Theoretical reflections on governance in health regions.
This article analyzes governance in health regions, through the contributions of two studies: one on a governance model and the other on duties in the management of public policies networks. The former conducted a meta-analysis of 137 case studies in the literature on collaborative governance aimed at preparing an explanatory and analytical model. Authors identified critical variables that will influence the results: a previous history of conflict or cooperation, incentives for participation, power imbalances, leadership and institutional design. They also identified key factors: face-to-face dialogue, trust building and development of commitment and shared vision. The latter study examined networks of public policies in the analytic tradition and the perspective of governance, incorporating concepts from the field of political science, economics and interorganizational relations, in order to support the management of public policies networks. The study identified network management as equivalent to a strategic game involving functions: network activation, framework of relations, intermediation, facilitation and consensus building and mediation and arbitration. The combination of the two reflections provides a conceptual reference for better understanding of governance in health regions.


Protein modifications induced in mouse epidermis by potent and weak tumor-promoting hyperplasiogenic agents.
Two-dimensional gel electrophoresis was used to compare the changes in mouse epidermal proteins induced by the potent tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), by the moderate promoter mechanical abrasion, and by the weakly promoting hyperplasiogenic agents mezerein and ethylphenylpropiolate. Evidence is presented which indicates that TPA caused many changes in the epidermal protein profiles especially related to the keratins which are the major differentiation product of the epidermis. The keratin modification progresses with time after TPA treatment, resulting in a keratin pattern which resembles that of newborn mouse epidermis. The criteria used for the identification of the keratins were extractability, isoelectric points, molecular weights, filament formation in vitro, immunological cross-reactivity, amino acid composition, and peptide mapping. Several other protein changes were evident in the more soluble epidermal proteins which were also prominent in the newborn epidermis. These protein alterations are observed not only early during the TPA induction of hyperplasia and inflammation at 48 and 72 hr but also in 1- and 2-week samples in which the morphology of the epidermis has returned to normal. Mezerein and abrasion produced protein changes similar to those induced by TPA. Ethylphenylpropiolate-induced protein modifications not only occurred at later times compared with either mezerein or TPA but also were less in magnitude. However, although many of the protein modifications induced by TPA appear to be associated with the hyperplasiogenic properties of TPA, the major difference between a potent promoter like TPA and a weak promoter like ethylphenylpropiolate appeared to be related to the magnitude of the response and the time of appearance of the protein changes.


Phosphoproteomic analysis of seed maturation in Arabidopsis, rapeseed, and soybean.
To characterize protein phosphorylation in developing seed, a large-scale, mass spectrometry-based phosphoproteomic study was performed on whole seeds at five sequential stages of development in soybean (Glycine max), rapeseed (Brassica napus), and Arabidopsis (Arabidopsis thaliana). Phosphopeptides were enriched from 0.5 mg of total peptides using a combined strategy of immobilized metal affinity and metal oxide affinity chromatography. Enriched phosphopeptides were analyzed by Orbitrap tandem mass spectrometry and mass spectra mined against cognate genome or cDNA databases in both forward and randomized orientations, the latter to calculate false discovery rate. We identified a total of 2,001 phosphopeptides containing 1,026 unambiguous phosphorylation sites from 956 proteins, with an average false discovery rate of 0.78% for the entire study. The entire data set was uploaded into the Plant Protein Phosphorylation Database (www.p3db.org), including all meta-data and annotated spectra. The Plant Protein Phosphorylation Database is a portal for all plant phosphorylation data and allows for homology-based querying of experimentally determined phosphosites. Comparisons with other large-scale phosphoproteomic studies determined that 652 of the phosphoproteins are novel to this study. The unique proteins fall into several Gene Ontology categories, some of which are overrepresented in our study as well as other large-scale phosphoproteomic studies, including metabolic process and RNA binding; other categories are only overrepresented in our study, like embryonic development. This investigation shows the importance of analyzing multiple plants and plant organs to comprehensively map the complete plant phosphoproteome.


Neighing, barking, and drumming horses-object related sounds help and hinder picture naming.
The study presented here investigated how environmental sounds influence picture naming. In a series of four experiments participants named pictures (e.g., the picture of a horse) while hearing task-irrelevant sounds (e.g., neighing, barking, or drumming). Experiments 1 and 2 established two findings, facilitation from congruent sounds (e.g., picture: horse, sound: neighing) and interference from semantically related sounds (e.g., sound: barking), both relative to unrelated sounds (e.g., sound: drumming). Experiment 3 replicated the effects in a situation in which participants were not familiarized with the sounds prior to the experiment. Experiment 4 replicated the congruency facilitation effect, but showed that semantic interference was not obtained with distractor sounds which were not associated with target pictures (i.e., were not part of the response set). The general pattern of facilitation from congruent sound distractors and interference from semantically related sound distractors resembles the pattern commonly observed with distractor words. This parallelism suggests that the underlying processes are not specific to either distractor words or distractor sounds but instead reflect general aspects of semantic-lexical selection in language production. The results indicate that language production theories need to include a competitive selection mechanism at either the lexical processing stage, or the prelexical processing stage, or both. (PsycINFO Database Record


[Adrenergic nature of vagal acceleration of heart beat in frogs].
Acceleration of heartbeats occurring during the irritation of vagal nerve was discovered as early as in 1846. Many hypotheses about mechanisms of its appearance proved to be not confirmed. Some authors (researchers) considered that the accelerated effect resulted from secreting a small amount of acetylcholine by vagal nerve. We discovered the cause of this groundless opinion and determined that vagal acceleration of heartbeats in frogs was accomplished, partially, by sympathetic fibres of vagal nerve, partially, by catecholamines secreted, probably, from chromaffin cells.


Photopolymerized microtips for sample preparation in proteomic analysis.
We demonstrate a novel method for the fabrication of disposable plastic microtips, which we name "EasyTip", by a photopolymerization technique. C18 reversed-phase (C18) and ion metal affinity chromatography (IMAC) beads were immobilized on a plastic pipette tip, made of polypropylene materials, by photo-initiated polymerization. The fabricated EasyTips can be manipulated using commercial pipettes for wash/elution of minute amount of biological samples (< 10 microL) and can be applied for mass spectrometry (MS)-based proteomic analysis, in which the detection sensitivity depends critically on the optimal sample preparation. The recovery of a sample of 25 fmol of tryptic hemoglobin digest loaded in a C18 EasyTip was near 100% and we estimated the loading capacity to be around 0.4-2.0 microg of total proteins or peptides, which is well above a sufficient quantity for MS analysis. The effectiveness of the C18 EasyTips in enhancing the detection sensitivity of matrix-assisted laser desorption/ionization (MALDI)-MS signal, and thus providing a greater sequence coverage, was also demonstrated by the analysis of hemoglobin digest and the in-gel digested epidermal growth factor receptor (EGFR) protein from A431 cell lysate. We also demonstrated the usefulness of the immobilized IMAC EasyTips in extracting the signal of tryptic phosphopeptides of beta-casein (10 pmol) having one and four phosphorylation sites by using an IMAC EasyTip prior to off-line analysis by MS. The combination of IMAC EasyTips and MALDI-MS allowed the unambiguous identification of phosphopeptides based on the phosphatase assay as well as the post-source decay. Compared to other miniaturized devices, this fabrication method is simple, cheap, and requires less human intervention. Moreover, the method of manipulating the EasyTips is straightforward and can be automated readily by a robotic system for high-throughput analysis.


Healthcare costs for abortions performed in ambulatory surgery centers vs office-based settings.
Several states require that abortions be provided in ambulatory surgery centers. Supporters of such laws argue that they make abortions safer, yet previous studies have found no differences in abortion-related morbidities or adverse events for abortions performed in ambulatory surgery centers versus office-based settings. However, little is known about how costs of abortions provided in ambulatory surgery centers differ from those provided in office-based settings. To compare healthcare expenditures for abortions performed in ambulatory surgery centers versus office-based settings using a large national private insurance claims database. A retrospective cohort study compared expenditures for abortions performed in ambulatory surgery centers versus office-based settings. Data on women who had abortions in an ambulatory surgery center or office-based setting between January 1, 2011, and December 31, 2014 were obtained from the MarketScan Commercial Claims and Encounters database. The sample was limited to women who were continuously enrolled in their insurance plans for at least 1 year before and at least 6 weeks after the abortion. Healthcare expenditures were assessed separately for the index abortion and the 6-week period after the abortion. Costs were measured from the perspective of the healthcare system and included all payments to the provider, including insurance company payments and any patient out-of-pocket payments. Overall, 49,287 beneficiaries who had 50,311 abortions met inclusion criteria. Of the included abortions, 47% were first-trimester aspiration, 27% first-trimester medication, and 26% second-trimester or later abortions. Most abortions (89%) were provided in office-based settings, with 11% provided in ambulatory surgery centers. Unadjusted mean index abortion costs were higher in ambulatory surgery centers than in office-based settings ($1704 versus $810; P < .001). After adjusting for patient clinical and demographic characteristics, costs of index abortions were $772 higher (95% confidence interval, $746-$797), total follow-up costs for abortions that had any follow-up care were $1099 higher (95% confidence interval, $1004-$1,195), and total follow-up costs for abortions that had an abortion-related morbidity or adverse event were not significantly different in ambulatory surgery centers compared to office-based settings. There were also no significant differences in the likelihood of having any follow-up care or abortion-related event follow-up care. Abortions performed at ambulatory surgery centers are significantly more costly than those performed in office-based settings, with no difference in the likelihood of receiving follow-up care. Laws requiring that abortions be provided in ambulatory surgery centers may only result in increased costs for abortions, with no effect on abortion safety.


Diversity of interaction types and ecological community stability.
Ecological theory predicts that a complex community formed by a number of species is inherently unstable, guiding ecologists to identify what maintains species diversity in nature. Earlier studies often assumed a community with only one interaction type, either an antagonistic, competitive, or mutualistic interaction, leaving open the question of what the diversity of interaction types contributes to the community maintenance. We show theoretically that the multiple interaction types might hold the key to understanding community dynamics. A moderate mixture of antagonistic and mutualistic interactions can stabilize population dynamics. Furthermore, increasing complexity leads to increased stability in a "hybrid" community. We hypothesize that the diversity of species and interaction types may be the essential element of biodiversity that maintains ecological communities.


Near-total thyroidectomy could be the best treatment for thyroid disease in endemic regions.
Near-total thyroidectomy, on the basis of its low morbidity rate, is an appropriate treatment option in the surgical management of various thyroid diseases in an endemic region in Turkey. Single-institution study of patients with various thyroid diseases treated by means of near-total thyroidectomy within 2 years in an endemic region, with comparison of the results vs the complication rates of bilateral subtotal and total thyroidectomy reported in the literature. Tertiary academic referral center. One hundred fifty-two patients who underwent near-total thyroidectomy for various thyroid diseases. Surgical treatments of various benign thyroid diseases were compared according to the complication rates and the achievable benefits of the procedures. In our clinic, near-total thyroidectomy was the principal surgical procedure performed for benign thyroid disease. The temporary recurrent laryngeal nerve palsy rate with respect to the nerves at risk was 3.3% (10 of 304 nerves), whereas temporary hypoparathyroidism was 7.2% (11 of 152 patients). Neither permanent recurrent laryngeal nerve palsy nor permanent hypoparathyroidism occurred. In 1 patient, wound hematoma developed and required re-exploration. Seroma in another patient needed no medical or surgical intervention. Neither wound infection nor mortality were noted. Near-total thyroidectomy achieves a lower complication rate of hypoparathyroidism and a similar complication rate of recurrent laryngeal nerve palsy and recurrence when compared with the rates reported in the literature for total thyroidectomy. It is an effective and safe surgical treatment option for various benign thyroid diseases.


Diagnosis of equine influenza.
During the summer months, there will be increased movement and, therefore, increased mixing of the horse population, leading to a higher risk of disease transmission and subsequent clinical cases. It is important that both vets and owners remain vigilant for equine influenza infection. Here, Adam Rash, of the Animal Health Trust, discusses the diagnosis of this disease.


Nurse autonomy, nurse job satisfaction and client satisfaction with nursing care: their place in nursing data sets.
Many changes have taken place in the healthcare system that have influenced nurse autonomy, job satisfaction and client satisfaction. Standardized language facilitates communication within the discipline of nursing. Examples of such language include the Nursing Intervention Classification (NIC) and the Nursing Outcomes Classification (NOC) systems as well as the Nursing Minimum Data Set (NMDS), which provides a formal structure for electronic data sets to support nursing care. The Nursing Management Minimum Data Set (NMMDS) was designed to identify variables to guide nurse managers in evaluating the impact of nursing interventions on client outcomes. Gaps within NMDS and NMMDS are discussed, and solutions are proposed.


Thermodynamic and Vibrational Aspects of Peptide Bond Hydrolysis and Their Potential Relationship to the Harmfulness of Infrared Radiation.
The primary physicochemical effect upon exposure to infrared radiation (IR) is the temperature increase of cells. The degradation of proteins via the hydrolysis of peptide bonds is related to cell malfunction. In this work, the degradation of proteins/peptides under the influence of IR radiation is theoretically studied. It is shown that the low value of enthalpy of peptide bond hydrolysis has two consequences: (a) the enthalpy of hydrolysis is sensitive to small variations in the bond strength, and the hydrolysis of weak peptide bonds is exothermic, while the hydrolysis of stronger bonds is endothermic; (b) the increase in temperature (e.g., due to IR exposure) changes the enthalpy of the reaction of some weak peptide bonds from exothermic to endothermic (that is, their hydrolysis will be favored upon further increase in temperature). Simple calculations reveal that the amount of absorbed energy during the overtone and hot band transitions of the H-O-H and C-N stretching vibrations is comparable to the activation energy of the (uncatalyzed) hydrolysis. A critical discussion is provided regarding the influence of different IR wavelengths on peptide bond hydrolysis.


[Premature twins: clinic comparison, reproductive assisted techniques versus spontaneous conception].
Twin pregnancies occur one out of every 800. Due to the increase in maternal age and infertility, the use of assisted reproductive technology (ART) has risen. Thirty five percent of pregnancies solved using ART are twin pregnancies. A RT consists of infertility treatments in which oocytes and sperm are manipulated. The objective of this study was to compare the clinical behavior of premature twins born through ART versus those born after spontaneous conception (SC). Retrolective cohort study. The data was obtained from the records of preterm twins born between August 2010 and August 2013. Preterm twins were categorized in two groups: the ones conceived using ART versus those conceived through SC. Demographic characteristics and neonatal outcomes were compared. The total was of 398 patients: 208 conceived in consequence of ART and 190 through SC. The primary outcome and neonatal morbidities showed no significant difference, only the incidence of necrotizing enterocolitis and the use of parenteral nutrition showed significant difference. The length of hospital stay between groups was similar. In our population of preterm twins, the mode of conception had no detectable effect on mortality or short term morbidities.


Uncertainty of pesticide residue concentration determined from ordinary and weighted linear regression curve.
Determination of pesticide residues is based on calibration curves constructed for each batch of analysis. Calibration standard solutions are prepared from a known amount of reference material at different concentration levels covering the concentration range of the analyte in the analysed samples. In the scope of this study, the applicability of both ordinary linear and weighted linear regression (OLR and WLR) for pesticide residue analysis was investigated. We used 782 multipoint calibration curves obtained for 72 different analytical batches with high-pressure liquid chromatography equipped with an ultraviolet detector, and gas chromatography with electron capture, nitrogen phosphorus or mass spectrophotometer detectors. Quality criteria of the linear curves including regression coefficient, standard deviation of relative residuals and deviation of back calculated concentrations were calculated both for WLR and OLR methods. Moreover, the relative uncertainty of the predicted analyte concentration was estimated for both methods. It was concluded that calibration curve based on WLR complies with all the quality criteria set by international guidelines compared to those calculated with OLR. It means that all the data fit well with WLR for pesticide residue analysis. It was estimated that, regardless of the actual concentration range of the calibration, relative uncertainty at the lowest calibrated level ranged between 0.3% and 113.7% for OLR and between 0.2% and 22.1% for WLR. At or above 1/3 of the calibrated range, uncertainty of calibration curve ranged between 0.1% and 16.3% for OLR and 0% and 12.2% for WLR, and therefore, the two methods gave comparable results.


A fast and safe non-bleaching method for forensic skeletal preparation.
Over the last three decades, forensic anthropologists increasingly have consulted on fleshed human remains cases in which the examination of skeletal elements is critical in answering questions of identification and the circumstances of death. This was certainly the case at the Human Identification Laboratory in Tucson, Arizona. As the caseload increased, it became clear that a method for defleshing human remains was needed in order to expeditiously expose the osseous surfaces for analysis, yet at the same time, preserving the evidentiary nature of the material. As a result, a fast, safe and economical method for defleshing human remains and producing high quality, degreased skeletal elements was developed. This non-bleaching cooking method utilizes chemicals that are easily obtained and inexpensive standard household ingredients that can be purchased at most grocery stores.


Dose Escalation of Desmoteplase for Acute Ischemic Stroke (DEDAS): evidence of safety and efficacy 3 to 9 hours after stroke onset.
Desmoteplase is a novel plasminogen activator with favorable features in vitro compared with available agents. This study evaluated safety and efficacy of intravenous (IV) desmoteplase in patients with perfusion/diffusion mismatch on MRI 3 to 9 hours after onset of acute ischemic stroke. DEDAS was a placebo-controlled, double-blind, randomized, dose-escalation study investigating doses of 90 microg/kg and 125 microg/kg desmoteplase. Eligibility criteria included baseline National Institute of Health Stroke Scale (NIHSS) scores of 4 to 20 and MRI evidence of perfusion/diffusion mismatch. The safety end point was the rate of symptomatic intracranial hemorrhage. Primary efficacy co-end points were MRI reperfusion 4 to 8 hours after treatment and good clinical outcome at 90 days. The primary analyses were intent-to-treat. Before unblinding, a target population, excluding patients violating specific MRI criteria, was defined. Thirty-seven patients were randomized and received treatment (intent-to-treat; placebo: n=8; 90 microg/kg: n=14; 125 microg/kg: n=15). No symptomatic intracranial hemorrhage occurred. Reperfusion was achieved in 37.5% (95% CI [8.5; 75.5]) of placebo patients, 18.2% (2.3; 51.8) of patients treated with 90 microg/kg desmoteplase, and 53.3% (26.6; 78.7) of patients treated with 125 microg/kg desmoteplase. Good clinical outcome at 90 days occurred in 25.0% (3.2; 65.1) treated with placebo, 28.6% (8.4; 58.1) treated with 90 microg/kg desmoteplase and 60.0% (32.3; 83.7) treated with 125 microg/kg desmoteplase. In the target population (n=25), the difference compared with placebo increased and was statistically significant for good clinical outcome with 125 microg/kg desmoteplase (P=0.022). Treatment with IV desmoteplase 3 to 9 hours after ischemic stroke onset appears safe. At a dose of 125 microg/kg desmoteplase appeared to improve clinical outcome, especially in patients fulfilling all MRI criteria. The results of DEDAS generally support the results of its predecessor study, Desmoteplase in Acute Ischemic Stroke (DIAS).


Robotic-Assisted vs Nonrobotic-Assisted Minimally Invasive Transforaminal Lumbar Interbody Fusion: A Cost-Utility Analysis.
Management of degenerative disease of the spine has evolved to favor minimally invasive techniques, including nonrobotic-assisted and robotic-assisted minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF). Value-based spending is being increasingly implemented to control rising costs in the US healthcare system. With an aging population, it is fundamental to understand which procedure(s) may be most cost-effective. To compare robotic and nonrobotic MIS-TLIF through a cost-utility analysis. We considered direct medical costs related to surgical intervention and to the hospital stay, as well as 1-yr utilities. We estimated costs by assessing all cases involving adults undergoing robotic surgery at a single institution and an equal number of patients undergoing nonrobotic surgery, matched by demographic and clinical characteristics. We adopted a willingness to pay of $50 000/quality-adjusted life year (QALY). Uncertainty was addressed by deterministic and probabilistic sensitivity analyses. Costs were estimated based on a total of 76 patients, including 38 undergoing robot-assisted and 38 matched patients undergoing nonrobot MIS-TLIF. Using point estimates, robotic surgery was projected to cost $21 546.80 and to be associated with 0.68 QALY, and nonrobotic surgery was projected to cost $22 398.98 and to be associated with 0.67 QALY. Robotic surgery was found to be more cost-effective strategy, with cost-effectiveness being sensitive operating room/materials and room costs. Probabilistic sensitivity analysis identified robotic surgery as cost-effective in 63% of simulations. Our results suggest that at a willingness to pay of $50 000/QALY, robotic-assisted MIS-TLIF was cost-effective in 63% of simulations. Cost-effectiveness depends on operating room and room (admission) costs, with potentially different results under distinct neurosurgical practices.


Placental passage of benzoic acid, caffeine, and glyphosate in an ex vivo human perfusion system.
Ex vivo perfusion of the human term placenta is a method to study placental transfer without extrapolation from animal to human and with no ethical concerns for mother and child. However, ex vivo placenta perfusion has a limited potential within chemical screening and testing as the method is time-consuming. This study was an attempt to construct data needed to develop quantitative structure-activity relationship (QSAR) models that are able to predict placental transfer of new compounds. Placental transfer is a biological activity that statistically may be related to the physiochemical properties of a given group of compounds. Benzoic acid, caffeine, and glyphosate were chosen as model compounds because they are small molecules with large differences in physiochemical properties. Caffeine crossed the placenta by passive diffusion. The initial transfer rate of benzoic acid was more limited in the first part of the perfusion compared to caffeine, but reached the same steady-state level by the end of perfusion. The transfer of glyphosate was restricted throughout perfusion, with a lower permeation rate, and only around 15% glyphosate in maternal circulation crossed to the fetal circulation during the study period.


Juvenile psoriatic arthritis: longterm outcome and differentiation from other subtypes of juvenile idiopathic arthritis.
To compare outcomes in patients with juvenile psoriatic arthritis (PsA) with those in patients with other juvenile idiopathic arthritis (JIA) subtypes, and to evaluate characteristics and genetic markers that may differentiate PsA from other subtypes of JIA. JIA patients first admitted between 1980 and 1985 were clinically examined after a median of 15 years. Health status was reassessed by the Short Form-36 Health Survey (SF-36) after a median of 23 years. Of 336 JIA patients, 31 (9%) had PsA. Predictors of PsA were psoriasis in the patient (OR 12.06, p = 0.004) or first-degree relative (OR 30.86, p < 0.001), dactylitis (OR 10.97, p < 0.001), and ankle/toe arthritis (OR 3.04, p = 0.038) within the first 6 months. HLA-DRB1*11/12 status (OR 2.69, p = 0.040) and onset after age 6 years (OR 4.41, p = 0.004) differentiated PsA from either oligoarthritis or polyarthritis. After 15 years, PsA patients had poorer physical health than healthy population controls (p = 0.001). After 23 years, the SF-36 physical scores were poorer in PsA patients than in those with either oligoarthritis or polyarthritis (p < 0.045). The need for disease-modifying antirheumatic drugs and/or anti-tumor necrosis factor agents was present in 33% of PsA versus 8% in oligoarthritis and 13% in either oligoarthritis or polyarthritis patients (p < 0.001 and p = 0.002, respectively). In addition to a history of psoriasis, dactylitis, ankle or toe arthritis, and DRB1*11/12 in children with JIA indicate the likelihood of PsA, a subtype associated with unfavorable outcome.


A retrospective and correlative analysis of academic and nonacademic predictors of COMLEX level 1 performance.
National licensing exams (NLEs) including the Comprehensive Osteopathic Medical Licensing Examination (COMLEX) Level 1 evaluate student achievement. Scores have historically been utilized to stratify medical student applicants for residency. Grade point average (GPA), number of practice questions completed, and performance on practice exams have been shown to be predictive of NLE performance. Test anxiety and acute stress have been shown to negatively impact NLE performance. The role of study behaviors and other nonacademic factors in COMLEX Level 1 performance is unknown. This study aims to evaluate academic and nonacademic factors and to correlate them with COMLEX Level 1 performance. Additional analysis is conducted to associate COMLEX Level 1 performance with academic and nonacademic factors when controlling for GPA. An anonymous online survey was administered to third- (OMS III) and fourth-year (OMS IV) osteopathic medical students at Kansas City University that had completed the COMLEX Level 1 examination. In total, 72 students responded to the survey. Survey results were linked to student records of GPA and COMLEX Level 1 scores, resulting in 59 complete responses for analysis. Independent-sample t-tests and linear ordinary least squares regression were utilized to analyze the results. The majority of participants are male (62.7%) and OMS III (98.3%) with an average age of 27.14 ± 2.58 (mean ± standard deviation). Further demographic data reveal hours per week spent for personal time during dedicated study (n=46, 19.7 ± 18.53), hours of sleep per night during dedicated study (7.34 ± 0.92), and money spent on board preparation ($1,319.12 ± $689.17). High ($1,600-$3,000), average ($1,000-$1,500), and low ($100-$900) spenders do not statistically differ and COMLEX Level 1 performance is not related to the number of resources utilized (F statistics <1; p>0.05). Pearson correlations reveal a statistically significant relationship between COMLEX Level 1 scores with GPA (0.73, p<0.001), number of practice exams completed (0.39, p<0.001), number of questions completed (0.46, p<0.001), number of weeks of study (0.55, p<0.001), and preparation cost (0.28, p<0.05). The regression analysis revealed that money spent on board preparation, number of questions completed, and time spent studying accounted for 75.8% of the variance in COMLEX Level 1 scores after controlling for GPA. The data show the association of money spent on board preparation, numbers of questions competed, and time spent studying with a student's COMLEX Level 1 score. Additionally, these results highlight the amount of money students spend on extracurricular materials to prepare for COMLEX Level 1, yet the data show that the number of resources that students utilized is not related to a student's COMLEX Level 1 performance.


A systematic review of factors affecting vaccine uptake in young children.
Many parents make a conscious decision not to vaccinate their child. Multiple beliefs and perceptions surround this choice. If uptake of routine child vaccination is to increase, public health communications about vaccines must be informed by evidence on the factors affecting uptake. We conducted a systematic review to investigate psychological, social and contextual factors associated with uptake of routine vaccines in young children. Studies were included if they reported analyses of the association between psychological factors and uptake or included parents' self-reported reasons for or against vaccination. Our search identified 9110 citations after deduplication. Sixty-eight citations describing sixty-four studies were included in the review. The quality of the studies was mixed. There is strong evidence for an association between vaccination uptake and: not perceiving vaccines to cause adverse effects; general positive attitudes towards vaccination; positive vaccine recommendations; and perceiving fewer practical difficulties of vaccination. While there was good evidence for an association between vaccination and perceived susceptibility to the illness, evidence for an association between perceived severity of an illness and vaccination was weak. Other factors associated with vaccination include knowledge about the vaccine, social influences and trust in the healthcare profession. Having increased information about the vaccine was associated with vaccination, but the influence of different sources of information needs more research. Understanding which factors are consistently associated with the decision to vaccinate one's child is important to identify messages which should be targeted by public health communications about routine child vaccinations.


Multienzyme isothermal rapid amplification and lateral flow dipstick combination assay for visible detection of chicken chaphamaparvovirus.
Chicken chaphamaparvovirus (CkChpV) is a newly emerging pathogen that is currently prevalent in chickens with diarrhea symptoms. To diagnose CkChpV more conveniently and rapidly, this study established a multienzyme isothermal rapid amplification (MIRA) assay, with a reaction time of only 15 min and optimal reaction temperature of 38°C. In combination with the lateral flow dipstick assay, the CkChpV-MIRA assay can be completed within 20 min. We revealed that the detection limit of the MIRA assay using standard plasmids as templates was as low as 21.3 copies, and its sensitivity was 100 times higher than that of nested PCR. Moreover, the designed primer set and probe could only detect CkChpV specifically, and there was no cross reaction with avian nephritis virus, rotavirus, chicken parvovirus virus, Newcastle disease virus, and infectious bronchitis virus, which may cause diarrhea. These findings demonstrated that the CkChpV-MIRA assay established in this study is convenient, sensitive, and specific and does not require sophisticated equipment. It is more suitable for the detection of CkChpV in clinical samples.


Dual functioning ovaries and atresia in chickens. Is it a coincidence?
During the course of a QTL study involving an intercross between White Plymouth Rock chickens and Red Jungle Fowl, certain reproductive anomalies such as atresia and double ovary-oviduct became evident. Observed in reciprocal pedigreed F1 crosses were 2 full-sib pullets with dual functioning ovaries. One also had complete functioning left and right oviducts. The other had asymmetrical reproductive tracts with a typical left oviduct and a rudimentary right oviduct. In addition, there were 3 females with atresia. Although they had different sires and dams, their grandparents were related, and all 5 traced to a common ancestor. Pedigrees, photos, and biological aspects of these conditions are presented and discussed.


A nomogram predicting the need for abdominal and pelvic computed tomography in blunt trauma patients: A retrospective cohort study.
Abdominal and pelvic computed tomography (APCT) has become the preferred means for the initial evaluation of blunt trauma patients. However, computed tomography examination has some disadvantages, such as radiation exposure, the requirement for intravenous iodinated contrast medium, high cost, and time. We aimed to develop a nomogram to predict the need for APCT scanning after the primary survey of blunt trauma patients. We conducted a retrospective observational cohort study at a single-center and reviewed medical records of 972 trauma patients admitted between January 2013 and June 2016. We enrolled 786 blunt trauma patients who had undergone APCT and were 16 years of age or older. A multivariate logistic regression model was used to determine independent predictors for trauma-related findings on APCT scans. A nomogram was constructed to predict injury on APCT scans based on each predictive factor. Of 786 patients, 355 (45%) patients had at least 1 injury on APCT scans. Results of multivariate logistic regression analysis showed that independent predictive factors of injuries on APCT scans were as follows: falls (≥3 m high); pain (abdominal, back, flank, or pelvic); positive peritoneal signs; abnormal findings on chest radiographs; abnormal findings on pelvic radiographs; and positive findings on focused assessment with ultrasonography for trauma. The nomogram was developed using these parameters. The area under a receiver operating characteristic curve of the multivariate model for discrimination was 0.865 (95% confidence interval, 0.840-0.892). The calibration plot showed good agreement between predicted and observed outcomes. The maximal Youden index was 0.59, corresponding to a cutoff value > 59 points, which was considered the optimal cutoff value for the probability that the injury would be detected on APCT scans. The nomogram, based on initial clinical findings in blunt trauma patients, will help clinicians be more selective in their use of APCT evaluations.


A Link is not Enough - Reproducibility of Data.
Although many works in the database community use open data in their experimental evaluation, repeating the empirical results of previous works remains a challenge. This holds true even if the source code or binaries of the tested algorithms are available. In this paper, we argue that providing access to the raw, original datasets is not enough. Real-world datasets are rarely processed without modification. Instead, the data is adapted to the needs of the experimental evaluation in the data preparation process. We showcase that the details of the data preparation process matter and subtle differences during data conversion can have a large impact on the outcome of runtime results. We introduce a data reproducibility model, identify three levels of data reproducibility, report about our own experience, and exemplify our best practices.


Antigenicity of cryopreserved corneas.
The antigenicity of fresh and cryopreserved corneal tissue was analyzed by performing 5-mm interlamellar xenografts in albino rabbits using dog corneas as the donor tissue. The cryopreservative solutions contained 10% rabbit albumin, dimethyl sulfoxide, and sucrose. Fifty-nine rabbits received interlamellar xenografts using donor tissue prepared by one of the three following methods: (1) fresh, (2) cryopreserved, and (3) immersed in the cryopreservative solutions, but not frozen. Approximately one third of the grafts became cloudy within six weeks, and no statistically significant difference was noted clinically or histologically between the three preparations of donor tissue used. The results indicate that cryopreservation has no significant effect on the antigenicity of corneal tissue as exemplified by the interlamellar xenograft reaction in rabbits.


Vicarious group-based rejection: creating a potentially dangerous mix of humiliation, powerlessness, and anger.
Rejection can convey that one is seen as inferior and not worth bothering with. Is it possible for people to feel vicariously rejected in this sense and have reactions that are similar to those following personal rejection, such as feeling humiliated, powerless, and angry? A study on personal rejection was followed by two main studies on vicarious group-based rejection. It was found that merely observing rejection of ingroup members can trigger feelings of humiliation that are equally intense as those experienced in response to personal rejection. Moreover, given that the rejection is explicit, vicariously experienced feelings of humiliation can be accompanied by powerlessness and anger. Potentially, this combination of emotions could be an important source of offensive action against rejecters.


Micromachining of bulk glass with bursts of femtosecond laser pulses at variable repetition rates.
Oscillator-only femtosecond laser micromachining enables the manufacturing of integrated optical components with circular transverse profiles in transparent materials. The circular profile is due to diffusion of heat accumulating at the focus. We control the heat diffusion by focusing bursts of femtosecond laser pulses at various repetition rates into sodalime glass. We investigate the effect the repetition rate and number of pulses on the size of the resulting structures. We identify the combinations of burst repetition rate and number of pulses within a burst for which accumulation of heat occurs. The threshold for heat accumulation depends on the number of pulses within a burst. The burst repetition rate and the number of pulses within a burst provide convenient control of the morphology of structures generated with high repetition rate femtosecond micromachining.


Standards for scalable clinical decision support: need, current and emerging standards, gaps, and proposal for progress.
Despite their potential to significantly improve health care, advanced clinical decision support (CDS) capabilities are not widely available in the clinical setting. An important reason for this limited availability of CDS capabilities is the application-specific and institution-specific nature of most current CDS implementations. Thus, a critical need for enabling CDS capabilities on a much larger scale is the development and adoption of standards that enable current and emerging CDS resources to be more effectively leveraged across multiple applications and care settings. Standards required for such effective scaling of CDS include (i) standard terminologies and information models to represent and communicate about health care data; (ii) standard approaches to representing clinical knowledge in both human-readable and machine-executable formats; and (iii) standard approaches for leveraging these knowledge resources to provide CDS capabilities across various applications and care settings. A number of standards do exist or are under development to meet these needs. However, many gaps and challenges remain, including the excessive complexity of many standards; the limited availability of easily accessible knowledge resources implemented using standard approaches; and the lack of tooling and other practical resources to enable the efficient adoption of existing standards. Thus, the future development and widespread adoption of current CDS standards will depend critically on the availability of tooling, knowledge bases, and other resources that make the adoption of CDS standards not only the right approach to take, but the cost-effective path to follow given the alternative of using a traditional, ad hoc approach to implementing CDS.


The More the Merrier? Should Antibiotics be Used for Rhinoplasty and Septorhinoplasty?-A Review.
With antimicrobial resistance a global threat, optimizing antibiotic usage across the surgical continuum is vital. The American Academy of Otolaryngology-Head and Neck Surgery Foundation recently published the first guidelines addressing management in rhinoplasty. The authors reviewed pertinent literature on the role of systemic antibiotics in rhinoplasty and septorhinoplasty. The authors performed a MEDLINE search through PubMed using the key terms rhinoplasty, septorhinoplasty, infection, antimicrobials, and antibiotics. Ten studies met criteria. Studies evaluating antibiotics perioperatively showed similar infection and/or bacteremia rates (0-13.3%) in those receiving or not receiving antibiotics. No patients experienced significant local/systemic infections regardless of antibiotic use. In the 3 studies evaluating antibiotics postoperatively, antibiotics decreased the infection rate from 27% to 8% in complex revision cases. In a study evaluating postoperative antibiotics in noncomplex cases, there were no significant differences in infection rates between those receiving only a preoperative dose and those receiving preoperative in addition to 7 days of postoperative antibiotics, with the latter experiencing higher rates of antibiotic-related adverse events and costs. Peri- and postoperative antibiotics in noncomplex rhinoplasty and septorhinoplasty are not beneficial in decreasing infection risk. Antibiotics, with a first-generation cephalosporin such as cefazolin (non-β-lactam, such as clindamycin, if β-lactam allergy), should be considered in patients with comorbidities/undergoing complex surgery. If perioperative antibiotics are used, antibiotics should be administered within 1 hour of incision and discontinued within 24 hours of the operation. Further research is warranted to evaluate the optimal duration of postoperative antibiotics in complex cases.


Identification of Db- and Kb-restricted subdominant cytotoxic T-cell responses in lymphocytic choriomeningitis virus-infected mice.
Antiviral cytotoxic T-cells are critical for control of lymphocytic choriomeningitis virus (LCMV) infection in mice. In H-2b mice, the antiviral response is directed against three Db-restricted epitopes in the viral nucleoprotein (NP396-404) and glycoprotein (GP276-286 and GP33-41). Our present data revealed a clear hierarchy among these three epitopes, in which NP396-404 is immunodominant, followed by GP33-41 and GP276-286, respectively. In order to identify additional CTL epitopes in the LCMV nucleoprotein and glycoprotein, we used the motifs for Db2- and Kb-binding peptides, combined with MHC class I-binding assays. Out of 23 Db motif-fitting peptides, we identified 4 Db binders, one of which (GP92-101) turned out to be a new CTL epitope. Among 28 Kb motif-fitting peptides, 12 bound Kb, and one of these (NP205-212) was a CTL epitope. Both newly identified CTL peptides were recognized by LCMV-immune splenocytes after secondary in vitro stimulation. Both peptides bound their MHC class I molecules with intermediate affinity (470 and 170 nM for GP92-101 and NP205-212, respectively). Responses against these peptides were weaker than the responses against the three major epitopes. None of the high affinity binders were new epitopes, suggesting that high affinity binders are either immunodominant epitopes or no epitopes at all. Thus, analysis of 51 Kb and Db motif-fitting peptides yielded 2 new, subdominant epitopes. Immunization of C57BL/6 mice with these peptides, or vaccinia virus recombinants expressing these epitopes as minigenes, protected against chronic LCMV infection, demonstrating that immunization with subdominant epitopes can confer protection against chronic viral infection.


Computation of light scattering by axisymmetric nonspherical particles and comparison with experimental results.
A laboratory prototype of a novel experimental apparatus for the analysis of spherical and axisymmetric nonspherical particles in liquid suspensions has been developed. This apparatus determines shape, volume, and refractive index, and this is the main difference of this apparatus from commercially available particle analyzers. Characterization is based on the scattering of a monochromatic laser beam by particles [which can be inorganic, organic, or biological (such as red blood cells and bacteria)] and on the strong relation between the light-scattering pattern and the morphology and the volume, shape, and refractive index of the particles. To keep things relatively simple, first we focus attention on axisymmetrical particles, in which case hydrodynamic alignment can be used to simplify signal gathering and processing. Fast and reliable characterization is achieved by comparison of certain properly selected characteristics of the scattered-light pattern with the corresponding theoretical values, which are readily derived from theoretical data and are stored in a look-up table. The data in this table were generated with a powerful boundary-element method, which can solve the direct scattering problem for virtually arbitrary shapes. A specially developed fast pattern-recognition technique makes possible the on-line characterization of axisymmetric particles. Successful results with red blood cells and bacteria are presented.


Rituximab to treat prolidase deficiency due to a novel pathogenic copy number variation in PEPD.
Prolidase deficiency (PD) is a rare autosomal recessive inborn error of immunity caused by biallelic homozygous or compound heterozygous loss-of-function mutations in PEPD, the gene that encodes prolidase. PD typically manifests with variable dysmorphic features, chronic cutaneous ulcers, recurrent infections and autoimmune features, including systemic lupus erythematosus. So far, there is no consensus regarding treatment of PD and its autoimmune manifestations. Here, we present a 28-year-old female patient with PD due to a novel homozygous intragenic deletion in PEPD, diagnosed at the age of 6 years and 7 months with an undifferentiated connective tissue disease that, apart from its very early onset, would be consistent with the diagnosis of Sjögren's syndrome. Steroids and diverse conventional synthetic disease-modifying antirheumatic drugs failed to control PD-associated vasculitis and mucocutaneous ulcerations and led to infectious complications, including cytomegalovirus colitis. Introduction of rituximab (RTX) treatment in this patient led to sustained recession of mucocutaneous ulceration, enabling tapering of steroids. High interleukin-1β (IL-1β) production by this patient's monocytes, together with the detection of both IL-1β and interleukin-18 (IL-18) in her serum, suggest enhanced inflammasome activation in PD, whereas the therapeutic efficacy of RTX implies a role for CD20 positive B cells in the complex immunopathogenesis of PD.


Automated quantitative determination of the new renin inhibitor CGP 60536 by high-performance liquid chromatography.
A fully automated high-performance liquid chromatography method with fluorescence detection for the determination of the renin inhibitor CGP 60536 in animal and human plasma and urine has been developed and validated. After addition of an internal standard, the compounds were automatically extracted from 400 microl of plasma or urine with methyl alcohol-acetic acid (99:1, v/v) on 100-mg Bond-Elut CN cartridges using the Gilson ASPEC system. The on-line chromatographic separation was performed on a LiChrospher 100 RP8 5-microm particle size packed analytical column (25x0.4 cm I.D.). The mobile phase consisted of acetonitrile-0.01 M potassium dihydrogenphosphate (65:35, v/v) at a flow-rate of 0.8 ml/min. The analytes were detected using a fluorescence detector at excitation and emission wavelengths of 280 and 330 nm, respectively. The limit of quantitation was established at 4.5 ng/ml in plasma (accuracy 106% and precision 1%), and 9.0 ng/ml in urine (accuracy 101% and precision 13%). The method was applied to the investigation of the pharmacokinetics of CGP 60536.


Stabilizing the production of nonnative consonant clusters with acoustic variability.
Previous research on the perception, recognition, and learning of sounds and words has identified diverse effects of phonetic variation. The present study examined how variation affects cross-language production of consonant clusters. American English speakers shadowed words beginning with nonnative clusters in low- and high-variability conditions. Shadowing responses in the low-variability condition were quite sensitive to fine-grained phonetic properties that were manipulated across the stimuli. Notably, longer stop bursts led to increased rates of epenthesis, lower burst amplitudes resulted in more feature change and deletion, and intense periods of voicing at cluster onset elicited prothetic responses. Sensitivity to the acoustic manipulations was substantially attenuated in the high-variability condition, which combined stimuli from the first condition with baseline productions of the same items from two additional talkers. Detailed analyses of the response patterns indicate that more stable production targets in the high-variability condition resulted from integration, or blending, of the multiple talker stimuli. Implications of these findings for language-specific speech processing and the role of phonetic variability in second language acquisition are discussed.


Reconstruction after soft tissue sarcoma resection in the setting of brachytherapy: a 10-year experience.
Management of recurrent soft tissue sarcomas often involves surgical resection and adjuvant brachytherapy. This study reviews our experience in the management of these patients and proposes a logical approach toward reconstruction. All patients who underwent soft tissue sarcoma resection, adjuvant brachytherapy, and soft tissue flap reconstruction (pedicled or free) during the 10-year period from 1991 to 2000 were included in this study. There were 17 patients (14 male, 3 female) with a mean age of 51 years (range, 16-80 years). Soft tissue sarcomas were distributed in the lower extremity (n = 9), upper extremity (n = 5), and trunk (n = 3). Reconstruction was accomplished by regional transposition flaps (n = 10) and free tissue transfer (n = 7). The average defect size was 143 cm. Patients received 5 to 12 (mean, 8) brachytherapy catheters. The brachytherapy dose delivered ranged from 1600 to 4500 cGy (mean, 3773 cGy). Brachytherapy catheters were loaded with radioactive sources between 5 and 7 days postoperatively. All flaps in this series survived. One patient required return to the operating room for revision of a venous thrombosis with flap salvage. Closed suction drainage tubes were left in place until after the brachytherapy catheters were removed to avoid dislodging the catheters. Two patients developed postradiation partial-thickness skin necrosis with delayed secondary wound healing. This study demonstrates that soft tissue reconstruction in the setting of sarcoma resection and brachytherapy catheter placement is safe and efficacious. Postoperative wound healing complications can be minimized through coordination among the ablative surgeon, reconstructive surgeon, and radiation oncologist. Specifically, placement of microvascular anastomoses well away from the radiation target area is indicated whenever possible. Finally, removal of closed suction drainage tubes should be deferred until after the brachytherapy catheters are removed to minimize complications resulting from catheter dislodgement.


Seed removal of Dipteryx alata Vog. (Leguminosae: Faboidae) in the edge and interior of Cerrado.
Post-dispersal seed agents of mortality include pathogens, invertebrates, and vertebrates, which may shape tropical tree populations and communities. In this study I experimentally evaluated Dipteryx alata endocarp removal both in the interior and edge of Cerrado vegetation. Specifically, I simulated primary dispersion of endocarps by bats and evaluated secondary removal intensity according to habitat, season, and endocarp number. This study was developed in the Pombo Natural Municipal Park, a large Cerrado remnant with an area of 9,000 ha, located in the Western Brazil. In each of the two habitat types, I set down 45 points in which D. alata piles of 1, 3, 7, 15, and 40 endocarps were positioned. On average, endocarp removal in the interior was higher than in the edge, so that the intensity of removal varied from positive density-dependent (interior: rains), to negative density-dependent (edge: both seasons). Also, at both habitat types and seasons non removed endocarps were intensely attacked by fungus. Indeed, seed mortality by fungus infestation was positive density-dependent. The results indicate high propensity of endocarps to escape from removal in the edge, in principle, a defaunated area. As Dipteryx species strongly rely on caviomorph rodents for their dispersal, the scarcity of these vectors might reduce recruitment chances of D. alata in remnants of Cerrado. This imply in future changes in the vegetation structure from the edge to the interior towards low tree diversity.


Periprosthetic proximal medial femoral cortical destruction caused by a femoral arterial pseudoaneurysm.
A femoral artery aneurysm is a very rare complication of hip surgery treatment. In this presentation, we report a rare case of bone destruction caused by the femoral artery aneurysm after a hip hemiarthroplasty operation as a late stage complication. A 77-year-old male patient presented with swelling and pain in the hip without any signs of ischemia. X-ray showed a 7 cm bone destruction in the medial cortex of the proximal femur. After ultrasound imaging, a femoral artery aneurysm was diagnosed. The patient was operated on by vascular surgeons, and a large amount of organized hematoma with artery aneurysm was removed. The diagnosis of femoral aneurysm after hip surgery treatment can be difficult. Femoral aneurysm is rare and usually secondary to trauma, infection, or iatrogenic injury. We assumed that this case occurred during the hemiarthroplasty operation, caused by a compressing effect to the bone causing destruction. The patient was followed for 2 years, during which time the preoperative symptoms reduced dramatically and Harris Hip Score improved. After a hip arthroplasty operation, it must be kept in mind that a femoral arterial aneurysm can cause persistent symptoms. This case illustrates the importance of considering a pseudoaneurysm as a possible cause of periprosthetic osteolysis, particularly if rapid progressive growth has occurred. If this vascular injury had not been correctly diagnosed, the method of treatment could have been much different, probably with catastrophic consequences.


Structural basis for species specific inhibition of 17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1): computational study and biological validation.
17β-Hydroxysteroid dehydrogenase type 1 (17β-HSD1) catalyzes the reduction of estrone to estradiol, which is the most potent estrogen in humans. Inhibition of 17β-HSD1 and thereby reducing the intracellular estradiol concentration is thus a promising approach for the treatment of estrogen dependent diseases. In the past, several steroidal and non-steroidal inhibitors of 17β-HSD1 have been described but so far there is no cocrystal structure of the latter in complex with 17β-HSD1. However, a distinct knowledge of active site topologies and protein-ligand interactions is a prerequisite for structure-based drug design and optimization. An elegant strategy to enhance this knowledge is to compare inhibition values obtained for one compound toward ortholog proteins from various species, which are highly conserved in sequence and differ only in few residues. In this study the inhibitory potencies of selected members of different non-steroidal inhibitor classes toward marmoset 17β-HSD1 were determined and the data were compared with the values obtained for the human enzyme. A species specific inhibition profile was observed in the class of the (hydroxyphenyl)naphthols. Using a combination of computational methods, including homology modelling, molecular docking, MD simulation, and binding energy calculation, a reasonable model of the three-dimensional structure of marmoset 17β-HSD1 was developed and inhibition data were rationalized on the structural basis. In marmoset 17β-HSD1, residues 190 to 196 form a small α-helix, which induces conformational changes compared to the human enzyme. The docking poses suggest these conformational changes as determinants for species specificity and energy decomposition analysis highlighted the outstanding role of Asn152 as interaction partner for inhibitor binding. In summary, this strategy of comparing the biological activities of inhibitors toward highly conserved ortholog proteins might be an alternative to laborious x-ray or site-directed mutagenesis experiments in certain cases. Additionally, it facilitates inhibitor design and optimization by offering new information on protein-ligand interactions.


Bacterial single-stranded DNA-binding proteins are phosphorylated on tyrosine.
Single-stranded DNA-binding proteins (SSBs) are required for repair, recombination and replication in all organisms. Eukaryotic SSBs are regulated by phosphorylation on serine and threonine residues. To our knowledge, phosphorylation of SSBs in bacteria has not been reported. A systematic search for phosphotyrosine-containing proteins in Streptomyces griseus by immunoaffinity chromatography identified bacterial SSBs as a novel target of bacterial tyrosine kinases. Since genes encoding protein-tyrosine kinases (PTKs) have not been recognized in streptomycetes, and SSBs from Streptomyces coelicolor (ScSSB) and Bacillus subtilis (BsSSB) share 38.7% identity, we used a B.subtilis protein-tyrosine kinase YwqD to phosphorylate two cognate SSBs (BsSSB and YwpH) in vitro. We demonstrate that in vivo phosphorylation of B.subtilis SSB occurs on tyrosine residue 82, and this reaction is affected antagonistically by kinase YwqD and phosphatase YwqE. Phosphorylation of B.subtilis SSB increased binding almost 200-fold to single-stranded DNA in vitro. Tyrosine phosphorylation of B.subtilis, S.coelicolor and Escherichia coli SSBs occured while they were expressed in E.coli, indicating that tyrosine phosphorylation of SSBs is a conserved process of post-translational modification in taxonomically distant bacteria.


A RECENT STUDY OF CHOLERA WITH REFERENCE TO AN OUTBREAK IN TAIWAN IN 1962.
During the summer of 1962, cholera broke out in Taiwan after an absence of 16 years. This paper gives a full description of the prompt and energetic measures that were instituted to control the outbreak; these included the designation of emergency areas, the immediate detection and isolation of patients and probable carriers, and mass vaccination.In all, over 160 000 rectal swabs and stool specimens were examined for cholera organisms. The author's egg-starch medium, described in detail, proved very satisfactory for this purpose. During the main epidemic (July-September) 383 confirmed cases of cholera and 380 carriers were identified. While the over-all carrier: case ratio was thus approximately 1:1 the ratios in different age-groups varied considerably, being as high as 4:1 in children aged 0-4 years and as low as 0.3:1 in persons aged 55-59 years. The author suggests possible reasons for the older persons' tendency to develop clinical disease rather than become carriers.There were only 24 deaths in the outbreak (a fatality rate of 6%). The efficacy of vaccination was difficult to determine precisely, but in general more deaths and more cases occurred among the non-vaccinated than among the vaccinated.


Pigmentary characteristics, UV radiation exposure, and risk of non-Hodgkin lymphoma: a prospective study among Scandinavian women.
UV radiation and pigmentary characteristics may be associated with non-Hodgkin lymphoma (NHL) risk, but few prospective studies exist. We investigated these associations in a Norwegian-Swedish cohort. The cohort included women ages 30 to 50 years at enrolment in 1991 to 1992. Host factors, and exposure to sun and artificial tanning devices in life-decades 0 to 50 years were collected by questionnaire. Relative risks (RR) with 95% confidence intervals (CI) were estimated by Poisson regression. Among 104,953 women with complete follow-up through 2006 (Sweden) and 2007 (Norway), 158 were diagnosed with NHL. Women with brown hair had an increased risk of NHL compared with dark brown-haired/black-haired women (RR, 1.72; 95% CI, 1.08-2.74); decreased risks were found among women with gray, green or mixed (RR, 0.50; 95% CI, 0.32-0.77), or blue (RR, 0.54; 95% CI, 0.35-0.81) eyes compared with those with brown eyes, and among those with high propensity to burn compared with those with low propensity (RR, 0.57; 95% CI, 0.36-0.91). Annual number of sunburns and bathing vacations in any age decade, or ever use of artificial tanning devices were not significantly associated with NHL risk. After exposure at ages 10 to 39 years, RRs for ever versus never exposed were 0.99 (95% CI, 0.65-1.50) for sunburn, 1.00 (95% CI, 0.64-1.54) for bathing vacations, and 0.99 (95% CI, 0.67-1.46) for artificial tanning device use. Whereas several pigmentary characteristics were associated with NHL risk, our results do not support an association between UV radiation and NHL. Studies of UV radiation and NHL are warranted for etiologic understanding and public health recommendations.


Automatic detection and quantification of tree-in-bud (TIB) opacities from CT scans.
This study presents a novel computer-assisted detection (CAD) system for automatically detecting and precisely quantifying abnormal nodular branching opacities in chest computed tomography (CT), termed tree-in-bud (TIB) opacities by radiology literature. The developed CAD system in this study is based on 1) fast localization of candidate imaging patterns using local scale information of the images, and 2) Möbius invariant feature extraction method based on learned local shape and texture properties of TIB patterns. For fast localization of candidate imaging patterns, we use ball-scale filtering and, based on the observation of the pattern of interest, a suitable scale selection is used to retain only small size patterns. Once candidate abnormality patterns are identified, we extract proposed shape features from regions where at least one candidate pattern occupies. The comparative evaluation of the proposed method with commonly used CAD methods is presented with a dataset of 60 chest CTs (laboratory confirmed 39 viral bronchiolitis human parainfluenza CTs and 21 normal chest CTs). The quantitative results are presented as the area under the receiver operator characteristics curves and a computer score (volume affected by TIB) provided as an output of the CAD system. In addition, a visual grading scheme is applied to the patient data by three well-trained radiologists. Interobserver and observer-computer agreements are obtained by the relevant statistical methods over different lung zones. Experimental results demonstrate that the proposed CAD system can achieve high detection rates with an overall accuracy of 90.96%. Moreover, correlations of observer-observer (R(2)=0.8848, and observer-CAD agreements (R(2)=0.824, validate the feasibility of the use of the proposed CAD system in detecting and quantifying TIB patterns.


Hydrogen sulfide protects against myocardial ischemia and reperfusion injury by activating AMP-activated protein kinase to restore autophagic flux.
How hydrogen sulfide (H2S) protects against myocardial ischemia-reperfusion (I/R) injury is poorly understood. By using a slow-releasing H2S donor, we investigated if H2S protected against myocardial I/R injury by activating AMPK and restoring I/R-impaired autophagic flux. Male rats received anterior descending coronary artery occlusion followed by reperfusion. The H2S donor ADT and/or the AMPK inhibitor, compound C (CC), were administered after occlusion. Infarction was analyzed histologically. AMPK activation was assessed in the ischemic heart by analyzing phosphorylation of AMPK and S6 ribosomal protein. Autophagy was assessed by analyzing the following markers: microtubule-associated protein 1 light chain 3 (LC3) I and II, lysosome associated membrane protein-2 (LAMP-2), P62 and beclin-1. We further investigated if blocking autophagic flux with chloroquine abolished ADT cardioprotection in vivo. Myocardial I/R reduced serum H2S levels, which was elevated by ADT. ADT enhanced AMPK activation and reduced infarction following I/R, and both effects were abolished by AMPK inhibition. Myocardial I/R induced autophagosome accumulation, as evidenced by the increased ratios of LC3-II/LC3-I, upregulation of beclin-1 and P62 and reduction in LAMP-2. ADT blunted these autophagic changes induced by I/R, indicating that ADT restored I/R-impaired autophagic flux. The AMPK inhibitor CC blocked ADT effects on restoring I/R-impaired autophagy flux. Moreover, chloroquine pretreatment abolished cardioprotection of ADT and increased autophagosome accumulation in the ADT-treated heart following I/R. In conclusion, AMPK activation and subsequent restoration of I/R-impaired autophagic flux are unrecognized mechanisms underlying cardioprotective effects conferred by H2S donors.


The natural history of inflammatory bowel disease and primary sclerosing cholangitis after liver transplantation--a single-centre experience.
To describe the natural history of primary sclerosing cholangitis (PSC) and inflammatory bowel disease (IBD) after liver transplant, the predictors of PSC and IBD recurrence, and the interaction of these disease processes. Data regarding patients who received liver transplants for PSC at the University of Alberta Hospital (Edmonton, Alberta) from 1989 to 2006 were retrospectively reviewed. Recurrent PSC (rPSC) was defined by the Mayo Clinic criteria. Cox proportional hazards modelling and Kaplan-Meier statistics were used. Fifty-nine patients were studied, with a median follow-up of 68 months. A total of 71.2% of patients were diagnosed with IBD pretransplant. Clinical IBD severity post-transplant compared with severity pretransplant was unchanged in 67%, worse in 26.5% and improved in 6.1% of patients. Twenty-five per cent of patients developed rPSC posttransplant. The occurrence of at least one episode of acute cellular rejection (hazard ratio 5.7; 95% CI 1.3 to 25.8) and cytomegalovirus mismatch (hazard ratio 4.2; 95% CI 1.1 to 15.4) were found to be significant predictors of rPSC. Although not statistically significant, there was no rPSC in patients without pre- or post-transplant IBD, and in only one patient with a colectomy. Actuarial patient survival rates at one, five and 10 years posttransplant were 97%, 86% and 79%, respectively. Although a significant proportion of patients experienced worsening IBD post-transplantation, the presence or severity of IBD did not influence rPSC or patient survival. Acute cellular rejection and cytomegalovirus mismatch were both identified as independent predictors of rPSC. The impact of steroids and the ideal immunosuppressive regimen for the control of both IBD and PSC post-transplant requires further examination in prospective studies.


Inactivation of GSK-3beta in okadaic acid-induced neurodegeneration: relevance to Alzheimer's disease.
Hyperphosphorylation of tau is a characteristic feature of the neurodegenerative pathology in Alzheimer's disease (AD). Okadaic acid is used as a research model of AD to increase the tau phosphorylation and neuronal death. Using Western blotting, we found that the amounts of activated PKB[pS-473] and inactivated GSK-3beta[pS-9] were increased in proportion to the progress of okadaic acid induced tau phosphorylation. Immunocytochemistry showed that PKB[pS-473] and GSK-3beta[pS-9] immunoreactivity increased in dystrophic neurites and cell bodies in degenerating neurons after okadaic acid treatment. Double staining with phosphospecific tau antibodies showed that PKB[pS-473] and GSK-3beta[pS-9] were colocalized with phosphospecific tau in response to okadaic acid. Taken together, our data suggest that inhibition of protein phosphatase results in the hyperphosphorylation of tau without GSK-3beta overactivation.


Antibody Responses to Immunizations in Children with Type I Diabetes Mellitus: a Case-Control Study.
Type I diabetes mellitus (DM) has been associated with abnormalities of T cells. Our objective was to assess whether antibody responses to T-cell-dependent and -independent antigens in children with DM are lower than those of children without DM. We performed a case-control study matching children with DM to children without DM by age and by assessing antibody levels to pneumococcal serotypes, Haemophilus influenzae, and tetanus and diphtheria toxoids and reassessing antibody levels in patients with antibody levels below protective thresholds after booster immunization. We recruited 36 children with DM and 36 age-matched controls. The mean age was 10 years. There was no difference between groups in antibody levels against the antigens tested. Pneumococcal antibody levels below the protective threshold were found in 35.9% of DM patients after conjugate pneumococcal vaccination with no difference between groups. Booster immunization with unconjugated pneumococcal vaccine resulted in a median level against pneumococcal serotypes of 2.3 μg/ml (range, 0.05 to 664.7 μg/ml) in children with DM and 6.1 μg/ml (0.12 to 203.36 μg/ml) in children without DM (P = 0.013). Over 85% of children had levels above the protective threshold after booster immunization with no difference between groups. There was no evidence for a reduced antibody response to T-cell-dependent antigens given during childhood immunizations in children with DM. There was a reduced antibody response to antigens of pneumococcal strains in children with DM given unconjugated pneumococcal polysaccharide vaccine compared to that of children without DM without being associated with a difference in percentage of antibody levels below the protective threshold between groups.


Characterization of the Antibacterial Activity of Quinone-Based Compounds Originating from the Alnumycin Biosynthetic Gene Cluster of a Streptomyces Isolate.
Bacteria of the genus Streptomyces produce various specialized metabolites. Single biosynthetic gene clusters (BGCs) can give rise to different products that can vary in terms of their biological activities. For example, for alnumycin and the shunt product K115, antimicrobial activity was described, while no antimicrobial activity was detected for the shunt product 1,6-dihydro 8-propylanthraquinone. To investigate the antibacterial activity of 1,6-dihydro 8-propylanthraquinone, we produced alnumycin and 1,6-dihydro 8-propylanthraquinone from a Streptomyces isolate containing the alnumycin BGC. The strain was cultivated in liquid glycerol-nitrate-casein medium (GN), and both compounds were isolated using an activity and mass spectrometry-guided purification. The structures were validated via nuclear magnetic resonance (NMR) spectroscopy. A minimal inhibitory concentration (MIC) test revealed that 1,6-dihydro 8-propylanthraquinone exhibits antimicrobial activity against E. coli ΔtolC, B. subtilis, an S. aureus type strain, and a vancomycin intermediate-resistance S. aureus strain (VISA). Activity of 1,6-dihydro 8-propylanthraquinone against E. coli ΔtolC was approximately 10-fold higher than that of alnumycin. We were unable to confirm gyrase inhibition for either compound and believe that the modes of action of both compounds are worth reinvestigating.


HIV testing strategies across European countries.
With universal access to effective combination antiretroviral therapy (ART), people in need can gain effective treatment but are only able to benefit from these advances if they are aware of their serostatus and have effectively accessed testing services. Despite the expectation that ART would lead individuals to seek earlier testing, this trend has not been observed in practice, with stable or even increasing rates of late diagnosis in Europe being witnessed. Ahead of a gathering of key European stakeholders in Brussels in November 2007, we reviewed testing strategies across European countries. We show differences in policy and practices. Moreover, HIV testing strategies are changing, in line with new global guidelines issued by World Health Organization headquarters, and a number of countries are promoting an expansion of routine and opt-out testing. However, gaps in our understanding of effective testing strategies remain and, as a consequence, national policies across Europe remain incoherent and often lack an evidence base. This is likely to have serious public health implications.


Emergence of human avian influenza A(H7N9) virus infections in Wenshan City in Southwest China, 2017.
The emergence of human infection with avian influenza A(H7N9) virus was reported in Wenshan City, southwestern China in 2017. The study describes the epidemiological and virological features of the outbreak and discusses the origin of the infection. Poultry exposure and timelines of key events for each patient were collected. Samples derived from the patients, their close contacts, and environments were tested for influenza A(H7N9) virus by real-time reverse transcription polymerase chain reaction. Genetic sequencing and phylogenetic analysis were also conducted. Five patients were reported in the outbreak. An epidemiological investigation showed that all patients had been exposed at live poultry markets. The A(H7N9) isolates from these patients had low pathogenicity in avian species. Both epidemiological investigations of chicken sources and phylogenetic analysis of viral gene sequences indicated that the source of infection was from Guangxi Province, which lies 100 km to the east of Wenshan City. In the study, a sudden emergence of human cases of H7N9 was documented in urban area of Wenshan City. Chickens were an important carrier in the H7N9 virus spreading from Guangxi to Wenshan. Hygienic management of live poultry markets and virological screening of chickens transported across regions should be reinforced to limit the spread of H7N9 virus.


Physiological and proteomic evidences that domestication process differentially modulates the immune status of juvenile Eurasian perch (Perca fluviatilis) under chronic confinement stress.
The current study aimed to evaluate the influence of domestication process on the stress response and subsequent immune modulation in Eurasian perch juveniles (Perca fluviatilis) submitted to chronic confinement. Briefly, F1 and F4 generations were confined into small-size tanks and sampled 7 and 55 days after stocking. Cortisol and glucose levels as well as lysozyme activity and immunoglobulin level were evaluated in the serum. Spleen Somatic Index and spleen ROS production were also measured. A proteomic analysis was performed on serum sampled on day 7. Finally, both generations were genetically characterized using a microsatellite approach. Globally, results revealed that chronic confinement did not elicit a typical stress response but resulted in a prolonged immune stimulation. Proteomic results suggested that domestication process influenced the immune status of perch submitted to chronic confinement as the F1 confined fish displayed lower abundance of C3 complement component, transferrin and Apolipoprotein E. Microsatellite data showed a strong genetic drift as well as reduced genetic diversity, allelic number and heterozygosity along with domestication process. The present work is the first to report that fish under domestication can develop an immune response, assessed by a combined approach, following recurrent challenges imposed by captive environment despite a reduced genetic variation.


[Oxygenation of glioblastomas and normal brain tissue during radiation therapy].
Eight patients underwent simultaneous polarographic studies of pO2 in the brain normal tissue and glial tumors under the conditions of inhaling 100% oxygen and a hypoxic gaseous mixture (10% of oxygen and 90% of nitrogen). It has been established that the inhalation of 100% oxygen causes an increase in pO2 in the brain normal tissue and tumor by 100-115%. A short-term moderate gaseous hypoxia leads to a decrease in pO2 both in the brain normal tissue and tumor by 30% on an average.


Candidacidal activity of myeloperoxidase: mechanisms of inhibitory influence of soluble cell wall mannan.
We have previously demonstrated the ability of human neutrophil myeloperoxidase to bind to mannan isolated from Candida albicans. Mannan may therefore be a primary component of the yeast cell wall which provides for binding of myeloperoxidase, a requirement potentially important for the candidacidal activity of the enzyme. In this report, we describe experiments to consider the relationship of the mannan-binding activity of myeloperoxidase to its candidacidal activity and the possibility that free mannan may inhibit myeloperoxidase-mediated candidacidal activity. We observed that binding of myeloperoxidase to the target yeasts was required for killing of C. albicans. We also observed that addition of soluble mannan significantly reduced myeloperoxidase-mediated killing of the yeasts in a dose-dependent manner by antagonizing binding of myeloperoxidase. Soluble mannan was demonstrated to have a similar dose-dependent inhibitory effect on neutrophil-mediated candidacidal activity without influencing phagocytosis of the organism. On the basis of these observations, we speculate that mannan solubilized in plasma and tissue fluid may interfere with neutrophil-mediated host defense against Candida infection.


Preoperative Multifidus Muscle Quality is Associated With Patient Reported Outcomes After Lateral Lumbar Interbody Fusion.
Retrospective cohort. Lateral lumbar interbody fusion (LLIF) commonly involves a transpsoas approach. Despite the association between LLIF, postoperative iliopsoas weakness, and iatrogenic neuropraxia, no study has yet examined the effect of psoas or multifidus muscle quality on patient-reported outcomes (PROs). This study retrospectively reviewed patients who underwent LLIF with 1-year minimum follow-up. Psoas and multifidus muscle qualities were graded on preoperative magnetic resonance imaging using two validated classification systems for muscle atrophy. Average muscle quality was calculated as the mean score from all levels (L1-2 through L5-S1). Univariate and multivariate statistics were utilized to investigate the relationship between psoas/multifidus muscle quality and preoperative, 6-weeks postoperative, and final postoperative PROs. 74 patients (110 levels) with a mean follow-up of 18.71 ± 8.02 months were included for analysis. Greater multifidus atrophy was associated with less improvement on ODI, SF12, and VR12 (P < .05) on univariate analysis. On multivariate analysis, worse multifidus atrophy predicted less improvement on SF12 and VR12 (P < .05). Despite the direct manipulation of the psoas muscle inherent to LLIF, preoperative psoas muscle quality did not affect postoperative outcomes. Rather, the extent of preoperative multifidus fatty infiltration and atrophy was more likely to predict postoperative pain and disability. These findings suggest that multifidus atrophy may be more pertinent than psoas atrophy in its association with patient-reported outcome measures after LLIF.


[Lateral cervical chemodectoma].
Diagnostic and surgical features of lateral cervical chemodectomas are discussed in relation to a personal series of 13 patients with carotid body tumors and 7 with tumors of the intravagal glomus. Clinically, these affections are poor in symptomatology, those of the carotid glomus presenting as an isolated cervical tumor, intravagal glomus tumors as parapharyngeal masses of the retrostyloid type. Arteriographic findings are discussed in detail, as they constitute the only pathognomonic signs, enable differentiation between the two tumor sites, and supply essential pre-operative data: location of the pedicles, possible invasion of arterial walls, and the limits of the tumor. Surgical approach should be wide to enable control of the distal end of the internal carotid artery, if necessary under the base of the cranium, excision of the tumor usually being possible by employing the subadventitial resection technique.


Coronary angiography after out-of-hospital cardiac arrest without ST-segment elevation: a systematic review and meta-analysis of randomised trials.
Out-of-hospital cardiac arrest (OHCA) has a poor prognosis. The optimal timing and role of early coronary angiography (CAG) in OHCA patients without ST-segment elevation remains unclear. The goal of this study is to compare an early CAG versus delayed CAG strategy in OHCA patients without ST elevation. We systematically searched PubMed, Embase and Cochrane databases, in June 2022, for randomised controlled trials (RCTs) comparing early versus delayed early CAG. A random effects meta-analysis was performed. A total of seven RCTs were included, providing a total of 1625 patients: 816 in an early strategy and 807 in a delayed strategy. In terms of outcomes assessed, our meta-analysis revealed a similar rate of all-cause mortality (pooled odds ratio [OR] 1.22 [0.99-1.50], P = 0.06, I 2 = 0%), neurological status (pooled OR 0.94 [0.74-1.21], = 0.65, I 2 = 0%), need of renal replacement therapy (pooled OR 1.11 [0.78-1.74], P = 0.47, I 2 = 0%) and major bleeding events (pooled OR 1.51 [0.95-2.40], P = 0.08, I 2 = 69%). According to our meta-analysis, in patients who experienced OHCA without ST elevation, early CAG is not associated with reduced mortality or an improved neurological status.


Novel 1-hydroxypyridin-2-one metal chelators prevent and rescue ubiquitin proteasomal-related neuronal injury in an in vitro model of Parkinson's disease.
Ubiquitin proteasome system (UPS) impairment, excessive cellular oxidative stress, and iron dyshomeostasis are key to substantia nigra dopaminergic neuronal degeneration in Parkinson's disease (PD); however, a link between these features remains unconfirmed. Using the proteasome inhibitor lactacystin we confirm that nigral injury via UPS impairment disrupts iron homeostasis, in turn increasing oxidative stress and promoting protein aggregation. We demonstrate the neuroprotective potential of two novel 1-hydroxy-2(1H)-pyridinone (1,2-HOPO) iron chelators, compounds C6 and C9, against lactacystin-induced cell death. We demonstrate that this cellular preservation relates to the compounds' iron chelating capabilities and subsequent reduced capacity of iron to form reactive oxygen species (ROS), where we also show that the ligands act as antioxidant agents. Our results also demonstrate the ability of C6 and C9 to reduce intracellular lactacystin-induced α-synuclein burden. Stability constant measurements confirmed a high affinity of C6 and C9 for Fe3+ and display a 3:1 HOPO:Fe3+ complex formation at physiological pH. Reducing iron reactivity could prevent the demise of nigral dopaminergic neurons. We provide evidence that the lactacystin model presents with several neuropathological hallmarks of PD related to iron dyshomeostasis and that the novel chelating compounds C6 and C9 can protect against lactacystin-related neurotoxicity.


The effect of phenylpyruvate on pyruvate metabolism in rat brain.
1. The effect of phenylalanine and phenylpyruvate on the metabolism of pyruvate by isolated mitochondria from rat brain was investigated. 2. Phenylpyruvate inhibited the fixation of H(14)CO(3) (-) in the presence of pyruvate by intact rat brain mitochondria, whereas phenylalanine and other metabolites of this amino acid had no inhibitory effect on this process. 3. Pyruvate carboxylase activity in freeze-dried rat brain mitochondrial preparations was also inhibited only by phenylpyruvate, and a ;mixed type' inhibition was observed. 4. The K(m) for pyruvate of rat brain pyruvate carboxylase was about 0.2mm. 5. The concentration of phenylpyruvate required for a 50% inhibition of H(14)CO(3) (-) fixation by the intact mitochondria and of pyruvate carboxylase activity was dependent on the concentration of pyruvate used in the incubation medium. 6. The possible significance of inhibition of pyruvate carboxylase activity by phenylpyruvate in the brains of phenylketonuric patients is discussed.


The nucleotide-bound/substrate-bound conformation of the Mycoplasma genitalium DnaK chaperone.
Hsp70 chaperones keep protein homeostasis facilitating the response of organisms to changes in external and internal conditions. Hsp70s have two domains-nucleotide binding domain (NBD) and substrate binding domain (SBD)-connected by a conserved hydrophobic linker. Functioning of Hsp70s depend on tightly regulated cycles of ATP hydrolysis allosterically coupled, often together with cochaperones, to the binding/release of peptide substrates. Here we describe the crystal structure of the Mycoplasma genitalium DnaK (MgDnaK) protein, an Hsp70 homolog, in the noncompact, nucleotide-bound/substrate-bound conformation. The MgDnaK structure resembles the one from the thermophilic eubacteria DnaK trapped in the same state. However, in MgDnaK the NBD and SBD domains remain close to each other despite the lack of direct interaction between them and with the linker contacting the two subdomains of SBD. These observations suggest that the structures might represent an intermediate of the protein where the conserved linker binds to the SBD to favor the noncompact state of the protein by stabilizing the SBDβ-SBDα subdomains interaction, promoting the capacity of the protein to sample different conformations, which is critical for proper functioning of the molecular chaperone allosteric mechanism. Comparison of the solved structures indicates that the NBD remains essentially invariant in presence or absence of nucleotide.


Serial deletion reveals structural basis and stability for the core enzyme activity of human glutaminase 1 isoforms: relevance to excitotoxic neurodegeneration.
Glutaminase 1 is a phosphate-activated metabolic enzyme that catalyzes the first step of glutaminolysis, which converts glutamine into glutamate. Glutamate is the major neurotransmitter of excitatory synapses, executing important physiological functions in the central nervous system. There are two isoforms of glutaminase 1, KGA and GAC, both of which are generated through alternative splicing from the same gene. KGA and GAC both transcribe 1-14 exons in the N-terminal, but each has its unique C-terminal in the coding sequence. We have previously identified that KGA and GAC are differentially regulated during inflammatory stimulation and HIV infection. Furthermore, glutaminase 1 has been linked to brain diseases such as amyotrophic lateral sclerosis, Alzheimer's disease, and hepatic encephalopathy. Core enzyme structure of KGA and GAC has been published recently. However, how other coding sequences affect their functional enzyme activity remains unclear. We cloned and performed serial deletions of human full-length KGA and GAC from the N-terminal and the C-terminal at an interval of approximately 100 amino acids (AAs). Prokaryotic expressions of the mutant glutaminase 1 protein and a glutaminase enzyme activity assay were used to determine if KGA and GAC have similar efficiency and efficacy to convert glutamine into glutamate. When 110 AAs or 218 AAs were deleted from the N-terminal or when the unique portions of KGA and GAC that are beyond the 550 AA were deleted from the C-terminal, KGA and GAC retained enzyme activity comparable to the full length proteins. In contrast, deletion of 310 AAs or more from N-terminal or deletion of 450 AAs or more from C-terminal resulted in complete loss of enzyme activity for KGA/GAC. Consistently, when both N- and C-terminal of the KGA and GAC were removed, creating a truncated protein that expressed the central 219 AA - 550 AA, the protein retained enzyme activity. Furthermore, expression of the core 219 AA - 550 AA coding sequence in cells increased extracellular glutamate concentrations to levels comparable to those of full-length KGA and GAC expressions, suggesting that the core enzyme activity of the protein lies within the central 219 AA - 550 AA. Full-length KGA and GAC retained enzyme activities when kept at 4 °C. In contrast, 219 AA - 550 AA truncated protein lost glutaminase activities more readily compared with full-length KGA and GAC, suggesting that the N-terminal and C-terminal coding regions are required for the stability KGA and GAC. Glutaminase isoforms KGA and GAC have similar efficacy to catalyze the conversion of glutamine to glutamate. The core enzyme activity of glutaminase 1 protein is within the central 219 AA - 550 AA. The N-terminal and C-terminal coding regions of KGA and GAC help maintain the long-term activities of the enzymes.


Pseudomonas aeruginosa exotoxin A-induced hepatotoxicity: an animal model in rats.
Pseudomonas aeruginosa exotoxin A (PEA) has been generally used to induce liver injury in mice for experimental study. No PEA-induced hepatotoxicity study has ever been conducted in rats, although rats are the most common rodents used in toxicologic bioassay and pharmacological evaluation. The present study was conducted in male Wistar rats that were injected (i.v.) with PEA at doses of 0, 10, 20, 30 or 40 microg/kg body weight and evaluated at 12, 24, 36, 48 and 60 hr post-exposure (HPE). Rats exposed to PEA at 40 microg/kg died before 36 HPE, and the mortality was dose and time dependent. Liver injury was noted as increases in serum enzymes, along with alterations of liver histology in the 40 microg/kg group at 12 HPE. TUNEL-positive staining indicative of hepatocyte apoptosis was observed in the 20 microg/kg group at 12 HPE. Significant levels of DNA fragmentation ladder were observed in the 30 microg/kg group starting at 24 HPE. Serum levels of TNF-alpha was increased in the 30 and 40 microg/kg groups at 48 and 24 HPE, respectively. Other cytokines, such as IL-2, IL-6, and IL-10 were also increased at various doses and times. Furthermore, the elevated serum hepatic index levels decreased significantly by dexamethasone pretreatment. In contrast, these markers were exacerbated by co-administration of a non-toxic dose LPS. In overall evaluation, the PEA-induced liver injury can be used as a model for study of hyperimmune-mediated hepatotoxicity.


Effects of Intrabrachial metacholine infusion on muscle capillary recruitment and forearm glucose uptake during physiological hyperinsulinemia in obese, insulin-resistant individuals.
Impairment of insulin-mediated capillary recruitment in skeletal muscle contributes to a hampered glucose uptake in obesity. The objective of this study was to evaluate whether metacholine (MCh), a nitric oxide vasodilator, potentiates muscle capillary recruitment and forearm glucose uptake (FGU) during physiological hyperinsulinemia. The double-forearm technique [i.e. infused vs. control (Ctrl) forearm] was combined with im microdialysis during an oral glucose tolerance test in 15 nondiabetic, obese subjects divided into a group of insulin-resistant (IR) (n = 7) and insulin-sensitive (n = 8) individuals. After the oral glucose tolerance test, forearm blood flow in the Ctrl forearm was unchanged, whereas it increased about 3-fold (P < 0.0001 vs. baseline) in response to MCh. Capillary permeability surface area product for glucose (PS(glu)) (capillary recruitment), FGU, and interstitial insulin concentrations increased significantly over time (P < 0.001) in both forearms. Compared with insulin-sensitive, the IR subjects exhibited lower PS(glu) (P < 0.001) and FGU (P < 0.01) in the Ctrl arm, whereas this difference was insignificant in the MCh arm despite the blunted forearm blood flow increase. Moreover, in IR individuals MCh significantly (P < 0.05) ameliorated the delayed onset of insulin action, i.e. the FGU response to hyperinsulinemia. Finally, we found PS(glu) to be a strong and independent predictor of FGU response (adjusted R(2) 0.72; P < 0.0001). MCh-induced vasodilation may improve the microvascular and metabolic responses to physiological hyperinsulinemia in obese, IR individuals. Further studies are required to unravel whether stimulation of nitric oxide production in skeletal muscle may represent an attractive therapeutic approach to bypassing cellular resistance to glucose disposal.


Use of artificial intelligence for decision-support to avoid high-risk behaviors during laparoscopic cholecystectomy.
Bile duct injuries (BDIs) are a significant source of morbidity among patients undergoing laparoscopic cholecystectomy (LC). GoNoGoNet is an artificial intelligence (AI) algorithm that has been developed and validated to identify safe ("Go") and dangerous ("No-Go") zones of dissection during LC, with the potential to prevent BDIs through real-time intraoperative decision-support. This study evaluates GoNoGoNet's ability to predict Go/No-Go zones during LCs with BDIs. Eleven LC videos with BDI (BDI group) were annotated by GoNoGoNet. All tool-tissue interactions, including the one that caused the BDI, were characterized in relation to the algorithm's predicted location of Go/No-Go zones. These were compared to another 11 LC videos with cholecystitis (control group) deemed to represent "safe cholecystectomy" by experts. The probability threshold of GoNoGoNet annotations were then modulated to determine its relationship to Go/No-Go predictions. Data is shown as % difference [99% confidence interval]. Compared to control, the BDI group showed significantly greater proportion of sharp dissection (+ 23.5% [20.0-27.0]), blunt dissection (+ 32.1% [27.2-37.0]), and total interactions (+ 33.6% [31.0-36.2]) outside of the Go zone. Among injury-causing interactions, 4 (36%) were in the No-Go zone, 2 (18%) were in the Go zone, and 5 (45%) were outside both zones, after maximizing the probability threshold of the Go algorithm. AI has potential to detect unsafe dissection and prevent BDIs through real-time intraoperative decision-support. More work is needed to determine how to optimize integration of this technology into the operating room workflow and adoption by end-users.


Capillary-driven desalination in a synthetic mangrove.
According to the cohesion-tension theory, mangrove trees desalinate salty water using highly negative pressure (or tension) that is generated by evaporative capillary forces in mangrove leaves. Here, we demonstrate a synthetic mangrove that mimics the main features of the natural mangrove: capillary pumping (leaves), stable water conduction in highly metastable states (stem), and membrane desalination (root). When using nanoporous membranes as leaves, the maximum osmotic pressures of saline feeds (10 to 30 bar) allowing pure water uptake precisely correspond to expected capillary pressures based on the Young-Laplace equation. Hydrogel-based leaves allow for stable operation and desalination of hypersaline solutions with osmotic pressures approaching 400 bar, fivefold greater than the pressure limits of conventional reverse osmosis. Our findings support the applicability of the cohesion-tension theory to desalination in mangroves, provide a new platform to study plant hydraulics, and create possibilities for engineered membrane separations using large, passively generated capillary pressures.