Cryopreservation of human platelets with polyvinylpyrrolidone.
Fourteen polymeric compounds were assessed for their abilities to protect human platelets from injury during freezing and thawing. The effect of polymer concentration and molecular weight, cooling rate, platelet concentration, and short-term storage on platelet recovery were studied. Polyvinylpyrrolidone, average molecular weight 10,000, was found to provide the greatest degree of protection to platelets. Recoveries of platelets of from 60 to 80 per cent were achieved using a 5 per cent final concentration of polyvinylpyrrolidone and a cooling rate of 8 C per minute. Methods of freezing small aliquots of platelets and platelet concentrates were developed. The frozen-thawed platelets were viable in vitro as shown by osmotic reversal, clot retraction, and aggregation by collagen and adenosine diphosphate. The techniques described could be developed readily to facilitate the long-term storage of platelets.


The Safety of Soy Leghemoglobin Protein Preparation Derived from Pichia pastoris Expressing a Soy Leghemoglobin Gene from Glycine max: In Vitro and In Vivo Studies.
Soy leghemoglobin (LegH) protein derived from soy (Glycine max) produced in Pichia pastoris (reclassified as Komagataella phaffii) as LegH Prep is a novel food ingredient that provides meat-like flavor and aroma to plant-derived food products. The safety of LegH Prep has been previously assessed in a battery of in vivo and in vitro testing and found no adverse effects under the conditions tested. In this new work, we present the results of new in vivo and in vitro tests evaluating the safety of LegH Prep. LegH Prep was nonmutagenic in a bacterial reverse mutation assay and nonclastogenic in an in vitro micronucleus assay in human lymphocytes. Systemic toxicity was evaluated in the 90 day dietary study in male and female Sprague-Dawley® rats that included a 28 day recovery period. The study resulted in no animal deaths associated with the administration of LegH Prep at the highest dose (90,000 ppm). There were no significant adverse clinical or physical changes attributed to LegH Prep administration, and no observed adverse effects on either male or female rats over the course of the 28 day recovery phase study. The new 90 day dietary toxicity study established a no observed adverse effect level (NOAEL) of 4798.3 and 5761.5 mg/kg/day, the maximum level tested for male and female rats, respectively. Thus, the results of the studies demonstrate that under the conditions tested, LegH Prep is not toxic for consumption in meat analog products.


[Anesthetic management of a 1.7-kg premature infant undergoing thoracoscopic thoracic duct ligation].
We report successful anesthetic management of a 1.7-kg premature infant who underwent thoracoscopic thoracic duct ligation under general anesthesia. She was born at 30 weeks gestation with birth weight of 1,546 g and was suffering from respiratory distress due to persistent right chylothorax for two months after birth. Chest tube drainage, fasting and intrapleural fibrin glue did not reduce her right chylothorax. Thoracoscopic thoracic duct ligation was scheduled on her day 64 under general anesthesia. The tracheal tube end was placed in the midtrachea and carbon dioxide was insufflated into the operative side of the thorax. During thoracoscopy her left lung was ventilated with the right lung pressed with spatulaes, but her respiratory status did not deteriorate so much despite of unilateral ventilation. We speculate that, due to massive right chylothorax, her pulmonary blood flow had already shifted to the left lung, therefore intraoperative substantial left unilateral lung ventilation exerted minimal effect on her respiratory status. The operation was successful and she was weaned from the ventilator on the following day.


Phytic Acid: A Novel Dentine Conditioning Agent for Glass Ionomer Cement Restorations: An In Vitro Scanning Electron Microscopic Study.
Various dentin conditioners have been recommended to enhance the bond strength of glass ionomer cement (GIC) to dentin. The purpose of the in vitro study was to determine the efficacy of three dentin conditioners on the microshear bond strength of GIC and the failure patterns of the restoration, as well as the efficacy of removing the smear layer using a scanning electron microscope (SEM). Eighty healthy mandibular premolars were cleaned, isolated, and divided into four groups (n = 20). Flat coronal dentinal surfaces were accomplished by removing the occlusal surface of the crowns. Each group received the following treatment: group 1 with 10% polyacrylic acid; group 2 with 10% ethylenediaminetetraacetic acid (EDTA); group 3 with 1% phytic acid; and group 4 with no conditioning (control). High-strength Fuji IX GIC was applied to the dentin, and samples were thermocycled and de-bonded with a crosshead speed of 0.5 mm/min employing a shear force. A stereomicroscope and a SEM were used to examine the dentin surface at magnifications of 2000× and 5000×. One-way analysis of variance (ANOVA), Tukey's post hoc test, and Chi-squared test were used to assess data on microshear bond strength, smear layer removal, and failure modes. The bond strength of group 4 was statistically highly significant (P ≤ 0.001) compared to the other groups. However, the differences between groups 2 and 3 were insignificant. Mixed pattern of failure was the least in group 3 (IP6). In group 3, the SEM photomicrograph revealed open dentinal tubules with complete removal of the smear layer. The results of the present study affirm that phytic acid enhances the bond strength of GIC restorations and removal of smear layer to a level comparable to EDTA as a dentin conditioner.


Effects of interferon-alpha on a reduced release of interleukin-8 from latently HIV-1-infected monocytic cell line U937 cells.
The effects of human interferon-alpha (IFN-alpha) on the release of an antimicrobial interleukin, interleukin-8 (IL-8), from human immunodeficiency virus type 1 (HIV-1)-infected myelomonocytic cell line, U937, were studied in vitro to evaluate the potential of IFN-alpha in the management of acquired immunodeficiency syndrome (AIDS)-associated opportunistic diseases. The latently HIV-1-infected U937 cells (U937/HIV-1(L)) showed a marked reduction of IL-8 secretion as compared to uninfected U937 cells, whereas IL-8 release from productively HIV-1-infected U937 cells was comparable to uninfected cells. The IFN-alpha recovered partially the reduced IL-8 level from U937/HIV-1(L) cells in a dose-dependent manner. Any significant inhibition of IFN-alpha-augmented IL-8 secrement by anti-IL-1 antibody was not observed, suggesting that the enhanced IL-8 secretion occurred without augmenting IL-1 production. The IFN-alpha-augmented IL-8 secretion from latently HIV-1-infected U937 cells may suggest a beneficial potential of IFN-alpha in a treatment of bacterial or fungal infection frequently seen in patients with progressive stages of HIV-1 infection.


Aerodynamic modelling of a Cretaceous bird reveals thermal soaring capabilities during early avian evolution.
Several flight modes are thought to have evolved during the early evolution of birds. Here, we use a combination of computational modelling and morphofunctional analyses to infer the flight properties of the raven-sized, Early Cretaceous bird Sapeornis chaoyangensis-a likely candidate to have evolved soaring capabilities. Specifically, drawing information from (i) mechanical inferences of the deltopectoral crest of the humerus, (ii) wing shape (i.e. aspect ratio), (iii) estimations of power margin (i.e. difference between power required for flight and available power from muscles), (iv) gliding behaviour (i.e. forward speed and sinking speed), and (v) palaeobiological evidence, we conclude that S. chaoyangensis was a thermal soarer with an ecology similar to that of living South American screamers. Our results indicate that as early as 125 Ma, some birds evolved the morphological and aerodynamic requirements for soaring on continental thermals, a conclusion that highlights the degree of ecological, functional and behavioural diversity that resulted from the first major evolutionary radiation of birds.


[Brazilian Protocol for Sexually Transmitted Infections 2020: epidemiological surveillance].
This article summarizes the chapter on epidemiological surveillance of sexually transmitted infections (STI) that forms part of the Clinical Protocol and Treatment Guidelines (PDCT) for Comprehensive Care for People with STI 2020), published by the Health Surveillance Secretariat of the Ministry of Health of Brazil. It presents some reflections on the new case definitions for surveillance of acquired syphilis, syphilis in pregnant women and congenital syphilis. The PCDT-IST 2020 was developed based on scientific evidence and validated in discussions with specialists. Epidemiological and clinical aspects are addressed, as well as guidelines for health service managers regarding programmatic and operational management of these diseases, guidelines for health professionals on screening, diagnosing and treating people with STIs and their sex partners, in addition to strategies for surveillance, prevention and control actions.


Cause and manner of death in drug-related fatality: an analysis of drug-related deaths recorded by coroners in England and Wales in 2000.
This study investigated causes and manner of drug-related fatalities recorded in 2000 in the United Kingdom, measuring the 'masked' manner of death in cases typically recorded as overdose. A retrospective cohort study was used of 1037 cases of accidental drug-related fatalities reported by coroners in England and Wales to the National Programme of Substance Abuse Deaths. Whilst 802 cases were identified as direct acute overdose, representing 77% of the total accidental deaths, 23% of 'overdose' fatalities were caused by asphyxiation (7%), drug-related medical conditions (7%), non-drug-related conditions (4%), traumatic accidents (3%) and infections (2%). Younger people show higher risk of overdose and asphyxiation; older people show higher risk from pre-existing medical conditions. This study not only confirmed the high risk of overdose associated with heroin and polydrug use, but it also identified other high fatality risk factors for heroin/morphine users such as contracting an acute infection leading to septicaemia or endocarditis, or contracting a chronic infection such as HIV, HBV or HCV. In contrast, stimulants particularly featured in traumatic accidents, with amphetamine use most associated with cardio-vascular fatality. These findings highlight the 'masked' manner of death in cases commonly recorded as overdose and demonstrate the need for a more-detailed and systematic method of recording drug-related deaths in order to inform drug education and harm reduction strategies.


[Diagnosis and long-term outcome after surgical therapy of tarsal tunnel syndrome].
In differential diagnosis of foot pain the tarsal tunnel syndrome has to be considered. Only few publications concerning clinical signs, diagnostic means, especially electrophysiological methods and postoperative long term results can be found in literature. In this study 21 of 32 patients who had a decompression operation of the posterior tibial nerve between 1972 and 1995 were reexamined at an average follow-up time of 12 years. Electrodiagnostic evaluation and clinical results by time of follow-up were compared to preoperative findings. Using the criteria described by Kaplan 10 postoperative results were rated as very good, 6 as good and 5 as poor. Sensible neurography has proven the most sensitive electrodiagnostic method. In the majority of cases the tarsal tunnel syndrome is still a clinical diagnosis.


Application of bedside ultrasound in predicting the outcome of weaning from mechanical ventilation in elderly patients.
With the increased ageing of society, more and more elderly people are admitted to the intensive care unit, How to accurately predict whether elderly patients can successfully wean from the ventilator is more complicated. Diaphragmatic excursion (DE) and diaphragm thickening fraction (DTF) were measured by bedside ultrasound to assess diaphragm function. The lung ultrasound score (LUS) and the rapid shallow breathing index (RBSI) were used as indices of diaphragm function to predict the outcome of weaning from mechanical ventilation. The aim of this study was to examine the clinical utility of these parameters in predicting extubation success. This prospective study included 101 consecutive elderly patients undergoing a trial of extubation in the ICU of Haidian Hospital between June 2017 and July 2020. Patients were divided into the successful weaning group (n = 69) and the failed weaning group (n = 32). Baseline characteristics, including RSBI, were recorded. Measurements of DE, DTF and LUS were made using ultrasound within 24 h before extubation. Median DE was greater in patients with extubation success than in those with extubation failure (1.64 cm vs. 0.78 cm, p = 0.001). Patients with extubation success had a greater DTF than those with extubation failure (49.48% vs. 27.85%, p = 0.001). The areas under the receiver operating curves for the RSBI, LUS, DE and DFT were 0.680, 0.764, 0.831 and 0.881, respectively. The best cut-off values for predicting successful weaning were DTF ≥ 30%, DE ≥ 1.3 cm, LUS ≤ 11, and RSBI ≤ 102. The specificity of DTF (84%) in predicting weaning outcome was higher than that of RBSI (53%), that of LUS (55%), and that of DE (62%). The sensitivity of DTF (94%) was greater than that of RBSI (85%), that of LUS (71%), and that of DE (65%). The combination of RSBI, LUS, DE, and DTF showed the highest AUC (AUC = 0.919), with a sensitivity of 96% and a specificity of 89%. DTF has higher sensitivity and specificity for the prediction of successful weaning in elderly patients than the other parameters examined. The combination of RSBI, LUS, DE and DFT performed well in predicting weaning outcome. This has potentially important clinical application and merits further evaluation.


Use of a passive hemagglutination inhibition test for monitoring levels of serum carcinoembryonic antigen following surgical therapy.
A technique of passive hemagglutination inhibition (PHI) has been used to monitor levels of carcinoembryonic antigen (CEA) in human sera following surgical therapy. CEA was coupled to human type O-negative erythrocytes in the presence of bis-diazotized benzidine. Pre-operative and post-operative sera from 11 patients with primary adenocarcinomas of the gastrointestinal tract and from one patient with ulcerative colitis were then tested for their capacity to inhibit the agglutination of the sensitized cells in the presence of a predetermined amount of goat anti-CEA serum. Positive sera were defined as those which inhibited agglutination at dilutions of greater than 1:8. The pre-operative sera from 11 of the 12 patients inhibited agglutination at dilutions of 1:16 or greater. The one negative serum was from a patient with primary adenocarcinoma of the colon in the stage of Dukes' C. At one month post-resection, the PHI titer of six patients with colon cancer and of the patient with ulcerative colitis was less than or equal to 1:8. However, by 4 months post-resection, all but 3 of the patients had PHI titers in the positive range. These elevated titers were accompanied by recurrence of tumor growth and/or metastatic dissemination. A radioimmunoassay was used to quantitate CEA in 22 of the sera which had been tested by PHI. When positive sera were defined as those which inhibited agglutination at dilutions of greater than 1:8 and contained CEA in excess of 5 ng per ml, the results of the two procedures were in agreement for 17 of the 22 specimens. Five sera, representative of 2 patients with colon cancer, were false negative by PHI.


Interaction of selenium compounds with zinc finger proteins involved in DNA repair.
As an essential element, selenium is present in enzymes from several families, including glutathione peroxidases, and is thought to exert anticarcinogenic properties. A remarkable feature of selenium consists of its ability to oxidize thiols under reducing conditions. Thus, one mode of action recently suggested is the oxidation of thiol groups of metallothionein, thereby providing zinc for essential reactions. However, tetrahedral zinc ion complexation to four thiolates, similar to that found in metallothionein, is present in one of the major classes of transcription factors and other so-called zinc finger proteins. Within this study we investigated the effect of selenium compounds on the activity of the formamidopyrimidine-DNA glycosylase (Fpg), a zinc finger protein involved in base excision repair, and on the DNA-binding capacity and integrity of xeroderma pigmentosum group A protein (XPA), a zinc finger protein essential for nucleotide excision repair. The reducible selenium compounds phenylseleninic acid, phenylselenyl chloride, selenocystine, ebselen, and 2-nitrophenylselenocyanate caused a concentration-dependent decrease of Fpg activity, while no inhibition was detected with fully reduced selenomethionine, methylselenocysteine or some sulfur-containing analogs. Furthermore, reducible selenium compounds interfered with XPA-DNA binding and released zinc from the zinc finger motif, XPAzf. Zinc release was even evident at high glutathione/oxidised glutathine ratios prevailing under cellular conditions. Finally, comparative studies with metallothionein and XPAzf revealed similar or even accelerated zinc release from XPAzf. Altogether, the results indicate that zinc finger motifs are highly reactive towards oxidizing selenium compounds. This could affect gene expression, DNA repair and, thus, genomic stability.


Pharmacokinetics of surotomycin from phase 1 single and multiple ascending dose studies in healthy volunteers.
Surotomycin, a novel, orally administered, cyclic, lipopeptide antibacterial in development for the treatment of Clostridium difficile-associated diarrhea, has demonstrated minimal intestinal absorption in animal models. Safety, tolerability, and plasma pharmacokinetics of single and multiple ascending oral doses (SAD/MAD) of surotomycin in healthy volunteers were characterized in two randomized, double-blind, placebo-controlled, phase 1 studies. Participants were sequentially enrolled into one of four SAD (500, 1000, 2000, 4000 mg surotomycin) or three MAD (250, 500, 1000 mg surotomycin twice/day for 14 days) cohorts. Ten subjects were randomized 4:1 into each cohort to receive surotomycin or placebo. Surotomycin plasma concentrations rose as dose increased (maximum plasma concentration [Cmax]: 10.5, 21.5, 66.6, and 86.7 ng/mL). Systemic levels were generally low, with peak median surotomycin plasma concentrations observed 6-12 h after the first dose. In the MAD study, surotomycin plasma concentrations were higher on day 14 (Cmax: 25.5, 37.6, and 93.5 ng/mL) than on day 1 (Cmax: 6.8, 11.0, and 21.1 ng/mL for increasing doses), indicating accumulation. In the SAD study, <0.01% of the administered dose was recovered in urine. Mean surotomycin stool concentration from the 1000 mg MAD cohort was 6394 μg/g on day 5. Both cohorts were well tolerated with all adverse events reported as mild to moderate. Both SAD and MAD studies of surotomycin demonstrated minimal systemic exposure, with feces the primary route of elimination following oral administration; consistent with observations with similar compounds, such as fidaxomicin. Results of these phase 1 studies support the continued clinical development of surotomycin for the treatment of Clostridium difficile-associated diarrhea. NCT02835118 and NCT02835105 . Retrospectively registered, July 13 2016.


Associations between skeletal muscle energetics and accelerometry-based performance fatigability: Study of Muscle, Mobility and Aging.
Performance fatigability is typically experienced as insufficient energy to complete daily physical tasks, particularly with advancing age, often progressing toward dependency. Thus, understanding the etiology of performance fatigability, especially cellular-level biological mechanisms, may help to delay the onset of mobility disability. We hypothesized that skeletal muscle energetics may be important contributors to performance fatigability. Participants in the Study of Muscle, Mobility and Aging completed a usual-paced 400-m walk wearing a wrist-worn ActiGraph GT9X to derive the Pittsburgh Performance Fatigability Index (PPFI, higher scores = more severe fatigability) that quantifies percent decline in individual cadence-versus-time trajectory from their maximal cadence. Complex I&II-supported maximal oxidative phosphorylation (max OXPHOS) and complex I&II-supported electron transfer system (max ETS) were quantified ex vivo using high-resolution respirometry in permeabilized fiber bundles from vastus lateralis muscle biopsies. Maximal adenosine triphosphate production (ATPmax ) was assessed in vivo by 31 P magnetic resonance spectroscopy. We conducted tobit regressions to examine associations of max OXPHOS, max ETS, and ATPmax with PPFI, adjusting for technician/site, demographic characteristics, and total activity count over 7-day free-living among older adults (N = 795, 70-94 years, 58% women) with complete PPFI scores and ≥1 energetics measure. Median PPFI score was 1.4% [25th-75th percentile: 0%-2.9%]. After full adjustment, each 1 standard deviation lower max OXPHOS, max ETS, and ATPmax were associated with 0.55 (95% CI: 0.26-0.84), 0.39 (95% CI: 0.09-0.70), and 0.54 (95% CI: 0.27-0.81) higher PPFI score, respectively. Our findings suggested that therapeutics targeting muscle energetics may potentially mitigate fatigability and lessen susceptibility to disability among older adults.


Altered protein profile of lymphocytes in an antigen-specific model of colitis: a comparative proteomic study.
Lymphocytes are deeply involved in the initiation and perpetuation of inflammatory response in inflammatory bowel disease (IBD) and lymphocyte-derived proteins are associated with the pathogenesis of the disease. The aim of this study was to identify the altered protein profiles of lymphocytes from rats with colitis. Colitis models were induced by colonic administration of trinitrobenzene sulfonic acid (TNBS) in 50% ethanol in male SD rats. Seven days after administration of TNBS/ethanol, lymphocytes were harvested from mesenteric lymph nodes (MLNs) and proteins were extracted. Two-dimensional polyacrylamide gel electrophoresis and PDQuest 2D-image-analysis software were used to display and analyze the protein spots. The differentially-expressed proteins were identified by tryptic in-gel digestion and mass spectrometry. Real-time RT-PCR was used for selected transcripts to validate the findings of the proteomics analysis. A total of 1,100 protein spots including 26 proteins with at least a two-fold difference in abundance between colitis and control groups were identified. Among all the detected spots, 17 were up-regulated and 9 were down-regulated. It was found that the altered proteins included the regulators of the cell cycle and cell proliferation, signal transduction factors, inflammatory factors, apoptosis-related proteins and metabolic enzymes. In lymphocytes of rats with TNBS-induced colitis, 26 altered proteins were identified. They involve inflammation, apoptosis, metabolism, and regulation of the cell cycle, cell proliferation, and signal transduction.


Well-being and Anticipation for Future Positive Events: Evidences from an fMRI Study.
Anticipation for future confers great benefits to human well-being and mental health. However, previous work focus on how people's well-being correlate with brain activities during perception of emotional stimuli, rather than anticipation for the future events. Here, the current study investigated how well-being relates to neural circuitry underlying the anticipating process of future desired events. Using event-related functional magnetic resonance imaging, 40 participants were scanned while they were performing an emotion anticipation task, in which they were instructed to anticipate the positive or neutral events. The results showed that bilateral medial prefrontal cortex (MPFC) were activated during anticipation for positive events relative to neutral events, and the enhanced brain activation in MPFC was associated with higher level of well-being. The findings suggest a neural mechanism by which the anticipation process to future desired events correlates to human well-being, which provide a future-oriented view on the neural sources of well-being.


[Ultrasound-guided percutaneous fine-needle puncture cytology in the diagnosis of malignant tumors of the liver].
Results of ultrasound guided percutaneous fine-needle puncture cytology in 142 cases of malignant tumors of the liver are reported. The positive cytology was noted in 118 (83.1%) (10 suspicious). The primary liver cancer comprised 99 cases. In 7 lesions, equal to or less than 3 cm in diameters, 6 were positive in cytology. In 20,3-5 cm in size, 18 were positive. In 15 false-negatives, 12 were larger than 5 cm in diameter. Among 43 cases of metastatic liver cancer, 34 showed positive cytology and 9 false-negative. Among 108 cases of benign hepatic diseases, in 49.1%, it was difficult to arrive at definitive diagnosis by ultrasonography only, but in 97.2% malignancy was excluded by ultrasound guided fine-needle puncture cytology. The suspicious false-positive result occurred only in 3 cases. In this series, there were 250 cases of malignant tumors and benign diseases. The overall accurate diagnostic rate was 89.2%. All the patients had been followed for more than 6 months. The differential diagnosis between malignant and benign tumors, causes of misdiagnosis and complications are discussed.


Fluorescent sensing of non-steroidal anti-inflammatory drugs naproxen and ketoprofen by dansylated squaramide-based receptors.
Bis-squaramide receptors L1-L4 bearing a dansyl moiety were synthesised and their potential applications as fluorescent probes towards non steroidal anti-inflammatory drugs naproxen and ketoprofen was investigated. A detailed photophysical characterization in CH3CN/DMSO solution (9 : 1 v/v) was conducted and demonstrated that the two macrocyclic receptors L1 and L2 show good sensitivity towards ketoprofen with an ON-OFF fluorescent response, while the two open chain receptors L3 and L4 behave similarly with the three guests considered. DFT theoretical calculations carried out on L2 and L4 as model receptors allowed to propose a possible coordination mode towards the guests. Finally, 1H-NMR spectroscopy in DMSO-d6/0.5% water solution demonstrated that the four receptors interact with the considered guests via H-bonds.


Phase behavior of elemental aluminum using monte carlo simulations.
Monte Carlo simulations are presented for two models of aluminum: an embedded-atom model and an explicit many-body model. Vapor/liquid coexistence curves are determined using Gibbs ensemble Monte Carlo simulations. The normal boiling points predicted by both models are somewhat higher (by about 10%) than the experimental value. Isothermal constant-stress simulations are used to simulate solid Al from 300 K to the triple point. The solid structures are at least metastable in the face-centered cubic configuration, and the specific heat is determined to be lower than the experimental value. The melting point for the embedded-atom model determined via thermodynamic integration along a pseudo-supercritical path is approximately 20% higher than the experimental value.


The relationship between lipocalin-2 and free testosterone levels in polycystic ovary syndrome.
Lipocalin-2 is an adipokine that is mainly produced from adipocytes and macrophages. Data related to PCOS and other obesity-associated disorders have shown divergent results. Here, we studied lipocalin-2 concentrations in women with PCOS and in healthy women, and investigated the potential contributors underlying lipocalin association with PCOS. Forty-four women with PCOS and 47 age- and BMI-matched healthy women were enrolled. Fasting serum glucose, insulin, homeostasis model assessment of insulin resistance (HOMA-IR), high-sensitivity C-reactive protein (hs-CRP), and free testosterone levels were measured. The body fat percentage was measured by bioelectrical impedance. Lipocalin-2 concentrations were significantly higher in the PCOS group than in the control group (55.74 ± 17.54 ng/mL vs. 36.46 ± 19.62 ng/mL, p = 0.011). There was a correlation between lipocalin-2 levels and free testosterone. In a multiple regression model, the body fat percentage, HOMA-IR, and hs-CRP were not associated with lipocalin-2. However, only free testosterone was associated with lipocalin-2. A "lipocalin-2 = 11.214 + (1.943 × free-testosterone)" equation was obtained. Serum lipocalin-2 levels were higher in women with PCOS, and only free testosterone was associated with lipocalin-2. Lipocalin-2 levels and their influencing factors have discrepant results in both PCOS and other obesity- or insulin resistance-related metabolic disorders. Thus, the potential role of lipocalin-2 in PCOS should be clarified. (Endokrynol Pol 2017; 68 (1): 7-12).


Quantum chemical studies of methadone.
Quantum chemical calculations were performed on the flexible methadone molecule to test the hypothesis that it structurally mimics the fused ring structure of morphine. In these calculations using the semiempirical, PCILO method, protonated and nonprotonated conformations were considered representative of different types of intramolecular interaction at the morphine receptor. Calculated energies for these conformations were compared to those calculated for protonated and nonprotonated extended chain and crystal structure conformers. Lowest energy conformations showed intramolecular bonding but the resultant molecular geometries were not very morphine-like. A comparison of the structure of methadone to that of meperidine seemed equally as valid.


Intramuscular Adeno-Associated Virus-Mediated Expression of Monoclonal Antibodies Provides 100% Protection Against Ebola Virus Infection in Mice.
The 2013-2016 West Africa outbreak demonstrated the epidemic potential of Ebola virus and highlighted the need for counter strategies. Monoclonal antibody (mAb)-based therapies hold promise as treatment options for Ebola virus infections. However, production of clinical-grade mAbs is labor intensive, and immunity is short lived. Conversely, adeno-associated virus (AAV)-mediated mAb gene transfer provides the host with a genetic blueprint to manufacture mAbs in vivo, leading to steady release of antibody over many months. Here we demonstrate that AAV-mediated expression of nonneutralizing mAb 5D2 or 7C9 confers 100% protection against mouse-adapted Ebola virus infection, while neutralizing mAb 2G4 was 83% protective. A 2-component cocktail, AAV-2G4/AAV-5D2, provided complete protection when administered 7 days prior to challenge and was partially protective with a 3-day lead time. Finally, AAV-mAb therapies provided sustained protection from challenge 5 months following AAV administration. AAV-mAb may be a viable alternative strategy for vaccination against emerging infectious diseases.


Local Spatial Analysis and Dynamic Simulation of Childhood Obesity and Neighbourhood Walkability in a Major Canadian City.
Body weight is an important indicator of current and future health and it is even more critical in children, who are tomorrow's adults. This paper analyzes the relationship between childhood obesity and neighbourhood walkability in Calgary, Canada. A multivariate analytical framework recognizes that childhood obesity is also associated with many factors, including socioeconomic status, foodscapes, and environmental factors, as well as less measurable factors, such as individual preferences, that could not be included in this analysis. In contrast with more conventional global analysis, this research employs localized analysis and assesses need-based interventions. The one-size-fit-all strategy may not effectively control obesity rates, since each neighbourhood has unique characteristics that need to be addressed individually. This paper presents an innovative framework combining local analysis with simulation modeling to analyze childhood obesity. Spatial models generally do not deal with simulation over time, making it cumbersome for health planners and policy makers to effectively design and implement interventions and to quantify their impact over time. This research fills this gap by integrating geographically weighted regression (GWR), which identifies vulnerable neighbourhoods and critical factors for childhood obesity, with simulation modeling, which evaluates the impact of the suggested interventions on the targeted neighbourhoods. Neighbourhood walkability was chosen as a potential target for localized interventions, owing to the crucial role of walking in developing a healthy lifestyle, as well as because increasing walkability is relatively more feasible and less expensive then modifying other factors, such as income. Simulation results suggest that local walkability interventions can achieve measurable declines in childhood obesity rates. The results are encouraging, as improvements are likely to compound over time. The results demonstrate that the integration of GWR and simulation modeling is effective, and the proposed framework can assist in designing local interventions to control and prevent childhood obesity.


Identification of kinases, phosphatases, and phosphorylation sites in human and porcine spermatozoa.
Multiple inter-connected signaling pathways, involving kinases and phosphatases, form a framework that controls sperm motility, function, and fertilizing ability. Methods that give a broad view of the proteomic landscape may prove valuable in uncovering new crosstalk connections, as well as in discovering new proteins within this regulatory framework. A multi-immunoblotting strategy was utilized to evaluate this concept on human and porcine spermatozoa samples. In human and porcine spermatozoa, a diversity of kinases were identified including protein kinase A (PKA), protein kinase B (PKB), isoforms of protein kinase C (PKC), calmodulin-dependent kinases (CAMK), casein kinase (CK), and isoforms of glycogen synthase kinase (GSK3). Several phosphatases, such as protein phosphatase (PP)-1, PP2A, PP2C, and mitogen activated protein kinase (MAPK) phosphatase (MKP-1), were identified in human spermatozoa. The phosphorylation epitopes recognized belonged to members of the MAPK family, in addition to α and β isoforms of GSK3 and cAMP response element binding protein (CREB). Proteomic approaches that allow a broad view may aid in understanding the crosstalk between signaling systems in spermatozoal physiology.


Dissociation dynamics of thiolactic acid at 193 nm: detection of the nascent OH product by laser-induced fluorescence.
Electronically excited thiolactic acid (2-mercaptopropionic acid), H(3)C-CH(SH)-COOH, undergoes the C-OH bond cleavage on excitation to the S(2) state at 193 nm, generating the primary product OH (v,J), which is detected by laser-induced fluorescence technique in a collisionless condition of flow system. The partitioning of the available energy between vibrational, rotational, and translational degrees of freedom of nascent photofragments is obtained from relative intensities of ro-vibronic lines in laser-induced fluorescence spectrum of OH, and their Doppler profiles. The rotational population of OH (v(")=0) is characterized by rotational temperature of 408+/-25 K. OH is produced in a vibrationally cold state, i.e., mostly in v(")=0. The average translational energy of OH (v(")=0,J(")) is found to be 21.5+/-2.0 kcal/mol, which implies 25.6 kcal/mol of energy in relative translation of photoproducts corresponding to the f(t) value of approximately 0.6. The observed high translational energy is due to the presence of a barrier in the exit channel, implying that the C-OH bond scission takes place on an electronically excited potential energy surface. The observed partitioning of the available energy between various degrees of the photofragments is theoretically modeled, and the hybrid model, with 26.0 kcal/mol of barrier in the exit channel, is found to explain the measured data quite well. The experimental results are also supported with ab initio molecular orbital calculations for both the ground and the excited electronic states. Time-dependent density functional theory is used to understand the nature of various electronic transitions connecting the lower excited states. Potential energy curves as a function of the C-OH bond length of thiolactic acid suggest distinct exit barriers in the S(1), T(1), and T(2) states. But, we could locate the transition state structure for OH formation in the S(1) state alone. Thus, although thiolactic acid is excited to the S(2) state at 193 nm, it undergoes internal conversion to S(1) where it dissociates to yield OH. In addition to the OH channel from excited electronic states, we studied theoretically all probable dissociation channels occurring on the ground electronic state of thiolactic acid.


The metabolic and ventilatory response to the infusion of stress hormones.
Sepsis and trauma result in increases in epinephrine, glucagon, and cortisol secretion as well as alterations in respiratory pattern that is characterized by increased minute ventilation, decreased tidal volume, and increased frequency. Six male subjects were infused for 5.5 hours with cortisol, epinephrine, and glucagon in amounts designed to simulate plasma levels seen in patients following trauma. During the initial 20 minutes of the hormone infusion, minute ventilation (VE), oxygen consumption (VO2), and carbon dioxide production (VCO2) increased above preinfusion values. VCO2 increased more than VO2 resulting in an increase in respiratory quotient (RQ) from 0.93 to 1.14. The increase in VE was due to increased tidal volume and not frequency (f). After 4.5 hours, the VE, VO2, and VCO2 were still above preinfusion levels but the RQ had decreased to 0.98 because of a decrease in VCO2. Frequency had increased from 19 +/- 4.8 breaths/min preinfusion to 22 +/- 4.7 after 4.5 hours. After 4.5 hours, VT was still above preinfusion levels while pH and PaCO2 had decreased below them. The latter was associated with an increase in serum lactate. At no time was a decrease in tidal volume observed. Therefore, the infusion of these hormones does not simulate all the alterations observed during trauma and sepsis.


Turbulence and entrainment length scales in large wind farms.
A number of large wind farms are modelled using large eddy simulations to elucidate the entrainment process. A reference simulation without turbines and three farm simulations with different degrees of imposed atmospheric turbulence are presented. The entrainment process is assessed using proper orthogonal decomposition, which is employed to detect the largest and most energetic coherent turbulent structures. The dominant length scales responsible for the entrainment process are shown to grow further into the wind farm, but to be limited in extent by the streamwise turbine spacing, which could be taken into account when developing farm layouts. The self-organized motion or large coherent structures also yield high correlations between the power productions of consecutive turbines, which can be exploited through dynamic farm control.This article is part of the themed issue 'Wind energy in complex terrains'.


Method to Solve Underwater Laser Weak Waves and Superimposed Waves.
With the rapid development of Lidar technology, the use of Lidar for underwater terrain detection has become feasible. There is still a challenge in the process of signal resolution: the underwater laser echo signal is different to propagating in the air, and it is easy to produce weak waves and superimposed waves. However, existing waveform decomposition methods are not effective in processing these waveform signals, and the underwater waveform signal cannot be correctly decomposed, resulting in subsequent data-processing errors. To address these issues, this study used a drone equipped with a 532 nm laser to detect a pond as the study background. This paper proposes an improved inflection point selection decomposition method to estimate the parameter. By comparing it with other decomposition methods, we found that the RMSE is 2.544 and R2 is 0.995975, which is more stable and accurate. After estimating the parameters, this study used oscillating particle swarm optimization (OPSO) and the Levenberg-Marquardt algorithm (LM) to optimize the estimated parameters; the final results show that the method in this paper is closer to the original waveform. In order to verify the processing effect of the method on complex waveform, this paper decomposes and optimizes the simulated complex waveforms; the final RMSE is 0.0016, R2 is 1, and the Gaussian component after decomposition can fully represent the original waveform. This method is better than other decomposition methods in complex waveform decomposition, especially regarding weak waves and superimposed waves.


Universal bounds on the time evolution of entanglement entropy.
Using relative entropy, we derive bounds on the time rate of change of geometric entanglement entropy for any relativistic quantum field theory in any dimension. The bounds apply to both mixed and pure states, and may be extended to curved space. We illustrate the bounds in a few examples and comment on potential applications and future extensions.


Differential Clinical Characteristics of Older Black and White Nursing Home Residents: A Pilot Study.
Older black patients have higher levels of medical morbidity yet utilize nursing home services at lower rates than white persons. The authors hypothesized that older patients residing in nursing homes may differ clinically by race in ways that suggest new hypotheses about these differences. They compared clinical characteristics of a biracial, inner-city nursing home sample. No racial differences were found in prevalence of dementia-spectrum diagnoses. Depression was typically diagnosed more than twice as often in white patients, whereas black patients showed higher chronic medical illness burden. These results suggest that cognitive, medical, and psychiatric disabilities may interact differently in black and white patients to affect nursing home placement.


The Gaussian Generalized Born model: application to small molecules.
This work presents a Generalized Born model for the computation of the electrostatic component of solvation energies which is based on volume integration. An analytic masking function is introduced to remove Coulombic singularities. This approach leads to analytic formulae for the computation of Born radii, which are differentiable to arbitrary order, and computationally straightforward to implement.


A biochemical characterization of human breast tumors.
The activities of PHI, LDH and LDH isoenzymes were examined in 45 samples of human breast tissue. There were 25 carcinomas, 10 fibroadenomas and 10 nontumor tissue samples. In the group of nontumor breast tissue, values were as follows: PHI (X = 574 SD = 69), LDH (X = 634 SD = 91), LDH5/LDH1 (X = 0.42 SD = 0.23); in the benign group, PHI (X = 425 SD = 95), LDH (X = 59 SD = 188), LDH5/LDH1 (X = 0.55 SD = 0.38); and in the malignant tumor group, PHI (X = 2871 SD = 1350), LDH (X = 3108 SD = 1417), LDH5/LDH1 (X = 3.33 SD = 0.95). The enzyme activities of nontumor and benign tissue were statistically similar, but when these parameters were compared with the enzyme values of malignant tissue, there was a significant statistical difference. Based on these results, it is possible to establish a biochemical criterion for malignancy with the PHI and LDH activity levels greater than 720 U/g and the LDH5/LDH1 ration greater than 1.3. By using this criterion, the examined specimens can be categorized as nontumor or benign in one group and malignant in another group. This corresponds to the histiopathologic system of classification.


Effects of interventional therapy on AQP4 gene expression and neuron apoptosis in rabbits with ischemic brain injury caused by carotid artery stenosis.
Objective: To explore the effects of interventional therapy (IT) on Aquaporin-4 (AQP4) gene expression and neuron apoptosis in rabbits with ischemic brain injury caused by carotid artery stenosis, and to further optimize the therapeutic regime of clinical ischemic brain injury (IBI). Methods: 30 New Zealand rabbits were randomly divided into three groups: Sham group (n=10), IBI group (n=10) and IBI+IT group (n=10). A rabbit model of carotid artery stenosis was established to induce IBI. 12 hours later, interventional therapy was achieved through percutaneous transluminal coronary angioplasty (PTCA) by pumping up the balloon to widen the catheter. The expression of AQP4 mRNA and protein were determined by reverse transcription polymerase chain reaction (RT-PCR) and western blot, respectively. The distributions of rabbit hippocampus and cerebral cortex were detected by Nissl staining. The neuronal function of rabbits in the three groups after surgery was assessed with mNSS score. TUNEL (Terminal Transferase-mediated dUTP Nick End-labelling) staining was used to observe neuron apoptosis in rabbit brain tissue. The expressions of Bax and Bcl-2 protein were detected by western blot. Markers of oxidative stress in rabbit brain tissues were detected by Reactive Oxygen Species ELISA Kits. Morphology of the organelles in rabbit brain tissues was observed with transmission electron microscopy (TEM). Results: The protein and mRNA expressions of AQP4 in rabbit brain tissues were significantly decreased in the IBI group. Nissl staining results showed that IT improved IBI in rabbit hippocampus. In addition, IT significantly ameliorated the neuronal function of rabbits, and reduced the apoptosis and oxidative stress level of neurons in brain tissues. Furthermore, we found that after IT, organelle damage was significantly reduced in rabbit neurons. Conclusion: After receiving IT, AQP4 gene level and neuron apoptosis were significantly reduced in rabbit brain tissues with ischemic brain injury from carotid artery stenosis.


The social worker as moral citizen: ethics in action.
Social workers today face some of the most complex ethical dilemmas in the history of the profession. This article presents a framework of moral citizenship to guide ethical social work practice. The framework includes the action philosophies of philosopher Hannah Arendt and Lutheran theologian Paul Tillich integrated with concepts of professional responsibility and the unique contributions of social work pioneer Charlotte Towle. Social conscience and social consciousness, including awareness, thinking, feeling, and action, are major components of the framework.


Is parental sense of coherence associated with child health?
Antonowsky's concept of sense of coherence (SOC) has, during recent years, gained increased attention as a salutogenic model on the relationship between health and disease. However, only sparse information connecting child chronic health conditions to parental SOC is yet available. This article presents results from a cross-sectional study of about 10,000 children aged 2-17 years in the five Nordic countries in 1996. Factors associated with parental SOC were analysed, with focus on child chronic health conditions. Sense of coherence was measured according to a short and condensed three-item instrument based on Antonovsky's original 29-item instrument. Overall, about 23% of parents in Nordic countries had a poor sense of coherence, the lowest proportion found among Icelandic parents. Compared to the higher social classes, poor SOC was more common in the lower social classes. The association of child chronic health conditions with parental poor SOC was found to be disability specific. Parents of children with diabetes, epilepsy or psychiatric/nervous problems had approximately 2-5 higher odds of having poor SOC compared to parents of children without a specific diagnosis. The overall effect of having a child with chronic health conditions was, however, low, lower than the effect of the parents' own health complaints.


Protein S-Nitrosylation Controls Glycogen Synthase Kinase 3β Function Independent of Its Phosphorylation State.
GSK-3β (glycogen synthase kinase 3β) is a multifunctional and constitutively active kinase known to regulate a myriad of cellular processes. The primary mechanism to regulate its function is through phosphorylation-dependent inhibition at serine-9 residue. Emerging evidence indicates that there may be alternative mechanisms that control GSK-3β for certain functions. Here, we sought to understand the role of protein S-nitrosylation (SNO) on the function of GSK-3β. SNO-dependent modulation of the localization of GSK-3β and its ability to phosphorylate downstream targets was investigated in vitro, and the network of proteins differentially impacted by phospho- or SNO-dependent GSK-3β regulation and in vivo SNO modification of key signaling kinases during the development of heart failure was also studied. We found that GSK-3β undergoes site-specific SNO both in vitro, in HEK293 cells, H9C2 myoblasts, and primary neonatal rat ventricular myocytes, as well as in vivo, in hearts from an animal model of heart failure and sudden cardiac death. S-nitrosylation of GSK-3β significantly inhibits its kinase activity independent of the canonical phospho-inhibition pathway. S-nitrosylation of GSK-3β promotes its nuclear translocation and access to novel downstream phosphosubstrates which are enriched for a novel amino acid consensus sequence motif. Quantitative phosphoproteomics pathway analysis reveals that nuclear GSK-3β plays a central role in cell cycle control, RNA splicing, and DNA damage response. The results indicate that SNO has a differential effect on the location and activity of GSK-3β in the cytoplasm versus the nucleus. SNO modification of GSK-3β occurs in vivo and could contribute to the pathobiology of heart failure and sudden cardiac death.


Osmotic and diffusio-osmotic flow generation at high solute concentration. II. Molecular dynamics simulations.
In this paper, we explore osmotic transport by means of molecular dynamics (MD) simulations. We first consider osmosis through a membrane and investigate the reflection coefficient of an imperfectly semi-permeable membrane, in the dilute and high concentration regimes. We then explore the diffusio-osmotic flow of a solute-solvent fluid adjacent to a solid surface, driven by a chemical potential gradient parallel to the surface. We propose a novel non-equilibrium MD (NEMD) methodology to simulate diffusio-osmosis, by imposing an external force on every particle, which properly mimics the chemical potential gradient on the solute in spite of the periodic boundary conditions. This NEMD method is validated theoretically on the basis of linear-response theory by matching the mobility with their Green-Kubo expressions. Finally, we apply the framework to more realistic systems, namely, a water-ethanol mixture in contact with a silica or a graphene surface.


Life With HIV in a Canadian Suburban Community: A Qualitative Inquiry of Health Care and Social Services Access.
HIV has been examined in urban and rural contexts, but the suburban gradient has not been sufficiently described, despite the fact that many Canadians live in suburbia. Using qualitative description, we investigated how people living with HIV in a suburban community in Ontario, Canada, accessed health care and social services. Posters at the regional AIDS Service Organization and snowball sampling were used to recruit and interview 13 adult participants with various experiences and perspectives. A content analysis identified three meta-themes in the interviews: (a) transportation cost and time: barriers to access, (b) isolation, and (c) defective primary care: unmet and deflected needs. The findings have implications for the (a) development of community-based groups, (b) the role of transportation in health care and social services utilization, (c) community-based, interprofessional health and social care services, and (d) aging with HIV.


Utility of serology in determining Helicobacter pylori eradication after therapy.
To determine the usefulness of four serological tests in confirming cure of H pylori infection before the previously reported six-month post-treatment delay. As part of a prospective, blinded, controlled trial, in which patients with duodenal ulcers were randomized to receive different combinations of antibiotics, serum samples were obtained in 89 patients before treatment, as well as on several occasions after treatment. Antibody titres were determined by ELISA with Bio-Rad immunoglobulin (Ig) A, Bio-Rad IgG, Pyloriset EIA-A for IgA and Pyloriset EIA-G for IgG. Eradication was confirmed with antral biopsy three months after therapy. The percentage drop in titre following treatment was significantly larger for the group of patients who were treated successfully with all four kits. Optimal cut-offs for identifying successful therapy were determined, and accuracy improved as the interval between testing and therapy was prolonged. Six months after therapy, the IgG test from Bio-Rad achieved 100% sensitivity and 80% specificity, and that from Pyloriset achieved 88% sensitivity and 100% specificity. At three months, however, test performance was quite good, with 90% sensitivity and 80% specificity when using a Pyloriset IgA titre drop of 20% or greater to predict successful eradication. Serology is a simple, easily available, noninvasive method that exhibits good positive predictive value in the confirmation of successful cure of H pylori infection three or six months after treatment.


Dispositional Awe and Online Altruism: Testing a Moderated Mediating Model.
Dispositional awe has a positive effect on prosociality. However, it has not been tested whether this disposition appears in online altruism. Using a large sample of 3,080 Chinese undergraduates, this study tested a moderated mediating model that takes self-transcendent meaning in life (STML) as a mediator and subjective socioeconomic status (SSES) as a moderator. As predicted, dispositional awe was positively correlated with online altruism, partly via the indirect effect of STML. SSES moderated both the direct and indirect effects. Specifically, the predictive effects of dispositional awe on both online prosocial behavior and STML were greater for lower rather than higher SSES. This study extends the prosociality of dispositional awe to cyberspace. Other implications are also discussed herein.


Numerical comparison of acoustic wedge models, with application to ultrasonic telemetry.
Ultrasonic telemetry imaging systems are used to monitor such immersed structures as main vessels of nuclear reactors. The interaction between acoustic beams and targets involves scattering phenomena, mainly specular reflection and tip diffraction. In order to assist in the design of imaging systems, a simulation tool is required for the accurate modeling of such phenomena. Relevant high-frequency scattering models have been developed in electromagnetic applications, in particular, the geometrical optics (GO), Geometrical Theory of Diffraction (GTD) and its uniform corrections (UAT and UTD), Kirchhoff approximation (KA) and Physical Theory of Diffraction (PTD). Before adopting any of them for simulation of scattering of acoustic waves by edged immersed rigid bodies, it is important to realize that in acoustics the characteristic dimension to the wave length ratio is usually considerably smaller than in electromagnetics and a further study is required to identify models' advantages, disadvantages and regions of applicability. In this paper their numerical comparison is carried out. As the result, the most suitable algorithm is identified for simulating ultrasonic telemetry of immersed rigid structures.


Characterization of human malignant melanoma cell lines. VII. Glycoprotein synthesis and shedding as revealed by [3H]glucosamine labeling.
We have established conditions for the study of membrane glycoprotein synthesis and turnover in cultured human malignant melanoma cell lines using the labeled precursor [3H]glucosamine. Uptake of label increased parallel with cell growth, reaching a steady state in resting cultures. Fifteen to 30% of incorporated label can be released from the cells by trypsin treatment depending on the conditions of exposure to the enzyme, and about 50% of the incorporated label is spontaneously shed from the cells within 96 hr of incubation. Labeling in exhausted medium gave a 5- to 8-fold increase in uptake which was inhibited by addition of glucose (2 mg per ml) into the culture medium. The percentage of trypsin-releasable material was identical in fresh and exhausted medium; however, the percentage shed was less in cells initially labeled in exhausted medium. These data provide background information for further studies on the antigenic composition of the glycoproteins of cultured melanoma cells.


Inhibition of biliary phospholipid and cholesterol secretion by bilirubin in the Sprague-Dawley and Gunn rat.
Bilirubin is one of several organic anions that selectively inhibit the biliary secretion of phospholipid and cholesterol without affecting bile salt secretion. To determine the site of this inhibition and gain insight into its mechanism, normal, Sprague-Dawley and homozygous Gunn rats were infused with unconjugated bilirubin and a water-soluble model conjugate, bilirubin ditaurate. Either unconjugated bilirubin or bilirubin ditaurate infused into Sprague-Dawley rats caused an increase in biliary bilirubin secretion from 2.1 +/- 0.3 to 315 +/- 15 nmol/min, whereas phospholipid and cholesterol secretion fell significantly, in parallel, to 25% of control. Bilirubin ditaurate infused into homozygous Gunn rats caused changes in biliary lipid and bilirubin secretion similar to those seen in Sprague-Dawley rats. Biliary bilirubin secretion rose from 0.9 +/- 0.6 to 303 +/- 9 nmol/min, whereas biliary phospholipid and cholesterol secretion fell to 28% of controls. In contrast, unconjugated bilirubin infused into Gunn rats caused only a slight increase in biliary bilirubin secretion, from 0.9 +/- 0.3 to 6.0 +/- 2.1 nmol/min, whereas biliary lipid secretion remained within 90% of control. These results indicate that the inhibition of biliary phospholipid and cholesterol secretion by bilirubin occurs after microsomal conjugation. It is possible that the more hydrophilic conjugates of bilirubin aggregate with intracellular phospholipid and cholesterol destined for bile. Consequently, micelle-forming bile salts could be prevented from recognizing or aggregating with these lipids and fail to promote their secretion into bile.


[A Case of Penile Calciphylaxis in a Hemodialysis Patient].
In the present case of a 56-year-old male, hemodialysis was introduced from December 20XX-2 due to chronic renal failure caused by diabetic nephropathy. In February 20XX, a glans penis ulcer was observed. It gradually expanded. Angiography conducted in April revealed complete occlusion of the left internal pudendal artery and poor visualization of the bilateral penile arteries. Given the high risk of obstruction, endovascular treatment was not conducted. The glans penis ulcer continued to expand, and maintenance dialysis became difficult due to intractable pain. Opioids were introduced, but the pain could not be controlled. In May 20XX, the patient was referred to our department for surgical treatment, and partial penile resection was performed. The patient was diagnosed with penile calciphylaxis based on clinical findings and pathological diagnosis. After the surgery, the pain subsided considerably, and the patient is being followed on an out-patient basis.


Association of the major ganglioside in sea urchin eggs with yolk lipoproteins.
The predominant ganglioside in sea urchin eggs, M5 (NeuGc alpha 2-6Glc beta 1-1'Cer), exists mainly in the cytoplasm before and after fertilization. Using immunofluorescence microscopy, M5 ganglioside has been shown to localize in the network of cortical endoplasmic reticulum and unidentified vesicles in the eggs. In this study, the subcellular localization of M5 ganglioside in unfertilized eggs was further examined by immunoelectronmicroscopy using preembedding and postembedding labeling techniques. In order to prevent lipid extraction during dehydration and embedding, fixed eggs were subjected to rapid-freezing and freeze-substitution, and then embedded in hydrophilic resin. When ultrathin sections were treated with the anti-M5 ganglioside monoclonal antibody, the labeling of M5 ganglioside localized in the endoplasmic reticula, yolk granules, and microvilli. M5 ganglioside in yolk granules was not restricted to the membrane, suggesting that M5 ganglioside associated with soluble proteins or components in yolk granules. Indeed, gel filtration of the soluble fraction of the yolk lysate showed that M5 ganglioside was coeluted with yolk lipoproteins which were one of the main components in yolk granules. Moreover, M5 ganglioside was extracted from the electrophoretically purified yolk lipoproteins. These results demonstrate that M5 ganglioside in yolk granules associate with yolk lipoproteins. These data suggest that much of M5 ganglioside may be taken up into growing oocytes during oogenesis and transported from yolk granules to other cellular components during embryogenesis. M5 ganglioside associating with yolk lipoproteins in yolk granules may be a significant stored material to be utilized for early embryogenesis.


KMT2C is a Potential Biomarker of Anti-PD-1 Treatment Response in Metastatic Melanoma.
Metastatic melanoma (MM) represents a common malignancy with poor prognosis. Immune checkpoint inhibition (ICI), including PD-1 blockade, has been emerging as the popular therapeutic in MM for its durable treatment effect, but its response rate is still limiting. We comprehensively analyzed the associations between KMT2C somatic mutation and the tumor microenvironment as well as the ICI response of MM patients based on three published cohorts. Gene differential expression analysis between tumor samples with mutated and wild-type KMT2C was performed by DESeq2 package. Functional enrichment analysis was conducted by using clusterProfiler package. Kaplan-Meier was used to perform overall survival probability estimate through survival package and rms package was applied for the construction of nomogram model. We report here that KMT2C is a potential biomarker for anti-PD-1 treatment in MM. This biomarker can be used for comprehensively analyzing its association with patients' prognosis, tumor microenvironment and genomic features. Mutations of KMT2C profoundly altered expression of immune- and DNA replication-related genes in MM tumors. MM patients harboring KMT2C mutations showed significantly better overall survival (OS) after treatment with PD-1 monoclonal antibody as compared to wild-type KMT2C. Although KMT2C mutation has no significant influence on immune cell infiltration into MM tumors, the tumor mutation load and neoantigen load are indeed elevated in KMT2C mutated MM samples. This might represent a possible pathway through which KMT2C regulates the response of MM patients to anti-PD-1 treatment. Finally, we constructed a nomogram model by combing the independent prognostic factors, including KMT2C mutation, which could effectively predict the 1-year survival probability of MM patients after anti-PD-1 treatment. In conclusion, we report the role of KMT2C in anti-PD-1 treatment response regulation in MM for the first time. This may consequently be helpful for KMT2C personalized application.


Telling stories of vaccine-preventable diseases: why it works.
In this paper, we explore the benefits of storytelling in health communication and, in particular, immunization education. During the mid-20th century polio epidemic, both personal stories and scientific information abounded in the media. However, as rates of vaccine-preventable diseases declined, narratives about the dangers of such diseases faded as did the public fear of them. Meanwhile, anti-vaccine advocates flooded the media and Internet with stories of injured children and tied those injuries, such as autism, to vaccines. Medical experts often counter anti-vaccine concerns with scientific information which can fail to persuade parents. Furthermore, evidence suggests that many people misunderstand quantitative information resulting in a misinterpretation of risk. Compared to scientific information, stories relate life lessons and values. They are effective because they are memorable and relatable. Evidence also suggests that storytelling can effectively improve health knowledge and behaviors. Inspired by In Harm's Way--True Stories of Uninsured Texas Children by the Children's Defense Fund and Faces of Influenza by the American Lung Association, we published Vaccine-Preventable Disease: The Forgotten Story, a collection of photographs and personal stories of families affected by vaccine-preventable diseases. We have found that the stories included in our booklet capture all the benefits of storytelling. Given the many benefits of storytelling, providers should strive to include stories along with medical facts in their daily practice.


Ubiquitinated Proteins Isolated From Tumor Cells Are Efficient Substrates for Antigen Cross-Presentation.
We have previously shown that inhibition of the proteasome causes defective ribosomal products to be shunted into autophagosomes and subsequently released from tumor cells as defective ribosomal products in Blebs (DRibbles). These DRibbles serve as an excellent source of antigens for cross-priming of tumor-specific T cells. Here, we examine the role of ubiquitinated proteins (Ub-proteins) in this pathway. Using purified Ub-proteins from tumor cells that express endogenous tumor-associated antigen or exogenous viral antigen, we tested the ability of these proteins to stimulate antigen-specific T-cell responses, by activation of monocyte-derived dendritic cells generated from human peripheral blood mononuclear cells. Compared with total cell lysates, we found that purified Ub-proteins from both a gp100-specific melanoma cell line and from a lung cancer cell line expressing cytomegalovirus pp65 antigen produced a significantly higher level of IFN-γ in gp100- or pp65-specific T cells, respectively. In addition, Ub-proteins from an allogeneic tumor cell line could be used to stimulate tumor-infiltrating lymphocytes isolated and expanded from non-small cell lung cancer patients. These results establish that Ub-proteins provide a relevant source of antigens for cross-priming of antitumor immune responses in a variety of settings, including endogenous melanoma and exogenous viral antigen presentation, as well as antigen-specific tumor-infiltrating lymphocytes. Thus, ubiquitin can be used as an affinity tag to enrich for unknown tumor-specific antigens from tumor cell lysates to stimulate tumor-specific T cells ex vivo or to be used as vaccines to target short-lived proteins.


Hypothesis: when is a seminoma not a seminoma?
A xenograft line, HX 53, has been established in immune-suppressed mice from a specimen of a lymph node metastasis in a patient with a histological diagnosis of seminoma but with markedly raised circulating levels of alpha-fetoprotein. Histological, immunocytochemical, and ultrastructural studies of this xenograft line have suggested that a solid variant of yolk sac carcinoma may exist, which morphologically resembles seminoma, or that a continuum of differentiation exists between seminoma and yolk sac carcinoma.


Invasion and persistence of Streptococcus dysgalactiae within bovine mammary epithelial cells.
Little is known about bacterial and host factors that contribute to the establishment and persistence of intramammary infection by Streptococcus dysgalactiae. Streptococcus dysgalactiae adheres to epithelial cells from the bovine mammary gland and to extracellular matrix proteins in vitro and invades mammary epithelial cell cultures, all of which can be potentially important pathogenic mechanisms. In this study, mechanisms involved in the invasion of Strep. dysgalactiae into epithelial cells from the bovine mammary gland were characterized. Studies were conducted to determine whether Strep. dysgalactiae invaded mammary epithelial cell cultures in a dose-dependent manner and whether mammary epithelial cells that harbored different numbers of Strep. dysgalactiae for varying times were damaged. Bacterial invasion increased as inoculum size increased; however, the number of intracellular bacteria was not proportional to the inoculum size, increased; however, the number of intracellular bacteria was not proportional to the inoculum size, indicating that a finite number of organisms is capable of invading epithelial cells. No net increase in intracellular organisms was detected at any bacterial density evaluated; however, Strep. dysgalactiae remained viable throughout the evaluation. In addition, Strep. dysgalactiae did not appear to cause cell injury at any bacterial density or time point evaluated. These data suggest that Strep. dysgalactiae can survive within mammary epithelial cells for an extended time without losing viability or damaging the eukaryotic cell. This feature may be associated with the development of persistent infection and protection of organisms from antimicrobial drugs and host defense mechanisms and may provide a route for bacterial colonization of subepithelial tissues.


A Novel Indigenously Developed Device to Measure Bite Force.
In stomatology, the evaluation of bite power is crucial. It is considered a significant objective approach to evaluating masticatory performance. Bite force has become a significant outcome analysis index for various therapies in dentistry research. Presently several devices being used globally have their graces and faults. They are costly and also not available easily to the general dental practitioner. Development of a novel indigenous instrument for the measurement of human bite force. This paper describes an indigenously developed and researched instrument to measure human bite force. The sensor data (change in electronic resistance under applied force) will be read by the microprocessor and converted to force values in newton. The bite force result will be instantly displayed on the screen of the instrument and the device with which it is connected. The developed instrument is handy and user-friendly and can measure bite force accurately and repeatedly. In this research paper, an economical, lightweight, user-friendly, accurate, and reproducible human bite force measurement device is explained, which has been developed indigenously.


Molecular and clinical analysis of predictive biomarkers in non-small-cell lung cancer.
Non-small-cell lung cancer (NSCLC) is the leading cause of cancer-specific death in the USA and Europe. Over the last two decades, the pathogenetic mechanisms and the molecular alterations of NSCLC have been investigated more intensively, a number of potential therapeutic targets have been identified and new agents against specific molecular targets have been introduced in the treatment of NSCLC. Acquired abnormalities in the genes encoding RAS, p53, KRAS, EGFR and ALK, are particularly important in this field. Whenever targetable mutations are not found, the research of other genetic abnormalities can be useful to personalize chemotherapy. The attention has been focused, in particular, on the endonuclease excision repair cross-complementing1 and BRCA1 status. The use of antimetabolite drugs and the level of expression of their cellular targets seem to be correlated and influence the clinical efficacy of those agents. This review will focus on the role of predictive biomarkers for the treatment of non-small cell lung cancer.


Preoperative ketamine nebulization attenuates the incidence and severity of postoperative sore throat: A randomized controlled clinical trial.
Endotracheal intubation is the prominent cause of airway mucosal injury which results in postoperative sore throat (POST), with an incidence of 21%-65%. Although this complication is minor, if left unresolved, it produces significant agony and annoyance to the patient. This study was conducted to evaluate the efficacy of nebulized ketamine in decreasing POST. After written informed consent, 96 patients of the American Society of Anesthesiologists physical status (PS) 1-2 between 18 and 60 years, of either sex undergoing general anesthesia (GA) with tracheal intubation were enrolled in this prospective, randomized, placebo-control, and double-blind controlled trial. Patients were randomized into two groups; Group 1 received ketamine 50 mg (1.0 ml) with 4.0 ml of saline nebulization, while Group 2 received saline nebulization 5.0 ml for 15 min. GA was administered 15 min after completing nebulization. On reaching postanesthesia care unit, POST monitoring was done at 0, 2, 4, 6, 12, and 24 h after extubation. POST was graded on a four-point scale (0-3). The overall incidence of POST in this study was 25%: POST was experienced by 7 patients (14.6%) in ketamine and 17 patients (35.4%) in saline group (Fisher's exact P = 0.018). There was statistically significant reduction in the incidence of POST in ketamine group when compared to saline, at 2, 4, 6,12, and 24 h postoperatively (P < 0.05*). Severity of sore throat was also higher in saline group when compared to ketamine at 4 h (P = 0.030*) and 6 h (P = 0.016*) postextubation. Preoperative ketamine nebulization effectively reduced the incidence and severity of POST, with no adverse effects.


Knowledge and Perceptions Regarding Climacteric Among Rural Women in Jammu District of UT of J and K, India: A Cross-Sectional Study.
The word Climacteric is shrouded in a culture of silence in our country, more so in the vast rural swathes of India. This survey was carried out to assess the knowledge and perceptions regarding climacteric in rural females of the Jammu district. The present study was conducted in the Kot-Bhalwal health block of Jammu district. A sample of 215 was calculated assuming a minimum prevalence of 65% of menopausal symptoms. Using a two-step simple random sampling technique, a total of 245 postmenopausal women from four villages of the health block were interviewed using a semi-structured questionnaire. Among the sources of information about climacteric, family and friends were prime sources followed by community elders. About one-third (35.10%) of the respondents had no discussion about climacteric with anyone. The majority of the participants had a positive perception of climacteric and described it as a feeling of relief. About 77.9% of study participants rated their health as poor to fair. However, about one-fourth of women had taken health checkups for menopause-related symptoms. None of the respondents had any clue about oncological screening. Despite positive perceptions, there is a need to dispel the misconceptions with rigorous awareness and counseling. The role of village-level health workers along with ANM at Sub Health Centers/Health and wellness centers is the game-changer in this context. Every opportunity including Antenatal clinics should be utilized to educate as well as screen menopausal females for various health problems.


Secretion of the lantibiotics epidermin and gallidermin: sequence analysis of the genes gdmT and gdmH, their influence on epidermin production and their regulation by EpiQ.
The closely related lantibiotics epidermin and gallidermin are produced by Staphylococcus epidermidis Tu3298 and S. gallinarum Tu3928, respectively. The epidermin biosynthetic genes involved in maturation, regulation, and immunity have been identified previously. How epidermin or gallidermin is secreted, however, has remained unclear. Here, we characterize two additional genes, epiH and epiT, as well as the homologous gallidermin genes gdmH and gdmT. EpiT and GdmT are similar to one-component ABC transporters that are involved in the secretion of proteins or peptides. EpiH and GdmH are hydrophobic proteins without conspicuous similarities to other proteins. Comparison of the gene sequences revealed that epiT is incomplete, having an internal deletion that causes a frame shift and a second deletion at the 3'-end, while gdmT is intact. Introduction of epiT and epiH into the heterologous host S. carnosus (pTepi14) bearing the maturation and regulation genes had no significant effect on the rather low level of epidermin production. The presence of the homologous gdmT and gdmH, however, resulted in a strong increase (seven- to tenfold) in the production level, which is very likely to be due to increased efficiency of epidermin secretion. Both gdmT and gdmH were necessary for this effect, indicating that the two gene products cooperate in some way. In the epidermin-producing wild-type strain Tu3298, which contains epiH and the disrupted epiT, the addition of gdmT alone led to a two-fold increase in epidermin production. Both gdmT and gdmH and the corresponding epi genes were activated by the transcriptional regulator EpiQ; this is in accordance with the presence of putative EpiQ operator sites in the promoter regions.


Characterization of raloxifene glucuronidation in vitro: contribution of intestinal metabolism to presystemic clearance.
Raloxifene, a selective estrogen receptor modulator used for the treatment of osteoporosis, undergoes extensive conjugation to the 6-beta- and 4'-beta-glucuronides in vivo. This paper investigated raloxifene glucuronidation by human liver and intestinal microsomes and identified the responsible UDP-glucuronosyltransferases (UGTs). UGT1A1 and 1A8 were found to catalyze the formation of both the 6-beta- and 4'-beta-glucuronides, whereas UGT1A10 formed only the 4'-beta-glucuronide. Expressed UGT1A8 catalyzed 6-beta-glucuronidation with an apparent K(m) of 7.9 microM and a V(max) of 0.61 nmol/min/mg of protein and 4'-beta-glucuronidation with an apparent K(m) of 59 microM and a V(max) of 2.0 nmol/min/mg. Kinetic parameters for raloxifene glucuronidation by expressed UGT1A1 could not be determined due to limited substrate solubility. Based on rates of raloxifene glucuronidation and known extrahepatic expression, UGT1A8 and 1A10 appear to be primary contributors to raloxifene glucuronidation in human jejunum microsomes. For human liver microsomes, the variability of 6-beta- and 4'-beta-glucuronide formation was 3- and 4-fold, respectively. Correlation analyses revealed that UGT1A1 was responsible for 6-beta- but not 4'-beta-glucuronidation in liver. Treatment of expressed UGTs with alamethicin resulted in minor increases in enzyme activity, whereas in human intestinal microsomes, maximal increases of 8-fold for the 6-glucuronide and 9-fold for the 4'-glucuronide were observed. Intrinsic clearance values in intestinal microsomes were 17 microl/min/mg for the 6-glucuronide and 95 microl/min/mg for the 4'-isomer. The corresponding values for liver microsomes were significantly lower, indicating that intestinal glucuronidation may be a significant contributor to the presystemic clearance of raloxifene in vivo.


Kallikrein stimulates prostacyclin production in bovine vascular endothelial cells.
Cultured endothelial cells isolated from bovine carotid aorta produce prostacyclin (prostaglandin I2) and a small amount of prostaglandin E2. The effects of kallikrein (EC 184.108.40.206) on the release of prostacyclin from the cells were studied with the radioimmunoassay technique. Kallikrein stimulated the release of prostacyclin in a dose-dependent manner. The maximal stimulation reached up to 9.2-fold at 0.1 micrograms/ml of kallikrein. The effect was not associated with the activation of the fatty acid cyclooxygenase, but with the stimulation of arachidonic acid release. But kallikrein itself did not have phospholipase activity. On the other hand, at the same doses, kallikrein failed to induce platelet aggregation or enhance platelet aggregation induced by collagen. Our findings suggest that the vasodilator effect of kallikrein is mediated in part by prostacyclin production. Furthermore, we investigated the possibility that the stimulatory effect of kallikrein on prostacyclin production in endothelial cells is associated with kinin formation. Bradykinin and lysylbradykinin (kallidin) also stimulated the release of prostacyclin, but the effects were far less than that of kallikrein. And the stimulation due to the addition of both kallikrein and bradykinin on prostacyclin and arachidonic acid release was not competitive or additive, but synergistic. Moreover, even if fetal calf serum was incubated with kallikrein, bradykinin was not detected at all. When kallikrein was pre-incubated with aporotinin, which is an inactivator of kallikrein, the effect of kallikrein was completely abolished. These findings suggest that the stimulatory effect of kallikrein on the release of prostacyclin from vascular cells is possibly not due to kinin formation, but to other substance(s) produced by this serine proteinase.


[Noninferiority and equivalence randomized trials: lessons and reservations--secondary publication].
A noninferiority trial seeks to determine, whether a new intervention is not inferior to a reference intervention within a prespecified noninferiority interval (-Delta to 0) for the primary outcome. An equivalence trial aims to determine whether two interventions have a similar effect within a prespecified interval (-Delta to +Delta). These trials are controversial and difficult to grasp and very often poorly reported and conducted. Any claims regarding efficacy of new treatments, based on these trials, should be interpreted with caution, and it should be noted that Delta is often far too large to be clinically meaningful.


Electrochemical Strategy for Eradicating Fluconazole-Tolerant Candida albicans Using Implantable Titanium.
A persistent problem in modern health care derives from the overwhelming presence of antibiotic-resistant microbes on biomaterials, more specifically, fungal growth on metal-based implants. This study seeks to investigate the antifungal properties of low-level electrochemical treatments delivered using titanium electrodes against Candida albicans. We show that C. albicans can be readily controlled with electrical currents/potentials, reducing the number of viable planktonic cells by 99.7% and biofilm cells by 96.0-99.99%. Additionally, this study explores the ability of the electrochemical treatments to potentiate fluconazole, a clinically used antifungal drug. We have found that electrochemical treatment substantially enhances fluconazole killing activity. While fluconazole alone exhibits a low efficiency against the stationary phase and biofilm cells of C. albicans, complete eradication corresponding to 7-log killing is achieved when the antifungal drug is provided subsequently to the electrochemical treatment. Further mechanistic analyses have revealed that the sequential treatment shows a complex multimodal action, including the disruption of cell wall integrity and permeability, impaired metabolic functions, and enhanced susceptibility to fluconazole, while altering the biofilm structure. Altogether, we have developed and optimized a new therapeutic strategy to sensitize and facilitate the eradication of fluconazole-tolerant microbes from implantable materials. This work is expected to help advance the use of electrochemical approaches in the treatment of infections caused by C. albicans in both nosocomial and clinical cases.


Virulence factors of Aeromonas salmonicida subsp. salmonicida strains associated with infections in turbot Psetta maxima.
Virulence factors for Aeromonas salmonicida subsp. salmonicida (ASS) strains isolated from cultured turbot Psetta maxima L. are unknown with regard to this host. The presence of virulence genes associated with different stages of ASS infection in salmonids (vapA, tapA, fla, ascV, ascC, aexT, satA and aspA) was analysed using a polymerase chain reaction (PCR) technique in ASS strains isolated from turbot. Other ASS strains isolated from salmonids and environmental A. salmonicida (AS) strains were included for comparison. The presence of the genes was evaluated with respect to ASS virulence in turbot based on intraperitoneal and bath challenges. The genetic profile, including all of the genes studied, that was linked to virulent behaviour after intraperitoneal challenge was significantly more frequent in strains isolated from turbot than in those from salmonids or the environment. The data prove that it is not possible to predict the virulence of ASS in turbot based only on the presence of all genes tested. Moreover, the combined PCR results of vapA, aexT, ascV and ascC were useful for separating most of the ASS from environmental A. salmonicida strains. An association between virulence or genetic profile and the geographical or facility origin of the strains was not found.


Soluble and insoluble solids contributions to high-solids enzymatic hydrolysis of lignocellulose.
The rates and extents of enzymatic cellulose hydrolysis of dilute acid pretreated corn stover (PCS) decline with increasing slurry concentration. However, mass transfer limitations are not apparent until insoluble solids concentrations approach 20% w/w, indicating that inhibition of enzyme hydrolysis at lower solids concentrations is primarily due to soluble components. Consequently, the inhibitory effects of pH-adjusted pretreatment liquor on the enzymatic hydrolysis of PCS were investigated. A response surface methodology (RSM) was applied to empirically model how hydrolysis performance varied as a function of enzyme loading (12-40 mg protein/g cellulose) and insoluble solids concentration (5-13%) in full-slurry hydrolyzates. Factorial design and analysis of variance (ANOVA) were also used to assess the contribution of the major classes of soluble components (acetic acid, phenolics, furans, sugars) to total inhibition. High sugar concentrations (130 g/L total initial background sugars) were shown to be the primary cause of performance inhibition, with acetic acid (15 g/L) only slightly inhibiting enzymatic hydrolysis and phenolic compounds (9 g/L total including vanillin, syringaldehyde, and 4-hydroxycinnamic acid) and furans (8 g/L total of furfural and hydroxymethylfurfural, HMF) with only a minor effect on reaction kinetics. It was also demonstrated that this enzyme inhibition in high-solids PCS slurries can be approximated using a synthetic hydrolyzate composed of pure sugars supplemented with a mixture of acetic acid, furans, and phenolic compounds, which indicates that generally all of the reaction rate-determining soluble compounds for this system can be approximated synthetically.


Anti-vascular endothelial growth factor therapy in retinopathy of prematurity: An updated literature review.
Laser photocoagulation can still be considered the gold standard for treatment for retinopathy of prematurity (ROP). However, anti-vascular endothelial growth factor (anti-VEGF) therapy has increasingly become an important option that plays a significant role in the treatment of ROP. Major clinical trials have been published regarding the anti-VEGF use in ROP, along with multiple other studies looking into the different agents, doses, techniques, and possible complications. Anti-VEGF therapies can be considered as a safe and effective option for managing ROP. More longitudinal randomized clinical trials are necessary to evaluate the preferred treatment agent, the appropriate dose, best follow-up protocol, and the long-term ocular and systemic outcomes following treatment.


Systemic and regional hemodynamics in normotensive and spontaneously hypertensive rats after slow-channel calcium blocker nitrendipine.
To determine the effects of the recently synthesized slow channel calcium blocker nitrendipine on systemic and regional hemodynamics, mean arterial pressure, heart rate, cardiac index and organ blood flow distribution were measured with the combined radioactive reference sample and microsphere techniques before and one hour after its oral administration (1 mg or 10 mg/kg). Twenty-week-old normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) conscious male rats were studied. At the 10 mg dose, nitrendipine exerted a hypotensive effect within 15 minutes, and this effect persisted throughout the study. After one hour, mean arterial pressure fell by 14 percent of pretreatment levels in WKY controls; it fell by 25 percent in SHR (p less than 0.001). This action was mediated by a fall in total peripheral resistance index (p less than 0.01) that was nonuniformly distributed to the organ circulations since blood flow was maintained or increased to brain, heart, kidneys, lungs, and adrenals, but decreased to skin. No change in cardiac index was observed.


Acyl-protein thioesterase 2 catalyzes the deacylation of peripheral membrane-associated GAP-43.
An acylation/deacylation cycle is necessary to maintain the steady-state subcellular distribution and biological activity of S-acylated peripheral proteins. Despite the progress that has been made in identifying and characterizing palmitoyltransferases (PATs), much less is known about the thioesterases involved in protein deacylation. In this work, we investigated the deacylation of growth-associated protein-43 (GAP-43), a dually acylated protein at cysteine residues 3 and 4. Using fluorescent fusion constructs, we measured in vivo the rate of deacylation of GAP-43 and its single acylated mutants in Chinese hamster ovary (CHO)-K1 and human HeLa cells. Biochemical and live cell imaging experiments demonstrated that single acylated mutants were completely deacylated with similar kinetic in both cell types. By RT-PCR we observed that acyl-protein thioesterase 1 (APT-1), the only bona fide thioesterase shown to mediate deacylation in vivo, is expressed in HeLa cells, but not in CHO-K1 cells. However, APT-1 overexpression neither increased the deacylation rate of single acylated GAP-43 nor affected the steady-state subcellular distribution of dually acylated GAP-43 both in CHO-K1 and HeLa cells, indicating that GAP-43 deacylation is not mediated by APT-1. Accordingly, we performed a bioinformatic search to identify putative candidates with acyl-protein thioesterase activity. Among several candidates, we found that APT-2 is expressed both in CHO-K1 and HeLa cells and its overexpression increased the deacylation rate of single acylated GAP-43 and affected the steady-state localization of diacylated GAP-43 and H-Ras. Thus, the results demonstrate that APT-2 is the protein thioesterase involved in the acylation/deacylation cycle operating in GAP-43 subcellular distribution.


[Antihypertensive therapy and renoprotection: do we really need to block the renin-angiotension system?].
Recent studies questioned the existence of a specific renoprotective effects of ACE-inhibitors (ACE-i) and angiotensin receptor blockers (ARBs) besides their blood pressure lowering effect. In the ALLHAT study patients were randomly assigned to receive chlorthalidone, amlodipine and lisinopril. Results showed that, even in patients with reduced GFR, neither lisinopril nor amlodipine was superior to chlorthalidone in reducing the rate of development of ESRD or a 50% or greater decrement in GFR. Because of inclusion criteria the ALLHAT population was selected as at low risk for renal outcomes. Moreover, over 50% of the patients who were randomized to lisinopril either never received the medication or received the lower possible dose. Casas et al selected RCT comparing ACE-i and ARBs with other regimens. They concluded that ACE-i and ARBs are not more renoprotective that can be explained by lowering of blood pressure (BP) in diabetic nephropathy, while in non diabetic kidney disease a blood pressure independent renoprotective effect is uncertain. They made a very heterogeneous selection of trials that was dominated by the ALLHAT study; the analysis was not based on individual patient data. The Benedict Study showed that in hypertensive, normoalbuminuric patients with type 2 diabetes, BP reduction and ACE-i therapy both independently may prevent microalbuminuria. ACE-i therapy is particularly effective when BP is poorly controlled. We conclude that the recommendation of the Guidelines to use ACE-i and/or ARBs as first-line antihypertensive drugs for renoprotection in patients with diabetic and non diabetic kidney disease is still valid.


The Use of Mouse Mammary Tumor Cells in an In Vitro Invasion Assay as a Measure of Oncogenic Cell Behavior.
The in vitro invasion assay uses a protein-rich matrix in a Boyden chamber to measure the ability of cultured cells to pass through the matrix and a porous membrane in a process analogous to the initial steps of cancer cell metastasis. The tested cells can be altered for the gene expression or treated with inhibitors to test for changes in the invasion potential. This experiment tests the aggressive phenotype of the mouse mammary tumor cells to discover and characterize the potential oncogenes that promote cell invasion. This technique, however, can be versatile and adapted to many different applications. The experiment itself can be done in one day and the results are acquired by light microscopy in less than a day. The results include counts of the number of invading cells for comparison and analysis. The in vitro invasion assay is a rapid, inexpensive, and clear-cut method for determining cell behavior in a culture that can be used as an initial assessment before more involved in vivo assays.


Vulnerability of networks of interacting Markov chains.
The concept of vulnerability is introduced for a model of random, dynamical interactions on networks. In this model, known as the influence model, the nodes are arranged in an arbitrary network, while the evolution of the status at a node is according to an internal Markov chain, but with transition probabilities that depend not only on the current status of that node but also on the statuses of the neighbouring nodes. Vulnerability is treated analytically and numerically for several networks with different topological structures, as well as for two real networks--the network of infrastructures and the EU power grid--identifying the most vulnerable nodes of these networks.


A Statistical Method to Distinguish Functional Brain Networks.
One major problem in neuroscience is the comparison of functional brain networks of different populations, e.g., distinguishing the networks of controls and patients. Traditional algorithms are based on search for isomorphism between networks, assuming that they are deterministic. However, biological networks present randomness that cannot be well modeled by those algorithms. For instance, functional brain networks of distinct subjects of the same population can be different due to individual characteristics. Moreover, networks of subjects from different populations can be generated through the same stochastic process. Thus, a better hypothesis is that networks are generated by random processes. In this case, subjects from the same group are samples from the same random process, whereas subjects from different groups are generated by distinct processes. Using this idea, we developed a statistical test called ANOGVA to test whether two or more populations of graphs are generated by the same random graph model. Our simulations' results demonstrate that we can precisely control the rate of false positives and that the test is powerful to discriminate random graphs generated by different models and parameters. The method also showed to be robust for unbalanced data. As an example, we applied ANOGVA to an fMRI dataset composed of controls and patients diagnosed with autism or Asperger. ANOGVA identified the cerebellar functional sub-network as statistically different between controls and autism (p < 0.001).


Evaluation of herb-drug interaction of ambrisentan with shikonin based on UPLC-MS/MS.
Ambrisentan is an oral endothelin-receptor antagonist (ERA). However, there is no report on the interaction between ambrisentan and shikonin. To investigate the effect of shikonin on ambrisentan metabolism in vivo and in vitro. This study developed an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for simultaneous determination of ambrisentan and (S)-4-hydroxymethyl ambrisentan in rat plasma. Twelve male Sprague-Dawley (SD) rats were divided into two groups (n = 6): the control group and shikonin (20 mg/kg) group. The pharmacokinetics of ambrisentan (2.5 mg/kg) were investigated after 30 min. Additionally, human and rat liver microsomes were used to investigate the herb-drug interaction. The UPLC-MS/MS method was shown to be accurate, precise and reliable, and was successfully applied to the herb-drug interaction study of ambrisentan with shikonin. When co-administrated with 20 mg/kg shikonin, the Cmax and AUC(0-∞) of ambrisentan were significantly increased by 44.96 and 16.65%, respectively (p < 0.05). In addition, there were modest decreases in (S)-4-hydroxymethyl ambrisentan Cmax and AUC(0-∞) in the presence of shikonin (p < 0.05), which indicated that these results were in accordance with the inhibition of shikonin on ambrisentan metabolism. Moreover, enzyme kinetic study indicated that shikonin had an inhibitory effect on human and rat microsomes where the IC50 values of shikonin were 5.865 and 6.358 μM, respectively. Our study indicated that shikonin could inhibit ambrisentan metabolism. Further studies need to be carried out to verify whether similar interaction truly apply in humans and whether this interaction has clinical significance.


Determinants of emergency contraception non-use among women in unplanned or ambivalent pregnancies.
Objective To analyze the determinants of emergency contraception non-use among women in unplanned and ambivalent pregnancies. Method Cross-sectional study with a probabilistic sample of 366 pregnant women from 12 primary health care units in the city of São Paulo, Brazil. A multinomial logistic regression was performed, comparing three groups: women who used emergency contraception to prevent ongoing pregnancies (reference); women who made no use of emergency contraception, but used other contraceptive methods; and women who made no use of any contraceptive methods at all. Results Cohabitation with a partner was the common determinant of emergency contraception non-use. No pregnancy risk awareness, ambivalent pregnancies and no previous use of emergency contraception also contributed to emergency contraception non-use. Conclusion Apart from what is pointed out in the literature, knowledge of emergency contraception and the fertile period were not associated to its use.