Diploid and triploid Chinook salmon (Oncorhynchus tshawytscha) have altered microRNA responses in immune tissues after infection with Vibrio anguillarum.
Production of sterile fishes through artificial retention of a third set of chromosomes (triploidy) is a sustainable alternative for aquaculture since it reduces escapee pressure on wild populations. However, these fishes have reduced survival in stressful conditions and in response to infection. In this study, the impact of Vibrio anguillarum infection on diploid and triploid Chinook salmon (Oncorhynchus tshawytscha) was investigated to identify if there was any significant immune regulation by microRNAs (miRNA). Small RNAs from hindgut, head kidney, and spleen were sequenced to determine if miRNA transcript abundance was altered due to ploidy and infection in nine-month old full-sibling diploids and triploids. All three tissues had differentially expressed miRNA prior to infection, indicating subtle changes in epigenetic regulation due to increased ploidy. Additionally, miRNA were altered by infection, but there was only a difference in spleen miRNA expression between diploids and triploids at three days of infection. Furthermore, one miRNA (ssa-miR-2188-3p) was confirmed as having an altered response to infection in triploids compared to diploids, implicating potential immune dysregulation due to increased ploidy. The miRNAs identified in this study are predicted to target immune pathways, providing evidence for their importance in regulating responses to pathogens. This study is the first to investigate how increased ploidy alters miRNA expression in response to infection. Additionally, it provides evidence for epigenetic dysregulation in triploid fishes, which may contribute to their poor performance in response to stress.


Comparison of Ketamine, Dexmedetomidine and Lidocaine in Multimodal Analgesia Management Following Sleeve Gastrectomy Surgery: A Randomized Double-Blind Trial.
The aim of this study was to compare the effects of ketamine, dexmedetomidine, and lidocaine infusions added to the multimodal analgesia regimen on pain scores and analgesic requirement in laparoscopic sleeve gastrectomy. A prospective randomized double-blind trial. Seventy-three patients aged 18 to 65 years (ASA II-III) undergoing laparoscopic sleeve gastrectomy were included. The patients were divided into 3 groups. Intravenous (IV) ketamine (0.5 mg/kg/h), dexmedetomidine (0.5 mcg/kg/h), and lidocaine (2 mg/kg/h) were administered to Groups K, D and L, respectively. Postoperative infusions were continued for 12 hours. Visual Analog Scale (VAS) scores (during rest and movement) in the admission to postanesthesia care unit, 1, 3, 6, 12, 24, 48 hours, and on day 15 were assessed postoperatively. Rescue analgesia requirement, the number of patients with nausea, retching, and vomiting, time to mobilization, and hospital length of stay (LOS) were recorded. VASrest values during all measurements in the first 24 hours, and VASmovement values in the first 6 hours and at 24 hours were lower in Group L when compared to Group K and Group D (P < .001, P < .001, P = .008, respectively). VASrest at 48 hours and VASmovement at 12 and 48 hours were lower in Group L when compared to Group K (P = .044, P = .001 and P = .011, respectively). There was no statistically significant difference between Group D compared to the other two groups at these times (P > .05). The requirement of rescue analgesia on postoperative day 1 was significantly higher in Group K (P < .001). Hospital LOS was shorter in Group L than in the other groups (P = .002). IV lidocaine added to multimodal analgesia provided better pain control in the early postoperative period compared to dexmedetomidine and ketamine and decreased the hospital LOS.


The validity of smoking self-reports by adolescents: a reexamination of the bogus pipeline procedure.
Cigarette smoking was measured in a naive tenth grade population under conditions expected to influence the student's willingness to admit smoking. All students were tested for smoking both by questionnaire and by expired-air carbon monoxide assessment. The carbon monoxide data were used to test the equivalence of the study groups and to partition the sample into smokers and nonsmokers. Of the smokers those who were advised in advance of the biological test were twice as likely to admit cigarette use in the past week compared to those who were advised of the testing procedure only after they had completed their questionnaire. A live explanation and demonstration of the biological testing procedure proved as effective as a videotaped message. These data support earlier reports of the 'bogus pipeline' effect. Several methodological issues are discussed which may explain previous failures to replicate this finding.


A simple scoring rule to predict survival to discharge after out of hospital cardiac arrest at the time of ED arrival.
It is important to be able to predict the chance of survival to hospital discharge upon ED arrival in order to determine whether to continue or terminate resuscitation efforts after out of hospital cardiac arrest. This study was conducted to develop and validate a simple scoring rule that could predict survival to hospital discharge at the time of ED arrival. This was a multicenter retrospective cohort study based on a nationwide registry (Korean Cardiac Arrest Research Consortium) of out of hospital cardiac arrest (OHCA). The study included adult OHCA patients older than 18 years old, who visited one of 33 tertiary hospitals in South Korea from September 1st, 2015 to June 30th, 2020. Among 12,321 screened, 5471 patients were deemed suitable for analysis after exclusion. Pre-hospital ROSC, pre-hospital witness, shockable rhythm, initial pH, and age were selected as the independent variables. The dependent variable was set to be the survival to hospital discharge. Multivariable logistic regression (LR) was performed, and the beta-coefficients were rounded to the nearest integer to formulate the scoring rule. Several machine learning algorithms including the random forest classifier (RF), support vector machine (SVM), and K-nearest neighbor classifier (K-NN) were also trained via 5-fold cross-validation over a pre-specified grid, and validated on the test data. The prediction performances and the calibration curves of each model were obtained. Pre-processing of the registry was done using R, model training & optimization using Python. A total of 5471 patients were included in the analysis. The AUROC of the scoring rule over the test data was 0.7620 (0.7311-0.7929). The AUROCs of the machine learning classifiers (LR, SVM, k-NN, RF) were 0.8126 (0.7748-0.8505), 0.7920 (0.7512-0.8329), 0.6783 (0.6236-0.7329), and 0.7879 (0.7465-0.8294), respectively. A simple scoring rule consisting of five, binary variables could aid in the prediction of the survival to hospital discharge at the time of ED arrival, showing comparable results to conventional machine learning classifiers.


Spontaneous flow created by active topological defects.
Topological defects are at the root of the large-scale organization of liquid crystals. In two-dimensional active nematics, two classes of topological defects of charges [Formula: see text] are known to play a major role due to active stresses. Despite this importance, few analytical results have been obtained on the flow-field and active-stress patterns around active topological defects. Using the generic hydrodynamic theory of active systems, we investigate the flow and stress patterns around these topological defects in unbounded, two-dimensional active nematics. Under generic assumptions, we derive analytically the spontaneous velocity and stall force of self-advected defects in the presence of both shear and rotational viscosities. Applying our formalism to the dynamics of monolayers of elongated cells at confluence, we show that the non-conservation of cell number generically increases the self-advection velocity and could provide an explanation for their observed role in cellular extrusion and multilayering. We finally investigate numerically the influence of the Ericksen stress. Our work paves the way to a generic study of the role of topological defects in active nematics, and in particular in monolayers of elongated cells.


L-leucine and its analogue: specific inhibitors for S-benzyl-L-cysteine-p-nitroanilide-hydrolyzing enzyme in Escherichia coli B.
An enzyme that catalyzes hydrolysis of S-benzyl-L-cysteine-p-nitroanilide was purified from E. coli B. The enzyme was a monomer with a molecular weight of 82,000. In addition to L-cysteinylglycine, the enzyme hydrolyzed various glycine-containing dipeptides most efficiently at pH 7.0. The enzyme required no metal ions for activity and was specifically inhibited by L-leucine and its analogue with free carboxyl group at the physiological concentrations.


Nontoxic suramin treatments enhance docetaxel activity in chemotherapy-pretreated non-small cell lung xenograft tumors.
We reported that nontoxic suramin treatments enhance the activity of chemotherapy in preclinical models, a finding supported by the results of subsequent phase I/II trials in chemotherapy-naive non-small cell lung cancer (NSCLC) patients who received/carboplatin (P/C) combination therapy. The present study evaluated whether suramin enhances the activity of docetaxel in human NSCLC xenografts. The in vitro effect of suramin on docetaxel activity was evaluated using 3-D histocultures of chemotherapy-naive A549 tumors. For in vivo activity evaluation, we first established the P/C pretreatment schedule that produced tumor growth inhibition, but not tumor eradication, and established the maximally tolerated docetaxel/suramin regimens. In the second study, P/C-treated animals received physiological saline, single-agent suramin (10 mg/kg), docetaxel (10 mg/kg), or the combination twice weekly for 3 weeks. The in vitro results showed that 20 microM suramin, which had no activity as single agent, enhanced the docetaxel activity (measured as 50% inhibition of DNA synthesis) by more than 10-fold. The in vivo studies showed reduced tumor growth by P/C (30% growth in 14 days vs. 75% in control). In contrast, docetaxel produced tumor regression (15% reduction) in P/C-treated animals, significantly reduced, on a cellular level, the viable cell density and the proliferating fraction (40% reduction for both measurements), and enhanced the apoptotic fraction 4-fold (p < 0.05 for all effects). Suramin had no activity or toxicity (measured as body weight loss) but significantly enhanced the docetaxel activity. Compared to docetaxel alone, the combination showed earlier onset of tumor size reduction, greater extent of tumor regression (31 vs. 15%), greater reduction of viable cell density and proliferating fraction (additional 15-25% reduction), and greater apoptotic fraction (additional 2.5-fold increase) (p < 0.05 for all parameters). Results of the present study indicate that nontoxic suramin treatments enhanced the activity of docetaxel in P/C-pretreated A549 xenograft tumors in mice without enhancing host toxicity. These encouraging results provided the basis for phase I/II trials of docetaxel plus low-dose suramin in patients with NSCLC in second-/third-line settings.


Overestimate of remediation efficiency due to residual sodium persulfate in PAHs contaminated soil and a solution.
Oxidation remediation is a commonly used technology for PAHs contaminated soil presently, but the overestimate of efficiency due to ongoing remediation by residual oxidants during extraction and testing has not been paid enough attention. In this study, persulfate was activated by Fe(II) to investigate the effects of residual oxidants on PAHs removal during detection process and the elimination effects of adding Na2SO3 and extending sampling time on residual oxidants. Results verified that the residual oxidants removed PAHs in extraction process, making the results lower than the actual values: the detection recovery rate η of ∑PAHs and 3-6 ring PAHs ranged from 24.3% (25% Na2S2O8 treatment) to 87.4% (5% Na2S2O8+4/4Fe2+ treatment), 20.1%-99.0%, 28.9%-87.9%, 20.8%-89.4%, and 18.6%-76.9%, respectively. After adding Na2SO3, the accuracy of detection results increased significantly: the η of ∑PAHs and 3-6 ring PAHs increased to 64.1%-96.5%, 58.8%-95.5%, 73.8%-114.4%, 60.6%-95.6%, and 45.4%-77.1%, respectively. After 49 days of adding oxidants, residual oxidants had no considerable effect on the detection of PAHs, indicating it was appropriate to start soil remediation verification sampling49 days after the remediation was completed. The observed results will help scientific evaluation of the remediation effects of chemical oxidation on organic contaminated soil.


Significance of elevated serum thyrotropin during treatment of congenital hypothyroidism.
Serum thyrotropin concentrations are frequently elevated during treatment of children with congenital hypothyroidism. It is unclear if elevated thyrotropin during early treatment indicates non-optimal treatment. In a cohort of 49 children with congenital hypothyroidism, we studied the decline in serum thyrotropin concentration after initiating L-thyroxine treatment, the relationship between elevated thyrotropin and treatment variables, and non-compliance with the treatment as a possible cause of elevated thyrotropin. The initial mean dose of thyroxine was 8.5 (SD 3.3) micrograms/kg body weight/day: 71% of the serum samples obtained 15-21 days after the start of treatment had serum thyrotropin concentrations < 10 mU/l. Six children had no samples with serum thyrotropin < 10 mU/l during the first 3 months of treatment. These children had a lower thyroxine dose prescribed, and serum thyrotropin was normalized when the dose was sufficiently increased. During treatment, from 6 weeks of age, serum thyrotropin > 10 mU/l was related to a lower dose of thyroxine and lower serum thyroxine, and was not due to non-compliance with treatment.


Effect of the whole seamless connection of nursing from WeChat interactive platform on stigma and quality of life in patients with urinary system cancer.
To evaluate the effects of intervention by "whole seamless connection of nursing from WeChat interactive platform" on stigma and quality of life of the patients with urinary system cancer. Overall, 80 patients with urinary cancer were randomly divided (40 cases per group) into control and observation groups. Routine nursing was provided to the control group, whereas positive psychological intervention was provided to the intervention group through a "whole seamless connection of nursing from the WeChat interactive platform" in addition to routine nursing. The Chinese version of social impact and cancer patients' quality of life scales were used to evaluate the effects before and after the intervention. After the intervention, the total score for stigma was significantly lower (p < 0.01), while that of quality of life was higher (p < 0.05) in the observation group relative to the control group. The whole seamless connection of nursing from the WeChat interactive platform could reduce stigma and improve the quality of life of patients with urinary cancer.


Treatment of acute promyelocytic leukemia: strategy toward further increase of cure rate.
Acute promyelocytic leukemia (APL) has become a curable disease by all-trans retinoic acid (ATRA)-based induction therapy followed by two or three courses of consolidation chemotherapy. Currently around 90% of newly diagnosed patients with APL achieve complete remission (CR) and over 70% of patients are curable. To further increase the CR and cure rates, detection and diagnosis of this disease at its early stage is very important, hopefully before the appearance of APL-associated coagulopathy. In induction therapy, concomitant chemotherapy is indispensable, except for patients with low initial leukocyte counts. Prophylactic use of intrathecal methotrexate and cytarabine should be done, particularly for patients with hyperleukocytosis. If patients relapse hematologically or even molecularly, arsenic trioxide will be the treatment of choice under careful electrocardiogram monitoring. Am80, liposomal ATRA, gemtuzumab ozogamicin or ATRA in combination with cytotoxic drugs may be used at this stage or later. Allogeneic SCT will be the treatment of choice after patients of age <50 years have relapsed, provided that they have HLA-identical family donors or DNA-identical unrelated donors.


Apoptosis in leukemia: from molecular pathways to targeted therapies.
Knowledge about apoptosis pathways developed in the last few years is already being translated into novel targeted therapies. Two illustrative approaches to this process are the development of small molecule BCL2 inhibitors and X-linked inhibitors of apoptosis proteins (XIAP) inhibitors. This work demonstrates both the development of new agents based on molecular mechanisms and how these agents further our understanding of the biology of apoptosis.


Stimulation of 2-deoxyglucose uptake in rat adipocytes by a human growth hormone fragment (hGH 4-15).
The in vivo and in vitro effects of a hypoglycaemic fragment of human growth hormone containing the sequence H2N-Ile-Pro-Leu-Ser-Arg-Leu-Phe-Asp-Asn-Ala-Met-Leu-COOH (hGH 4-15) on 2-deoxy-[1-14C]-D-glucose uptake in adipocytes were studied. The isolated cells from rats after a single intravenous injection of hGH 4-15 (1 mg/kg) significantly increased uptake of 2-deoxyglucose (p less than 0.005). Adipocytes from untreated rats pre-incubated with the hGH fragment (10 micrograms/ml) at 37 degrees C for 30 min also clearly showed an elevated uptake of 2-deoxyglucose in the absence and the presence of exogenous insulin. The effect of hGH 4-15 was concentration-dependent, steadily increasing with the maximum effect at 10 micrograms/ml. The present findings suggest that the enhancement of glucose transport in target tissues may be a major contributing factor to the hypoglycaemic action of the amino-terminal fragments of human growth hormone.


Statins Reduce Hepatocellular Carcinoma Risk in Patients with Chronic Kidney Disease and End-Stage Renal Disease: A 17-Year Longitudinal Study.
Hepatocellular carcinoma (HCC) is the most common cancer in end-stage renal disease (ESRD) patients in Taiwan. Whether statin therapy associated with the HCC risk in hyperlipidemic patients with chronic kidney disease (CKD) and ESRD is unclear. Using population-based insurance claim data from Taiwan, we identified from hyperlipidemic patients taking statins or not (677,364 versus 867,707) in 1999-2015. Among them, three pairs of propensity score matched statin and non-statin cohorts were established by renal function: 413,867 pairs with normal renal function (NRF), 46,851 pairs with CKD and 6372 pairs with ESRD. Incidence rates of HCC were compared, by the end of 2016, between statin and non-statin cohorts, between hydrophilic statins (HS) and lipophilic statins (LS) users, and between statin-ezetimibe combination therapy (SECT) and statin monotherapy (SM) users. The HCC incidence increased progressively from NRF to CKD and ESRD groups, was lower in the statin cohort than in the non-statin cohort, with the differences of incidence per 10,000 person-years increased from (7.77 vs. 21.4) in NRF group to (15.8 vs. 37.1) in CKD group to (19.1 vs. 47.8) in ESRD group. The incidence increased with age, but the Cox method estimated hazard ratios showed a greater statin effectiveness in older patients. Among statin users, the HCC incidence was lower in HS users than in LS users, and lower in SECT users than in SM users, but the difference was significant only in the NRF group. Hyperlipidemic patients with CKD and ESRD receiving statins are at reduced HCC risks; the treatment effectiveness is superior for HS users than for LS users, and for SECT users than for SM users, but not significant.


Modelling social readers: novel tools for addressing reception from online book reviews.
Social reading sites offer an opportunity to capture a segment of readers' responses to literature, while data-driven analysis of these responses can provide new critical insight into how people 'read'. Posts discussing an individual book on the social reading site, Goodreads, are referred to as 'reviews', and consist of summaries, opinions, quotes or some mixture of these. Computationally modelling these reviews allows one to discover the non-professional discussion space about a work, including an aggregated summary of the work's plot, an implicit sequencing of various subplots and readers' impressions of main characters. We develop a pipeline of interlocking computational tools to extract a representation of this reader-generated shared narrative model. Using a corpus of reviews of five popular novels, we discover readers' distillation of the novels' main storylines and their sequencing, as well as the readers' varying impressions of characters in the novel. In so doing, we make three important contributions to the study of infinite-vocabulary networks: (i) an automatically derived narrative network that includes meta-actants; (ii) a sequencing algorithm, REV2SEQ, that generates a consensus sequence of events based on partial trajectories aggregated from reviews, and (iii) an 'impressions' algorithm, SENT2IMP, that provides multi-modal insight into readers' opinions of characters.


Impact of Overtraining on Cognitive Function in Endurance Athletes: A Systematic Review.
Endurance sports require significant training loads to elicit the desired training effects on an athlete's body. However, if adequate recovery is not provided, overtraining may occur, with corresponding decrements in performance. As such, there is a need for measures that can be tracked, to monitor athlete adaptation to training loads, and provide early warning of possible maladaptation. The aim of this review was to determine if a relationship exists between overtraining and cognitive function in endurance athletes. A systematic search of AMED, MEDLINE, SPORTDiscus and APA PsycINFO was conducted. Eligibility criteria included original peer reviewed research, written in English, published between January 2000 and May 2022, and using human participants. Of the 221 articles screened, a total of seven studies met the inclusion criteria and were included in this review. The findings of the review suggest that there is a relationship between overtraining and cognitive function with all seven studies finding that cognitive performance declined in response to athletes becoming overreached or overtrained. These studies found that reaction time (as measured by Stroop colour test) increased in response to both overreaching and overtraining. Cognitive function tests such as the Stroop Colour Test could be included as part of a broader programme for monitoring athlete adaptation to training.


Estrogens, androgens and generalized behavioral arousal in gonadectomized female and male C57BL/6 mice.
General arousal has been operationally defined as an enhanced motor activity and enhanced intensity of response to sensory stimuli. Even though the effects of gonadal hormones on mating behavior have been much studied, their potential effect on generalized arousal, as defined above, has never been evaluated. In the present study we employed a thoroughly validated assay of general arousal to determine the effects of estradiol (E) and testosterone (T) in gonadectomized female and male mice, respectively. The steroids were administered in three different ways: A fast-acting, water soluble preparation given intraperitoneally, an oil solution given subcutaneously, and an oil solution in a subcutaneous Silastic capsule. Motor activity and responses to sensory stimuli were recorded for 24h, 91h, and seven days following hormone administration, respectively. All measures of arousal varied according to the day/night cycle. The water soluble steroid preparation had no reliable effect. When the same doses of estradiol and testosterone were administered subcutaneously in an oil vehicle no effect of either treatment on arousal was observed. The subcutaneously implanted capsule containing estradiol or testosterone had a delayed effect on motor activity in females (four to seven days) but no effect in males. The long time required by the gonadal hormones for affecting arousal would be consistent with, but does not prove, a genomic action. The limited effects of E and T in our arousal assay suggest to us that the strongest actions of these hormones on arousal occur in the context of sequences of responses to sexually relevant stimuli.


Morphological and functional evidence for sexual dimorphism in neurokinin B signalling in the retrochiasmatic area of sheep.
Neurokinin B (NKB) is critical for fertility in humans and stimulates gonadotrophin-releasing hormone/luteinising hormone (LH) secretion in several species, including sheep. There is increasing evidence that the actions of NKB in the retrochiasmatic area (RCh) contribute to the induction of the preovulatory LH surge in sheep. In the present study, we determined whether there are sex differences in the response to RCh administration of senktide, an agonist to the NKB receptor (neurokinin receptor-3 [NK3R]), and in NKB and NK3R expression in the RCh of sheep. To normalise endogenous hormone concentrations, animals were gonadectomised and given implants to mimic the pattern of ovarian steroids seen in the oestrous cycle. In females, senktide microimplants in the RCh produced an increase in LH concentrations that lasted for at least 8 hours after the start of treatment, whereas a much shorter increment (approximately 2 hours) was seen in males. We next collected tissue from gonadectomised lambs 18 hours after the insertion of oestradiol implants that produce an LH surge in female, but not male, sheep for immunohistochemical analysis of NKB and NK3R expression. As expected, there were more NKB-containing neurones in the arcuate nucleus of females than males. Interestingly, there was a similar sexual dimorphism in NK3R-containing neurones in the RCh, NKB-containing close contacts onto these RCh NK3R neurones, and overall NKB-positive fibres in this region. These data demonstrate that there are both functional and morphological sex differences in NKB-NK3R signalling in the RCh and raise the possibility that this dimorphism contributes to the sex-dependent ability of oestradiol to induce an LH surge in female sheep.


Simultaneous estimation of the intermediate correlation matrix for arbitrary marginal densities.
A popular approach to the simulation of multivariate, non-normal data in the social sciences is to define a multivariate normal distribution first, and then alter its lower-dimensional marginals to achieve the shape of the distribution intended by the researchers. A consequence of this process is that the correlation structure is altered, so further methods are needed to specify an intermediate correlation matrix in the multivariate normal distribution step. Most of the techniques available in the literature estimate this intermediate correlation matrix bivariately (i.e., correlation by correlation), risking the possibility of generating a non-positive definite matrix. The present article addresses this issue by offering an algorithm that estimates all elements of the intermediate correlation matrix simultaneously, through stochastic approximation. A small simulation study demonstrates the feasibility of the present method to induce the correlation structure both in simulated and empirical data.


Presentation of histoplasmosis as mononucleosis syndrome in an immunocompetent patient.
Histoplasmosis is a fungal disease usually occurring in endemic areas that can affect immuno-impaired patients in whom pulmonary involvement is the rule. We present the case of an 18 year-old immunocompetent, male patient, resident of the State of Florida, who showed signs of mononucleosis syndrome that included odynophagia, cervical adenomegaly, sporadic fever and rash; however, no pulmonary involvement or visceromegaly were present. Faced with this atypical and unexpected clinical picture, histoplasmosis infection was eventually diagnosed following cervical lymph-node biopsy. Disseminated histoplasmosis may have unexpected manifestations, as is pointed out in the case described below.


Codon Optimization, Cloning, Expression, Purification, and Secondary Structure Determination of Human ETS2 Transcription Factor.
Transcription factor ETS2 regulates genes involved in development, differentiation, angiogenesis, proliferation, and apoptosis. In addition, it is one of the core reprogramming factors responsible for the generation of human cardiomyocytes from adult somatic cells. In this study, we report the heterologous expression of human ETS2 in E. coli to produce a highly pure recombinant protein. To accomplish this, the codon-optimized 1507 bp coding sequence of the human ETS2 gene in fusion with a His-tag, a cell-penetrating peptide, and a nuclear localization sequence was cloned in the protein expression vector and transformed into E. coli strain BL21(DE3) for expression. The recombinant protein was purified to homogeneity under native conditions using immobilized metal ion affinity chromatography, and its identity was confirmed by Western blotting with an ETS2 antibody. Using far-UV circular dichroism spectroscopy, we have demonstrated that the recombinant protein has retained its secondary structure, predominantly comprising of random coils and β-sheets. Prospectively, this biological recombinant ETS2 protein can substitute viral and genetic forms of ETS2 in a cell reprogramming process to facilitate the generation of clinical-grade cells. It can also be used to investigate its molecular role in various biological processes and diseases and for biochemical and structural studies.


Profiles of Little-Known Medicinal Polypores: Haploporus odorus (Agaricomycetes).
The purpose of this study was to compile a comprehensive characterization of little-known polypores, which have recently been found to possess anticancer activity and thus can also be used in cancer target therapy. Haploporus odorus is a polypore of Holarctic distribution and has been found by harvesters working in taiga floodlands and broadleaf forests of the Northern Hemisphere. A substance known as haploporic acid A was determined in methylene chloride extract from the dried basidiomata of H. odorus. This substance can be used in cancer therapy; more details of its health benefits could be used in mycotherapy.


A Developmental Approach to Pediatric Oral Health.
Orofacial growth and development is a complex process spanning the life course. This article provides an oral health overview in the context of overall growth, physical and social development from infancy through adolescence. It reviews oral health-specific developmental milestones during childhood (0-12 years) and adolescence (≥13 years). It examines issues particular to each age category or spanning multiple ages (eg, pediatric overweight and obesity, tobacco use, and dental trauma) in relation to oral health and development. In addition, the oral microbiome and its potential role in informing personalized oral health care across the life course is discussed.


Transperineal minimally invasive abdomino-perineal resection: preliminary outcomes and future perspectives.
Abdomino-perineal resection (APR) for rectal cancer is challenging, due to the difficult exposure of the surgical field. Many investigations proved worst results in terms of circumferential resection margin (CRM) involvement compared to rectal anterior resection (RAR) with total mesorectal excision (TME). Extralevator abdomino-perineal excision (ELAPE) improved oncologic outcomes, but is burdened by important limitations (positioning, wound closure). Applying the concept of transanal minimally invasive surgery (TAMIS) and the experience in transanal TME (TaTME) to the perineal phase of APR could overcome these limitations. A series of consecutive cases of transperineal minimally invasive APR was matched with an historical series of standard laparoscopic APR. Primary endpoints were global complication rate and CRM involvement, secondary endpoints were operative time, time to flatus and oral feeding and length of hospital stay. Fifteen patients underwent a transperineal minimally invasive APR for cancer, median age was 65 (49-88) years, median distance from the anal verge was 3 (2-5) cm and six patients underwent neoadjuvant chemo-radiotherapy. No intraoperative complications occurred in both groups; need of post-operative blood transfusions was significantly higher in the traditional laparoscopic APR group. No differences were reported in terms of wound dehiscence between the two groups. Positive CRM was reduced in transperineal minimally invasive APR versus standard laparoscopic APR, but the difference was not statistically significant. Transperineal minimally invasive APR appears to be safe and could improve post-operative and oncologic outcomes by means of better vision and reduced surgical trauma.


Management of fatigue in cancer patients.
Cancer-related fatigue (CRF) is either a symptom or a syndrome depending on criteria for diagnosis. CRF is present in 20% to 30% of long-term cancer survivors and 80% to 90% during treatment and at the end of life. Assessment requires determining the presence, severity, and interference with daily activities. Different descriptors for fatigue (eg, tiredness, lack of vigor) measure different patient experiences. Associated factors such as depression, pain, insomnia, dyspnea, anemia, and deconditioning worsen CRF and should be treated if present. Associated factors that contribute to the severity of fatigue differ depending on the stage of cancer. Pharmacologic interventions include recombinant erythropoietin, psychostimulants, corticosteroid, anti-inflammatory drugs other than steroids, and L-carnitine. Advances in the management of CRF will require an understanding of the underlying mechanism before target-specific therapies can be developed.


Agrobacterium- tumefaciens-mediated transformation of Helminthosporium turcicum, the maize leaf-blight fungus.
Agrobacterium tumefaciens has the ability to transfer its T-DNA to plants, yeast, filamentous fungi, and human cells and integrate it into their genome. Conidia of the maize pathogen Helminthosporium turcicum were transformed to hygromycin B resistance by a Agrobacterium- tumefaciens-mediated transformation system using a binary plasmid vector containing the hygromycin B phosphotransferase ( hph) and the enhanced green fluorescent protein ( EGFP) genes controlled by the gpd promoter from Agaricus bisporus and the CaMV 35S terminator. Agrobacterium- tumefaciens-mediated transformation yielded stable transformants capable of growing on increased concentrations of hygromycin B. The presence of hph in the transformants was confirmed by PCR, and integration of the T-DNA at random sites in the genome was demonstrated by Southern blot analysis. Agrobacterium- tumefaciens-mediated transformation of Helminthosporium turcicum provides an opportunity for advancing studies of the molecular genetics of the fungus and of the molecular basis of its pathogenicity on maize.


Effect of night temperature on daytime stomatal conductance in early and late successional plants.
Sixteen annuals, biennials, and herbaceous and woody perennials characteristic of early and late successional old field ecosystems and upland and floodplain habitats were analyzed for their response of stomatal conductance to changes in night temperature. Early successional species that germinate in early spring when temperatures are low, but above freezing are insensitive to cool nights, i.e., their conductance in the following days is unaffected by low night temperature. Later spring and summer-emerging species' stomatal conductance is inhibited by low temperatures. Tree species show the same effects and in some an enhancement of stomatal conductance by low night temperatures was observed. However, adaptive differences in response to night temperatures appear related to both phenology of germination and growth and habitat types.


Successful off pump coronary artery bypass grafting in a patient with an undiagnosed pheochromocytoma.
We report a successful off pump coronary artery bypass case in a patient with a pre-operatively undiagnosed pheochromocytoma. The patient had no signs of ischemia intra- or post-operatively due to aggressive antihypertensive treatment. We would also like to emphasize the importance of using an adequate stabilizing device in order to safely perform anastomoses in a situation like this.


Uptake and retention of moulting hormones by the integument of crayfishes in vitro: I. Influence of temperature, hormone concentration and hormone structure.
Uptake kinetics of [3H]ecdysone into crayfish hypodermis in vitro reveals three different components: a very rapid (within 1 min), highly temperature-dependent (Q10 = 3) and high uptake rate, a second much lower uptake which is only slightly temperature-dependent (Q10 = 1-1.3), and a plateau which is reached after about 20 min at 20 degrees C. Preincubation of tissue with unlabelled hormone reduces the initial uptake rate to a value characteristic for the second, less steep uptake phase. The activation energy for the initial uptake is 45 kJ/mole. At equilibrium the hormone is 4-fold enriched as compared to the medium, the delta G for this enrichment being 3 kJ/mole. The efflux is much lower than the influx and is linear. There is no linear relation between external hormone concentration and initial uptake rate. The initial uptake can be significantly reduced by an excess of diverse ecdysteroids with moulting hormone activity but not by hydrocortisone.


Fatal hemorrhage from a gastric cirsoid aneurysm.
Gastric cirsoid aneurysm has been reported as a cause of massive bleeding from the stomach in less than 50 cases. It is an abnormally wide and tortuous submucosal gastric artery that is manifested clinically only subsequent to erosion and hemorrhage. A patient is described in whom the source of hemorrhage was unrecognized despite angiographic and gastroscopic examinations. Medical management was unsuccessful, and the source of fatal hemorrhage was not determined until postmortem examination.


Colonic bleeding due to rupture of an isolated iliac artery aneurysm into a caecal carcinoma.
We describe a case of a fistula between an atherosclerotic aneurysm of the right common iliac artery and a caecal carcinoma, causing massive colonic bleeding. A fistula between a true aneurysm and a carcinoma of the colon has not to our knowledge been previously reported. Various aspects of this case are discussed from a pathogenetic point of view.


Dataset of lower extremity joint angles, moments and forces in distance running.
This study presents a database of joint angles, moments, and forces of the lower extremity from distance running at a submaximal speed in recreational runners. Twenty recreational runners participated in two experimental sessions, specifically pre and post a 5k treadmill run, with a synchronous collection of markers trajectories and ground reaction forces for both limbs in walking and running trials. The raw data in C3D files could be used for musculoskeletal modelling. Extra datasets of joint angles, moments, and forces are presented ready-for-use in MAT files, which could be as reference for study of biomechanical alterations from distance running. Applying advanced data processing techniques (Machine Learning algorithms) to these datasets ( C3D & MAT ), such as Principal Component Analysis, could extract key features of variation, thus potentially being applied for correlation with accelerometric and gyroscope parameters from wearable sensors during field running. Dataset of multi-segmental foot could be another contribution for the investigation of foot complex biomechanics from distance running. The dataset from Asian males may also be used for population-based studies of running biomechanics.


Stimulation and maintenance by nerve growth factor of phenylethanolamine-N-methyltransferase in superior cervical ganglia of adult rats.
Treatment of newborn rats with nerve growth factor (NGF) results in a striking increase in phenylethanolamine-N-methyltransferase (PNMT) in the superior cervical ganglia. Between one and three weeks of age there is normally a 10-fold decrease in PNMT activity in ganglia of infant rats. NGF treatment maintains the PNMT in ganglia at levels 10-fold greater than in untreated controls, but the levels of enzyme in the ganglion show the same magnitude of decrease with age. Epinephrine levels are markedly increased in ganglia of NGF-treated rats younger than one week of age, but at older ages the levels of the catecholamine are only slightly greater than the controls. Dexamethasone is less effective than NGF in increasing the levels of PNMT in ganglia of infant rats and, unlike NGF, becomes ineffective by 44 days of age. These results suggest that there may be two types of PNMT-containing cells in ganglia of newborn rats.


CALX-CBD1 Ca2+-Binding Cooperativity Studied by NMR Spectroscopy and ITC with Bayesian Statistics.
The Na+/Ca2+ exchanger of Drosophila melanogaster, CALX, is the main Ca2+-extrusion mechanism in olfactory sensory neurons and photoreceptor cells. Na+/Ca2+ exchangers have two Ca2+ sensor domains, CBD1 and CBD2. In contrast to the mammalian homologs, CALX is inhibited by Ca2+ binding to CALX-CBD1, whereas CALX-CBD2 does not bind Ca2+ at physiological concentrations. CALX-CBD1 consists of a β-sandwich and displays four Ca2+-binding sites at the tip of the domain. In this study, we used NMR spectroscopy and isothermal titration calorimetry (ITC) to investigate the cooperativity of Ca2+ binding to CALX-CBD1. We observed that this domain binds Ca2+ in the slow exchange regime at the NMR chemical shift timescale. Ca2+ binding restricts the dynamics in the Ca2+-binding region. Experiments of 15N chemical exchange saturation transfer and 15N R2 dispersion allowed the determination of Ca2+ dissociation rates (∼30 s-1). NMR titration curves of residues in the Ca2+-binding region were sigmoidal because of the contribution of chemical exchange to transverse magnetization relaxation rates, R2. Hence, a novel, to our knowledge, approach to analyze NMR titration curves was proposed. Ca2+-binding cooperativity was examined assuming two different stoichiometric binding models and using a Bayesian approach for data analysis. Fittings of NMR and ITC binding curves to the Hill model yielded nHill ∼2.9, near maximal cooperativity (nHill = 4). By assuming a stepwise model to interpret the ITC data, we found that the probability of binding from 2 up to 4 Ca2+ is approximately three orders of magnitude higher than that of binding a single Ca2+. Hence, four Ca2+ ions bind almost simultaneously to CALX-CBD1. Cooperative Ca2+ binding is key to enable this exchanger to efficiently respond to changes in the intracellular Ca2+ concentration in sensory neuronal cells.


Dietary restriction differentially protects from neurodegeneration in animal models of excitotoxicity.
Reduced caloric intake obtained through long-term dietary restriction has been found beneficial in some animal models of neurodegeneration. We report here that rats maintained under dietary restriction from the second to the eighth month of age are fully protected towards degeneration of GABAergic neurons in the hippocampus and the olfactory-entorhinal cortex caused by systemic administration of the convulsant toxin, kainic acid. However, in a different model of excitotoxic neurodegeneration, injection of ibotenic acid in the forebrain magnocellular basal nucleus, the decrease of a cholinergic marker in the target areas of the cortex was only partially protected by dietary restriction. Thus, in different experimental models neurodegeneration can be differentially rescued by dietary restriction. Analysis of alterations in the expression of relevant genes in different experimental conditions, could help in better understanding these differences.


Laparoscopic simulator performance and learning curves under different optical angles.
Deviated optical angles create visuospatial and psychomotor challenges during laparoscopic procedures, resulting in delayed operative time and possibly adverse events. If it is possible to train the skills needed to work under these deviated optical angles, this could benefit procedure time and patient safety. This study investigates the influence of the optical angle on development of basic laparoscopic surgical skills. A total of 58 medical students performed a four-session laparoscopic training course on a Virtual Reality Simulator. During each session, they performed an identical task under optical angles of 0°, 45° and - 45°. Performance parameters of task duration and damage were compared between the optical angles to investigate the effect of optical angle on performance development. The 4th session performance was compared to the 2nd session performance for each angle to determine improvement. Participants performed the task significantly faster under the 0° optical angle compared to the plus and minus 45° optical angles during the last three sessions (z between - 2.95 and - 2.09, p < .05). Participants improved significantly and similarly for task duration during the training course under all optical angles. At the end of the training course however significant performance differences between the zero and plus/minus 45 optical angles remained. Performance for damage did not improve and was not affected by optical angle throughout the course. Dedicated virtual reality training improves laparoscopic basic skills performance under deviated optical angles as it leads to shorter task duration, however a lasting performance impairment compared to the 0° optical angle remained. Training for performing under deviating optical angles can potentially shorter the learning curve in the operating room.


Involvement of Arabidopsis thaliana ribosomal protein S27 in mRNA degradation triggered by genotoxic stress.
A recessive Arabidopsis mutant with elevated sensitivity to DNA damaging treatments was identified in one out of 800 families generated by T-DNA insertion mutagenesis. The T-DNA generated a chromosomal deletion of 1287 bp in the promoter of one of three S27 ribosomal protein genes (ARS27A) preventing its expression. Seedlings of ars27A developed normally under standard growth conditions, suggesting wild-type proficiency of translation. However, growth was strongly inhibited in media supplemented with methyl methane sulfate (MMS) at a concentration not affecting the wild type. This inhibition was accompanied by the formation of tumor-like structures instead of auxiliary roots. Wild-type seedlings treated with increasing concentrations of MMS up to a lethal dose never displayed such a trait, neither was this phenotype observed in ars27A plants in the absence of MMS or under other stress conditions. Thus, the hypersensitivity and tumorous growth are mutant-specific responses to the genotoxic MMS treatment. Another important feature of the mutant is its inability to perform rapid degradation of transcripts after UV treatment, as seen in wild-type plants. Therefore, we propose that the ARS27A protein is dispensable for protein synthesis under standard conditions but is required for the elimination of possibly damaged mRNA after UV irradiation.


Randomized controlled trial of an energy conservation course for persons with multiple sclerosis.
To assess the short-term efficacy and effectiveness of a six-week energy conservation course on fatigue impact, quality of life and self-efficacy for persons with multiple sclerosis (MS). In this randomized controlled trial, we randomly assigned 169 persons with MS to an immediate intervention group or a delayed control group using a crossover design. The outcome measures: Fatigue Impact Scale, SF-36 Health Survey and Self-Efficacy for Performing Energy Conservation Strategies were measured before and after courses and no intervention control periods. We performed intent-to-treat analysis and compliers-only analyses using mixed effects analysis of variance models. Taking the energy conservation course had significant effects on reducing the physical and social subscales of Fatigue Impact Scale and on increasing the Vitality subscale of the SF-36 scores compared with not taking the course. Additional subscales were significant depending on methods of analyses. Self-Efficacy for Performing Energy Conservation Strategies Assessment increased significantly (P <0.05) postcourse compared to precourse. Results support the efficacy and effectiveness of the energy conservation course to decrease fatigue impact, and to increase self-efficacy and some aspects of quality of life.


"The sun keeps rising but darkness surrounds us": a qualitative exploration of the lived experiences of women with obstetric fistula in Ethiopia.
Obstetric fistula is a hole between the vagina and bladder, and/or between the vagina and rectum, triggered by prolonged obstructed labor. The World Health Organization has estimated that at least 50,000 to 100,000 cases of obstetric fistula occur every year, and that over two million women with obstetric fistula in developing countries remain untreated. Research on women's lived experiences of obstetric fistula is limited. This study aimed to explore the lived experience of women with obstetric fistula at Bahir Dar Hamlin Fistula Center, Amhara Regional State, Ethiopia. A qualitative study design, drawing from a phenomenological approach, was employed to explore the lived experience of purposively-selected sample of ten women with obstetric fistula. In-depth interviews were conducted in the local language (Amharic) using an interview guide. Interviews were transcribed and translated into English, and transcripts were entered as primary documents into Atlas.ti 7 software. Thematic categories were identified, and transcripts were coded accordingly. Participants perceived that the contributing factors to obstetric fistula were: instrument-assisted delivery; inappropriate physical examination and care; early marriage; and long duration of labour. As a result of obstetric fistula, the patients suffered from uncontrolled dripping of urine and/or faeces (and associated offensive odours), ostracization by their family and community members, and feeling hopeless and isolation from the community. Patients used different coping mechanisms, including frequent washing of clothes and changing of underwear; they also expressed that they preferred to be alone. Women with obstetric fistula experienced urine incontinence and associated bad odour; social and psychological problems like isolation, divorce and fears were commonly reported. Our findings from perspectives of Ethiopian setting suggest that integrated services for women with obstetric fistula are warranted, including physical therapy, psychological support and social reintegration.


Microvascular complications burden (nephropathy, retinopathy and peripheral polyneuropathy) affects risk of major vascular events and all-cause mortality in type 1 diabetes: a 10-year follow-up study.
Microvascular complications (MC) have been claimed to increase the risk for cardiovascular disease in diabetic subjects. However, the effect of MC burden on the risk of major vascular outcomes and all-cause mortality in type 1 diabetes is still poorly explored. We evaluated the relationship between microvascular complications burden and incidence of major cardiovascular events and all-cause mortality in subjects with type 1 diabetes. We recruited 774 participants with type 1 diabetes in a single-center observational study over a follow-up of 10.8 ± 2.5 years. Hazard ratios (HR) for cardiovascular outcomes and all-cause death associated with microvascular complications were determined by unadjusted and adjusted Cox regression analysis. Out of 774 individuals, 54.9% had no-MC, 32.3% 1 MC, 9.7% 2 MC and 3.1% 3 MC. A total of 54 deaths (7.0%) occurred. Death rate increased from no-MC 2.1% (Ref) to 1 MC 7.2% (HR 3.54 [95% CI 1.59-7.87]), 2 MC 14.7% (HR 6.41 [95% CI 2.65-15.49]) and 3 MC 66.7% (HR 41.73 [95% CI 18.42-94.57], p < 0.0001). After adjustments, HRs were: 1 MC 2.05 (95% CI 0.88-4.76), 2 MC 1.98 (95% CI 0.75-5.21), 3 MC 7.02 (95% CI 2.44-20.20, p = 0.002). Forty-nine subjects (6.7%) had at least one cardiovascular event, and cumulative incidence went from no-MC 2.2% (Ref) to 1 MC 5.0%; (HR 2.27 [95% CI 0.96-5.38]), 2 MC 26.8% (HR 12.88 [95% CI 5.82-28.50]) and 3 MC 40.9% (HR 29.34 [95% CI 11.59-74.25], p < 0.0001). Upon adjustments, HRs were: 1 MC 1.59 (95% CI 0.65-3.88), 2 MC 4.33 (95% CI 1.75-10.74), 3 MC 9.31 (95% CI 3.18-27.25, p < 0.0001). Thirty-five individuals (4.8%) had at least one coronary event, which cumulative incidence increased with MC burden (p < 0.0001). In type 1 diabetes, microvascular complications burden increases in an independent dose-dependent manner the risk of major cardiovascular outcomes and all-cause mortality. The presence and number of microvascular complications should be considered in stratifying overall cardiovascular risk in type 1 diabetes.


Early stage cost-effectiveness analysis of a BRCA1-like test to detect triple negative breast cancers responsive to high dose alkylating chemotherapy.
Triple negative breast cancers (TNBC) with a BRCA1-like profile may benefit from high dose alkylating chemotherapy (HDAC). This study examines whether BRCA1-like testing to target effective HDAC in TNBC patients can be more cost-effective than treating all patients with standard chemotherapy. Additionally, we estimated the minimum required prevalence of BRCA1-like and the required positive predictive value (PPV) for a BRCA1-like test to become cost-effective. Our Markov model compared 1) the incremental costs; 2) the incremental number of respondents; 3) the incremental number of Quality Adjusted Life Years (QALYs); and 4) the incremental cost-effectiveness ratio (ICER) of treating TNBC women with personalized HDAC based on BRCA1-like testing vs. standard chemotherapy, from a Dutch societal perspective and a 20-year time horizon, using probabilistic sensitivity analysis. Furthermore, we performed one-way sensitivity analysis (SA) to all model parameters, and two-way SA to prevalence and PPV. Data were obtained from a current trial (NCT01057069), published literature and expert opinions. BRCA1-like testing to target effective HDAC would presently not be cost-effective at a willingness-to-pay threshold of €80.000/QALY (€81.981/QALY). SAs show that PPV drives the ICER changes. Lower bounds for the prevalence and the PPV were found to be 58.5% and 73.0% respectively. BRCA1-like testing to target effective HDAC treatment in TNBC patients is currently not cost-effective at a willingness-to-pay of €80.000/QALY, but it can be when a minimum PPV of 73% is obtained in clinical practice. This information can help test developers and clinicians in decisions on further research and development of BRCA1-like tests.


MRI Utilization and the Associated Use of Sedation and Anesthesia in a Pediatric ACO.
MRI is commonly used in the pediatric population and often requires sedation or general anesthesia to complete. This study used data from a pediatric accountable care organization (ACO) to investigate trends in MRI utilization and in the requirement for anesthesia to complete MRI examinations. The Partners for Kids (PFK) ACO claims database was queried for MRI examination encounters involving patients 0 to 18 years old from 2009 to 2014, with utilization expressed as encounters per 10,000 PFK members-months. Data were limited to 2011 to 2014 to ensure consistent billing of anesthesia services. Encounters were classified according to the presence of procedure codes for anesthesia or sedation. MRI utilization was approximately constant over the study period at 11 to 12 encounters per 10,000 member-months. The need for anesthesia increased from 21% to 28% of encounters over 2011 to 2014. The latter increase was shared across 1- to 6-year-old, 7- to 12-year-old, and 12- to 18-year-old subgroups. In multivariable regression analysis of monthly utilization, increasing need for anesthesia could not be attributed to secular trends in patient demographics or types of examinations ordered. Paid cost data were available for outpatient MRIs, and MRIs with sedation accounted for an increasing share of these costs (from 22% in 2011 to 33% in 2014). There was an increasing need for anesthesia services to complete MRI examinations in this pediatric population, resulting in increasing cost of MRI examinations and presenting a challenge to ACO cost containment.


Total synthesis of a tyrosine suppressor transfer RNA gene. XVI. Enzymatic joinings to form the total 207-base pair-long DNA.
The total synthesis of a 207-base pair-long DNA, which is biologically functional as a tyrosine suppressor transfer RNA gene, has been completed. The synthesis involved the enzymatic joining of the previously synthesized duplexes. Thus, the duplex corresponding to the promoter region [P] (Sekiya, T., Brown, E.L., Ramamoorthy, B., Fritz, H.-J., Gait, M.J., Lees, R.G., Ryan, M.J., Khorana, H.G., and Norris, K.E. (1979) J. Biol. Chem. 254, 5781-5786) was jointed to Duplex [I] (Caruthers, M.H., Kleppe, R., Kleppe, K., and Khorana, H.G. (1976) J. Biol Chem. 251, 658-666) to form [P + I]. Separatively, Duplex [III + IV + Vb] was prepared from the previously described Duplexes [III], [IV], and [Vb]. (Loewen, P.C., Miller, R.C., Panet, A., Sekiya, T., and Khorana, H.G. (1976) J. Biol. Chem. 251, 642-650; Sekiya, T., Besmer, P., Takeya, T., and Khorana, H.G. 1976) J. Biol. Chem. 251, 634-641; Ramamoorthy, B., Lees, R.G., Kleid, D., and Khorana, H.G., (1976) J. Biol Chem. 251, 676-694). The product [P + I], was joined to Duplex [II] (Panet, A., Kleppe, R., Kleppe, K., and Khorana, H.G. (1976) J. Biol. Chem. 251, 651-657) and then to [III + IV + Vb] without isolation of the intermediates. In all the above joinings, the duplexes carried 32P-labeled phosphate groups at the appropriate 5'-ends. The total DNA and the intermediate duplexes were all characterized by their relative mobilities in electrophoresis on polyacrylamide gel slabs, by nearest neighbor analysis, and by degradation to 5'-nucleotides of radioactively labeled joined products. Two succeeding papers describe the transcription in vitro and the suppressor activity in vivo, of the synthetic gene now described.


Infusion of packed erythrocytes: an in vitro study of hemolysis.
Blood infusion through standard 20- and 22-gauge spinal needles, with varying hematocrit and rates of 1-6 mL/minute, did not result in significant amounts of erythrocyte hemolysis.


No relationship between right-to-left shunt with the severity of white matter hyperintensities.
Studies have shown that right-to-left shunt (RLS) is closely related to the occurrence of white matter hyperintensities (WMHs). Therefore, the detection of RLS is of great significance for the diagnosis and treatment of cerebral small vessel disease, especially for the prevention and treatment of WMHs. In this study, the c-TCD foaming experiment was selected to screen RLS, and evaluate the correlation between RLS and the severity of WMHs. We enrolled 334 migraineurs from a multicentre study from July 1 2019 and January 31 2020. Participants were all evaluated using contrast-enhanced transcranial Doppler, magnetic resonance imaging (MRI), and completed a questionnaire covering demographics, the main risk factors of vascular disease, and migraine status. RLS was classified into four grades (Grade 0 = Negative; Grade I = 1 ≤microbubbles (MBs)≤ 10; Grade II = MBs > 10 and no curtain; Grade III = curtain). Silent brain ischemic infarctions (SBI) and white matter hyperintensities (WMHs) were evaluated on MRI. In the incidence of WMHs, we found a significant difference between patients with RLS and no RLS (p < 0.05). There is no relationship between different grades of RLS and the severity of WMHs (p > 0.05). Overall, the positive rate of RLS is related to the incidence of WMHs. The different grades of RLS have no-relationship to do with the severity of WMHs.


Low-Carbon Fuel and Chemical Production by Anaerobic Gas Fermentation.
World energy demand is expected to increase by up to 40% by 2035. Over this period, the global population is also expected to increase by a billion people. A challenge facing the global community is not only to increase the supply of fuel, but also to minimize fossil carbon emissions to safeguard the environment, at the same time as ensuring that food production and supply is not detrimentally impacted. Gas fermentation is a rapidly maturing technology which allows low carbon fuel and commodity chemical synthesis. Unlike traditional biofuel technologies, gas fermentation avoids the use of sugars, relying instead on gas streams rich in carbon monoxide and/or hydrogen and carbon dioxide as sources of carbon and energy for product synthesis by specialized bacteria collectively known as acetogens. Thus, gas fermentation enables access to a diverse array of novel, large volume, and globally available feedstocks including industrial waste gases and syngas produced, for example, via the gasification of municipal waste and biomass. Through the efforts of academic labs and early stage ventures, process scale-up challenges have been surmounted through the development of specialized bioreactors. Furthermore, tools for the genetic improvement of the acetogenic bacteria have been reported, paving the way for the production of a spectrum of ever-more valuable products via this process. As a result of these developments, interest in gas fermentation among both researchers and legislators has grown significantly in the past 5 years to the point that this approach is now considered amongst the mainstream of emerging technology solutions for near-term low-carbon fuel and chemical synthesis.


Pediatricians' screening urinalysis practices.
To determine pediatricians' routine screening urinalysis practices. This was a survey of a nationally representative sample of pediatricians practicing in the U.S. regarding their screening urinalysis practices in childhood. Of the 1502 pediatricians sampled, 653 eligible subjects participated, for an estimated response rate of 49.5%. The vast majority of participants (78%) routinely screen asymptomatic children with urinalysis in at least 1 age group. Pediatricians' screening urinalysis practice varies based on age group: 9% screen during infancy (<1 year), 60% screen during early childhood (1 up to 5 years), 55% screen during late childhood (5 to 12 years), and 58% screen during adolescence (13 to 20 years). The majority of pediatricians (58%) routinely screen more than 1 age group. Some 38% of the pediatricians surveyed believe that the overall health of children is improved by screening all asymptomatic children with urinalysis. Many pediatricians routinely conduct screening urinalysis during childhood, frequently at ages not recommended by the American Academy of Pediatrics.


Possible enzymatic mechanism underlying chemical tolerance and characteristics of tolerant population in Scapholeberis kingi.
To determine the potential effects of pesticides on aquatic organisms inhabiting a realistic environment, we explored the characteristics and mechanisms of chemical tolerance in Scapholeberis kingi(Cladocera). We established a chemical-tolerant population via continuous exposure to pirimicarb, an acetylcholinesterase (AChE) inhibitor, and examined the effects of pirimicarb concentration on the intrinsic growth rates (r) of tolerant cladocerans. We also explored the association between r and feeding rate and tested the involvement of antioxidant enzymes [peroxidase (PO) and superoxide dismutase] and AChE in pirimicarb sensitivity. S. kingi was continuously exposed to lethal and sublethal pirimicarb concentrations (0, 2.5, 5, and 10 µg/L) for 15 generations, and changes (half maximal effective concentration at 48 h, 48 h-EC50) in chemical sensitivity were investigated. In the F14 generation, the sensitivity of the 10 µg/L group was three times lower than that of the control group, suggesting the acquisition of chemical tolerance. Moreover, r was significantly and negatively correlated with 48 h-EC50, suggesting a fitness cost for tolerance. Surprisingly, there was no significant correlation between r and feeding rate. There was a weak but significant positive correlation between each enzyme activity and the 48 h-EC50 value (p < 0.05). Thus, oxidative stress regulation and enhanced AChE may be involved in the acquisition of chemical tolerance in cladocerans. These findings will help elucidate the characteristics and mechanisms of chemical tolerance in aquatic organisms.


Venous-Left Atrial Extracorporeal Membrane Oxygenation Configuration Use in Right Ventricular Failure and Tricuspid Ring.
Mechanical circulatory support devices are used to support the heart in cardiogenic shock. We present a case of demonstrating the feasible use of left ventricular assistive device with reverse configuration to support severe right ventricular failure in a patient with recent tricuspid annuloplasty ring.


Short-term association between outdoor temperature and the hydration-marker copeptin: a pooled analysis in five cohorts.
Whereas outdoor temperature is linked to both mortality and hydration status, the hormone vasopressin, measured through the surrogate copeptin, is a marker of cardiometabolic risk and hydration. We recently showed that copeptin has a seasonal pattern with higher plasma concentration in winter. Here, we aimed to investigate the association between outdoor temperature and copeptin. Copeptin was analysed in fasting plasma from five cohorts in Malmö, Sweden (n = 26,753, 49.7% men, age 18-86 years). We utilized a multivariable adjusted non-linear spline model with four knots to investigate the association between short-term temperature (24 h mean apparent) and log copeptin z-score. We found a distinct non-linear association between temperature and log copeptin z-score, with both moderately low and high temperatures linked to higher copeptin concentration (p < 0.0001). Between 0 °C and nadir at the 75th temperature percentile (corresponding to 14.3 °C), log copeptin decreased 0.13 z-scores (95% CI 0.096; 0.16), which also inversely corresponded to the increase in z-score log copeptin between the nadir and 21.3 °C. The J-shaped association between short-term temperature and copeptin resembles the J-shaped association between temperature and mortality. Whereas the untangling of temperature from other seasonal effects on hydration warrants further study, moderately increased water intake constitutes a feasible intervention to lower vasopressin and might mitigate adverse health effects of both moderately cold and hot outdoor temperatures. Swedish Research Council, Å Wiberg, M Stephen, A Påhlsson, Crafoord and Swedish Heart-Lung Foundations, Swedish Society for Medical Research and Swedish Society of Medicine.


Toluidine blue staining in the detection of oral precancerous and malignant lesions.
One hundred thirty-two consecutive patients suspected of having malignant or precancerous oral lesions were studied by comparing toluidine blue dye uptake clinically with a simultaneous biopsy. The histopathologic diagnosis confirmed 57 squamous carcinomas, 42 epithelial dysplasias, and 33 benign mucosal changes. Overall accuracy of the toluidine blue uptake was 91%. In the dysplastic and malignant lesions the false negatives were 2%, and there were 30% false positives in the benign lesions. It was concluded that toluidine blue staining is a useful adjunct to careful examination, clinical judgment, and biopsy.


The structure of the Early Rehabilitation Barthel Index (ERBI) should be modified: evidence from a Rasch analysis study.
The Early Rehabilitation Barthel Index (ERBI) comprises seven items of the Early Rehabilitation Index and ten items of the Barthel Index. The ERBI is usually used to measure functional changes in patients with severe acquired brain injury (sABI), but its measurement properties have yet to be extensively assessed. To study the unidimensionality and internal construct validity (ICV) of the ERBI through Confirmatory Factor Analysis (CFA), Mokken Analysis (MA), and Rasch Analysis (RA). Multicenter prospective study. Inpatients from five intensive rehabilitation centers. Two hundred and forty-seven subjects with sABI. ERBI was administered on admission and discharge to study its unidimensionality through CFA and MA and its ICV, reliability, and targeting through RA. The preliminary analyses showed a lack of unidimensionality (RMSEA=0.460 >0.06; SRMR=0.176 >0.06; CFI=1.000 >0.950; TLI=1.000 >0.950). According to CFA, "Confusional state" and "Behavioral disturbance" items showed low factor loadings (<0.40), whereas these two items composed a separate scale within the MA. Furthermore, the baseline RA showed that three items misfitted ("Mechanical ventilation," "Confusional state," "Behavioral disturbances") and a lack of conformity of several ICV requirements. After deletion of three misfitting items and further non-structural modifications (i.e., testlets creation to absorb local dependence between items and item misfit), the solution obtained showed adequate ICV, adequate reliability for measurements at the individual level (PSI>0.85), although with a frank floor effect. This final solution was successfully replicated in a total sample of the subjects. After post-hoc modifications of the score structure of two out of three misfitting items, the subsequent CFA (RMSEA=0.044 <0.06; SRMR=0.056 <0.06; CFI=1.000 >0.950 TLI=1.000 >0.950) and MA showed the resolution of the unidimensional issues. Although the ERBI is a potentially valuable tool for measuring functioning in the coma-to-community continuum, our analyses suggested its lack of ICV, partly due to an incorrect scoring design of some items. A new perspective multicenter study is proposed to validate a modified version of the ERBI that overcomes the problems highlighted in this analysis. Our results do not support the use of the original structure of the ERBI in clinical practice and research, as a lack of ICV was highlighted.


The added value of the selective SuperPolymyxin™ medium in detecting rectal carriage of Gram-negative bacteria with acquired colistin resistance in intensive care unit patients receiving selective digestive decontamination.
The objective of this study was to determine the value of using SuperPolymyxin™ selective medium (ELITech Group, Puteaux, France) in addition to conventional non-selective inoculation methods in the detection of acquired colistin resistance in a Dutch intensive care unit (ICU) that routinely uses selective decontamination of the digestive tract (SDD). We performed a cross-sectional study with prospective data collection in a tertiary-care ICU. All consecutive surveillance rectal swabs of ICU-patients receiving SDD were included and cultured in an observer-blinded approach using (1) a conventional culture method using non-selective media and (2) SuperPolymyxin™ selective medium. MIC values for colistin of non-intrinsically colistin-resistant Gram-negative isolates were determined with broth microdilution (BMD) using Sensititre™ and colistin resistance was confirmed using BMD according to EUCAST guidelines. One thousand one hundred five rectal swabs of 428 unique ICU-patients were inoculated using both culture methods, yielding 346 and 84 Gram-negative isolates for BMD testing with the conventional method and SuperPolymyxin™ medium, of which 308 and 80 underwent BMD, respectively. The number of identified rectal carriers of isolates with acquired colistin resistance was 3 (0.7%) for the conventional method, 4 (0.9%) for SuperPolymyxin™, and 5 (1.2%) for both methods combined. The number of isolates with acquired colistin resistance was 4 (1.0%) for the conventional method, 8 (2.1%) for SuperPolymyxin™ and 9 (2.3%) for both methods combined. In a surveillance setting of low prevalence of acquired colistin resistance in patients that receive SDD in a Dutch tertiary-care ICU, SuperPolymyxin™ had a higher diagnostic yield than conventional inoculation methods, but the combination of both had the highest diagnostic yield.


Disease patterns of microbial keratitis in Singapore: A retrospective case series.
To investigate the disease patterns of Microbial Keratitis(MK) in patients seen in a tertiary referral hospital, to evaluate the clinical outcomes of MK and the risk factors for poorer visual outcomes. This is a retrospective case series of all culture-positive corneal scrapings between April 2012 and October 2016. A total of 230 patients(n = 230) were included into this study. Patient demographics, clinical information and microbiological characteristics of organisms are collected. 64.3% of patients with MK are contact lens(CL) users. Among CL users, there is a preponderance of females(68.9%) and they tend to be younger (27.1 ± 10.6 years). The most frequently isolated organism in this study is Pseudomonas aeruginosa(51.7%) with 69.6% of cases belonging to CL users. MK in non-CL users tend to involve other organisms, such as coagulase-negative Staphylococci, Staphylococcus aureus and Streptococcus pneumoniae. Pseudomonas aeruginosa exhibits good sensitivity rates to ciprofloxacin, levofloxacin and gentamicin. Non-Pseudomonas organisms display similar sensitivities to ciprofloxacin, levofloxacin and gentamicin. MK in non-CL users is related to predisposing factors of prior ocular trauma and concomitant ocular pathology. They tend to have worse visual acuity(VA) on presentation and after treatment compared to CL users. Poorer VA outcome is associated with larger ulcers, increasing age, trauma and non-CL wearers. Successful clinical outcome is achieved in 97.8% of patients, with only 2.2% requiring further surgical intervention. CL use alters the disease patterns of MK as well as the underlying microbiological etiology. Fluoroquinolones and aminoglycosides are good empirical antibiotics for MK treatment. Early referral to a tertiary centre will likely allow for earlier treatment, which can result in better VA outcome, especially so in patients who are older, non-CL wearers and have larger ulcers with associated trauma.


Early exercise in blunt chest wall trauma: protocol for a mixed-methods, multicentre, parallel randomised controlled trial (ELECT2 trial).
Chronic pain and disability are now well-recognised long-term complications of blunt chest wall trauma. Limited research exists regarding therapeutic interventions that can be used to address these complications. A recent feasibility study was completed testing the methods of a definitive trial. This protocol describes the proposed definitive trial, the aim of which is to investigate the impact of an early exercise programme on chronic pain and disability in patients with blunt chest wall trauma. This mixed-methods, multicentre, parallel randomised controlled trial will run in four hospitals in Wales and one in England over 12-month recruitment period. Patients will be randomised to either the control group (routine physiotherapy input) or the intervention group (routine physiotherapy input plus a simple exercise programme completed individually by the patient). Baseline measurements including completion of two surveys (Brief Pain Inventory and EuroQol 5-dimensions, 5-Levels) will be obtained on initial assessment. These measures and a client services receipt inventory will be repeated at 3-month postinjury. Analysis of outcomes will focus on rate and severity of chronic pain and disability, cost-effectiveness and acceptability of the programme by patients and clinicians. Qualitative feedback regarding acceptability will be obtained through patient and clinician focus groups. London Riverside Research Ethics Committee (Reference number: 21/LO/0782) and the Health Research Authority granted approval for the trial in December 2021. Patient recruitment will commence in February 2022. Planned dissemination is through publication in a peer-reviewed Emergency Medicine Journal, presentation at appropriate conferences and to stakeholders at professional meetings. ISRCTN65829737; Pre-results.


Endoscopic versus operative gastrostomy: final results of a prospective randomized trial.
This study compared operative gastrostomy (OG) (by surgeons) with endoscopic gastrostomy (PEG) (by physicians) in a prospective randomized fashion to determine whether one technique was superior. PEG (Sachs-Vine) and OG (Stamm) were done using local anesthesia. Patients were assessed for complications, mortality, tube function, and cost. Groups were equally matched for indications and underlying disease. Fifty-seven had OG and 64 had attempted PEG. Complications occurred in 26% of OG patients and 9% died. Complications occurred in 25% of PEG patients and 12% died. Tube feeding was initiated in both groups within a mean of 29 (24 to 72) hours of the gastrostomy placement. OG cost $1675 and PEG $979 to perform. Twenty-one PEG patients required endoscopic tube change which raised their total cost to $1574. We conclude there is no difference between OG (using local anesthesia) and PEG with regard to morbidity, mortality, or tube function. The endoscopic technique does appear to have economic advantage.


App-based guided problem-solving intervention for adolescent mental health: a pilot cohort study in Indian schools.
This paper describes the pilot evaluation of 'POD Adventures', a lay counsellor-guided problem-solving intervention delivered via a smartphone app in Indian secondary schools. To test the feasibility and acceptability of POD Adventures for adolescents with a felt need for psychological support, and to explore the intervention's effects on self-reported mental health symptoms, prioritised problems, stress and well-being. We used a mixed-methods pre-post cohort design. Participants were self-referred from grades 9-12 in two coeducational government-aided secondary schools in Goa, India. The intervention was delivered in two formats, 'mixed' (comprising individual and small group sessions) and 'group' (small group sessions only). 248 participants enrolled in the study and 230 (92.7%) completed the intervention. Outcomes at 4 weeks showed significant improvements on all measures that were maintained at 12 weeks. Large effects were observed on problem severity scores (4 weeks, d=1.47; 12 weeks, d=1.53) while small to moderate effects were seen on mental health symptoms, stress and well-being. 22 students completed qualitative interviews about their experience of the intervention. Participants found POD Adventures easy to use, engaging and helpful in solving their problems. They were satisfied with the guidance provided by the counsellor irrespective of delivery format. POD Adventures was feasible to deliver with guidance from lay counsellors in Indian schools, acceptable to participants and associated with large improvements in problem severity and mental health symptom severity. POD Adventures has promise as an early intervention for adolescents with a felt need for psychological support in low-resource settings.


Early Impacts of a Healthy Food Distribution Program on the Availability and Price of Fresh Fruits and Vegetables in Small Retail Venues in Los Angeles.
Healthy food distribution programs that allow small retailers to purchase fresh fruits and vegetables at wholesale prices may increase the profitability of selling produce. While promising, little is known about how these programs affect the availability of fresh fruits and vegetables in underserved communities. This study examined the impacts of a healthy food distribution program in Los Angeles County over its first year of operation (August 2015-2016). Assessment methods included: (1) a brief survey examining the characteristics, purchasing habits, and attitudes of stores entering the program; (2) longitudinal tracking of sales data examining changes in the volume and variety of fruits and vegetables distributed through the program; and (3) the collection of comparison price data from wholesale market databases and local grocery stores. Seventeen stores participated in the program over the study period. One-fourth of survey respondents reported no recent experience selling produce. Analysis of sales data showed that, on average, the total volume of produce distributed through the program increased by six pounds per week over the study period (95% confidence limit: 4.50, 7.50); trends varied by store and produce type. Produce prices offered through the program approximated those at wholesale markets, and were lower than prices at full-service grocers. Results suggest that healthy food distribution programs may reduce certain supply-side barriers to offering fresh produce in small retail venues. While promising, more work is needed to understand the impacts of such programs on in-store environments and consumer behaviors.


Hemolytic porcine intestinal Escherichia coli without virulence-associated genes typical of intestinal pathogenic E. coli.
Testing 1,666 fecal or intestinal samples from healthy and diarrheic pigs, we obtained hemolytic Escherichia coli isolates from 593 samples. Focusing on hemolytic E. coli isolates without virulence-associated genes (VAGs) typical for enteropathogens, we found that such isolates carried a broad variety of VAGs typical for extraintestinal pathogenic E. coli.


Synthesis and biofilm inhibition studies of 2-(2-amino-6-arylpyrimidin-4-yl)quinazolin-4(3H)-ones.
Synthesis of novel 4(3H)-quinazolinonyl aminopyrimidine derivatives has been achieved via quinazolinonyl enones which in turn were obtained from 2-acyl-4(3H)-quinazolinone. They have been assayed for biofilm inhibition against Gram-positive (methicillin-resistant Staphylococcus aureus (MRSA)) and Gram-negative bacteria (Acinetobacter baumannii). The analogues with 2,4,6-trimethoxy phenyl, 4-methylthio phenyl, and 3-bromo phenyl substituents (5h, 5j & 5k) have been shown to inhibit biofilm formation efficiently in MRSA with IC50 values of 20.7-22.4 μM). The analogues 5h and 5j have demonstrated low toxicity in human cells in vitro and can be investigated further as leads.


Structure-Activity Relationship Studies of 2,4,5-Trisubstituted Pyrimidine Derivatives Leading to the Identification of a Novel and Potent Sirtuin 5 Inhibitor against Sepsis-Associated Acute Kidney Injury.
Sepsis-associated acute kidney injury (AKI) is a serious clinical problem without effective drugs. Inhibition of sirtuin 5 (SIRT5) has been confirmed to protect against AKI, suggesting that SIRT5 inhibitors might be a promising therapeutic approach for AKI. Herein, structural optimization was performed on our previous compound 1 (IC50 = 3.0 μM), and a series of 2,4,5-trisubstituted pyrimidine derivatives have been synthesized. The structure-activity relationship (SAR) analysis led to the discovery of three nanomolar level SIRT5 inhibitors, of which the most potent compound 58 (IC50 = 310 nM) was demonstrated to be a substrate-competitive and selective inhibitor. Importantly, 58 significantly alleviated kidney dysfunction and pathological injury in both lipopolysaccharide (LPS)- and cecal ligation/perforation (CLP)-induced septic AKI mice. Further studies revealed that 58 regulated protein succinylation and the release of proinflammatory cytokines in the kidneys of septic AKI mice. Collectively, these results highlighted that targeting SIRT5 has a therapeutic potential against septic AKI.


Crystallization of the complex of actin with gelsolin segment 1.
Crystals of a 1:1 complex between human gelsolin segment 1 and actin have been grown from solutions containing polyethylene glycol 6000. The crystals are orthorhombic, space group P2(1)2(1)2(1); the axes are a = 57.4 A, b = 70.4 A, c = 184.5 A. They are moderately stable to X-rays and diffract to beyond 2.5 A. There is one molecule of complex in the asymmetric unit.


Endogenous abscisic acid is involved in methyl jasmonate-induced reactive oxygen species and nitric oxide production but not in cytosolic alkalization in Arabidopsis guard cells.
We recently demonstrated that endogenous abscisic acid (ABA) is involved in methyl jasmonate (MeJA)-induced stomatal closure in Arabidopsis thaliana. In this study, we investigated whether endogenous ABA is involved in MeJA-induced reactive oxygen species (ROS) and nitric oxide (NO) production and cytosolic alkalization in guard cells using an ABA-deficient Arabidopsis mutant, aba2-2, and an inhibitor of ABA biosynthesis, fluridon (FLU). The aba2-2 mutation impaired MeJA-induced ROS and NO production. FLU inhibited MeJA-induced ROS production in wild-type guard cells. Pretreatment with 0.1 μM ABA, which does not induce stomatal closure in the wild type, complemented the insensitivity to MeJA of the aba2-2 mutant. However, MeJA induced cytosolic alkalization in both wild-type and aba2-2 guard cells. These results suggest that endogenous ABA is involved in MeJA-induced ROS and NO production but not in MeJA-induced cytosolic alkalization in Arabidopsis guard cells.


A longitudinal study of immunological status in Chinese haemophiliacs: importance of the heat viral inactivation of factor concentrates. II. Improvements of CD4/CD8 ratio after treatments with heat-inactivated factor concentrates.
Screened and heated clotting factor concentrates of intermediate purity have been used in Taiwanese haemophiliacs since the end of 1986. A significant improvement of CD4/CD8 ratio during the years 1987-1989 as compared with those during the years 1984-1986 was observed in haemophilia A patients [mean +/- SD (median), 1.191 +/- 0.495 (1.163) vs. 0.880 +/- 0.325 (0.838), P = 0.0020] who were seronegative for human immunodeficiency virus. Almost all patients received an increased amount of factor VIII concentrates and total plasma products since 1987. Multiple linear regression analysis for the association of CD4/CD8 ratio with changes in dosage of plasma products revealed that there was a significant positive association of CD4/CD8 ratio measured during 1987-1989 with dosage of factor VIII concentrate administered during 1984-1986 (P = 0.0230), which is an indicator for changes in viral load, but not with changes in dosage of plasma products, which are indicators for changes in plasma protein intake. Our data indicate that immunological abnormalities after replacement therapy observed in haemophiliacs are mainly attributed to virus infection through infusion of factor concentrates, not to allogeneic proteins existing in plasma products.


Decreased plasma fibronectin levels in children with hemolytic-uremic syndrome.
Fibronectin is a normal plasma glycoprotein thought to have an important role in platelet aggregation and clot formation. Because of the participation of the coagulation system in hemolytic-uremic syndrome, the present study sought to determine if fibronectin plays a role in the pathogenesis of this disease. With this purpose in mind, plasma fibronectin levels were measured in 17 children with the clinical diagnosis of hemolytic-uremic syndrome and in 22 age-matched control subjects. Fibronectin levels were significantly depressed in 13 of 17 (76 percent) patients with hemolytic-uremic syndrome during the acute phase of their illness. The levels did not correlate with age, sex, serum creatinine level, platelet count, or hemoglobin concentration. Serial plasma samples were available in eight of these patients: fibronectin remained depressed from two to 10 days and then returned toward the normal range concomitant with increasing platelet counts and improvement in renal function. During remission, fibronectin levels were normal in all nine patients tested. To try to determine if fibronectin is deposited in the kidney during hemolytic-uremic syndrome, kidney biopsy specimens from six patients with hemolytic-uremic syndrome were examined by immunofluorescence for the presence of fibronectin, fibrinogen, and platelet antigens. Extensive deposition of all three antigens was demonstrated along the glomerular capillary wall in all biopsy specimens. In conclusion, plasma fibronectin levels are decreased during the acute phases of hemolytic-uremic syndrome in the majority of patients. Kidney biopsy findings suggest that fibronectin is intimately involved in the activation of the coagulation system in this disease and that decreased plasma fibronectin levels in hemolytic-uremic syndrome may be due, at least in part, to accelerated consumption of the protein at the sites of injury.


Antibodies against Kaposi sarcoma-associated herpes virus (KSHV) complement control protein (KCP) in infected individuals.
Kaposi sarcoma-associated herpesvirus (KSHV) is the most important etiopathological factor of Kaposi's sarcoma (KS) and some specific types of malignant lymphomas. One of the viral lytic genes encodes the KSHV complement control protein (KCP), which functionally mimics human complement inhibitors. Although this protein provides an advantage for evading the complement attack, it can serve as target for adaptive immune response. Herein, we identified anti-KCP IgG antibodies in patients with KS and KSHV-related lymphomas. KCP-specific antibodies were only detected in sera of those patients who had high titres of antibodies against lytic or latent KSHV antigens. Complement control protein domain 2 (CCP2) was found to be the most immunogenic part of the KCP protein. Furthermore, pre-incubation of KCP-expressing CHO cells with patient sera containing anti-KCP antibodies resulted in an increased complement deposition when incubated with human serum.


Flow Cytometric Monitoring for Residual Disease in B Lymphoblastic Leukemia Post T Cell Engaging Targeted Therapies.
The use of targeted therapy is growing in the setting of hematopoietic neoplasms. Flow cytometry is a cornerstone of residual disease monitoring post therapy in this group of malignancies. Often, there is overlap between antigens targeted by immunotherapies and gating reagents utilized for population identification by flow cytometry. Such overlap can render a previously excellent gating reagent inadequate for disease detection. Recently, several anti-CD19 T cell-engaging immunotherapeutic agents and an anti-CD22 immunotoxin have been FDA approved for use in B lymphoblastic leukemia (B-LL), with an anti-CD22 T cell-engaging agent in development. In the setting of such targeted therapies, CD19 and CD22 expression may be altered, compromising the use of these reagents for identification of abnormal blasts. We describe herein a strategy for flow cytometric monitoring for residual disease in patients with B-LL post T cell-engaging anti-CD19 and anti-CD22 therapies. © 2018 by John Wiley & Sons, Inc.


Successful pregnancies from cryopreserved human embryos produced by in vitro fertilization.
Sixty-three embryos produced after in vitro fertilization in 30 infertile couples were frozen and stored. Dimethylsulfoxide was used as a cryoprotectant and embryos were frozen from the two-cell stage to early blastocyst development. Replacement occurred during spontaneous ovulatory cycles 2 to 15 months after embryo freezing. Embryo replacement was performed 3 to 6 days following identification of the luteinizing hormone surge in the spontaneous cycle. Thirty-five embryos were replaced into 25 women and two viable pregnancies resulted.


Barrier protective activities of curcumin and its derivative.
Curcumin, a poly-phenolic compound, possesses diverse pharmacologic activities. However, the barrier protective functions of curcumin or its derivative have not yet been studied. The objective of this study was to investigate the barrier protective activities of curcumin and its derivative (bisdemethoxycurcumin, BDMC) on lipopolysaccharide (LPS) barrier disruption in human umbilical vein endothelial cells (HUVECs) were investigated. The barrier protective effects of curcumin and BDMC such as permeability, expression of cell adhesion molecules, monocytes adhesion and migration toward HUVECs were tested. Curcumin and BDMC inhibited LPS-induced barrier permeability, monocyte adhesion and migration; inhibitory effects were significantly correlated with inhibitory functions of curcumin and BDMC on LPS-induced cell adhesion molecules (vascular cell adhesion molecules, intracellular cell adhesion molecule, E-selectin). Furthermore, LPS-induced nuclear factor-κB (NF-κB) activation and tumor necrosis factor-α (TNF-α) release from HUVECs were inhibited by curcumin and BDMC. Surprisingly, the barrier protective activities of BDMC were better than those of curcumin, indicating that the methoxy group in curcumin negatively regulated barrier protection function of curcumin. Given these results, curcumin or its derivative, BDMC, showed barrier protective activities and they could be a therapeutic candidates for various systemic inflammatory diseases.


The potential of antioxidant-rich Maoberry (Antidesma bunius) extract on fat metabolism in liver tissues of rats fed a high-fat diet.
Obesity and dyslipidemia are major risk factors associated with non-alcoholic fatty liver disease (NAFLD). NAFLD refers to the accumulation of fat in more than 5% of the liver without alcohol consumption. NAFLD is the most common liver disease and is rapidly becoming a global public health problem. Maoberry (Antidesma bunius) is a fruit rich in antioxidants, especially phenolic compounds, which are reported to have benefits for patients with NAFLD. We evaluated the effect of Maoberry extract on fat metabolism in liver tissues of high fat diet-induced rats. Five (5) groups (n = 12) of male Sprague-Dawley (SD) rats were divided into those given a high fat diet with no treatment (HF), different dosages of Maoberry extracts (0.38 [ML], 0.76 [MM) and 1.52 [MH] g/kg body weight) and 10 mg/kg statin (STAT). The rats were fed a high fat diet for 4 weeks to induce obesity and subsequently continued more for 12 weeks with treatments of Maoberry extracts or STAT. The levels of triglyceride, liver enzymes, oxidative stress and inflammation markers, triglyceride synthesis regulators, and pathology of the liver in high fat diet-induced rats were investigated. Feeding Maoberry extract in MH groups resulted in decreasing levels of serum alanine aminotransferase (ALT), liver triglyceride, liver thiobarbituric acid reactive substances (TBARS) and mRNA expression of tumour necrosis factor (TNF)-α, interleukin (IL)-6, glycerol-3-phosphate acyltransferase (GPAT)-1 and acetyl-coenzyme A carboxylase (ACC) compared with the HF group (P < 0.05). Moreover, histopathological study of the liver showed reduced fat droplets in the Maoberry extract treatment groups, especially in MH groups and STAT treatment groups. The improvements of fat metabolism in liver tissues of rats fed a high-fat diet were observed in Maoberry extracts treatment groups. The underline mechanism that link to fat metabolism might be through the process accompanied with down-regulated the gene expression of key enzymes of lipid production, antioxidant activity, and anti-inflammation properties of Maoberry extracts which contains high levels of phenolic and flavonoid compounds.


Orexins and feeding: special occasions or everyday occurrence?
Neurons expressing prepro-orexin, the precursor of orexin-A and -B, are found in the lateral hypothalamic area, a region classically implicated in driving feeding. Orexin-A induces feeding transiently when injected centrally, and food intake can be decreased when orexin action is disrupted by immunoneutralization of orexin-A, or by pharmacological blockade of orexin receptors, or by transgenic knockout of orexin. Here, we argue that orexin neurons may act to stimulate feeding in the short term, and that important regulatory signals may be a fall in plasma glucose (stimulatory), countered by satiety signals generated by eating, such as gastric distention (inhibitory).


A simple method of drying virus on inanimate objects for virucidal testing.
A simple method for drying virus on inanimate objects (cover slips) under vacuum in the cold is described. Following this procedure virus maintains high titers (10(6-7)) for periods of 1-3 wk at -70 degrees C depending on the virus. For virucidal assay of disinfectants, cover slips are exposed to medium simulating the disinfectant (virus control) or disinfectant in an upright position in an Ultra-Vu cuvette. Cover slips are readily removed and placed in tissue culture medium for dilution of virus and determination of virus titer. Cytotoxicity of disinfectant is determined by exposing cover slip without virus to disinfectant, then placing it in medium, diluting the medium and incubating with the indicator cells. The use of this technique results in high titers of virus on cover slips, which are inanimate objects requiring minimal manipulation. The titration of virus or cytotoxicity in microplates is cell, medium, serum, and labware economical.