Identification of four trp1 gene variants murine pancreatic beta-cells.
Insulin secretion is stimulated by glucose, hormones and neurotransmitters. Both activation of a non-selective cation current and activation of a Ca2+ current in response to depletion of intracellular Ca2+ stores have been suggested to play a role in this stimulation. The properties of these currents resemble those reported for the Drosophila genes trp and trpl. Using the reverse transcription polymerase chain reaction and Northern blot analysis we found that of the six mammalian trp-related genes (trp1-6), only trp1 was expressed at high levels in the mouse insulinoma cell line MIN6. We cloned the murine homologue of human trp1 from MIN6 cells and identified four variants (alpha, beta, gamma and delta), generated by alternative splicing near the N-terminus of the protein. In vitro translation showed that only the alpha and beta splice variants are efficiently expressed. The beta variant is the dominant form in MIN6 cells (and probably in mouse pancreatic islets), whereas the alpha variant is the major type in the mouse brain. The beta variant showed 99% identity to the human homologue at the amino acid level.


Ice hockey injuries: a 4-year prospective study of a Swedish élite ice hockey team.
In this prospective study, we have investigated the incidence of injuries of different severity, types of injury, and mechanisms of injury during ice hockey practice and games. One Swedish élite hockey team was closely observed during four seasons (1986-1990). There were 376 injuries, of which 148 resulted in absence from practice or games. The incidence of injury (injuries associated with later absence) during practice was 2.6 per 1000 player-practice hours and 74.1 per 1000 player-game hours. Nuisance injuries (without any later absence) and minor injuries (absence < 1 week) constituted the vast majority (95.2%) and only 4.8% (18 cases) were classified as moderate or major injuries (absence > 1 week). Of the injuries 85% were caused by trauma and 15% by overuse. Injuries were most often localized to the lower limb (37.8%) and head/face (31.4%). The commonest injuries were contusions, lacerations/wounds, strains and sprains. Most injuries resulted from stick or player contact (predominantly checking). The results are in close agreement with those of a previous investigation of another Swedish élite hockey team covering the years 1982-1985. It should be possible to reduce the number of injuries by stricter enforcement of the hockey rules, especially against stick violations, and a more widespread use of visors.


Graded regulation of the Kv2.1 potassium channel by variable phosphorylation.
Dynamic modulation of ion channels by phosphorylation underlies neuronal plasticity. The Kv2.1 potassium channel is highly phosphorylated in resting mammalian neurons. Activity-dependent Kv2.1 dephosphorylation by calcineurin induces graded hyperpolarizing shifts in voltage-dependent activation, causing suppression of neuronal excitability. Mass spectrometry-SILAC (stable isotope labeling with amino acids in cell culture) identified 16 Kv2.1 phosphorylation sites, of which 7 were dephosphorylated by calcineurin. Mutation of individual calcineurin-regulated sites to alanine produced incremental shifts mimicking dephosphorylation, whereas mutation to aspartate yielded equivalent resistance to calcineurin. Mutations at multiple sites were additive, showing that variable phosphorylation of Kv2.1 at a large number of sites allows graded activity-dependent regulation of channel gating and neuronal firing properties.


Health care disconnect: gaps in coverage and care for minority adults. Findings from the Commonwealth Fund Biennial Health Insurance Survey (2005).
Analysis of the Commonwealth Fund Biennial Health Insurance Survey (2005) finds that uninsured rates for Hispanic and African American adults are one and-a-half to three times greater than the rate for white adults. Nearly two-thirds (62%) of working-age Hispanics and one-third (33%) of African Americans were uninsured at some point during 2005, compared with 20 percent of working-age whites. Hispanics are particularly disconnected from the health care system, being substantially less likely than whites to have a regular doctor, to have visited a doctor in the past year, or to feel confident about their ability to manage their health problems. African Americans are significantly more likely than whites to visit the emergency room for non-urgent care and to experience serious problems with medical bills and medical debt. Along with expanded insurance coverage, policies promoting continuity in patients' relationships with health care providers also are needed to reduce disparities in access.


Hybrid equation/agent-based model of ischemia-induced hyperemia and pressure ulcer formation predicts greater propensity to ulcerate in subjects with spinal cord injury.
Pressure ulcers are costly and life-threatening complications for people with spinal cord injury (SCI). People with SCI also exhibit differential blood flow properties in non-ulcerated skin. We hypothesized that a computer simulation of the pressure ulcer formation process, informed by data regarding skin blood flow and reactive hyperemia in response to pressure, could provide insights into the pathogenesis and effective treatment of post-SCI pressure ulcers. Agent-Based Models (ABM) are useful in settings such as pressure ulcers, in which spatial realism is important. Ordinary Differential Equation-based (ODE) models are useful when modeling physiological phenomena such as reactive hyperemia. Accordingly, we constructed a hybrid model that combines ODEs related to blood flow along with an ABM of skin injury, inflammation, and ulcer formation. The relationship between pressure and the course of ulcer formation, as well as several other important characteristic patterns of pressure ulcer formation, was demonstrated in this model. The ODE portion of this model was calibrated to data related to blood flow following experimental pressure responses in non-injured human subjects or to data from people with SCI. This model predicted a higher propensity to form ulcers in response to pressure in people with SCI vs. non-injured control subjects, and thus may serve as novel diagnostic platform for post-SCI ulcer formation.


Amount and distribution of fume contaminants in the lungs of an arc welder post mortem.
This study describes various post-mortem examinations of the lungs of a shipyard arc welder. These investigations comprised morphological methods, including gross pathology and radiography, histological and ultrastructural examinations, chemical analysis of lung tissue, and sensitive magnetic measurements, which can now be used to detect magnetic contaminants, such as welding fume, in human tissue. Such methods can also show the distribution of contaminants. The amount of welding-fume contaminants was low, only 110 mg. The contaminant iron seemed to represent only 10% of the total iron chemically analysed in lung tissue. The concentration of the total iron was 4-10 times higher than that in control tissue. Contaminants tended to collect in the central areas of the lungs. Two concentration centres could be observed in each lung, one anterior and one posterior. There was slight fibrosis around pigmented areas. Histological findings were similar to those of previous studies. Transmission electron micrographs showed that enlarged lysosomes of macrophages contained electron-dense granules, 450 nm to less than 5 nm in diameter, which formed aggregates and short chains, and resembled welding-fume particles in the breathing zone. Our results show the value of studying the same material by a number of inter-related methods.


Continuity of medication management and continuity of care: Conceptual and operational considerations.
Continuity of care is considered foundational to high-quality care. Traditional continuity of care constructs may adequately characterize care quality in general populations, but may merit reconceptualization for patients with multiple chronic conditions. Specifically, interactions between multiple chronic condition patients and providers involve complex medication management; therefore care continuity measurement may be more relevant if focused on the provider subset who prescribes essential medications for chronic conditions-a construct we call continuity of medication management. Our objective was to explore conceptual distinctions between continuity of medication management and continuity of care, survey existing evidence in this area, and discuss implications of our findings for future research and intervention development. In this topical review, we discuss conceptual distinctions between continuity of medication management and continuity of care, review the limited continuity of medication management-related empirical evidence, and discuss implications for future research and interventions. Continuity of medication management represents a potential conceptual and measurement advance by reflecting interpersonal continuity and management continuity, and may provide a means of identifying patients at high-risk of adverse events. Empirical evidence also establishes support for continuity of medication management as a meaningful measure of care continuity. Finally, continuity of medication management may also be a potential target for future intervention to improve care delivery among multiple chronic condition patients. If continuity of medication management is validated in diverse populations, correlated with patient outcomes, and responsive to change, then it may be an important target for improving the health and health care of multiple chronic condition patients.


Antineutrophil antibody: a test for autoimmune primary sclerosing cholangitis in childhood?
The detection of an antineutrophil antibody which is highly sensitive and specific for adult primary sclerosing cholangitis using indirect immunoalkaline phosphatase has been previously described. In this study, the diagnostic potential of this method in childhood primary sclerosing cholangitis is described. A range of 72 blinded children's sera (36 boys), aged six months to 21 years (10 primary sclerosing cholangitis, eight autoimmune chronic active hepatitis, 10 alpha-1 antitrypsin deficiency, 12 extrahepatic bile duct atresia, 11 ulcerative colitis and 21 normal subjects) was assayed. Eight of the 10 primary sclerosing cholangitis patients were correctly identified. Three patients with chronic active hepatitis also showed the characteristic primary sclerosing cholangitis pattern of staining. No ulcerative colitis patients or any other patients showed this pattern of staining. All normal subjects were negative. As in adult primary sclerosing cholangitis, there is a specific antineutrophil antibody in childhood primary sclerosing cholangitis and this provides further evidence towards an autoimmune aetiology of this condition. The test may have diagnostic potential.


Remote, proactive, telephone based management of toxicity in outpatients during adjuvant or neoadjuvant chemotherapy for early stage breast cancer: pragmatic, cluster randomised trial.
To evaluate the effectiveness of remote proactive management of toxicities during chemotherapy for early stage breast cancer. Pragmatic, cluster randomised trial. 20 cancer centres in Ontario, Canada, allocated by covariate constrained randomisation to remote management of toxicities or routine care. All patients starting adjuvant or neoadjuvant chemotherapy for early stage breast cancer at each centre. 25 patients from each centre completed patient reported outcome questionnaires. Proactive, standardised, nurse led telephone management of common toxicities at two time points after each chemotherapy cycle. The primary outcome, cluster level mean number of visits to the emergency department or admissions to hospital per patient during the whole course of chemotherapy treatment, was evaluated with routinely available administrative healthcare data. Secondary patient reported outcomes included toxicity, self-efficacy, and quality of life. Baseline characteristics of participants were similar in the intervention (n=944) and control arms (n=1214); 22% were older than 65 years. Penetration (that is, the percentage of patients who received the intervention at each centre) was 50-86%. Mean number of visits to the emergency department or admissions to hospital per patient was 0.91 (standard deviation 0.28) in the intervention arm and 0.94 (0.40) in the control arm (P=0.94); 47% (1014 of 2158 patients) had at least one visit to the emergency department or a hospital admission during chemotherapy. Among 580 participants who completed the patient reported outcome questionnaires, at least one grade 3 toxicity was reported by 48% (134 of 278 patients) in the intervention arm and by 58% (163 of 283) in the control arm. No differences in self-efficacy, anxiety, or depression were found. Compared with baseline, the functional assessment of cancer therapy trial outcome index decreased by 6.1 and 9.0 points in the intervention and control participants, respectively. Proactive, telephone based management of toxicities during chemotherapy did not result in fewer visits to the emergency department or hospital admissions. With the rapid rise in remote care because of the covid-19 pandemic, identifying scalable strategies for remote management of patients during cancer treatment is particularly relevant. ClinicalTrials.gov NCT02485678.


Flexible human collective wisdom.
Group decisions typically outperform individual decisions. But how do groups combine their individual decisions to reach their collective decisions? Previous studies conceptualize collective decision making using static combination rules, be it a majority-voting rule or a weighted-averaging rule. Unknown is whether groups adapt their combination rules to changing information environments. We implemented a novel paradigm for which information obeyed a mixture of distributions, such that the optimal Bayesian rule is nonlinear and often follows minority opinions, while the majority rule leads to suboptimal but above chance performance. Using perceptual (Experiment 1) and cognitive (Experiment 2) signal-detection tasks, we switched the information environment halfway through the experiments to a mixture of distributions without informing participants. Groups gradually abandoned the majority rule to follow any minority opinion advocating signal presence with high confidence. Furthermore, groups with greater ability to abandon the majority rule achieved higher collective-decision accuracies. It is important to note that this abandonment was not triggered by performance loss for the majority rule relative to the first half of the experiment. Our results propose a new theory of human collective decision making: Humans make inferences about how information is distributed across individuals and time, and dynamically alter their joint decision algorithms to enhance the benefits of collective wisdom.


Nonhelical inverse transfer of a decaying turbulent magnetic field.
In the presence of magnetic helicity, inverse transfer from small to large scales is well known in magnetohydrodynamic (MHD) turbulence and has applications in astrophysics, cosmology, and fusion plasmas. Using high resolution direct numerical simulations of magnetically dominated self-similarly decaying MHD turbulence, we report a similar inverse transfer even in the absence of magnetic helicity. We compute for the first time spectral energy transfer rates to show that this inverse transfer is about half as strong as with helicity, but in both cases the magnetic gain at large scales results from velocity at similar scales interacting with smaller-scale magnetic fields. This suggests that both inverse transfers are a consequence of universal mechanisms for magnetically dominated turbulence. Possible explanations include inverse cascading of the mean squared vector potential associated with local near two dimensionality and the shallower k^{2} subinertial range spectrum of kinetic energy forcing the magnetic field with a k^{4} subinertial range to attain larger-scale coherence. The inertial range shows a clear k^{-2} spectrum and is the first example of fully isotropic magnetically dominated MHD turbulence exhibiting weak turbulence scaling.


On approximation of smooth functions from samples of partial derivatives with application to phase unwrapping.
This paper addresses the problem of approximating smooth bivariate functions from the samples of their partial derivatives. The approximation is carried out under the assumption that the subspace to which the functions to be recovered are supposed to belong, possesses an approximant in the form of a principal shift-invariant (PSI) subspace. Subsequently, the desired approximation is found as the element of the PSI subspace that fits the data the best in the (2)-sense. In order to alleviate the ill-posedness of the process of finding such a solution, we take advantage of the discrete nature of the problem under consideration. The proposed approach allows the explicit construction of a projection operator which maps the measured derivatives into a stable and unique approximation of the corresponding function. Moreover, the paper develops the concept of discrete PSI subspaces, which may be of relevance for several practical settings where one is given samples of a function instead of its continuously defined values. As a final point, the application of the proposed method to the problem of phase unwrapping in homomorphic deconvolution is described.


Protein side-chain resonance assignment and NOE assignment using RDC-defined backbones without TOCSY data.
One bottleneck in NMR structure determination lies in the laborious and time-consuming process of side-chain resonance and NOE assignments. Compared to the well-studied backbone resonance assignment problem, automated side-chain resonance and NOE assignments are relatively less explored. Most NOE assignment algorithms require nearly complete side-chain resonance assignments from a series of through-bond experiments such as HCCH-TOCSY or HCCCONH. Unfortunately, these TOCSY experiments perform poorly on large proteins. To overcome this deficiency, we present a novel algorithm, called NASCA: (NOE Assignment and Side-Chain Assignment), to automate both side-chain resonance and NOE assignments and to perform high-resolution protein structure determination in the absence of any explicit through-bond experiment to facilitate side-chain resonance assignment, such as HCCH-TOCSY. After casting the assignment problem into a Markov Random Field (MRF), NASCA: extends and applies combinatorial protein design algorithms to compute optimal assignments that best interpret the NMR data. The MRF captures the contact map information of the protein derived from NOESY spectra, exploits the backbone structural information determined by RDCs, and considers all possible side-chain rotamers. The complexity of the combinatorial search is reduced by using a dead-end elimination (DEE) algorithm, which prunes side-chain resonance assignments that are provably not part of the optimal solution. Then an A* search algorithm is employed to find a set of optimal side-chain resonance assignments that best fit the NMR data. These side-chain resonance assignments are then used to resolve the NOE assignment ambiguity and compute high-resolution protein structures. Tests on five proteins show that NASCA: assigns resonances for more than 90% of side-chain protons, and achieves about 80% correct assignments. The final structures computed using the NOE distance restraints assigned by NASCA: have backbone RMSD 0.8-1.5 Å from the reference structures determined by traditional NMR approaches.


Childhood chromium exposure and neuropsychological development in children living in two polluted areas in southern Spain.
This study aimed to assess the association between exposure to chromium and neuropsychological development among children. A cross-sectional study was conducted with 393 children aged 6-11 years old randomly selected from State-funded schools in two provinces in Southern Spain (Almeria and Huelva), in 2010 and 2012. Chromium levels in urine and hair samples were analyzed by inductively coupled plasma mass spectrometry with an octopole reaction system. Neuropsychological development was evaluated using the Wechsler Intelligence Scale for Children-Fourth Edition (WISC-IV) and three computerized tests from the Behavioural Assessment and Research System (BARS): Reaction Time Test (RTT), Continuous Performance Test (CPT) and Selective Attention Test (SAT). Multivariable linear regression models adjusted for potential confounders, including heavy metals, were applied to examine the association between chromium levels and neuropsychological outcomes. A 10-fold increase in urine chromium levels was associated with a decrease of 5.99 points on the WISC-IV Full-Scale IQ (95% CI: 11.98 to -0.02). Likewise, a 10-fold increase in urine chromium levels in boys was associated with a decrease of 0.03 points in the percentage of omissions (95% CI: 0.0 to 0.05) in the SAT, with an increase of 68.35 points in latency (95% CI: 6.60 to 130.12) in the RTT, and with an increase in the number of trials with latencies > 1000 ms (β = 37.92; 95% CI: 2.73 to 73.12) in the RTT. An inverse significant association was detected between chromium levels in hair and latency in the SAT in boys (β = -50.53; 95% CI: 86.86 to -14.22) and girls (β = -55.95; 95% CI: 78.93 to -32.97). Excluding trials with latencies >1000 ms in the RTT increased latency scores by 29.36 points in boys (95% CI: 0.17 to 58.57), and 39.91 points in girls (95% CI: 21.25 to 58.59). This study is the first to show the detrimental effects of postnatal chromium exposure on neuropsychological development in school-aged children.


Efficacy of single-dose antibiotic against early-onset pneumonia in comatose patients who are ventilated.
Comatose patients present a high risk of early-onset ventilator-associated pneumonia (EO-VAP) for which antibiotic prophylaxis has been proposed. Comatose patients were studied to evaluate the efficacy of a single-dose of antibiotic prophylaxis at intubation against EO-VAP. A prospective cohort of comatose patients (Glasgow Coma Score ≤ 8) who were admitted in 2009-2010 and administered a single-dose of antibiotic within 4 h of intubation was compared with comatose patients (admitted ≥ 4 h after intubation in 2009-2010 or admitted in 2007-2008) who did not receive antibiotic prophylaxis. We analyzed the incidence of EO-VAP, late-onset VAP, and ventilator-associated tracheobronchitis in both groups. Propensity scores for receiving antibiotic prophylaxis were derived on the basis of patients' characteristics (eg, age and severity) to assess its impact on EO-VAP development. We included 129 patients (71 in the prophylaxis group and 58 in the control group). The global incidence of VAP and incidence of EO-VAP were lower in the prophylaxis group: 10.8 vs 28.4 episodes/1,000 days on mechanical ventilation (P = .015) and 4.4 vs 23.1 episodes/1,000 days on mechanical ventilation (P = .02), respectively. The incidence of late-onset VAP did not differ. The prophylaxis group tended toward lower incidence of ventilator-associated tracheobronchitis (15.5% vs 25.9%, P = .14). No differences in mortality were found between groups. The propensity-score regression analysis confirmed that a single dose of antibiotic prophylaxis was independently associated with lower incidence of EO-VAP (OR, 0.11; 95% CI, 0.02-0.58; P = .009). A single dose of antibiotic prophylaxis at intubation might lower the incidence of EO-VAP. However, a randomized clinical trial should be conducted to confirm our findings.


T-Coffee: a web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension.
This article introduces a new interface for T-Coffee, a consistency-based multiple sequence alignment program. This interface provides an easy and intuitive access to the most popular functionality of the package. These include the default T-Coffee mode for protein and nucleic acid sequences, the M-Coffee mode that allows combining the output of any other aligners, and template-based modes of T-Coffee that deliver high accuracy alignments while using structural or homology derived templates. These three available template modes are Expresso for the alignment of protein with a known 3D-Structure, R-Coffee to align RNA sequences with conserved secondary structures and PSI-Coffee to accurately align distantly related sequences using homology extension. The new server benefits from recent improvements of the T-Coffee algorithm and can align up to 150 sequences as long as 10,000 residues and is available from both http://www.tcoffee.org and its main mirror http://tcoffee.crg.cat.


Family pain history predicts child health status in children with chronic rheumatic disease.
To examine the relationships of parental and family pain history on the pain experience of children with chronic rheumatic disease. The aims of the study were as follows: 1) to describe the pain history of parents and families of children with rheumatic disease, 2) to examine relationships between parental and family pain history and the pain report and physician-rated health status of children with chronic rheumatic disease, and 3) to determine whether child coping mediates the relationship between family pain history and the child's pain and physician-rated health status. Parents of 100 children were recruited from a pediatric rheumatology clinic during routine visits. Parents completed questionnaires assessing parental pain history and family characteristics. Children in the study completed a series of questionnaires to assess pain and pain coping strategies, including the Coping Strategies Questionnaire and parts of the Pediatric Pain Questionnaire. A pediatric rheumatologist provided a global assessment of disease severity on a 100-mm visual analog scale as an index of child health status. A high number of parents of children seen in a pediatric rheumatology clinic described a personal pain history. More than 90% of parents reported having at least 1 chronic pain condition, with an equal proportion reporting an episode of pain in the past month. The most commonly reported pain conditions were lower back pain, shoulder/neck pain, and migraine headache pain. On average, this group of parents reported a history of 3.5 chronic pain conditions (standard deviation: 2.3) and reported having sought treatment for 1.7 (standard deviation: 2.3) of these conditions. Additionally, 93% of all parents reported extended family members experiencing at least 1 chronic pain condition. Correlational analyses indicated that parents reporting higher levels of current pain and higher mean levels of pain during the past month were more likely to have children reporting higher levels of current pain (r = 0.23 and r = 0.27). In addition, parents who sought more treatment for their own pain were more likely to have children reporting higher levels of pain (r = 0.22) and presenting with poorer health status (r = 0.22). Similarly, parents reporting higher levels of pain-related interference with activity were more likely to have children reporting higher levels of current pain (r = 0.23). Correlational analyses also indicated that children whose extended families reported a history of multiple pain conditions were more likely to report higher levels of current pain (r = 0.24) and more pain locations (r = 0.23). Finally, a series of mediational statistical models confirmed that child use of the pain coping strategy, catastrophizing, partially accounted for the relationship between several parent and family pain history variables and the child's own current pain ratings and physician global assessment. Specifically, child catastrophizing mediated the relationships between the total number of treated pain conditions and children's current pain ratings and physician global assessment. In addition, child catastrophizing was shown to mediate the relationship between parental mean level of pain in the past month and children's current pain rating and the relationship between total number of family pain conditions and children's current pain rating. Taken together, our results suggest that parental and familial pain experiences predict children's use of catastrophizing to cope with pain, which in turn predicts physician global assessment and children's current pain. The results from the present study indicate that many of the parents of children seen in a pediatric rheumatology clinic have a personal pain history and highlight the potential impact of parental pain history on children's pain experiences. Specifically, parents who were more likely to seek treatment for their own pain or more likely to report interference with recreational activities because of pain had children with higher pain ratings and poorer health status as measured by the physician global assessment. Additionally, a series of mediational models showed that child catastrophizing serves as a specific mechanism through which parental and familial pain history variables influence child ratings of current pain and physician ratings of health status. Future studies are needed to determine exactly how children living in families with painful conditions become more reliant on catastrophizing to cope with their pain. In addition, more research is needed to identify other potential mediators, such as positive ways parents may influence children's pain coping. There are several important clinical implications of our findings. First, our results suggest that by gathering information from parents about their own pain histories, health care providers may be able to identify children at risk for developing maladaptive pain coping strategies and higher levels of disease-related pain and disability. Second, our results indicate that intervention programs should focus specifically on reducing children's use of catastrophizing to cope with their pain. Perhaps most importantly, our results highlight the need to include parents in interventions aimed at reducing children's pain and improving children's abilities to cope with pain.


Improving the Ability of Deep Neural Networks to Use Information from Multiple Views in Breast Cancer Screening.
In breast cancer screening, radiologists make the diagnosis based on images that are taken from two angles. Inspired by this, we seek to improve the performance of deep neural networks applied to this task by encouraging the model to use information from both views of the breast. First, we took a closer look at the training process and observed an imbalance between learning from the two views. In particular, we observed that layers processing one of the views have parameters with larger gradients in magnitude, and contribute more to the overall loss reduction. Next, we tested several methods targeted at utilizing both views more equally in training. We found that using the same weights to process both views, or using modality dropout, leads to a boost in performance. Looking forward, our results indicate improving learning dynamics as a promising avenue for improving utilization of multiple views in deep neural networks for medical diagnosis.


Neurons and neuroendocrine cells contain chromogranin: detection of the molecule in normal bovine tissues by immunochemical and immunohistochemical methods.
Gel-eluted bovine chromogranin (CG), the 75,000 dalton acidic protein abundantly present in adrenal chromaffin granules, was used as immunogen to prepare anti-CG serum. The specificity of the antiserum was demonstrated in immunoblots of electrophoresed bovine CG and in immunohistochemical studies of bovine adrenal medulla. In the immunoblots, the predominant immunoreactive band had a molecular weight of 75,000 daltons. Bands with a higher or lower molecular weight were also immunoreactive and may represent CG precursors or breakdown products. In the adrenal gland, only adrenal chromaffin cells contained CG immunoreactivity. Immunoblots and immunohistochemistry were also used to characterize the distribution of CG in bovine tissues. CG was expressed by cells of the diffuse neuroendocrine system (DNS) including: adrenal chromaffin cells, enterochromaffin cells, pancreatic islet cells, cells of the adenohypophysis, thyroid C cells, parathyroid cells, and submandibular gland. CG was also seen in four locations not previously recognized to express this antigen: thymic epithelial cells, neurons, the inner segment of rods and cones, and the submandibular gland. We demonstrate a wider distribution of CG than previously recognized and that the molecule detected in tissue by immunohistochemistry is indeed CG. We conclude that CG is expressed by neurons, cells of the DNS, and by a few other cells that may or may not be related to the DNS. The antiserum described here should prove valuable in developing an understanding of the function(s) of CG.


Lipid tail protrusions mediate the insertion of nanoparticles into model cell membranes.
Recent work has demonstrated that charged gold nanoparticles (AuNPs) protected by an amphiphilic organic monolayer can spontaneously insert into the core of lipid bilayers to minimize the exposure of hydrophobic surface area to water. However, the kinetic pathway to reach the thermodynamically stable transmembrane configuration is unknown. Here, we use unbiased atomistic simulations to show the pathway by which AuNPs spontaneously insert into bilayers and confirm the results experimentally on supported lipid bilayers. The critical step during this process is hydrophobic-hydrophobic contact between the core of the bilayer and the monolayer of the AuNP that requires the stochastic protrusion of an aliphatic lipid tail into solution. This last phenomenon is enhanced in the presence of high bilayer curvature and closely resembles the putative pre-stalk transition state for vesicle fusion. To the best of our knowledge, this work provides the first demonstration of vesicle fusion-like behaviour in an amphiphilic nanoparticle system.


Mechanical properties of resting and active isolated coronary arteries.
Coronary arterial smooth muscle is myogenically active, is acted upon by a variety of modulating agents, and is subjected, in situ, to compression and distension by the myocardium. Its mechanical properties thus play a key role in the regulation of coronary blood flow. To describe these, we applied phase-plane analysis of shortening velocity vs. length and load clamping to strips of isolated coronary arteries made to contract by an increase in extracellular potassium concentration. At any load, length was larger in relaxed than in "resting" preparations. In addition, preloaded shortening was smaller than would be expected from the relation between active isometric force and muscle length. These suggest the occurrence of stretch-activation and shortening-inactivation. To judge from both phase-plane analysis and quick release experiments, shortening velocity depended on load as well as on time. Velocity decreased with increasing duration of contraction. Shortened coronary arteries resisted lengthening induced by loading and could transiently bear loads that considerably exceeded isometric force. This load-bearing capacity increased with increasing shortening. In conclusion, coronary arterial smooth muscle displays the classical relationship between length, force, and velocity. However, the nature of this relationship changes with duration of activity. In addition, it is greatly affected during changes in length or load, such as expected when the arterial wall is exposed to pulsatile blood flow and is surrounded by mechanically active muscle.


A systematic review and network meta-analysis of post-imatinib therapy in advanced gastrointestinal stromal tumour.
The standard first-line systemic therapy for advanced gastrointestinal stromal tumour (gist) is imatinib. However, most gists develop imatinib resistance, highlighting the need for new agents in the imatinib-refractory setting. Currently, no randomized studies have directly compared the available post-first-line treatments. In a systematic review, the medline, embase, and central databases, and American Society of Clinical Oncology abstracts to July 2014 were searched to identify randomized controlled trials that included gist patients treated with post-first-line therapies. Hazard ratios (hrs) for progression-free (pfs) and overall survival (os) were extracted. Direct pairwise meta-analyses and indirect comparisons using the Butcher method were performed. Four studies were identified for the systematic review. One study showed that sunitinib in the second-line setting (vs. placebo) was associated with improved pfs, but not improved os. Three studies examined the third-line setting (imatinib resumption vs. placebo, regorafenib vs. placebo, nilotinib vs. best supportive care). In the third-line settings, the two placebo-controlled and the non-placebo-controlled trials showed significant heterogeneity (I2 = 98%). Indirect comparisons of imatinib resumption and regorafenib suggested that the hr for pfs was 0.59 (95% confidence interval: 0.31 to 1.12; p = 0.10), trending in favour of regorafenib. Indirect comparisons found that toxicities were higher in the regorafenib group, with a risk difference of 27.8% for any-grade toxicities and 19.5% for grades 3 and 4 toxicities. Because a head-to-head study of imatinib resumption compared with regorafenib is unlikely ever to be conducted, our study suggests that, in terms of pfs, regorafenib might be the preferred treatment. However, given the increased toxicity observed with regorafenib, clinicians should interpret that evidence with caution at an individual patient level.


Definition of a conjugation of dysfunction in Gilbert's syndrome: studies of the handling of bilirubin loads and of the pattern of bilirubin conjugates secreted in bile.
1. Intravenous doses of bilirubin (3.4 mumol/kg) were given to normal subjects and patients with Gilbert's syndrome. Both groups displayed an identical initial disappearance of a substantial proportion of the bilirubin but, late in time, the Gilbert's patients exhibited reduced clearance with a sustained elevation of the plasma bilirubin and no reflux into the plasma space of conjugated bilirubin. Increasing the dose in normal subjects (by factors of 3 and 6) failed to reproduce the response found in the Gilbert's patients. 2. In the the bile-containing duodenal aspirates of Gilbert's patients the average proportion of bilirubin found as bilirubin diglucuronide was 68% (normal 88%) and of bilirubin monoglucuronide, 23% (normal 7%). Both differences were significant at the P less than 0.001 level. In the Gilbert's patients restriction of caloric intake to 1569 kJ/day for 2 days characteristically raised the serum bilirbuin with no modification of the biliary pigment pattern; phenobarbital (180 mg/day for 2 weeks) decreased the plasma bilirubin to the normal range with, concomitantly, a reversion of the biliary pigment pattern towards normal. 3. We conclude that there is no hepatic uptake defect in Gilbert's syndrome but that there is decreased activity in the conjugation process underlying the addition of the second glucuronic acid moiety to bilirubin, to form bilirubin diglucuronide.


Balking blood pressure "control" by older persons of Bambuí, Minas Gerais State, Brazil: an ethno-epidemiological inquiry.
This ethno-epidemiological inquiry aims to comprehend hypertension-related experiences in the elderly population of Bambuí, in the State of Minas Gerais, Brazil. It combines ethnographic descriptions with statistical data. The subjective significance of factors associated with adequate arterial pressure control is explored. A baseline cohort of 26 people with hypertension, randomly selected from a total number of 1,494 residents over the age of 60, was interviewed utilizing signs, meanings and actions methodology. Multivariate analysis shows an association (p < 0.001) between female gender and monthly household income and treatment of hypertension and adequate blood pressure control. The number of doctor visits is associated with treatment but not with adequate blood pressure control. Conflicting cultural construction of "blood pressure problems" contributes to "non-adherence" to treatment. There is a fine line between blood pressure "control" and what is perceived as health professionals "controlling" patients' lives. Doctor-prescribed regimes are perceived as "prohibiting life's pleasures" and "controlling" personal liberty and free choice. Giving elderly people a voice regarding their social context can promote autonomy, well-being and happiness in later life.


Self-action of continuous laser radiation and Pearcey diffraction in a water suspension with light-absorbing particles.
Water suspension of light-absorbing nano-sized particles is an example of a medium in which non-linear effects are present at moderate light intensities favorable for optical treatment of organic and biological objects. We study experimentally the phenomena emerging in a thin layer of such a medium under the action of inhomogeneous light field formed due to the Pearcey diffraction pattern near a microlens focus. In this high-gradient field, the light energy absorbed by the particles induces inhomogeneous distribution of the medium refraction index, which results in observable self-diffraction of the incident light, here being strongly sensitive to the medium position with respect to the focus. This technique, based on the complex spatial structure of both the incident and the diffracted fields, can be employed for the detection and measurement of weak non-linearities.


Accelerometery and Heart Rate Responses of Professional Fast-Medium Bowlers in One-Day and Multi-Day Cricket.
The physical demands of fast-medium bowling are increasingly being recognised, yet comparative exploration of the differing demands between competitive formats (i.e. one-day [OD] versus multi-day [MD] matches) remain minimal. The aim of this study was to describe in-match physiological profiles of professional fast-medium bowlers from England across different versions of competitive matches using a multivariable wearable monitoring device. Seven professional cricket fast-medium bowlers wore the BioharnessTM monitoring device during matches, over three seasons (>80 hours in-match). Heart Rate (HR) and Acceleromety (ACC) was compared across match types (OD, MD) and different in-match activity states (Bowling, Between over bowling, Fielding). Peak acceleration during OD bowling was significantly higher in comparison to MD cricket ([OD vs. MD] 234.1 ± 57.9 vs 226.6 ± 32.9 ct·episode-1, p < 0.05, ES = 0.11-0.30). Data for ACC were also higher during OD than MD fielding activities (p < 0.01, ES = 0.11-.30). OD bowling stimulated higher mean HR responses (143 ± 14 vs 137 ± 16 beats·min-1, p < 0.05, ES = 0.21) when compared to MD matches. This increase in OD cricket was evident for both between over (129 ± 9 vs 120 ± 13 beats·min-1,p < 0.01, ES = 0.11-0.50) and during fielding (115 ± 12 vs 106 ± 12 beats·min-1, p < 0.01, ES = 0.36) activity. The increased HR and ACC evident in OD matches suggest greater acute physical loads than MD formats. Therefore, use of wearable technology and the findings provided give a valuable appreciation of the differences in match loads, and thus required physiological preparation and recovery in fast-medium bowlers.


Characterization of the FKBP12-Encoding Genes in Aspergillus fumigatus.
Invasive aspergillosis, largely caused by Aspergillus fumigatus, is responsible for a growing number of deaths among immunosuppressed patients. Immunosuppressants such as FK506 (tacrolimus) that target calcineurin have shown promise for antifungal drug development. FK506-binding proteins (FKBPs) form a complex with calcineurin in the presence of FK506 (FKBP12-FK506) and inhibit calcineurin activity. Research on FKBPs in fungi is limited, and none of the FKBPs have been previously characterized in A. fumigatus. We identified four orthologous genes of FKBP12, the human FK506 binding partner, in A. fumigatus and designated them fkbp12-1, fkbp12-2, fkbp12-3, and fkbp12-4. Deletional analysis of the four genes revealed that the Δfkbp12-1 strain was resistant to FK506, indicating FKBP12-1 as the key mediator of FK506-binding to calcineurin. The endogenously expressed FKBP12-1-EGFP fusion protein localized to the cytoplasm and nuclei under normal growth conditions but also to the hyphal septa following FK506 treatment, revealing its interaction with calcineurin. The FKBP12-1-EGFP fusion protein didn't localize at the septa in the presence of FK506 in the cnaA deletion background, confirming its interaction with calcineurin. Testing of all deletion strains in the Galleria mellonella model of aspergillosis suggested that these proteins don't play an important role in virulence. While the Δfkbp12-2 and Δfkbp12-3 strains didn't show any discernable phenotype, the Δfkbp12-4 strain displayed slight growth defect under normal growth conditions and inhibition of the caspofungin-mediated "paradoxical growth effect" at higher concentrations of the antifungal caspofungin. Together, these results indicate that while only FKBP12-1 is the bona fide binding partner of FK506, leading to the inhibition of calcineurin in A. fumigatus, FKBP12-4 may play a role in basal growth and the caspofungin-mediated paradoxical growth response. Exploitation of differences between A. fumigatus FKBP12-1 and human FKBP12 will be critical for the generation of fungal-specific FK506 analogs to inhibit fungal calcineurin and treat invasive fungal disease.


Comparison of biofilms formed by Candida albicans and Candida parapsilosis on bioprosthetic surfaces.
Little is known about fungal biofilms, which may cause infection and antibiotic resistance. In this study, biofilm formation by different Candida species, particularly Candida albicans and C. parapsilosis, was evaluated by using a clinically relevant model of Candida biofilm on medical devices. Candida biofilms were allowed to form on silicone elastomer and were quantified by tetrazolium (XTT) and dry weight (DW) assays. Formed biofilm was visualized by using fluorescence microscopy and confocal scanning laser microscopy with Calcofluor White (Sigma Chemical Co., St. Louis, Mo.), concanavalin A-Alexafluor 488 (Molecular Probes, Eugene, Oreg.), and FUN-1 (Molecular Probes) dyes. Although minimal variations in biofilm production among invasive C. albicans isolates were seen, significant differences between invasive and noninvasive isolates (P < 0.001) were noted. C. albicans isolates produced more biofilm than C. parapsilosis, C. glabrata, and C. tropicalis isolates, as determined by DW assays (P was <0.001 for all comparisons) and microscopy. Interestingly, noninvasive isolates demonstrated a higher level of XTT activity than invasive isolates. On microscopy, C. albicans biofilms had a morphology different from that of other species, consisting of a basal blastospore layer with a dense overlying matrix composed of exopolysaccharides and hyphae. In contrast, C. parapsilosis biofilms had less volume than C. albicans biofilms and were comprised exclusively of clumped blastospores. Unlike planktonically grown cells, Candida biofilms rapidly (within 6 h) developed fluconazole resistance (MIC, >128 microg/ml). Importantly, XTT and FUN-1 activity showed biofilm cells to be metabolically active. In conclusion, our data show that C. albicans produces quantitatively larger and qualitatively more complex biofilms than other species, in particular, C. parapsilosis.


S-nitrosylated l-serine-modified dendrimer as a kidney-targeting nitric oxide donor for prevention of renal ischaemia/reperfusion injury.
Nitric oxide (NO) deficiency is known to play a role in renal ischaemia/reperfusion injury; therefore, kidney-targeting NO donor is expected to prevent renal ischaemia/reperfusion injury. We therefore developed an S-nitrosylated L-serine-modified polyamidoamine dendrimer (SNO-Ser-PAMAM), in which multiple S-nitrosothiols (NO donors) were covalently bound to L-serine-modified dendrimer, as a kidney-targeting NO donor. In the pharmacokinetic study, approximately 76% of 111In-SNO-Ser-PAMAM accumulated in the kidney after intravenous injection in mice. Furthermore, single photon emission computed tomography/computed tomography (SPECT/CT) imaging study showed that 111In-SNO-Ser-PAMAM specifically accumulated in the renal cortex after intravenous injection. SNO-Ser-PAMAM gradually released NO over a day in plasma, indicating that SNO-Ser-PAMAM would show sustained release of NO in vivo. In a mouse model of renal ischaemia/reperfusion injury, increased plasma creatinine, a kidney damage marker, and histological changes were effectively inhibited by intravenous administration of SNO-Ser-PAMAM. These results indicate that SNO-Ser-PAMAM is a promising kidney-targeting NO donor for the efficient prevention of renal ischaemia/reperfusion injury.


JS-K, a nitric oxide prodrug, has enhanced cytotoxicity in colon cancer cells with knockdown of thioredoxin reductase 1.
The selenoenzyme thioredoxin reductase 1 has a complex role relating to cell growth. It is induced as a component of the cellular response to potentially mutagenic oxidants, but also appears to provide growth advantages to transformed cells by inhibiting apoptosis. In addition, selenocysteine-deficient or alkylated forms of thioredoxin reductase 1 have also demonstrated oxidative, pro-apoptotic activity. Therefore, a greater understanding of the role of thioredoxin reductase in redox initiated apoptotic processes is warranted. The role of thioredoxin reductase 1 in RKO cells was evaluated by attenuating endogenous thioredoxin reductase 1 expression with siRNA and then either inducing a selenium-deficient thioredoxin reductase or treatment with distinct redox challenges including, hydrogen peroxide, an oxidized lipid, 4-hydroxy-2-nonenol, and a nitric oxide donating prodrug. Thioredoxin redox status, cellular viability, and effector caspase activity were measured. In cells with attenuated endogenous thioredoxin reductase 1, a stably integrated selenocysteine-deficient form of the enzyme was induced but did not alter either the thioredoxin redox status or the cellular growth kinetics. The oxidized lipid and the nitric oxide donor demonstrated enhanced cytotoxicity when thioredoxin reductase 1 was knocked-down; however, the effect was more pronounced with the nitric oxide prodrug. These results are consistent with the hypothesis that attenuation of the thioredoxin-system can promote apoptosis in a nitric oxide-dependent manner.


A comparison between whole mung bean and decorticated mung bean: beneficial effects on the regulation of serum glucose and lipid disorders and the gut microbiota in high-fat diet and streptozotocin-induced prediabetic mice.
The aim of this study is to investigate the beneficial effects of whole mung bean (WMB) and decorticated mung bean (DMB) on the regulation of serum glucose and lipid disorders in high-fat diet (HFD) and streptozotocin (STZ)-induced prediabetic mice, and to further explore their gut microbiota modulatory effects. In the present study, the ability of mung bean-based diets to combat prediabetes-related metabolic disorders was determined by assessing the changes in the physiological, biochemical, and histological parameters, and the gut microbiota composition of prediabetic mice. The supplementation of both WMB and DMB can effectively alleviate HFD and STZ-induced impaired glucose tolerance (P < 0.05), which was accompanied by improvements in pancreatic β-cell damage and hepatic steatosis. However, only WMB supplementation significantly decreased the fasting blood glucose and fasting serum insulin levels by sensitizing insulin action (P < 0.05), and reduced the serum lipid profiles and glycosylated serum protein levels (P < 0.05). Furthermore, high-throughput pyrosequencing of the 16S rRNA gene revealed that WMB and DMB supplementation could prevent HFD and STZ-induced gut microbiota dysbiosis, especially for the enrichment of some benign bacteria, such as Bifidobacterium and Akkermansia, and the reduction of some harmful bacteria (Staphylococcus and Enterococcus). Overall, although decortication processing had an impact on the beneficial effects of mung bean, it did not cause the loss of all health benefits.


Hand hygiene instruction decreases illness-related absenteeism in elementary schools: a prospective cohort study.
Illness-related absences have been shown to lead to negative educational and economic outcomes. Both hand washing and hand sanitizer interventions have been shown to be effective in reducing illness-related absences. However, while the importance of hand hygiene in schools is clear, the role of instruction in use is less obvious. The purpose of this study was to compare absenteeism rates among elementary students given access to hand hygiene facilities versus students given both access and short repetitive instruction in use, particularly during influenza season when illness-related absences are at a peak. A hand hygiene intervention was implemented from October to May during the 2009/2010 academic year, including peak flu season, in two Chicago Public Elementary Schools among students grades pre-kindergarten to eighth grade (ages 4-14). Classrooms were systematically assigned to an intervention or control group by grade (cluster design). Hand hygiene facilities (sanitizer and soap) were made available to all students. Students in the intervention group also received short repetitive instruction in hand hygiene every 2 months. Only absences as a result of respiratory or gastrointestinal illness were used to establish illness-related absenteeism rates. Percent absent days were calculated and bivariate analyses were performed to compare percent absent days among students given access to hand hygiene facilities versus students given both access and instruction. Prior to the intervention, teachers' perceptions of students' hand hygiene were also evaluated. Teacher perceptions were analysed to describe attitudes and beliefs. Data were collected and analysed for 773 students reporting 1,886 absences during the study period (1.73% of total school days). Both the percent total absent days and percent illness-related absent days were significantly lower in the group receiving short instruction during flu season (P = 0.002, P < 0.001, respectively). This difference peaked during the influenza season (when intervention began) and declined in the following months. Teachers (n = 23) agreed that hand hygiene is not performed properly among students and reported time constraints as a barrier to frequent hand washing. Adding hand hygiene instruction to existing hand hygiene practices improved attendance at public elementary schools during the flu season. Standardized and brief repetitive instruction in hand hygiene holds potential to significantly reduce absenteeism.


Metagenomics-Based Proficiency Test of Smoked Salmon Spiked with a Mock Community.
An inter-laboratory proficiency test was organized to assess the ability of participants to perform shotgun metagenomic sequencing of cold smoked salmon, experimentally spiked with a mock community composed of six bacteria, one parasite, one yeast, one DNA, and two RNA viruses. Each participant applied its in-house wet-lab workflow(s) to obtain the metagenomic dataset(s), which were then collected and analyzed using MG-RAST. A total of 27 datasets were analyzed. Sample pre-processing, DNA extraction protocol, library preparation kit, and sequencing platform, influenced the abundance of specific microorganisms of the mock community. Our results highlight that despite differences in wet-lab protocols, the reads corresponding to the mock community members spiked in the cold smoked salmon, were both detected and quantified in terms of relative abundance, in the metagenomic datasets, proving the suitability of shotgun metagenomic sequencing as a genomic tool to detect microorganisms belonging to different domains in the same food matrix. The implementation of standardized wet-lab protocols would highly facilitate the comparability of shotgun metagenomic sequencing dataset across laboratories and sectors. Moreover, there is a need for clearly defining a sequencing reads threshold, to consider pathogens as detected or undetected in a food sample.


A Novel Truncating Mutation in PAX1 Gene Causes Otofaciocervical Syndrome Without Immunodeficiency.
Otofaciocervical syndrome (OTFCS) is a rare genetic disorder of both autosomal recessive and autosomal dominant patterns of inheritance. It is caused by biallelic or monoallelic mutations in PAX1 or EYA1 genes, respectively. Here, we report an OTFCS2 female patient of 1st consanguineous healthy parents. She manifested facial dysmorphism, hearing loss, intellectual disability (ID), and delayed language development (DLD) as the main clinical phenotype. The novel homozygous variant c.1212dup (p.Gly405Argfs*51) in the PAX1 gene was identified by whole exome sequencing (WES), and family segregation confirmed the heterozygous status of the mutation in the parents using the Sanger sequencing. The study recorded a novel PAX1 variant representing the sixth report of OTFCS2 worldwide and the first Egyptian study expanding the geographic area where the disorder was confined.


Spruce budworm mortality as a function of aerial spray droplet size.
The size and number of aerial spray drops impinging on spruce budworm in its conifer forest habitat were determined by means of a new tracer method that uses fluorescent particles in a liquid spray. Examination of 1113 larvae affected by an experimental insecticide that had been applied to a 5000-acre (2024-ha) test area in Montana showed that 93 percent had not been contacted by any droplets larger than 50 micro in diameter. Small numbers of droplets 50 to 100 micro in diameter were found on 7 percent of the larvae, along with lethal numbers of smaller drops. No evidence was found that significant numbers of drops larger than 100 micro reached the target insects. Because about 95 percent of the spray applied to forests by current methods consist of droplets larger than 50 micro, the biologically effective portion of the drop spectrum is only a few percent. The data foreshadow a major potential reduction in insecticide requirements for the successful control of spruce budworm.


Comparative genome analysis reveals the evolution of chloroacetanilide herbicide mineralization in Sphingomonas wittichii DC-6.
The environmental fate of the extensively used chloroacetanilide herbicides (CH) has been a cause of increasing concern in the past decade because of their carcinogenic properties. Although microbes play important roles in CH degradation, Sphingomonas wittichii DC-6 was the first reported CH-mineralizing bacterium. In this study, the complete genome of strain DC-6 was sequenced and comparative genomic analysis was performed using strain DC-6 and other three partial CH-degrading bacteria, Sphingobium quisquiliarum DC-2, Sphingobium baderi DE-13, and Sphingobium sp. MEA3-1. 16S rDNA phylogenetic analysis indicated that strain DC-2, MEA3-1, and DE-13 are closely related and DC-6 has relatively distant genetic relationship with the other three strains. The identified CH degradation genes responsible for the upstream and downstream pathway, including cndA, cmeH, meaXY, and meaAB, were all located in conserved DNA fragments (or genetic islands) in the vicinity of mobile element proteins. Protein BLAST in the NCBI database showed that cndA and cmeH were present in the genomes of other sequenced strains isolated from various habitats; however, the gene compositions in these host strains were completely different from those of other sphingomonads, and codon usage of genes for upstream pathway were also different from that of downstream pathway. These results showed that the upstream and downstream pathways of CH degradation in strain DC-6 have evolved by horizontal gene transfer and gene combination. In addition, the genes of the ring-cleavage pathway were not conserved and may have evolved directly from bacterial degradation of hydroxyquinol. The present study provides insights into the evolutionary strategy and microbial catabolic pathway of CH mineralization.


Virologic Response to First-line Efavirenz- or Nevirapine-based Antiretroviral Therapy in HIV-infected African Children.
Poorer virologic response to nevirapine- versus efavirenz-based antiretroviral therapy (ART) has been reported in adult systematic reviews and pediatric studies. We compared drug discontinuation and viral load (VL) response in ART-naïve Ugandan/Zimbabwean children ≥3 years of age initiating ART with clinician-chosen nevirapine versus efavirenz in the ARROW trial. Predictors of suppression <80, <400 and <1000 copies/mL at 36, 48 and 144 weeks were identified using multivariable logistic regression with backwards elimination (P = 0.1). A total of 445 (53%) children received efavirenz and 391 (47%) nevirapine. Children receiving efavirenz were older (median age, 8.6 vs. 7.5 years nevirapine, P < 0.001) and had higher CD4% (12% vs. 10%, P = 0.05), but similar pre-ART VL (P = 0.17). The initial non-nucleoside-reverse-transcriptase-inhibitor (NNRTI) was permanently discontinued for adverse events in 7 of 445 (2%) children initiating efavirenz versus 9 of 391 (2%) initiating nevirapine (P = 0.46); at switch to second line in 17 versus 23, for tuberculosis in 0 versus 26, for pregnancy in 6 versus 0 and for other reasons in 15 versus 5. Early (36-48 weeks) virologic suppression <80 copies/mL was superior with efavirenz, particularly in children with higher pre-ART VL (P = 0.0004); longer-term suppression was superior with nevirapine in older children (P = 0.05). Early suppression was poorer in the youngest and oldest children, regardless of NNRTI (P = 0.02); longer-term suppression was poorer in those with higher pre-ART VL regardless of NNRTI (P = 0.05). Results were broadly similar for <400 and <1000 copies/mL. Short-term VL suppression favored efavirenz, but long-term relative performance was age dependent, with better suppression in older children with nevirapine, supporting World Health Organization recommendation that nevirapine remains an alternative NNRTI.


Study protocol for a phase III multicentre, randomised, open-label, blinded-end point trial to evaluate the efficacy and safety of immunoglobulin plus cyclosporin A in patients with severe Kawasaki disease (KAICA Trial).
Kawasaki disease (KD) is an acute, self-limited vasculitis of unknown aetiology that predominantly affects infants and young children. We hypothesise that cyclosporin A (CsA) may be effective in treating KD by regulating the Ca(2+)/NFAT signalling pathway. This trial compares the current standard therapy of intravenous immunoglobulin (IVIG) and the combined IVIG+CsA therapy in paediatric patients with severe KD. This trial is a phase III, multicentre, randomised, open-label, blinded-end point trial that evaluates the efficacy and safety of IVIG+CsA therapy. Patients with severe KD who satisfy the eligibility criteria are randomised (1:1) to receive either CsA (5 mg/kg/day for 5 days; Neoral) plus high-dose IVIG (2 g/kg for 24 h and aspirin 30 mg/kg/day), or high-dose IVIG alone (2 g/kg for 24 h and aspirin 30 mg/kg/day). The primary end point is the frequency of occurrence of coronary artery abnormalities during the trial period. An independent end point review committee will be in charge of the trial assessment. The protocol was approved by the Institutional Review Board of each institution. The trial was notified and registered at the Pharmaceutical and Medical Devices Agency, in Japan. The trial is currently on-going and is scheduled to finish in April 2017. The findings will be disseminated through peer-reviewed publications and conference presentations. JMA-IIA00174; Pre-results.


The Genus Xanthagaricus: An Updated Global Species Distribution and Phylogeny with the Description of Two New Species from Oman.
Xanthagaricus is a saprotrophic mushroom genus with small-sized basidiomata in the family Agaricaceae (Agaricales). Prior to this study, 26 species belonging to this genus have been described and published. In this study, we reported Xanthagaricus for the first time from Oman with the description of two new species. Basidiomata of the new species Xanthagaricus appendiculatus and X. omanicus were collected during the monsoon rains of summer in 2018 in the southern coastal region of Oman. Species descriptions are based on morphological and molecular characterization. Phylogenetic analyses based on internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS) of the nuclear ribosomal DNA clustered the new species in the Xanthagaricus clade with strong statistical support. The new species Xanthagaricusappendiculatus can be distinguished from other species by its purplish pileus with umbonate disc and X. omanicus with the largest pileus (70-90 mm diameter) among the known species of the genus. A detailed description, photographs, line drawings, and a phylogenetic tree showing the position of both new species are provided. A dichotomous key to the known taxa of Xanthagaricus is proposed. Morphological comparisons of new species with known Xanthagaricus taxa are provided. Our observations highlight the diversity of Xanthagaricus and other lepiotaceous mushrooms in southern Oman and further document the need for additional systematic focus on the region's fungi.


Delayed pacemaker lead perforations: Why unusual presentations should prompt an early multidisciplinary team approach.
Our first case is an 84-year-old female diagnosed with sick sinus syndrome. She underwent implantation of dual chamber permanent pacemaker without complications. On the 8th day status-postimplantation, she returned to the emergency department (ED) with moderately severe left anterior chest pain and significant ecchymosis. She was given an initial diagnosis of shingles and discharged. Two days later, she returned to the ED with increasing chest pain, dyspnea, nausea, and vomiting. Lead migration and cardiac perforation was confirmed by chest X-ray and computed tomography (CT), respectively. She was taken to the operating room (OR) for lead repositioning, and she was discharged the next day. Our second case is a 64-year-old female with a diagnosis of 2:1 high-grade third-degree atrioventricular block. A dual chamber permanent pacemaker system was implanted without initial complication. Five days after implantation, she presented to the ED following an episode of syncope due to hypotension (67/46), shortness of breath, left flank pain, and fatigue. The initial diagnosis was sepsis. A chest CT was obtained, noting lead perforation and hemothorax. The patient was taken to the OR for lead repositioning.


Wear and surface cracking in early retrieved highly cross-linked polyethylene acetabular liners.
A higher degree of cross-linking has been shown to improve the tribological properties of ultra-high molecular weight polyethylene in laboratory studies; however, its effect on in vivo behavior has not been well established. We investigated in vivo wear mechanisms in retrieved highly cross-linked polyethylene acetabular liners in order to determine if early in vivo wear behavior is accurately predicted by hip-simulator studies. A total of twenty-four liners (twenty-one explanted and one unimplanted highly cross-linked liners and two explanted ethylene-oxide-sterilized non-cross-linked liners) were examined for this study. The average age of the patients was 59.9 years, and the average time in vivo was 10.1 months. Articular surface damage on the front and back sides of the liners was assessed with an optical scoring system. Surface quadrants were assigned a grade from 0 to 3 according to the observed wear mechanisms and the percentage of surface affected. The micromechanisms of liner damage were evaluated with use of scanning electron microscopy. The average front and back-side explant damage scores were 11 (range, 2 to 26.5) and 6.7 (range, 3.7 to 13.3), respectively. There was consistent evidence of early surface deformation and cracking. All explants exhibited some form of surface change, including surface cracking, abrasion, pitting, or scratching. The original machining marks on the liner surface were observed to be either unaltered, drastically distorted, or absent. Highly cross-linked ultra-high molecular weight polyethylene acetabular liners that were retrieved at an average of ten months after implantation exhibited signs of surface damage that had not been predicted by in vitro hip-simulator studies. These devices had not failed clinically as a result of wear. The discrepancy between in vitro and in vivo wear surfaces may be due to variability in terms of in vivo lubrication and cyclic loading or may represent early surface damage mechanisms that are not well demonstrated by long-term simulator studies.


Pediatric organ donation and transplantation.
There is increasing unmet need for solid organ donation. Alternative donor sources, such as donation after circulatory determination of death (DCDD), are needed. The objective of this study was to examine the impact of DCDD on trends in pediatric organ donation and transplantation. Data were obtained from the Organ Procurement and Transplantation Network for US organ recipients and donors from 2001 to 2010 stratified according to age, organ, and deceased donor type (DCDD or donation after neurologic determination of death). Additional data included transplant wait-list removals due to death. From 2001 to 2010, pediatric organ transplant recipients increased from 1170 to 1475. Organs from DCDD donors were transplanted into children infrequently but increased from 1 to 31. Pediatric donation after neurologic determination of death decreased by 13% whereas DCDD increased by 174% (50 to 137). Recipients of pediatric grafts decreased from 3042 to 2751. Adults receiving grafts from pediatric donors decreased from 2243 to 1780; children receiving pediatric grafts increased from 799 to 971. Transplant recipients receiving pediatric DCDD grafts were few but increased annually from 50 to 128 adults and 0 to 9 children. Pediatric candidates dying waiting for an organ decreased from 262 to 110. From 2001 to 2010, children received more solid organ transplants and fewer children died waiting. Organ recovery from pediatric and adult DCDD donors increased. The number of pediatric recipients of DCDD grafts remains small. Adults primarily receive the direct benefit from pediatric DCDD but other changes in organ allocation have directly benefited children.


A systematic proteomic analysis of Listeria monocytogenes house-keeping protein secretion systems.
Listeria monocytogenes is a firmicute bacterium causing serious infections in humans upon consumption of contaminated food. Most of its virulence factors are secretory proteins either released to the medium or attached to the bacterial surface. L. monocytogenes encodes at least six different protein secretion pathways. Although great efforts have been made in the past to predict secretory proteins and their secretion routes using bioinformatics, experimental evidence is lacking for most secretion systems. Therefore, we constructed mutants in the main housekeeping protein secretion systems, which are the Sec-dependent transport, the YidC membrane insertases SpoIIIJ and YqjG, as well as the twin-arginine pathway, and analyzed their secretion and virulence defects. Our results demonstrate that Sec-dependent secretion and membrane insertion of proteins via YidC proteins are essential for viability of L. monocytogenes. Depletion of SecA or YidC activity severely affected protein secretion, whereas loss of the Tat-pathway was without any effect on secretion, viability, and virulence. Two-dimensional gel electrophoresis combined with protein identification by mass spectrometry revealed that secretion of many virulence factors and of enzymes synthesizing and degrading the cell wall depends on the SecA route. This finding was confirmed by SecA inhibition experiments using sodium azide. Analysis of secretion of substrates typically dependent on the accessory SecA2 ATPase in wild type and azide resistant mutants of L. monocytogenes revealed for the first time that SecA2-dependent protein secretion also requires the ATPase activity of the house-keeping SecA protein.


EFFECTS OF RELIGIOUS VERSUS STANDARD COGNITIVE-BEHAVIORAL THERAPY ON OPTIMISM IN PERSONS WITH MAJOR DEPRESSION AND CHRONIC MEDICAL ILLNESS.
We compared the effectiveness of religiously integrated cognitive behavioral therapy (RCBT) versus standard CBT (SCBT) on increasing optimism in persons with major depressive disorder (MDD) and chronic medical illness. Participants aged 18-85 were randomized to either RCBT (n = 65) or SCBT (n = 67) to receive ten 50-min sessions remotely (94% by telephone) over 12 weeks. Optimism was assessed at baseline, 12 and 24 weeks by the Life Orientation Test-Revised. Religiosity was assessed at baseline using a 29-item scale composed of religious importance, individual religious practices, intrinsic religiosity, and daily spiritual experiences. Mixed effects growth curve models were used to compare the effects of treatment group on trajectory of change in optimism. In the intention-to-treat analysis, both RCBT and SCBT increased optimism over time, although there was no significant difference between treatment groups (B = -0.75, SE = 0.57, t = -1.33, P = .185). Analyses in the highly religious and in the per protocol analysis indicated similar results. Higher baseline religiosity predicted an increase in optimism over time (B = 0.07, SE = 0.02, t = 4.12, P < .0001), and higher baseline optimism predicted a faster decline in depressive symptoms over time (B = -0.61, SE = 0.10, t = -6.30, P < .0001), both independent of treatment group. RCBT and SCBT are equally effective in increasing optimism in persons with MDD and chronic medical illness. While baseline religiosity does not moderate this effect, religiosity predicts increases in optimism over time independent of treatment group.


Genotype-phenotype correlation in 18 Egyptian patients with glutaric acidemia type I.
Glutaric acidemia I (GAI) is an autosomal recessive metabolic disease caused by a deficiency of glutaryl-CoA dehydrogenase enzyme (GCDH). Patients with GAI are characterized by macrocephaly, acute encephalitis-like crises, dystonia and frontotemporal atrophy. In this study, we investigated 18 Egyptian patients that were diagnosed with GAI based on their clinical, neuroradiological, and biochemical profiles. Of the 18 patients, 16 had developmental delay and/or regression, dystonia was prominent in 75% of the cases, and three patients died. Molecular genetics analysis identified 14 different mutations in the GCDH gene in the 18 patients, of the 14 mutations, nine were missense, three were in the 3'-Untranslated Region (3'-UTR), one was nonsense, and one was a silent mutation. Four novel mutations were identified (c.148 T > A; p.Trp50Arg, c.158C > A; p.Pro53Gln, c.1284C > G; p.Ile428Met, and c.1189G > T; p.Glu397*) that were all absent in 300 normal chromosomes. The 3'-UTR mutation (c.*165A > G; rs8012), was the most frequent mutation observed (0.5; 18/36), followed by the most common mutation among Caucasian patients (p.Arg402Trp; rs121434369) with allele frequency of 0.36 (13/36), and the 3'-UTR mutation (c.*288G > T; rs9384, 0.22; 8/16). The p.Arg257Gln mutation was found with allele frequency of ~0.17 (6/36). The marked homozygosity observed in our patients is probably due to the high level of consanguinity that is observed in 100% of the cases. We used nine in silico prediction tools to predict the pathogenicity (SIFT, PhD-SNP, SNAP, Meta-SNP, PolyPhen2, and Align GVGD) and protein stability (I-Mutant2.0, Mupro, and istable) of the nine missense mutants. The mutant p.Arg402Trp was predicted to be most deleterious by all the six pathogenicity prediction tools and destabilizing by all the three-stability prediction tools, and highly conserved by the ConSurf server. Using the clinical, biochemical, family history of the 18 patients, and the in silico analysis of the missense mutations, our study showed a mix of conclusive and inconclusive genotype-phenotype correlations among our patient's cohort and suggests the usefulness of using various sophisticated computational analysis to be utilized for future variant classifications in the genetic clinics.


Factors associated with late Human Immunodeficiency Virus (HIV) diagnosis among peoples living with it, Northwest Ethiopia: hospital based unmatched case-control study.
Early HIV diagnosis and access to treatment is one of the most effective ways to prevent its further spread and to protect the health of those living with the virus. However, delay in diagnosis is the major risk factor for uptake of and response to antiretroviral therapy. Institution-based unmatched case-control study design was used in the study. The study was conducted in Debre-Markos and Finote-Selam Hospitals, Northwest Ethiopia. Cases were people living with HIV who had CD4 count <350cells/mm3 or WHO clinical stage III and IV regardless of the CD4 count at first presentation and controls were those who had CD4 count ≥350cells/mm3 or WHO clinical stage I and II. If both criteria were available, the CD4 count was used in the study as World Health Organization recommended. A total of 392 respondents (196 cases and 196 controls) were recruited and selected systematically. The data were collected by trained nurses using chart review and interviewer administered structured questionnaire. Binary Logistic Regression Model was used to identify the factors associated with late HIV diagnosis. About 95.9 % of study participants provided complete response. Having no understanding, compared to having understanding, about HIV/AIDS (AOR = 1.7, 95 %CI = 1.08-2.79) and ART (AOR = 2.1, 95 %CI: 1.25-3.72), being tested as a result of symptoms/ illness, compared to being tested for risk exposure (inverted AOR =2.5, 95 %CI: 1.64-4.76), and acquiring HIV through sexual contact, compared to acquiring it through other modes (AOR = 2.5, 95 %CI = 1.52-4.76) were positively and independently associated with late HIV diagnosis. Unlike perceived HIV stigma, having no understanding about HIV and ART, being tested for presence of symptoms/illness, and acquiring HIV through sexual contact were independent and significant factors for late HIV diagnosis.


Source strengths for indoor human activities that resuspend particulate matter.
A mathematical model was applied to continuous indoor and outdoor particulate matter (PM) measurements to estimate source strengths for a variety of prescribed human activities that resuspend house dust in the home. Activities included folding blankets, folding clothes, dry dusting, making a bed, dancing on a rug, dancing on a wood floor, vacuuming, and walking around and sitting on upholstered furniture. Although most of the resuspended particle mass from these activities was larger than 5 microm in diameter, the resuspension of PM2.5 and PM5 was substantial, with source strengths ranging from 0.03 to 0.5 mg min(-1) for PM2.5 and from 0.1 to 1.4 mg min(-1) for PM5. Source strengths for PM > 5 microm could not be quantified due to instrument limitations. The source strengths were found to be a function of the number of persons performing the activity, the vigor of the activity, the type of activity, and the type of flooring.


The interrelationship between anti-Müllerian hormone, ovarian follicular populations and age in mares.
Anti-Müllerian hormone (AMH) is a granulosa-cell-derived glycoprotein, which plays an important inhibitory role during folliculogenesis. Concentrations of AMH are highly correlated with antral follicle counts (AFCs) in other species, which in turn are related to follicular reserve. Relatively little is known about AMH and AFC in the mare. To determine plasma AMH concentrations and AFCs in mares of different ages, to measure the repeatability of AMH concentrations and AFCs within and across oestrous cycles and to assess the relationship between plasma AMH concentrations and AFCs with regard to mare age and follicle size. An observational study examining the relationship between AMH, AFC and age in 45 mares. Young (3-8 years), middle-aged (9-18 years) and old mares (19-27 years) were examined by transrectal ultrasonography over 2 or 3 oestrous cycles. Plasma AMH concentrations and AFCs were determined, and antral follicles were classified by size into different groups. Plasma AMH concentrations varied widely between mares within similar age groups. Antral follicle counts were significantly lower in old mares than in young and middle-aged mares, and AMH concentrations were significantly lower in old than in middle-aged mares. A positive relationship was detected between AFC and AMH, and this relationship varied by mare age with a strong correlation in older mares (ρ = 0.86; P<0.0001), a moderate correlation in middle-aged mares (ρ = 0.60; P = 0.01) and no correlation in young mares (ρ = 0.40, P<0.4). The AMH concentrations were significantly related to the number of antral follicles between 6 and 20 mm in diameter, and the repeatability of AFCs and AMH concentrations was high within and between oestrous cycles. Our findings indicate that the relationship between AMH and AFC varies across age groups, and concentrations of AMH might be a better reflection of reproductive age than calendar age.


Analysis of the analytical performance of laboratories participating in two major U.S. bulk asbestos proficiency testing programs.
Quality assurance and proficiency testing programs have been available for 27 years for U.S. laboratories using polarized light microscopy to analyze bulk building materials for asbestos. Total enrollment in the two principal bulk asbestos proficiency testing programs conducted by RTI International peaked at nearly 900 laboratories in the mid-1990s. Enrollment has stabilized in the last 5 years at approximately 475 laboratories, and rates of return of analysis results currently exceed 95%. More than 115,000 test samples have been sent worldwide by RTI to laboratories in these two programs. A review of more than 109,000 analysis results was undertaken to determine the frequencies of various qualitative error types, the types of bulk building materials most responsible for those errors, and the accuracy of precision of semiquantitative results. This assessment revealed that a small number of bulk building materials cause the majority of difficulties in each program, although error frequencies vary between the two programs. Overall laboratory performance has improved since the programs first began in the late 1980s, with a significant decline in qualitative errors and a much less dramatic decrease in errors associated with semiquantitative estimation of the amount of asbestos present.


Infiltration performance of engineered surfaces commonly used for distributed stormwater management.
Engineered porous media are commonly used in low impact development (LID) structures to mitigate excess stormwater in urban environments. Differences in infiltrability of these LID systems arise from the wide variety of materials used to create porous surfaces and subsequent maintenance, debris loading, and physical damage. In this study, the infiltration capacity of six common materials was tested by multiple replicate experiments with automated mini-disk infiltrometers. The tested materials included porous asphalt, porous concrete, porous brick pavers, flexible porous pavement, engineered soils, and native soils. Porous asphalt, large porous brick pavers, and curb cutout rain gardens showed the greatest infiltration rates. Most engineered porous pavements and soils performed better than the native silt loam soils. Infiltration performance was found to be related more to site design and environmental factors than material choice. Sediment trap zones in both pavements and engineered soil rain gardens were found to be beneficial to the whole site performance. Winter chloride application had a large negative impact on poured in place concrete, making it a poor choice for heavily salted areas.


Steroid receptor and molecular chaperone encounters in the nucleus.
Steroid hormone receptors interact with several different molecular chaperones. DeFranco and Csermely discuss how the molecular chaperones p23 and Hsp90 may serve to regulate the activity of the ligand-bound steroid receptors within the nucleus. The authors hypothesize that these chaperone proteins may have a proactive role in promoting recycling of receptors once they have interacted with chromatin and in allowing rebinding of ligand once the receptors have been recycled.


Manipulation of orbital angular momentum beams based on space diffraction compensation.
We put forward a technique to manipulate the size of orbital angular momentum (OAM) beams based on space diffraction compensation. Paraxial Fresnel diffraction which carries a negative spatial quadratic phase distribution can be regarded as a negative diffractive effect. To compensate the negative diffraction, we employ a 4f Fourier lens system containing a phase mask to generate an inverse quadratic phase. The size of OAM beams can be easily controlled by designing the phase mask profile without changing the OAM. The applications of space diffraction compensation in OAM demultiplexing, ring fiber coupling for OAM beams and optical manipulation of micro particles are also discussed.


Alteration of cholinergic system in keratinocytes cells produces acantholysis: a possible use of cholinergic drugs in pemphigus vulgaris.
Human epidermis shows a non-neuronal cholinergic system including keratinocyte (kc) acetylcholine (Ach) axis which is composed by enzymes and two families of Ach receptors (muscarinic and nicotinic receptors). The activity of these two receptors can regulate the interkeratinocytes and kcs-extracellular matrix adhesion modifying the regulation of intercellular adhesion molecules like cadherins and integrins. Some authors demonstrate that acantholysis in pemphigus depends not only on anti desmogleins antibodies (abs) (mostly IgG) but even on other abs directed against kc membrane antigens (e.g. anti Ach receptors Abs). In the early phase of pemphigus pathogenesis, anti Ach receptors Abs block Ach signaling essential for cell shape and intercellular adhesion and increase the phosphorylation of adhesion molecules. Combined with the action of abs antidesmogleins, anti Ach receptors Abs cause the acantholytic phenomenon. In vitro experiments show that high doses of Ach in acantholytic kcs can rapidly reverse this pathologic event. In vivo experiments using neonatal mice model of Pemphigus have demonstrated that cholinergic agonists reduce these lesions. Therapy with pyridostigmine bromide and Nicotinamide per os or pilocarpine used topically, drugs that present cholinomimetic effects, has lead to encouraging results in patients affected by Pemphigus disease. Cholinergic agents could have a strategic role in the therapy of pemphigus since they could be responsible for the early stage of acantholytic diseases.


A Case-Matched Gender Comparison Transcriptomic Screen Identifies eIF4E and eIF5 as Potential Prognostic Markers in Male Breast Cancer.
Purpose: Breast cancer affects both genders, but is understudied in men. Although still rare, male breast cancer (MBC) is being diagnosed more frequently. Treatments are wholly informed by clinical studies conducted in women, based on assumptions that underlying biology is similar.Experimental Design: A transcriptomic investigation of male and female breast cancer was performed, confirming transcriptomic data in silico Biomarkers were immunohistochemically assessed in 697 MBCs (n = 477, training; n = 220, validation set) and quantified in pre- and posttreatment samples from an MBC patient receiving everolimus and PI3K/mTOR inhibitor.Results: Gender-specific gene expression patterns were identified. eIF transcripts were upregulated in MBC. eIF4E and eIF5 were negatively prognostic for overall survival alone (log-rank P = 0.013; HR = 1.77, 1.12-2.8 and P = 0.035; HR = 1.68, 1.03-2.74, respectively), or when coexpressed (P = 0.01; HR = 2.66, 1.26-5.63), confirmed in the validation set. This remained upon multivariate Cox regression analysis [eIF4E P = 0.016; HR = 2.38 (1.18-4.8), eIF5 P = 0.022; HR = 2.55 (1.14-5.7); coexpression P = 0.001; HR = 7.04 (2.22-22.26)]. Marked reduction in eIF4E and eIF5 expression was seen post BEZ235/everolimus, with extended survival.Conclusions: Translational initiation pathway inhibition could be of clinical utility in MBC patients overexpressing eIF4E and eIF5. With mTOR inhibitors that target this pathway now in the clinic, these biomarkers may represent new targets for therapeutic intervention, although further independent validation is required. Clin Cancer Res; 23(10); 2575-83. ©2016 AACR.


Blood pressure measurements during intraoperative pediatric scoliosis surgery.
Intraoperative hypotension is frequently encountered during surgery and it can be associated with adverse outcomes. Blood pressure monitoring is critical during surgery, but there are no universally agreed upon standards for interpreting values of hypotension and no consensus regarding interventions. We performed a retrospective chart review of pediatric patients who underwent idiopathic scoliosis surgery by a single surgeon. We used the arterial line for all measures. Intraoperative hypotension was defined as 20% decrease of the baseline systolic blood pressure (SBP), 30% decrease of baseline SBP, or mean arterial pressure less than 60 mmHg. Use of vasopressor agents was also recorded and correlated with blood pressure definitions. Seventy idiopathic scoliosis patients were retrospectively evaluated. There was a significant correlation between the three measures of hypotension. Sixty percent of the patients received vasopressors. There was a significant correlation between a drop of mean arterial pressure to less than 60 mmHg and the use of the ephedrine. We did not find any changes on neuromonitoring measures during the case and there were no intraoperative or one-month postoperative complications. Blood pressure is only one of the measures anesthesiologists look to for good perfusion during surgery. Pediatric anesthesiologists and orthopedics agree in trying tight blood pressure control during surgery to decrease blood loss, but what the exact definition of that blood pressure number is, is still unclear. We propose that using mean arterial pressure less than 60 mmHg is perhaps a better definition. We provide recommendations for future studies.


Skeletal muscle protein anabolic response to resistance exercise and essential amino acids is delayed with aging.
Skeletal muscle loss during aging leads to an increased risk of falls, fractures, and eventually loss of independence. Resistance exercise is a useful intervention to prevent sarcopenia; however, the muscle protein synthesis (MPS) response to resistance exercise is less in elderly compared with young subjects. On the other hand, essential amino acids (EAA) increase MPS equally in both young and old subjects when sufficient EAA is ingested. We hypothesized that EAA ingestion following a bout of resistance exercise would stimulate anabolic signaling and MPS similarly between young and old men. Each subject ingested 20 g of EAA 1 h following leg resistance exercise. Muscle biopsies were obtained before and 1, 3, and 6 h after exercise to measure the rate of MPS and signaling pathways that regulate translation initiation. MPS increased early in young (1-3 h postexercise) and later in old (3-6 h postexercise). At 1 h postexercise, ERK1/2 MNK1 phosphorylation increased and eIF2alpha phosphorylation decreased only in the young. mTOR signaling (mTOR, S6K1, 4E-BP1, eEF2) was similar between groups at all time points, but MNK1 phosphorylation was lower at 3 h and AMP-activated protein kinase-alpha (AMPKalpha) phosphorylation was higher in old 1-3 h postexercise. We conclude that the acute MPS response after resistance exercise and EAA ingestion is similar between young and old men; however, the response is delayed with aging. Unresponsive ERK1/2 signaling and AMPK activation in old muscle may be playing a role in the delayed activation of MPS. Notwithstanding, the combination of resistance exercise and EAA ingestion should be a useful strategy to combat sarcopenia.


Randomised comparison of ceftriaxone and cefamandole therapy in lower respiratory tract infections in an elderly population.
Patients with pneumonia or bronchitis were randomized to receive ceftriaxone or cefamandole. A total of 30 of 38 patients were evaluable, 16 in the ceftriaxone group (average age 66.3 years) and 14 in the cefamandole group (average age 69.4 years). All but one had underlying diseases. Patients usually received 1 g of ceftriaxone intravenously every 12 h (mean duration 8.7 days) or 1.5 g of cefamandole intravenously every 6 h (mean duration 8.2 days). Adverse experiences attributable to the drugs were confined to one episode of discomfort at the infusion site in each group. Bacteriological results with ceftriaxone were 83% cured, 11% superinfected after eradication of pretherapy isolate, and 6% failed. Bacteriological results with cefamandole were 76% cured, 24% failed. Clinical results with ceftriaxone were 38% cured, 56% improved, 6% failed. Clinical results with cefamandole were 57% cured, 21% improved, 21% failed. Emergence of a resistant Serratia marcescens was seen in a ceftriaxone-treated patient. Disc diffusion susceptibility testing identified six of the seven pretherapy nonfastidious Gram-negative isolates as susceptible; however, two of the six could not be eradicated with the assigned drug and another two were eradicated with ensuing super-infection with susceptible isolates of Pseudomonas aeruginosa. In contrast, MBCs were an accurate guide to clinical outcome with nonfastidious Gram-negative bacilli.


Lung directed antibody gene transfer confers protection against SARS-CoV-2 infection.
The COVID-19 pandemic continues to be a worldwide threat and effective antiviral drugs and vaccines are being developed in a joint global effort. However, some elderly and immune-compromised populations are unable to raise an effective immune response against traditional vaccines. We hypothesised that passive immunity engineered by the in vivo expression of anti-SARS-CoV-2 monoclonal antibodies (mAbs), an approach termed vectored-immunoprophylaxis (VIP), could offer sustained protection against COVID-19 in all populations irrespective of their immune status or age. We developed three key reagents to evaluate VIP for SARS-CoV-2: (i) we engineered standard laboratory mice to express human ACE2 via rAAV9 in vivo gene transfer, to allow in vivo assessment of SARS-CoV-2 infection, (ii) to simplify in vivo challenge studies, we generated SARS-CoV-2 Spike protein pseudotyped lentiviral vectors as a simple mimic of authentic SARS-CoV-2 that could be used under standard laboratory containment conditions and (iii) we developed in vivo gene transfer vectors to express anti-SARS-CoV-2 mAbs. A single intranasal dose of rAAV9 or rSIV.F/HN vectors expressing anti-SARS-CoV-2 mAbs significantly reduced SARS-CoV-2 mimic infection in the lower respiratory tract of hACE2-expressing mice. If translated, the VIP approach could potentially offer a highly effective, long-term protection against COVID-19 for highly vulnerable populations; especially immune-deficient/senescent individuals, who fail to respond to conventional SARS-CoV-2 vaccines. The in vivo expression of multiple anti-SARS-CoV-2 mAbs could enhance protection and prevent rapid mutational escape.


Nocardia keratitis: A case report.
To describe a case of Nocardia keratitis resistant to 2% amikacin, with a toxic-allergic reaction to fortified topical 5% amikacin, and recurrence of the infection with topical corticosteroids. Nocardia was diagnosed from a smear and positive culture and identified as Nocardia asteroides by gas chromatography and quantitative fatty acid analysis using the Microbial Identification System. Treatment was started with topical 2% amikacin, which was subsequently raised to 5% because of clinical resistance. A toxic-allergic reaction was observed after 5% amikacin so the drug was discontinued and commercially available drugs combining 1% chloramphenicol, 0.5% tetracycline, and 18 mil IU colistin with 0.3% ofloxacin were given. These were well tolerated and the infection improved quickly. After 1 month the antibiotics were discontinued and topical 0.1% clobetasone was given to reduce scar formation. The infection recurred after 1 week but responded to 3 months of the previous antibiotic combination and its sensitivity was checked with the Epsilometer test. Nocardia keratitis may not respond to 2% topical amikacin and fortified topical 5% amikacin may cause a strong toxic-allergic reaction. A commercially available combination of chloramphenicol, tetracycline, and colistin, with ofloxacin, may be effective but the treatment must be continued for several months. Topical steroids should only be used with considerable caution since they can lead to relapse of the infection. (Eur J Ophthalmol 2006; 16: 164-7).