redesign, adjusting for respondent characteristics and clustering of within-clinic responses. We found that both physicians and nonphysician staff reported higher levels of engagement and teamwork after implementing redesigns. However, they also experienced higher levels of burnout and perceptions of the workplace as stressful. Trends were the same for both occupational groups, but the increased reports of stress were greater among physicians. Additionally, members of all clinics, except for the pilot site that developed the new workflows, reported higher burnout, while perceptions of workplace stress increased in all clinics after the redesign. Our findings partially align with expectations of work redesign as a route to improving physician and staff experiences in delivering care. Although teamwork and engagement increased, the redesigns in our study were not enough to moderate long-standing challenges facing primary care. Yet higher levels of empowerment and engagement, as observed in the pilot clinic, may be particularly effective in facilitating improvements while combating fatigue. To help practices cope with increasing burdens, interventions must directly benefit healthcare professionals without overtaxing an already overstretched workforce.


Evaluation and Management of Testosterone Deficiency: AUA Guideline.
There has been a marked increase in testosterone prescriptions in the past decade resulting in a growing need to give practicing clinicians proper guidance on the evaluation and management of the testosterone deficient patient. A systematic review utilized research from the Mayo Clinic Evidence Based Practice Center and additional supplementation by the authors. Evidence-based statements were based on body of evidence strength Grade A, B, or C and were designated as Strong, Moderate, and Conditional Recommendations with additional statements presented in the form of Clinical Principles or Expert Opinions (table 1 in supplementary unabridged guideline, http://jurology.com/). This guideline was developed by a multi-disciplinary panel to inform clinicians on the proper assessment of patients with testosterone deficiency and the safe and effective management of men on testosterone therapy. Additional statements were developed to guide the clinician on the appropriate care of patients who are at risk for or have cardiovascular disease or prostate cancer as well as patients who are interested in preserving fertility. The care of testosterone deficient patients should focus on accurate assessment of total testosterone levels, symptoms, and signs as well as proper on-treatment monitoring to ensure therapeutic testosterone levels are reached and symptoms are ameliorated. Future longitudinal observational studies and clinical trials of significant duration in this space will improve diagnostic techniques and treatment of men with testosterone deficiency as well as provide more data on the adverse events that may be associated with testosterone therapy.


The gene encoding bone morphogenetic protein 8B is required for the initiation and maintenance of spermatogenesis in the mouse.
Bone morphogenetic protein 8B (BMP8B) is a member of the TGFbeta superfamily of growth factors. In the mouse, Bmp8b is expressed in male germ cells of the testis and trophoblast cells of the placenta, suggesting that it has a role in spermatogenesis and reproduction. To investigate these possibilities, we have generated mice with a targeted mutation in Bmp8b. Here, we show that homozygous Bmp8b(tm1blh) mutant males exhibit variable degrees of germ-cell deficiency and infertility. Detailed analysis reveals two separable defects in the homozygous mutant testes. First, during early puberty (2 weeks old or younger) the germ cells of all homozygous mutants either fail to proliferate or show a marked reduction in proliferation and a delayed differentiation. Second, in adults, there is a significant increase in programmed cell death (apoptosis) of spermatocytes, leading to germ-cell depletion and sterility. Sertoli cells and Leydig cells appear relatively unaffected in mutants. This study therefore provides the first genetic evidence that a murine germ cell-produced factor, BMP8B, is required for the resumption of male germ-cell proliferation in early puberty, and for germ-cell survival and fertility in the adult.


Oceans apart: the higher health costs of women in the U.S. compared to other nations, and how reform is helping.
An estimated 18.7 million U.S. women ages 19 to 64 were uninsured in 2010, up from 12.8 million in 2000. An additional 16.7 million women had health insurance but had such high out-of-pocket costs relative to their income that they were effectively underinsured in 2010. This issue brief examines the implications of poor coverage for women in the United States by comparing their experiences to those of women in 10 other industrialized nations, all of which have universal health insurance systems. The analysis finds that women in the United States--both with and without health insurance--are more likely to go without needed health care because of cost and have greater difficulty paying their medical bills than women in the 10 other countries. In 2014, the Affordable Care Act will substantially reduce health care cost exposure for all U.S. women by significantly expanding and improving health insurance coverage.


IL-1R1-Dependent Signals Improve Control of Cytosolic Virulent Mycobacteria In Vivo.
Mycobacterium tuberculosis infections claim more than a million lives each year, and better treatments or vaccines are required. A crucial pathogenicity factor is translocation from phagolysosomes to the cytosol upon phagocytosis by macrophages. Translocation from the phagolysosome to the cytosol is an ESX-1-dependent process, as previously shown in vitro Here, we show that in vivo, mycobacteria also translocate to the cytosol but mainly when host immunity is compromised. We observed only low numbers of cytosolic bacilli in mice, armadillos, zebrafish, and patient material infected with M. tuberculosis, M. marinum, or M. leprae In contrast, when innate or adaptive immunity was compromised, as in severe combined immunodeficiency (SCID) or interleukin-1 receptor 1 (IL-1R1)-deficient mice, significant numbers of cytosolic M. tuberculosis bacilli were detected in the lungs of infected mice. Taken together, in vivo, translocation to the cytosol of M. tuberculosis is controlled by adaptive immune responses as well as IL-1R1-mediated signals.IMPORTANCE For decades, Mycobacterium tuberculosis has been one of the deadliest pathogens known. Despite infecting approximately one-third of the human population, no effective treatment or vaccine is available. A crucial pathogenicity factor is subcellular localization, as M. tuberculosis can translocate from phagolysosome to the cytosol in macrophages. The situation in vivo is more complicated. In this study, we establish that high-level cytosolic escape of mycobacteria can indeed occur in vivo but mainly when host resistance is compromised. The IL-1 pathway is crucial for the control of the number of cytosolic mycobacteria. The establishment that immune signals result in the clearance of cells containing cytosolic mycobacteria connects two important fields, cell biology and immunology, which is vital for the understanding of the pathology of M. tuberculosis.


Comparison of a Ca(2+)-gated conductance and a second-messenger-gated conductance in rat olfactory neurons.
The effect of a rise in intracellular Ca(2+) concentration was analyzed in isolated rat olfactory neurons using a whole-cell patch-clamp technique. Intracellular dialysis of 1 mmol l(-)(1) Ca(2+) in a standard-K(+), low-Cl(-) internal solution (E(Cl)=-69 mV) from the patch pipette into the olfactory neurons induced a sustained outward current of 49+/-5 pA (N=13) at -50 mV in all the cells examined. The outward currents were inhibited by external application of 100 micromol l(-)(1) 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB). External application of a Ca(2+) ionophore, 3 micromol l(-)(1) ionomycin, induced an inward current in three of eight cells whose voltages were clamped using the gramicidin-perforated technique, but ionomycin elicited an outward current in the other five cells, suggesting that natural intracellular Cl(-) concentration in the olfactory neurons was heterogeneous. While intracellular dialysis of 50 micromol l(-)(1) inositol 1,4,5-trisphosphate (1,4,5-InsP(3)) in the standard-K(+), low-Cl(-) internal solution induced the NPPB-sensitive outward current in 31 % of cells, and 500 micromol l(-)(1) cAMP induced it in 21 % of cells, a large proportion of the cells displayed an inward current in response to 1,4,5-InsP(3) and cAMP. The results suggest that 1,4,5-InsP(3) and cAMP can elicit Ca(2+)-dependent Cl(-) conductance and Ca(2+)-independent cation conductance in rat olfactory neurons.


Network-guided modeling allows tumor-type independent prediction of sensitivity to all-trans-retinoic acid.
All-trans-retinoic acid (ATRA) is a differentiating agent used in the treatment of acute-promyelocytic-leukemia (APL) and it is under-exploited in other malignancies despite its low systemic toxicity. A rational/personalized use of ATRA requires the development of predictive tools allowing identification of sensitive cancer types and responsive individuals. RNA-sequencing data for 10 080 patients and 33 different tumor types were derived from the TCGA and Leucegene datasets and completely re-processed. The study was carried out using machine learning methods and network analysis. We profiled a large panel of breast-cancer cell-lines for in vitro sensitivity to ATRA and exploited the associated basal gene-expression data to initially generate a model predicting ATRA-sensitivity in this disease. Starting from these results and using a network-guided approach, we developed a generalized model (ATRA-21) whose validity extends to tumor types other than breast cancer. ATRA-21 predictions correlate with experimentally determined sensitivity in a large panel of cell-lines representative of numerous tumor types. In patients, ATRA-21 correctly identifies APL as the most sensitive acute-myelogenous-leukemia subtype and indicates that uveal-melanoma and low-grade glioma are top-ranking diseases as for average predicted responsiveness to ATRA. There is a consistent number of tumor types for which higher ATRA-21 predictions are associated with better outcomes. In summary, we generated a tumor-type independent ATRA-sensitivity predictor which consists of a restricted number of genes and has the potential to be applied in the clinics. Identification of the tumor types that are likely to be generally sensitive to the action of ATRA paves the way to the design of clinical studies in the context of these diseases. In addition, ATRA-21 may represent an important diagnostic tool for the selection of individual patients who may benefit from ATRA-based therapeutic strategies also in tumors characterized by lower average sensitivity.


Assessing Expectations: Towards a Toolbox for an Ethics of Emerging Technologies.
In recent years, several authors have argued that the desirability of novel technologies should be assessed early, when they are still emerging. Such an ethical assessment of emerging technologies is by definition focused on an elusive object. Usually promises, expectations, and visions of the technology are taken as a starting point. As Nordmann and Rip have pointed out in a recent article, however, ethicists should not take for granted the plausibility of such expectations and visions. In this paper, we explore how the quality of expectations on emerging technologies might be assessed when engaging in a reflection on the desirability of emerging technologies. We propose that an assessment of expectations' plausibility should focus on statements on technological feasibility, societal usability, and desirability of the expected technology. Whereas the feasibility statement and, to a lesser extent, the usability statements are frequently quite futuristic, the claims on desirability, by contrast, often display a conservative stance towards the future. Assessing the quality of expectations and visions on behalf of emerging technologies requires, then, a careful and well-directed use of both skepticism and imagination. We conclude with a brief overview of the tools and methods ethicists could use to assess claims made on behalf of emerging technologies and improve the ethical reflection on them.


Hypoxemia during aeromedical evacuation of the walking wounded.
Hypobaric hypoxemia is a well-known risk of aeromedical evacuation (AE). Validating patients as safe to fly includes assessment of oxygenation status as well as oxygen-carrying capability (hemoglobin). The incidence and severity of hypoxemia during AE of noncritically injured casualties have not been studied. Subjects deemed safe to fly by the validating flight surgeon were monitored with pulse oximetry from the flight line until arrival at definitive care. All subjects were US military personnel or contractors following traumatic injuries. Noninvasive oxygen saturation (SpO2), pulse rate, and noninvasive hemoglobin were measured every 5 seconds and recorded to electronic memory. Patient demographics and physiologic data were collected by chart abstraction from the Air Force Form 3899, patient movement record. The incidence and duration of hypoxemic events (SpO2 < 90%) and critical hypoxemic events were determined (SpO2 < 85%). Sixty-one casualties were evaluated during AE from Bagram Air Base to Landstuhl Regional Medical Center. The mean (SD) age was 26.2 (6) years, Injury Severity Score (ISS) was 8 (11), and mean SpO2 before AE was 96% (2%). The mean (SD) transport time was 9.3 (1.3) hours. Patients were monitored before AE for a brief period, yielding a total recording time of 10.28 hours. The mean (SD) hemoglobin at the time of enrollment was 13.2 (3.5) g/dL (9.4-18.0 g/dL). Hypoxemia (SpO2 < 90%) was seen in 55 (90%) of 61 subjects. The mean duration of SpO2 less than 90% was 44 minutes. The mean (SD) change in SpO2 from baseline to mean in-flight SpO2 was 4% (1.2%). Thirty-four patients (56%) exhibited an SpO2 less than 85% for 11.7 (15) minutes. Hypoxemia is a common event during AE of casualties. In patients with infection and concussion or mild traumatic brain injury, this could have long-term consequences. Epidemiologic/prognostic study, level V.


Evidence Suggests That The ACA's Tobacco Surcharges Reduced Insurance Take-Up And Did Not Increase Smoking Cessation.
To account for tobacco users' excess health care costs and encourage cessation, the Affordable Care Act (ACA) allowed insurers to impose a surcharge on tobacco users' premiums for plans offered on the health insurance exchanges, or Marketplaces. Low-income tax credits for Marketplace coverage were based on premiums for non-tobacco users, which means that these credits did not offset any surcharge costs. Thus, this policy greatly increased out-of-pocket premiums for many tobacco users. Using data for 2011-14 from the Behavioral Risk Factor Surveillance System, we examined the effect of tobacco surcharges on insurance status and smoking cessation in the first year of the exchanges' implementation, among adults most likely to purchase insurance from them. Relative to smokers who faced no surcharges, smokers facing medium or high surcharges had significantly reduced coverage (reductions of 4.3 percentage points and 11.6 percentage points, respectively), but no significant differences in smoking cessation. In contrast, those facing low surcharges showed significantly less smoking cessation. Taken together, these findings suggest that tobacco surcharges conflicted with a major goal of the ACA-increased financial protection-without increasing smoking cessation. States should consider these potential effects when deciding whether to limit surcharges to less than the federal maximum.


Automated quantification of synapses by fluorescence microscopy.
The quantification of synapses in neuronal cultures is essential in studies of the molecular mechanisms underlying synaptogenesis and synaptic plasticity. Conventional counting of synapses based on morphological or immunocytochemical criteria is extremely work-intensive. We developed a fully automated method which quantifies synaptic elements and complete synapses based on immunocytochemistry. Pre- and postsynaptic elements are detected by their corresponding fluorescence signals and their proximity to dendrites. Synapses are defined as the combination of a pre- and postsynaptic element within a given distance. The analysis is performed in three dimensions and all parameters required for quantification can be easily adjusted by a graphical user interface. The integrated batch processing enables the analysis of large datasets without any further user interaction and is therefore efficient and timesaving. The potential of this method was demonstrated by an extensive quantification of synapses in neuronal cultures from DIV 7 to DIV 21. The method can be applied to all datasets containing a pre- and postsynaptic labeling plus a dendritic or cell surface marker.


Participant informed consent in cluster randomized trials: review.
The Nuremberg code defines the general ethical framework of medical research with participant consent as its cornerstone. In cluster randomized trials (CRT), obtaining participant informed consent raises logistic and methodologic concerns. First, with randomization of large clusters such as geographical areas, obtaining individual informed consent may be impossible. Second, participants in randomized clusters cannot avoid certain interventions, which implies that participant informed consent refers only to data collection, not administration of an intervention. Third, complete participant information may be a source of selection bias, which then raises methodological concerns. We assessed whether participant informed consent was required in such trials, which type of consent was required, and whether the trial was at risk of selection bias because of the very nature of participant information. We systematically reviewed all reports of CRT published in MEDLINE in 2008 and surveyed corresponding authors regarding the nature of the informed consent and the process of participant inclusion. We identified 173 reports and obtained an answer from 113 authors (65.3%). In total, 23.7% of the reports lacked information on ethics committee approval or participant consent, 53.1% of authors declared that participant consent was for data collection only and 58.5% that the group allocation was not specified for participants. The process of recruitment (chronology of participant recruitment with regard to cluster randomization) was rarely reported, and we estimated that only 56.6% of the trials were free of potential selection bias. For CRTs, the reporting of ethics committee approval and participant informed consent is less than optimal. Reports should describe whether participants consented for administration of an intervention and/or data collection. Finally, the process of participant recruitment should be fully described (namely, whether participants were informed of the allocation group before being recruited) for a better appraisal of the risk of selection bias.


Alterations in corticotropin-releasing factor-like immunoreactivity in discrete rat brain regions after acute and chronic stress.
Corticotropin releasing factor (CRF) may regulate endocrine, autonomic, and behavioral responses to stress. Evidence indicates that CRF-like immunoreactivity (CRF-LI) is widely distributed throughout the CNS. In this study, the distribution of CRF-LI was determined in 36 rat brain regions by combined radioimmunoassay-micropunch dissection techniques and the effect of stress on CRF-LI was investigated, using a chronic stress model that induces endocrine changes in rats similar to those seen in depressed humans. A control group of rats was handled daily. An acute stress group was subjected to 3 hr of immobilization at 4 degrees C, while a chronic stress group was exposed to unpredictable stressors. Thirty-six brain regions were microdissected by the technique of Palkovits and assayed for CRF-LI, using a specific antiserum to ovine CRF. CRF-LI was detected in most regions. In controls, the highest concentrations were found in the arcuate nucleus/median eminence, the hypothalamic paraventricular (PVN) nucleus, and the periventricular nucleus. The next highest levels were found in the raphe nuclei and dorsal vagal complex. CRF-LI was well represented in the locus coeruleus (LC); in the central, cortical, and medial amygdaloid nuclei; and in the bed nucleus of the stria terminalis. Low concentrations occurred in the hippocampus and cerebrocortical regions. Appreciable concentrations were detected in midbrain and brain stem regions. Acute stress reduced CRF-LI in the arcuate nucleus/median eminence (ME) (by 52%) and in the median preoptic (MPO) nucleus (by 32%) and doubled its concentration in the locus coeruleus.(ABSTRACT TRUNCATED AT 250 WORDS)


Immobilization of proteolytic enzymes on replica-molded thiol-ene micropillar reactors via thiol-gold interaction.
We introduce rapid replica molding of ordered, high-aspect-ratio, thiol-ene micropillar arrays for implementation of microfluidic immobilized enzyme reactors (IMERs). By exploiting the abundance of free surface thiols of off-stoichiometric thiol-ene compositions, we were able to functionalize the native thiol-ene micropillars with gold nanoparticles (GNPs) and these with proteolytic α-chymotrypsin (CHT) via thiol-gold interaction. The micropillar arrays were replicated via PDMS soft lithography, which facilitated thiol-ene curing without the photoinitiators, and thus straightforward bonding and good control over the surface chemistry (number of free surface thiols). The specificity of thiol-gold interaction was demonstrated over allyl-rich thiol-ene surfaces and the robustness of the CHT-IMERs at different flow rates and reaction temperatures using bradykinin hydrolysis as the model reaction. The product conversion rate was shown to increase as a function of decreasing flow rate (increasing residence time) and upon heating of the IMER to physiological temperature. Owing to the effective enzyme immobilization onto the micropillar array by GNPs, no further purification of the reaction solution was required prior to mass spectrometric detection of the bradykinin hydrolysis products and no clogging problems, commonly associated with conventional capillary packings, were observed. The activity of the IMER remained stable for at least 1.5 h (continuous use), suggesting that the developed protocol may provide a robust, new approach to implementation of IMER technology for proteomics research. Graphical abstract.


Understanding the Adolescent Gap in HIV Testing Among Clients of Antenatal Care Services in West and Central African Countries.
New HIV infections among children have declined significantly more slowly in West and central African countries (WCA) than in eastern and southern African countries between 2009 and 2015. Since adolescent fertility is particularly high in WCA countries, frequent mother-to-child transmission (MTCT) of HIV may in part be due to low coverage of HIV testing among adolescents during antenatal care (ANC). We investigated this adolescent gap in HIV testing using survey data from the demographic and health surveys and multiple indicators cluster surveys collected in 21 WCA countries since 2009. We found significant adolescent gaps in HIV testing in 12 out of 21 WCA countries, with the largest gap observed in Nigeria. In countries with a significant adolescent gap in HIV testing, we used Fairlie decompositions to assess what proportion of these gaps were explained by age-related differences in the distribution of (a) marital status, (b) socioeconomic status (SES), (c) MTCT-related knowledge, and (d) patterns of ANC utilization. Differences in SES and MTCT-related knowledge were the most consistent determinants of adolescent gaps in HIV testing during ANC. Differences in ANC utilization (e.g., fewer and possibly delayed ANC visits) also contributed to the adolescent gap in 8 out of 12 countries. Interventions that improve knowledge of MTCT risks, and/or promote the sustained use of ANC services, could help engage HIV-infected adolescents who become pregnant in PMTCT services. Targeting these interventions at the most disadvantaged households will be crucial in further reducing HIV infections among children.


Protecting Agriculture: The Legal Basis of Regulatory Action in Florida.
The Division of Plant Industry of the Department of Agriculture and Consumer Services of Florida is empowered to protect the state's agriculture from both introduction and intrastate movement of pests. This responsibility is shared by similar regulatory agencies of the federal government and other states. The legal basis of such regulatory action, police power and eminent domain, is reviewed. The regulatory processes by the State of Florida of two important citrus pests, the burrowing nematode and Asiatic citrus canker, are presented. It was concluded that Florida's regulatory power to take property is of great importance to the agricultural industry and to Florida's economy. This power must be exercised within the constitutional limitation that "no person shall be deprived of life, liberty, or property without due processes of law." However, the imminence of an emergency may justify state action without a prior hearing or just compensation of valueless items.


Immunophenotypic analysis of erythroid dysplasia in myelodysplastic syndromes. A report from the IMDSFlow working group.
Current recommendations for diagnosing myelodysplastic syndromes endorse flow cytometry as an informative tool. Most flow cytometry protocols focus on the analysis of progenitor cells and the evaluation of the maturing myelomonocytic lineage. However, one of the most frequently observed features of myelodysplastic syndromes is anemia, which may be associated with dyserythropoiesis. Therefore, analysis of changes in flow cytometry features of nucleated erythroid cells may complement current flow cytometry tools. The multicenter study within the IMDSFlow Working Group, reported herein, focused on defining flow cytometry parameters that enable discrimination of dyserythropoiesis associated with myelodysplastic syndromes from non-clonal cytopenias. Data from a learning cohort were compared between myelodysplasia and controls, and results were validated in a separate cohort. The learning cohort comprised 245 myelodysplasia cases, 290 pathological, and 142 normal controls; the validation cohort comprised 129 myelodysplasia cases, 153 pathological, and 49 normal controls. Multivariate logistic regression analysis performed in the learning cohort revealed that analysis of expression of CD36 and CD71 (expressed as coefficient of variation), in combination with CD71 fluorescence intensity and the percentage of CD117+ erythroid progenitors provided the best discrimination between myelodysplastic syndromes and non-clonal cytopenias (specificity 90%; 95% confidence interval: 84-94%). The high specificity of this marker set was confirmed in the validation cohort (92%; 95% confidence interval: 86-97%). This erythroid flow cytometry marker combination may improve the evaluation of cytopenic cases with suspected myelodysplasia, particularly when combined with flow cytometry assessment of the myelomonocytic lineage.


'Gender is not even a side issue…it's a non-issue': career trajectories and experiences from the perspective of male and female healthcare managers in Kenya.
Women comprise a significant proportion of the health workforce globally but remain under-represented in the higher professional categories. Concern about the under-representation of women in health leadership positions has resulted in increased research on the topic, although this research has focused primarily on high-income countries. An improved understanding of the career trajectories and experiences of healthcare leaders in low- and middle-income countries (LMICs), and the role of gender, is therefore needed. This qualitative case study was undertaken in two counties in coastal Kenya. Drawing on the life-history approach, 12 male and 13 female healthcare leaders were interviewed between August 2015 and July 2016 on their career progression and related experiences. Although gender was not spontaneously identified as a significant influence, closer exploration of responses revealed that gendered factors played an important role. Most fundamentally, women's role as child bearers and gendered societal expectations including child nurturing and other domestic responsibilities can influence their ability to take up leadership opportunities, and their selection and appointment as leaders. Women's selection and appointment as leaders may also be influenced by positive discrimination policies (in favour of women), and by perceptions of women and men as having different leadership styles (against women, who some described as more emotive and reactive). These gendered influences intersect in relatively invisible ways with other factors more readily identified by respondents to influence their progression and experience. These factors included: professional cadre, with doctors more likely to be selected into leadership roles; and personal and professional support systems ranging from family support and role models, through to professional mentorship and continuing education. We discuss the implications of these findings for policy, practice and research, including highlighting the need for more in-depth intersectionality analyses of leadership experience in LMICs.


Temperature dependence of the reflectivity in absorbing Bragg reflectors.
The reflectivity of absorbing Bragg reflectors consisting of a GaAs/AlAs Bragg mirror and a InGaAs/InGaAsP multiple-quantum-well cavity layer was studied as a function of temperature. An absorption dip in the stop band due to the optical confinement of the Fabry-Perot resonance was observed in the reflectivity spectra. The absorption intensity of the dip increased with temperature and was explained by the resonant coincidence of the Fabry-Perot cavity mode and the quantum-well absorption. The temperature-dependent reflectivity spectra were successfully reproduced using the transfer matrix method and the linear dependence of the refractive index on temperature.


Brain-derived neurotrophic factor as an indicator of chemical neurotoxicity: an animal-free CNS cell culture model.
Recent changes to the legislation on chemicals and cosmetics testing call for a change in the paradigm regarding the current 'whole animal' approach for identifying chemical hazards, including the assessment of potential neurotoxins. Accordingly, since 2004, we have worked on the development of the integrated co-culture of post-mitotic, human-derived neurons and astrocytes (NT2.N/A), for use as an in vitro functional central nervous system (CNS) model. We have used it successfully to investigate indicators of neurotoxicity. For this purpose, we used NT2.N/A cells to examine the effects of acute exposure to a range of test chemicals on the cellular release of brain-derived neurotrophic factor (BDNF). It was demonstrated that the release of this protective neurotrophin into the culture medium (above that of control levels) occurred consistently in response to sub-cytotoxic levels of known neurotoxic, but not non-neurotoxic, chemicals. These increases in BDNF release were quantifiable, statistically significant, and occurred at concentrations below those at which cell death was measureable, which potentially indicates specific neurotoxicity, as opposed to general cytotoxicity. The fact that the BDNF immunoassay is non-invasive, and that NT2.N/A cells retain their functionality for a period of months, may make this system useful for repeated-dose toxicity testing, which is of particular relevance to cosmetics testing without the use of laboratory animals. In addition, the production of NT2.N/A cells without the use of animal products, such as fetal bovine serum, is being explored, to produce a fully-humanised cellular model.


C-terminus of heat shock protein 70-interacting protein-dependent GTP cyclohydrolase I degradation in lambs with increased pulmonary blood flow.
We showed that nitric oxide (NO) signaling is decreased in the pulmonary vasculature before the development of endothelial dysfunction in a lamb model of congenital heart disease and increased pulmonary blood flow (Shunt). The elucidation of the molecular mechanism by which this occurs was the purpose of this study. Here, we demonstrate that concentrations of the endogenous NO synthase (NOS) inhibitor, asymmetric dimethylarginine (ADMA), are elevated, whereas the NOS cofactor tetrahydrobiopterin (BH(4)) is decreased in Shunt lambs. Our previous studies demonstrated that ADMA decreases heat shock protein-90 (Hsp90) chaperone activity, whereas other studies suggest that guanosine-5'-triphosphate cyclohydrolase 1 (GCH1), the rate-limiting enzyme in the generation of BH(4), may be a client protein for Hsp90. Thus, we determined whether increases in ADMA could alter GCH1 protein and activity. Our data demonstrate that ADMA decreased GCH1 protein, but not mRNA concentrations, in pulmonary arterial endothelial cells (PAECs) because of the ubiquitination and proteasome-dependent degradation of GCH1. We also found that Hsp90-GCH1 interactions were reduced, whereas the association of GCH1 with Hsp70 and the C-terminus of Hsp70-interacting protein (CHIP) increased in ADMA-exposed PAECs. The overexpression of CHIP potentiated, whereas a CHIP U-box domain mutant attenuated, ADMA-induced GCH1 degradation and reductions in cellular BH(4) concentrations. We also found in vivo that Hsp90/GCH1 interactions are decreased, whereas GCH1-Hsp70 and GCH1-CHIP interactions and GCH1 ubiquitination are increased. Finally, we found that supplementation with l-arginine restored Hsp90-GCH1 interactions and increased both BH(4) and NO(x) concentrations in Shunt lambs. In conclusion, increased concentrations of ADMA can indirectly alter NO signaling through decreased cellular BH(4) concentrations, secondary to the disruption of Hsp90-GCH1 interactions and the CHIP-dependent proteasomal degradation of GCH1.


Heparan sulfate mediates neuroprotection from degeneration in experimental glutaric aciduria.
Glutaric aciduria type 1 (GA1) is a childhood metabolic disorder associated with crises that lead to striatal necrosis. Although the disorder can be controlled with diet, there is no current treatment to ameliorate the neurodegeneration following a metabolic crisis. We hypothesized that heparan sulfate (HS) administration would stimulate neural stem cell proliferation by dimerizing with FGF-2 and binding to the FGF-2 receptor on neural stem cells, thus enhancing the number of newly generated neurons to repair damage following a metabolic crisis. In addition, FGF-2 is known to exert neuroprotective effects independent of neurogenesis, so HS may also have neuroprotective activities. To test these hypotheses, ibotenic acid was injected into the striatum of adult mice, mimicking the metabolic crisis and damage caused by glutaric aciduria. Daily doses of HS and bromodeoxyuridine (BrdU) or BrdU alone were administered starting 1 day after the ibotenic acid lesion. BrdU was used to label dividing cells. Fluorescent immunohistochemistry was used to quantify the lesion size and evaluate the phenotype of BrdU-positive cells. Intrastriatal administration of ibotenic acid resulted in a substantial striatal lesion that occupied 18.5% of the ipsilateral brain hemisphere. In contrast, animals treated with HS exhibited a lesion volume representing <1% of the ipsilateral brain hemisphere (ANOVA; p < 0.0001). Increased neurogenesis, however, was not observed in this group. These results suggest that HS administration 2 days after a "metabolic crisis" can ameliorate brain injury in an animal model of GA1. The neuroprotective mechanisms of HS, however, remain to be elucidated but may exert their actions indirectly through binding with FGF-2.


Five Years of Experimental Warming Increases the Biodiversity and Productivity of Phytoplankton.
Phytoplankton are key components of aquatic ecosystems, fixing CO2 from the atmosphere through photosynthesis and supporting secondary production, yet relatively little is known about how future global warming might alter their biodiversity and associated ecosystem functioning. Here, we explore how the structure, function, and biodiversity of a planktonic metacommunity was altered after five years of experimental warming. Our outdoor mesocosm experiment was open to natural dispersal from the regional species pool, allowing us to explore the effects of experimental warming in the context of metacommunity dynamics. Warming of 4°C led to a 67% increase in the species richness of the phytoplankton, more evenly-distributed abundance, and higher rates of gross primary productivity. Warming elevated productivity indirectly, by increasing the biodiversity and biomass of the local phytoplankton communities. Warming also systematically shifted the taxonomic and functional trait composition of the phytoplankton, favoring large, colonial, inedible phytoplankton taxa, suggesting stronger top-down control, mediated by zooplankton grazing played an important role. Overall, our findings suggest that temperature can modulate species coexistence, and through such mechanisms, global warming could, in some cases, increase the species richness and productivity of phytoplankton communities.


Effect of Brussels sprouts on oxidative DNA-damage in man.
The effect of consumption of Brussels sprouts on the excretion of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) into human urine was investigated. Ten non-smoking volunteers (five males and five females) were randomly assigned to two groups. Five volunteers started on a diet of 300 g of glucosinolate-free vegetables whereas the other five consumed 300 g of Brussels sprouts per day. After 1 week dietary regimes were reversed. Levels of 8-oxodG in 24-h urine samples were measured by high-performance liquid chromatography. In four of five males a reduction in 8-oxodG was found, whereas in the fifth male the 8-oxodG excretion was high in the control period and was even much higher in the sprouts period. In females no effect of consumption of Brussels sprouts on excretion of 8-oxodG was found. Our previous and present findings support the results of epidemiologic studies that consumption of brassica vegetables may diminish cancer risk.


Understanding the Representation and Computation of Multilayer Perceptrons: A Case Study in Speech Recognition.
Despite the recent success of deep learning, the nature of the transformations they apply to the input features remains poorly understood. This study provides an empirical framework to study the encoding properties of node activations in various layers of the network, and to construct the exact function applied to each data point in the form of a linear transform. These methods are used to discern and quantify properties of feedforward neural networks trained to map acoustic features to phoneme labels. We show a selective and nonlinear warping of the feature space, achieved by forming prototypical functions to account for the possible variation of each class. This study provides a joint framework where the properties of node activations and the functions implemented by the network can be linked together.


Plexitis as a predictive factor of early postoperative clinical recurrence in Crohn's disease.
A previous study suggested that the presence of myenteric plexitis in the proximal resection margins could be predictive of early endoscopic recurrence after ileocolonic or ileal resection for Crohn's disease (CD). The aim of the present study was to assess the predictive value of plexitis for early clinical CD recurrence. All consecutive patients with ileocolonic or ileal resection for active CD in Lariboisière Hospital (Paris) between 1995 and 2006 were included. Clinical, surgical, histological and follow-up data were extracted from medical charts. Early clinical recurrence was defined as the reappearance of CD clinical manifestations requiring a specific treatment within 2 years postsurgery. The proximal resection margin was analysed using haematein eosin saffron (HES) staining and immunochemistry targeting mastocytes (anti-CD117 antibody) and lymphocytes (anti-CD3 antibody). Eosinophils were detected by HES staining. Ten cases of ileocolonic resections for caecal carcinoma served as controls. Data were available from 171 postoperative follow-up periods in 164 patients with CD. Early clinical recurrence of CD occurred in 28.1%. In multivariate analysis, factors associated with postoperative recurrence were active smoking (hazard ratio (HR) = 1.94; 95% CI 1.06 to 3.60; p = 0.033), submucosal plexitis with >or=3 mastocytes (HR = 1.87; 95% CI 1.00 to 3.46; p = 0.048) and a disease-free resection margin <5 cm (HR = 0.52; 95% CI 0.27 to 1.02; p = 0.059). Submucosal plexitis is associated with early clinical recurrence and could be taken into account in studies searching for new treatment strategies in the immediate postoperative period.


Increase of RT-related transmitted drug resistance in non-CRF01_AE among HIV type 1-infected men who have sex with men in the 7 cities of China.
To elucidate new features in the prevalence of HIV-1 transmitted drug resistance (TDR) in men who have sex with men (MSM) in China. A total of 441 HIV-1-positive subjects were recruited from high-risk MSM populations in 7 cities across China between 2012 and 2013. Nucleotide sequences of 1.1-kb pol-RT regions were amplified and sequenced from 367 of the 441 samples. Phylogenetic and genetic drug-resistant analyses were performed. The overall distribution of HIV-1 genotypes was as the following: CRF01_AE, 52.3%; CRF07_BC, 33.2%; 01/B recombinants, 6.0%; subtype B (United States-European), 3.8%; subtype B', 3.8%; and CRF08_BC, 0.8%. About 91.3% of the sequences clustered together. An overall 4.6% TDR rate was found. Reverse transcriptase (RT)-related TDR reached 2.7%. TDR of 2.2% was detected in protease region. Most of RT-related TDRs were detected in non-CRF01_AE subtypes (90.0%, 9/10), including T215A/S, K101E, K103N, V106M, and E138G. Most of the strains with TDRs (88.2%, 15/17) were presented in the clusters. TDR strains against non-nucleoside reverse transcriptase inhibitors in non-CRF01_AE subtypes also formed some subclusters (70.0%, 7/10). One CRF07_BC subject with K103N in Kunming had a very close genetic distance with one that received highly active antiretroviral therapy locally (bootstrap = 99%). Two CRF55_01B subjects carrying K103N in Changsha and Nanjing also had a very close genetic distance (bootstrap = 100%). RT-related TDR of non-CRF01_AE became the main TDR among MSM in China. There is an increasing trend and a potential transmission risk for the RT-related TDR among MSM throughout China. Some TDRs could have already been transmitted among different cities. Intervention efforts should be strengthened among MSM to prevent further transmission of HIV and the proliferation of the strains with TDR.


Does Day of Surgery Affect Hospital Length of Stay and Charges Following Minimally Invasive Transforaminal Lumbar Interbody Fusion?
Retrospective Cohort. To determine if an association exists between surgery day and length of stay or hospital costs after minimally invasive transforaminal lumbar interbody fusion (MIS TLIF). Length of inpatient stay after orthopedic procedures has been identified as a primary cost driver, and previous research has focused on determining risk factors for prolonged length of stay. In the arthroplasty literature, surgery performed later in the week has been identified as a predictor of increased length of stay. However, no such investigation has been performed for MIS TLIF. A surgical registry of patients undergoing MIS TLIF between 2008 and 2016 was retrospectively reviewed. Patients were grouped based on day of surgery, with groups including early surgery and late surgery. Day of surgery group was tested for an association with demographics and perioperative variables using the student t test or χ analysis. Day of surgery group was then tested for an association with direct hospital costs using multivariate linear regression. In total, 438 patients were analyzed. In total, 51.8% were in the early surgery group, and 48.2% were in the late surgery group. There were no differences in demographics between groups. There were no differences between groups with regard to operative time, intraoperative blood loss, length of stay, or discharge day. Finally, there were no differences in total hospital charges between early and late surgery groups (P=0.247). The specific day on which a MIS TLIF procedure occurs is not associated with differences in length of inpatient stay or total hospital costs. This suggests that the postoperative course after MIS TLIF procedures is not affected by the differences in hospital staffing that occurs on the weekend compared with weekdays.


Stereoselective N-glycosylation of 2-deoxythioribosides for fluorescent nucleoside synthesis.
An efficient method for the N-2-deoxyribosylation of modified nucleobases by 2-deoxythioriboside donors is reported. In the presence of an in situ silylated nucleobase, thioglycosides can be activated with NIS/HOTf to give nucleosides in high yields and with good β-selectivity. By tuning the protecting groups on the C3 and C5 hydroxyls, α/β ratios ranging from 1.0:4.0 to 4.5:1.0 can be obtained. This strategy is applicable to the synthesis of various nucleosides, including ring-expanded pyrimidine derivatives containing sulfur that have previously been reported in low yields. The utility of this approach is further demonstrated by the synthesis of fluorescent nucleosides analogues such as quinazoline and oxophenothiazine that should find broad utility in DNA-folding and recognition studies.


DNA damage-induced cell cycle regulation and function of novel Chk2 phosphoresidues.
Chk2 kinase is activated by DNA damage to regulate cell cycle arrest, DNA repair, and apoptosis. Phosphorylation of Chk2 in vivo by ataxia telangiectasia-mutated (ATM) on threonine 68 (T68) initiates a phosphorylation cascade that promotes the full activity of Chk2. We identified three serine residues (S19, S33, and S35) on Chk2 that became phosphorylated in vivo rapidly and exclusively in response to ionizing radiation (IR)-induced DNA double-strand breaks in an ATM- and Nbs1-dependent but ataxia telangiectasia- and Rad3-related-independent manner. Phosphorylation of these residues, restricted to the G(1) phase of the cell cycle, was induced by a higher dose of IR (>1 Gy) than that required for phosphorylation of T68 (0.25 Gy) and declined by 45 to 90 min, concomitant with a rise in Chk2 autophosphorylation. Compared to the wild-type form, Chk2 with alanine substitutions at S19, S33, and S35 (Chk2(S3A)) showed impaired dimerization, defective auto- and trans-phosphorylation activities, and reduced ability to promote degradation of Hdmx, a phosphorylation target of Chk2 and regulator of p53 activity. Besides, Chk2(S3A) failed to inhibit cell growth and, in response to IR, to arrest G(1)/S progression. These findings underscore the critical roles of S19, S33, and S35 and argue that these phosphoresidues may serve to fine-tune the ATM-dependent response of Chk2 to increasing amounts of DNA damage.


HIV-1 Envelope Mimicry of Host Enzyme Kynureninase Does Not Disrupt Tryptophan Metabolism.
The HIV-1 envelope protein (Env) has evolved to subvert the host immune system, hindering viral control by the host. The tryptophan metabolic enzyme kynureninase (KYNU) is mimicked by a portion of the HIV Env gp41 membrane proximal region (MPER) and is cross-reactive with the HIV broadly neutralizing Ab (bnAb) 2F5. Molecular mimicry of host proteins by pathogens can lead to autoimmune disease. In this article, we demonstrate that neither the 2F5 bnAb nor HIV MPER-KYNU cross-reactive Abs elicited by immunization with an MPER peptide-liposome vaccine in 2F5 bnAb VHDJH and VLJL knock-in mice and rhesus macaques modified KYNU activity or disrupted tissue tryptophan metabolism. Thus, molecular mimicry by HIV-1 Env that promotes the evasion of host anti-HIV-1 Ab responses can be directed toward nonfunctional host protein epitopes that do not impair host protein function. Therefore, the 2F5 HIV Env gp41 region is a key and safe target for HIV-1 vaccine development.


A novel thromboxane synthetase inhibitor, DP-1904, inhibits human blood eosinophil degranulation.
Eosinophils have been recognized to be associated with various immune responses and disease processes including bronchial asthma. Eosinophils release a number of cytotoxic and neurotoxic mediators. However, the factors regulating such release and the underlying mechanisms are unclear. In this study, we investigated the effect of a selective and potent thromboxane synthase inhibitor, DP-1904, on the release of eosinophil cationic protein (ECP) in platelet activating factor (PAF) and IgG-stimulated human blood eosinophils. PAF (1 microM) and IgG both released ECP which constituted about 25-30% of the total ECP content. The control protein, ovalbumin, did not release any ECP over the basal values. DP-1904 in two different concentrations, 10 microM and 100 microM, significantly attenuated the release of ECP in response to PAF or IgG. The mean percent inhibition by 10 microM DP-1904 was 49 +/- 10 and 31 +/- 2 against PAF and IgG-induced ECP release, respectively. However, at 100 microM DP-1904 the percent inhibition was 76 +/- 14 and 67 +/- 2, respectively. These data suggest that TXA2 is an important mediator in the regulation of eosinophil degranulation, and DP-1904 thus might prove beneficial in the treatment of bronchial asthma.


Predicting clinical states in individual patients.
A probability model expresses the relation between the presence of clinical findings (input or independent variables) and the probability that a clinical state will occur (the dependent variable); for example, it expresses the probability that a disease is present or will develop or the probability that an outcome state will be reached. Probability models are developed by using selected study groups. Although these models are most often used to make predictions for groups of patients, they can also predict clinical states for individual patients. The following seven criteria provide a basis for the critical appraisal of probability models. In particular, physicians can use these criteria to decide when a specific probability model should be used to make a prediction in an individual patient. Five of the criteria are concerned with the applicability of a model to a particular patient: 1) the comparability of the patient and the study group used to develop the model; 2) the congruence between the clinical state of interest to patient and physician and the model's outcome; 3) the availability of all input variables where and when the prediction is to be made; 4) the usefulness of a quantitative estimate of the predicted clinical state; and 5) the degree of uncertainty in the probability estimate. The other two criteria are concerned with how well the probability model "works": 6) the fit of probabilities calculated from the model to the outcomes actually observed and 7) the model's ability to discriminate between outcome states relative to chance and to other, more traditional, prediction methods. We illustrate the use of these criteria by applying them, in the form of questions, to a convenient, tabular version of a model that estimates a patient's chances of surviving for 10 years after having definitive surgical therapy for primary cutaneous melanoma.


Mechanical properties of calvarial bones in a mouse model for craniosynostosis.
The mammalian cranial vault largely consists of five flat bones that are joined together along their edges by soft fibrous tissues called sutures. Premature closure of the cranial sutures, craniosynostosis, can lead to serious clinical pathology unless there is surgical intervention. Research into the genetic basis of the disease has led to the development of various animal models that display this condition, e.g. mutant type Fgfr2C342Y/+ mice which display early fusion of the coronal suture (joining the parietal and frontal bones). However, whether the biomechanical properties of the mutant and wild type bones are affected has not been investigated before. Therefore, nanoindentation was used to compare the elastic modulus of cranial bone and sutures in wild type (WT) and Fgfr2C342Y/+mutant type (MT) mice during their postnatal development. Further, the variations in properties with indentation position and plane were assessed. No difference was observed in the elastic modulus of parietal bone between the WT and MT mice at postnatal (P) day 10 and 20. However, the modulus of frontal bone in the MT group was lower than the WT group at both P10 (1.39±0.30 vs. 5.32±0.68 GPa; p<0.05) and P20 (5.57±0.33 vs. 7.14±0.79 GPa; p<0.05). A wide range of values was measured along the coronal sutures for both the WT and MT samples, with no significant difference between the two groups. Findings of this study suggest that the inherent mechanical properties of the frontal bone in the mutant mice were different to the wild type mice from the same genetic background. These differences may reflect variations in the degree of biomechanical adaptation during skull growth, which could have implications for the surgical management of craniosynostosis patients.


Quantification of anatomic, geometric, and load transfer characteristics of the wrist joint.
The work reported here is an assimilation of 8 years of research, the purpose of which was to gain a better understanding of the normal and abnormal workings of the wrist joint. The results are summarized in three major areas of concentration: descriptive anatomic studies, which include direct measurements of cadaver specimens, biomechanical loading studies to define load distribution through normal and abnormal wrists, and three-dimensional (3D) anatomy studies using solid models derived from computed tomographic (CT) images of in vivo and cadaver wrists. The descriptive anatomic studies used 393 cadaver wrists to evaluate the incidence and distribution of anatomic features, arthroses, chondromalacia, and soft-tissue lesions. The data were analyzed for any statistically significant associations among different variables. The biomechanical studies characterized the biomechanics of the human wrist in a variety of normal, simulated traumatic, and surgically treated conditions. The results of the load studies have provided clinically relevant information on the normal anatomy and functional mechanics of the wrist as well as guidelines for the treatment of a number of different fractures and ligament injuries. The 3D anatomic studies use quantitative analysis of 3D reconstructions of CT scan data to build a normative database of carpal bone morphology. Thirty-five wrists were imaged in a CT scanner. Quantitative measurements of volume, surface area, maximum length, and intercarpal distances were than assessed. A reliable 3D carpal height ratio (3DCHR) was calculated by dividing the capitate maximum length by the carpal height, which is the minimum distance between the fourth metacarpal and the radius.


Biocompatible fluorinated polyglycerols for droplet microfluidics as an alternative to PEG-based copolymer surfactants.
In droplet-based microfluidics, non-ionic, high-molecular weight surfactants are required to stabilize droplet interfaces. One of the most common structures that imparts stability as well as biocompatibility to water-in-oil droplets is a triblock copolymer surfactant composed of perfluoropolyether (PFPE) and polyethylene glycol (PEG) blocks. However, the fast growing applications of microdroplets in biology would benefit from a larger choice of specialized surfactants. PEG as a hydrophilic moiety, however, is a very limited tool in surfactant modification as one can only vary the molecular weight and chain-end functionalization. In contrast, linear polyglycerol offers further side-chain functionalization to create custom-tailored, biocompatible droplet interfaces. Herein, we describe the synthesis and characterization of polyglycerol-based triblock surfactants with tailored side-chain composition, and exemplify their application in cell encapsulation and in vitro gene expression studies in droplet-based microfluidics.


Phytotoxicity of four photosystem II herbicides to tropical seagrasses.
Coastal waters of the Great Barrier Reef (GBR) are contaminated with agricultural pesticides, including the photosystem II (PSII) herbicides which are the most frequently detected at the highest concentrations. Designed to control weeds, these herbicides are equally potent towards non-target marine species, and the close proximity of seagrass meadows to flood plumes has raised concerns that seagrasses may be the species most threatened by herbicides from runoff. While previous work has identified effects of PSII herbicides on the photophysiology, growth and mortality in seagrass, there is little comparative quantitative toxicity data for seagrass. Here we applied standard ecotoxicology protocols to quantify the concentrations of four priority PSII herbicides that inhibit photochemistry by 10, 20 and 50% (IC10, IC20 and IC50) over 72 h in two common seagrass species from the GBR lagoon. The photosystems of seagrasses Zosteramuelleri and Haloduleuninervis were shown to be generally more sensitive to the PSII herbicides Diuron, Atrazine, Hexazinone and Tebuthiuron than corals and tropical microalgae. The herbicides caused rapid inhibition of effective quantum yield (∆F/F m '), indicating reduced photosynthesis and maximum effective yields (Fv/Fm ) corresponding to chronic damage to PSII. The PSII herbicide concentrations which affected photosynthesis have been exceeded in the GBR lagoon and all of the herbicides inhibited photosynthesis at concentrations lower than current marine park guidelines. There is a strong likelihood that the impacts of light limitation from flood plumes and reduced photosynthesis from PSII herbicides exported in the same waters would combine to affect seagrass productivity. Given that PSII herbicides have been demonstrated to affect seagrass at environmental concentrations, we suggest that revision of environmental guidelines and further efforts to reduce PSII herbicide concentrations in floodwaters may both help protect seagrass meadows of the GBR from further decline.


Comparison of Costs of Care for Medicare Patients Hospitalized in Teaching and Nonteaching Hospitals.
Little empirical work has been performed on whether teaching hospitals are more expensive when considering total costs of care for an acute care episode. To compare total standardized costs at 30 days by hospital teaching status for common conditions. This cross-sectional study assessed the costs of hospitalizations among US Medicare beneficiaries 65 years and older at major, minor, and nonteaching hospitals from January 1, 2014, to November 30, 2015, for 15 medical conditions and 6 surgical procedures. Data analysis was performed from February 26, 2019, to April 16, 2019. Hospital teaching status (major, minor, and nonteaching hospitals). The primary outcome was 30-day total standardized costs to Medicare for hospitalizations for all 21 conditions in aggregate as well as stratified by medical and surgical condition categories. Secondary outcomes included 30-day spending for individual components of care (index hospitalization, physician services, readmission, and outpatient and post-acute care services) as well as total standardized costs at 90 days. The sample consisted of 1 249 006 hospitalizations at 3064 hospitals (232 [7.6%] major teaching, 837 [27.3%] minor teaching, and 1995 [65.1%] nonteaching hospitals). Treatment at a major teaching hospital was associated with lower total 30-day adjusted standardized costs ($18 605 vs $18 793 at minor teaching hospitals and $18 873 at nonteaching hospitals; difference between major and nonteaching hospitals: -$268; 95% CI, -$456 to -$80; P = .005). Treatment at a major teaching hospital was associated with higher spending for the index hospitalization ($8529 vs $8370 at minor teaching hospitals and $8180 at nonteaching hospitals; difference between major and nonteaching hospitals: $349; 95% CI, $308-$390; P < .001) but lower physician costs ($677 vs $725 at minor teaching hospitals and $728 at nonteaching hospitals; difference: -$50; 95% CI, -$60 to -$41; P < .001). Furthermore, post-acute care costs at 30 days were lowest at major teaching hospitals ($6015 vs $6239 for minor teaching hospitals and $6260 for nonteaching hospitals; difference: -$245; 95% CI, -$375 to -$115; P < .001). Thirty-day total costs were lower at major teaching hospitals compared with nonteaching hospitals for 12 of the 21 individual conditions examined. There was no difference in costs by teaching status at 90 days ($24 982 at major teaching hospitals vs $24 959 at minor teaching hospitals vs $25 044 at nonteaching hospitals; difference: -$61; 95% CI, -$310 to $188; P = .63). Medicare patients treated at major teaching hospitals had lower Medicare spending at 30 days and similar costs at 90 days compared with Medicare patients at nonteaching hospitals. These findings appear to raise doubts that care at teaching hospitals is necessarily more expensive than care at nonteaching hospitals.


Exploring associations between physical activity and perceived and objective measures of the built environment.
The built environment may be responsible for making nonmotorized transportation inconvenient, resulting in declines in physical activity. However, few studies have assessed both the perceived and objectively measured environment in association with physical activity outcomes. The purpose of this study was to describe the associations between perceptions and objective measures of the built environment and their associations with leisure, walking, and transportation activity. Perception of the environment was assessed from responses to 1,270 telephone surveys conducted in Forsyth County, NC and Jackson, MS from January to July 2003. Participants were asked if high-speed cars, heavy traffic, and lack of crosswalks or sidewalks were problems in their neighborhood or barriers to physical activity. They were also asked if there are places to walk to instead of driving in their neighborhood. Speed, volume, and street connectivity were assessed using Geographic Information Systems (GIS) for both study areas. Locations of crashes were measured using GIS for the NC study area as well. Objective and perceived measures of the built environment were in poor agreement as calculated by kappa coefficients. Few associations were found between any of the physical activity outcomes and perception of speed, volume, or presence of sidewalks as problems in the neighborhood or as barriers to physical activity in regression analyses. Associations between perceptions of having places to walk to and presence of crosswalks differed between study sites. Several associations were found between objective measures of traffic volume, traffic speed, and crashes with leisure, walking, and transportation activity in Forsyth County, NC; however, in Jackson, MS, only traffic volume was associated with any of the physical activity outcomes. When both objective and perceived measures of the built environment were combined into the same model, we observed independent associations with physical activity; thus, we feel that evaluating both objective and perceived measures of the built environment may be necessary when examining the relationship between the built environment and physical activity.


pH-Switchable vitamin B9 gels for stoichiometry-controlled spherical co-crystallization.
Vitamin B9 gels were found to be pH-switchable by adding triethylamine or acetic acid. Such recyclable gels can be used as novel co-crystallization media, resulting in four stoichiometric vitamin C co-crystals. Under highly supersaturated conditions, uniform microspheres of (VC)·(NA) with different particle sizes were obtained through a spherical crystallization process.


Evaluation of cytokine responses against novel Mtb antigens as diagnostic markers for TB disease.
We investigated the accuracy of host markers detected in Mtb antigen-stimulated whole blood culture supernatant in the diagnosis of TB. Prospectively, blood from 322 individuals with presumed TB disease from six African sites was stimulated with four different Mtb antigens (Rv0081, Rv1284, ESAT-6/CFP-10, and Rv2034) in a 24 h whole blood stimulation assay (WBA). The concentrations of 42 host markers in the supernatants were measured using the Luminex multiplex platform. Diagnostic biosignatures were investigated through the use of multivariate analysis techniques. 17% of the participants were HIV infected, 106 had active TB disease and in 216 TB was excluded. Unstimulated concentrations of CRP, SAA, ferritin and IP-10 had better discriminating ability than markers from stimulated samples. Accuracy of marker combinations by general discriminant analysis (GDA) identified a six analyte model with 77% accuracy for TB cases and 84% for non TB cases, with a better performance in HIV uninfected patients. A biosignature of 6 cytokines obtained after stimulation with four Mtb antigens has moderate potential as a diagnostic tool for pulmonary TB disease individuals and stimulated marker expression had no added value to unstimulated marker performance.


Current global status of female genital mutilation: a review.
To provide an overview of the current global status of female genital mutilation (FGM) or female circumcision practised in various countries. Major published series of peer reviewed journals writing about female genital mutilation (FGM) over the last two decades were reviewed using the index medicus and medline search. A few earlier publications related to the FGM ritual as practised earlier were also reviewed including the various techniques and tools used, the "surgeons or perpetrators" of the FGM ritual and the myriad of medical and sexual complications resulting from the procedure. Global efforts to abolish the ritual and why such efforts including legislation has resulted in little or no success were also critically reviewed. FGM remains prevalent in many countries including African countries where over 136 million women have been 'circumcised' despite persistent and consistent efforts by various governments, WHO and other bodies to eradicate the ritual by the year 2000 AD. This is as a result of deep rooted cultures, traditions and religions. Although FGM should be abolished globally, it must involve gradual persuasion which should include sensitisation and adequate community-based educational and medical awareness campaign. Mere repression through legislation has not been successful, and women need to be provided with other avenues for their expression of social status approval and respectability other than through FGM.


Examining Multilevel Environmental Correlates of Physical Activity Among Older Adults With Multiple Sclerosis.
Three hundred and sixty-three older adults with multiple sclerosis completed a cross-sectional study examining hierarchical correlates of physical activity using a social cognitive theory perspective within a social ecological model (i.e., built environment, social environment, and individual social cognitive theory variables). Hierarchical linear regression analyses were conducted, wherein significant associations were noted for built environment (i.e., land-use mix diversity and aesthetics) and physical activity in Step 1 (R2 = .09). Social and built environment were significant correlates in Step 2 (R2 = .15). Finally in Step 3, individual social cognitive theory variables (i.e., self-efficacy and outcome expectations) were the only significant correlates of total physical activity (R2 = .38). Results were comparable for health-promoting physical activity; however, self-efficacy was the only significant correlate in Step 3 (R2 = .36). This study provides guidance for researchers and practitioners on relevant targets for tailoring interventions for older adults with multiple sclerosis and supports an emphasis on self-efficacy as a primary predictor of health behavior change.


Pitfalls in processing home electronic spirometric data in asthma.
Electronic spirometers offer the prospect of paperless home monitoring, but data quality is not automatically better than from conventional monitoring. The aim of this study was to determine the extent to which the quality and processing of self-recorded spirometric data from patients with asthma complied with international guidelines for spirometry. Data were from 33 subjects with poorly controlled asthma who had completed the first 9 weeks of a clinical budesonide trial. MicroMedical DiaryCard electronic spirometers were used to record three spirometric manoeuvres in twice-daily sessions. Confounding events were recorded in a paper diary. Within-session reproducibility was calculated for forced expiratory volume in one second (FEV1), forced vital capacity (FVC) and peak expiratory flow (PEF) during the first week of run-in and week 9 of budesonide treatment. Geometric means of within-session reproducibility (mean difference between highest and second-highest value from each session over a one-week period) for FEV1, FVC and PEF were 76 mL, 116 mL and 18 L x min(-1), respectively, during run-in. Times of spirometric sessions varied widely, with some overlap between morning and evening session times. Manoeuvre-induced falls in PEF and FEV1 occurred only as often as expected by chance. Nonasthma events including equipment faults and painful conditions caused changes in spirometric results. Home spirometric monitoring can be carried out with excellent reproducibility in patients with asthma. However, quality-control issues are complex and an accompanying paper diary remains essential.


Hormones and cardiovascular health in women.
Cardiovascular diseases (CVDs) may have their origin before birth: the combination of being small at birth and having an overly rich post-natal diet increases the likelihood of obesity and of acquiring a specific metabolic syndrome in adulthood that carries an increased risk of CVD. The incidence of CVD and mortality is very low in women of reproductive age but rises to a significant level in older women. In this article, we discuss CVD in relation to hormonal contraception, pregnancy and polycystic ovarian syndrome (PCOS) in younger women and menopause in older women. Women with PCOS have a higher risk of diabetes and hypertension, but studies to date have not shown an effect on CVD events. Use of combined hormonal contraception has only small effects on CVD because of the low baseline incidence of myocardial infarction (MI), stroke and venous thromboembolism (VTE) among young women. Women with existing risk factors or existing CVD, however, should consider alternative contraception. In pregnancy, CVD is rare, although, in the West, it now accounts for a significant proportion of maternal mortality as the frequency of obstetrical causes of mortality has substantially declined. The frequency of VTE is 15 per 10,000 during pregnancy and the post-partum period. In older women, menopause causes a slightly higher risk of MI after allowing for age, although there is substantial heterogeneity in the results of studies on menopause and age at menopause and MI. A larger effect might have been expected, because estrogen reduces the risk of developing atherosclerosis in premenopausal women, whereas in post-menopausal women who may have established atherosclerotic disease, estrogen increases the risk of myocardial disease through the effects on plaque stability and clot formation. Recent trial results indicate that hormone treatment in menopause does not favourably affect the risk of MI, stroke or other vascular disease. Thus, prevention of CVD should rely on diet and fitness, low-dose aspirin and treatment of hypertension, hyperglycaemia and hyperlipidaemia.


A comparison of proton therapy and IMRT treatment plans for prostate radiotherapy.
Proton therapy (PT) is becoming a more widely available treatment option on the world stage and there is some interest in investment in this treatment option in Australia. The benefit of PT has been shown for a number of tumour sites, particularly for paediatric patients. The workload from these patients may not completely fill the maximum yearly workload of a machine. This work aims to ascertain if prostate cancer would be a suitable candidate to fill the rest of the workload at an Australian PT facility. Passive and intensity modulated proton therapy (IMPT) plans were generated for a prostate patient. These were compared to 7 field sliding window and step and shoot IMRT plans. All plans used a prescription dose of 78 CGE. IMRT and IMPT plans used inverse planning for optimisation. Homogeneity in the PTV was best for the IMPT plan. IMPT also gave the best rectal sparing. The bladder and femoral heads were exposed to less dose in both proton plans. Proton plans exposed normal tissue outside the PTV to less than 50% of the dose given by the IMRT plans. PT, particularly IMPT, is a suitable treatment option for the prostate cancer patient presented here.


Poly(ester amide)-based hybrid hydrogels for efficient transdermal insulin delivery.
Transdermal drug delivery is an attractive, non-invasive treatment. It can avoid first-pass hepatic metabolism and provides the possibility of self-administration. Hydrogels are promising biomaterials due to their important qualities such as biocompatibility and biodegradability. Recently, there has been tremendous growth in the area of hydrogels for transdermal drug delivery. In this work, a new kind of arginine-based poly(ester amide) (Arg-PEA) and polyethylene glycol diacrylamide (PEG-DA) hybrid hydrogel was developed for transdermal drug delivery. The hydrogels not only possess excellent swelling capacity, but also have good mechanical properties, which were then evaluated as drug delivery agents using insulin as a model system. Cytotoxicity testing and in vivo skin irritation tests demonstrated that the hydrogels were biocompatible. Finally, the results indicated that the prepared hydrogels could not only perform transdermal drug delivery, but also might regulate blood glucose levels in a mouse model with streptozotocin-induced diabetes.


Human brown adipose tissue depots automatically segmented by positron emission tomography/computed tomography and registered magnetic resonance images.
Reliably differentiating brown adipose tissue (BAT) from other tissues using a non-invasive imaging method is an important step toward studying BAT in humans. Detecting BAT is typically confirmed by the uptake of the injected radioactive tracer 18F-Fluorodeoxyglucose (18F-FDG) into adipose tissue depots, as measured by positron emission tomography/computed tomography (PET-CT) scans after exposing the subject to cold stimulus. Fat-water separated magnetic resonance imaging (MRI) has the ability to distinguish BAT without the use of a radioactive tracer. To date, MRI of BAT in adult humans has not been co-registered with cold-activated PET-CT. Therefore, this protocol uses 18F-FDG PET-CT scans to automatically generate a BAT mask, which is then applied to co-registered MRI scans of the same subject. This approach enables measurement of quantitative MRI properties of BAT without manual segmentation. BAT masks are created from two PET-CT scans: after exposure for 2 hr to either thermoneutral (TN) (24 °C) or cold-activated (CA) (17 °C) conditions. The TN and CA PET-CT scans are registered, and the PET standardized uptake and CT Hounsfield values are used to create a mask containing only BAT. CA and TN MRI scans are also acquired on the same subject and registered to the PET-CT scans in order to establish quantitative MRI properties within the automatically defined BAT mask. An advantage of this approach is that the segmentation is completely automated and is based on widely accepted methods for identification of activated BAT (PET-CT). The quantitative MRI properties of BAT established using this protocol can serve as the basis for an MRI-only BAT examination that avoids the radiation associated with PET-CT.


Distribution and elimination of 2-[14C]-acetone in mice after inhalation exposure.
This study was undertaken to determine the tissue distribution and elimination of acetone and its metabolic radioactive fragments in mice after exposure to about 1,200 mg/m3 (500 ppm) of 2-[14C]-acetone vapor. The tissue concentrations of acetone seemed to reach steady state plateaus within 6 h of exposure. In the adipose tissue the maximal concentration was about one-third of that in the highly perfused nonadipose tissues, in which acetone was rather evenly distributed. The contents of radioactivity also reached a plateau within 6 h of exposure in all tissues except the liver and brown adipose tissue. In these tissues the radioactivity increased during exposures up to 24 h. Prolonging the exposure time from 6 h to 6 h/d for three and five consecutive days gave no or only a small additional accumulation of radioactivity in all tissues except adipose tissue. The half-times of acetone after 6 h of exposure were between 2 and 5 h in all tissues. Almost equal amounts of acetone were excreted via the lungs unmetabolized or metabolized to carbon dioxide. In all tissues endogenous levels of acetone were reached within 24 h after exposure. Thus, acetone did not accumulate after prolonged or repeated exposure to concentrations of 1,200 mg/m3.


Transforming Atrial Fibrillation Research to Integrate Social Determinants of Health: A National Heart, Lung, and Blood Institute Workshop Report.
Only modest attention has been paid to the contributions of social determinants of health to atrial fibrillation (AF) risk factors, diagnosis, symptoms, management, and outcomes. The diagnosis of AF provides unique challenges exacerbated by the arrhythmia's often paroxysmal nature and individuals' disparate access to health care and technologies that facilitate detection. Social determinants of health affect access to care and management decisions for AF, increasing the likelihood of adverse outcomes among individuals who experience systemic disadvantages. Developing effective approaches to address modifiable social determinants of health requires research to eliminate the substantive inequities in health care delivery and outcomes in AF. The National Heart, Lung, and Blood Institute convened an expert panel to identify major knowledge gaps and research opportunities in the field of social determinants of AF. The workshop addressed the following social determinants: (1) socioeconomic status and access to care; (2) health literacy; (3) race, ethnicity, and racism; (4) sex and gender; (5) shared decision-making in systemically disadvantaged populations; and (6) place, including rurality, neighborhood, and community. Many individuals with AF have multiple adverse social determinants, which may cluster in the individual and in systemically disadvantaged places (eg, rural locations, urban neighborhoods). Cumulative disadvantages may accumulate over the life course and contribute to inequities in the diagnosis, management, and outcomes in AF. Workshop participants identified multiple critical research questions and approaches to catalyze social determinants of health research that address the distinctive aspects of AF. The long-term aspiration of this work is to eradicate the substantive inequities in AF diagnosis, management, and outcomes across populations.


Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study.
Gram-negative Enterobacteriaceae with resistance to carbapenem conferred by New Delhi metallo-beta-lactamase 1 (NDM-1) are potentially a major global health problem. We investigated the prevalence of NDM-1, in multidrug-resistant Enterobacteriaceae in India, Pakistan, and the UK. Enterobacteriaceae isolates were studied from two major centres in India--Chennai (south India), Haryana (north India)--and those referred to the UK's national reference laboratory. Antibiotic susceptibilities were assessed, and the presence of the carbapenem resistance gene bla(NDM-1) was established by PCR. Isolates were typed by pulsed-field gel electrophoresis of XbaI-restricted genomic DNA. Plasmids were analysed by S1 nuclease digestion and PCR typing. Case data for UK patients were reviewed for evidence of travel and recent admission to hospitals in India or Pakistan. We identified 44 isolates with NDM-1 in Chennai, 26 in Haryana, 37 in the UK, and 73 in other sites in India and Pakistan. NDM-1 was mostly found among Escherichia coli (36) and Klebsiella pneumoniae (111), which were highly resistant to all antibiotics except to tigecycline and colistin. K pneumoniae isolates from Haryana were clonal but NDM-1 producers from the UK and Chennai were clonally diverse. Most isolates carried the NDM-1 gene on plasmids: those from UK and Chennai were readily transferable whereas those from Haryana were not conjugative. Many of the UK NDM-1 positive patients had travelled to India or Pakistan within the past year, or had links with these countries. The potential of NDM-1 to be a worldwide public health problem is great, and co-ordinated international surveillance is needed.