Tumor MICA status predicts the efficacy of immunotherapy with cytokine-induced killer cells for patients with gastric cancer.
In this study, we determine the relationship between the expression of major histocompatibility complex class I chain-related gene A (MICA) in gastric cancer tumors after D2 gastrectomy and the clinical outcome of a CIK-containing adjuvant therapy. Ninety-five consecutive patients with gastric cancer after D2 gastrectomy who received adjuvant chemotherapy combined with CIK cell therapy were enrolled. The MICA expression of their tumors was determined by immunohistochemistry (IHC). High expression of MICA protein was documented by IHC in 38 of 95 tumor samples (40.0 %). The MICA status was significantly associated with the age and stage, p = 0.008 and 0.023, respectively. Analysis of NKG2D on in vitro expanded CIK cells showed that the percentages of NKG2D+ in CD3+/CD56+, CD3-/CD56+, and CD3+/CD8+ cells populations were 97.2 ± 1.4, 97.9 ± 1.8, and 95.6 ± 2.1 %, respectively. For patient with high MICA-expressing tumors, the median DFS and OS were longer than for the patients with tumors with low expression of MICA; 46.0 versus 41.0 months (p = 0.027), and 48.0 versus 42.0 months (p = 0.031), respectively. In a multivariate analysis, stage and MICA expression were independent prognostic factors for DFS and OS. Our findings show that adjuvant chemotherapy plus CIK therapy treatment is a promising modality for treating gastric cancer patients after D2 gastrectomy. Especially, those who have tumors with high expression of MICA were more likely to benefit from such a treatment strategy. Subsequent studies in clinical trial cohorts will be required to confirm the clinical utility of these markers.


Role of opioid receptors in modulation of P2X receptor-mediated cardiac sympathoexcitatory reflex response.
Myocardial ischemia evokes powerful reflex responses through activation of vagal and sympathetic afferents in the heart through the release of ischemic metabolites. We have demonstrated that extracellular ATP stimulates cardiac sympathetic afferents through P2 receptor-mediated mechanism, and that opioid peptides suppress these afferents' activity. However, the roles of both P2 receptor and endogenous opioids in cardiac sympathoexcitatory reflex (CSR) responses remain unclear. We therefore hypothesized that activation of cardiac P2 receptor evokes CSR responses by stimulating cardiac sympathetic afferents and these CSR responses are modulated by endogenous opioids. We observed that intrapericardial injection of α,β-methylene ATP (α,β-meATP, P2X receptor agonist), but not ADP (P2Y receptor agonist), caused a graded increase in mean arterial pressure in rats with sinoaortic denervation and vagotomy. This effect of α,β-meATP was abolished by blockade of cardiac neural transmission with intrapericardial procaine treatment and eliminated by intrapericardial A-317491, a selective P2X2/3 and P2X3 receptor antagonist. Intrapericardial α,β-meATP also evoked CSR response in vagus-intact rats. Furthermore, the P2X receptor-mediated CSR responses were enhanced by intrapericardial naloxone, a specific opioid receptor antagonist. These data suggest that stimulation of cardiac P2X2/3 and P2X3, but not P2Y receptors, powerfully evokes CSR responses through activation of cardiac spinal afferents, and that endogenous opioids suppress the P2X receptor-mediated CSR responses.


BORC/kinesin-1 ensemble drives polarized transport of lysosomes into the axon.
The ability of lysosomes to move within the cytoplasm is important for many cellular functions. This ability is particularly critical in neurons, which comprise vast, highly differentiated domains such as the axon and dendrites. The mechanisms that control lysosome movement in these domains, however, remain poorly understood. Here we show that an ensemble of BORC, Arl8, SKIP, and kinesin-1, previously shown to mediate centrifugal transport of lysosomes in nonneuronal cells, specifically drives lysosome transport into the axon, and not the dendrites, in cultured rat hippocampal neurons. This transport is essential for maintenance of axonal growth-cone dynamics and autophagosome turnover. Our findings illustrate how a general mechanism for lysosome dispersal in nonneuronal cells is adapted to drive polarized transport in neurons, and emphasize the importance of this mechanism for critical axonal processes.


Skin grafting of the anterior stomach wall following pharyngo-gastric anastomosis.
A simple fast reliable technique for closure of an anterior neck defect is described. The technique involves skin grafting directly onto the serosal layer of the anterior stomach wall following pharyngo-laryngo-oesophagectomy and pharyngo-gastric anastomosis.


Alcohol-use disorders and suicide: Results from a psychological autopsy study in Australia.
People who die by suicide have a higher risk of an alcohol-use disorder (AUD) at the time of death. The present study aims to compare 1) suicide cases with and without AUD, and 2) suicide and sudden-death controls with AUD. The psychological autopsy method was utilized to investigate suicide and sudden death in Australia (QLD and NSW). Initial information was gathered from coroners' offices. Potential informants were approached and semi-structured interviews were conducted. Univariate and multivariate logistic regression were applied. People with AUD who died by suicide were significantly more likely to have another substance-use disorder, history of suicide attempt, recent serious arguments with spouse/partner and other family members, been unfaithful to partner/spouse, be victims of a crime, and were less likely to be from a non-English speaking background. They were also younger and had higher levels of aggression compared to non-AUD suicides. AUD suicides were more likely to have mood disorders, previous suicide attempt, expressing hopelessness, higher scores in aggression towards self, romantic relationship breakup, and serious arguments with other family members than AUD sudden deaths. Aggressive behavior, having another substance-use disorder, and history of serious arguments with family members remained significant in the final model comparing suicides with and without AUD. Our findings support that aggressive behavior, comorbidity with other psychiatric disorders as predisposing factors, and recent interpersonal conflicts such as breakup and family conflicts can trigger suicide in people with AUD. There is a need for proper diagnosis, risk assessment, and treatment in suicidal people with AUD.


Prevalence of meeting physical activity guidelines for cancer prevention in Alberta.
Guidelines for recommended physical activity (PA) levels have been developed by the Canadian Society for Exercise Physiology (CSEP) and the U.S. Department of Health and Human Services (USDHHS) for health benefits and by the American Cancer Society (ACS) and the World Cancer Research Fund/American Institute for Cancer Research (WCRF/AICR) for cancer prevention benefits. We examined if these guidelines were met using a sample of 14 294 Albertan participants of the Tomorrow Project, aged 35 to 64 years, enrolled from 2001 to 2005. We used logistic regression to examine correlates of leisure PA behaviour. An estimated 55%, 42%, 26% and 23% of participants met CSEP, ACS, USDHHS, and WCRF/AICR guidelines, respectively. Women were less likely than men to meet ACS (Odds Ratio [OR] = 0.72, 95% confidence interval [CI]: 0.55-0.93), USDHHS (OR = 0.67, 95% CI: 0.50-0.89) and WCRF/AICR (OR = 0.63, 95% CI: 0.47-0.85) guidelines, and being obese was correlated with not meeting USDHHS (OR = 0.45, 95% CI: 0.32-0.65) and WCRF/AICR guidelines (OR = 0.79, 95% CI: 0.63-0.98). Albertans, particularly women and obese individuals, are not sufficiently active for cancer prevention benefits.


Chest pain caused by myocardial ischemia or not: sometimes the old handgrip test can solve the dilemma.
We describe the case of a patient with typical chest pain but negative maximal bicycle exercise-electrocardiogram test and for whom significant coronary artery disease was hypothesized by a positive handgrip exercise test and demonstrated by coronary angiography. Despite negative exercise stress test, handgrip as well as other provocative tests have to be considered when the pretest probability of coronary artery disease is intermediate and the symptoms are typical for angina.


Changtai granule, a traditional Chinese drug, protects hapten-induced colitis by attenuating inflammatory and immune dysfunctions.
The study was aimed to investigate the effects and mechanism of action of Changtai granule (CT), a traditional compound Chinese medicinal formula, in rodent 2,4,6-trinitrobenzene sulfonic acid (TNBS) colitis. Rats with TNBS/ethanol-induced colitis were used. The colonic wet weight, myeloperoxidase (MPO) activity, macroscopic and histological colon injury was observed. Inflammation cytokines were determined by ELISA methods and semi-quantitative RT-PCR. When dosed orally once daily, CT markedly attenuated TNBS-induced colitis. CT significantly attenuated colonic wet weight, macroscopic and histological colon injury. CT decreased mucosal mRNA levels for several inflammatory mediators: inducible nitric oxide synthase, cyclooxygenase 2, and macrophage inflammatory protein 2. CT also decreased mucosal mRNA and protein levels of T effectors cytokines: tumor necrosis factor-alpha (TNF-alpha), interleukin-2 (IL-2) and interferon-gamma (IFN-gamma). Systemic levels of these cytokines were also dramatically attenuated. CD3/CD28-mediated costimulation of T helper 1 effector cytokines release in lamina propria mononuclear cells (LPMC) was markedly inhibited by CT ex vivo and in vitro. Also CT prevented cytokines production by nuclear factor-kappaB (NF-kappaB). The potential anti-inflammatory and immunomodulatory effect of CT in TNBS colitis suggests that CT may be an effective treatment approach for inflammatory bowel disease.


Efficacy of early intravenous bolus oxycodone or fentanyl in emergence from general anaesthesia and postoperative analgesia following laparoscopic cholecystectomy: A randomized trial.
To compare prospectively the efficacy of early intravenous bolus of oxycodone or fentanyl in providing analgesia at emergence from general anaesthesia following laparoscopic cholecystectomy. Patients were randomly assigned to receive either 0.08 mg/kg oxycodone (Group O) or 1 µg/kg fentanyl (Group F), 20 min before the end of surgery. Postoperative pain was evaluated using a visual analogue scale (VAS). The time to first postoperative analgesic dose, requirement for analgesia and side-effects were assessed in the postanaesthesia care unit (PACU). The VAS scores at 0 min and 30 min and requirement for analgesia were significantly lower in Group O (n = 28) than in Group F (n = 26). The time to first analgesia dose was significantly longer in Group O than Group F. There were no significant between-group differences in the incidence of side-effects. Oxycodone relieves immediate postoperative pain significantly better than fentanyl, and is not associated with an increase in side-effects in patients undergoing laparoscopic cholecystectomy.


Peptide-containing nerves in human urinary bladder.
Nerves containing immunoreactive vasoactive intestinal polypeptide (VIP), substance P and two newly discovered peptides, neuropeptide tyrosine (NPY) and PHI (peptide having N-terminal histidine and C-terminal isoleucine), have been found in the human urinary bladder by immunocytochemistry and radioimmunoassay. Somatostatin immunoreactivity was detected by radioimmunoassay. The VIP-immunoreactive nerves were widely distributed in all regions, but were particularly dense beneath the epithelium and in the muscle layer. Scattered intramural ganglia were found to be reactive to VIP antiserum. Higher concentrations of extractable VIP were detected in the trigone than in the dome. VIP- and PHI-immunoreactive nerves were similarly distributed, the latter being less numerous. NPY-immunoreactive nerves were seen mainly in the muscle layer, particularly in the trigonal area. The distribution patterns of VIP- and NPY-immunoreactive nerves resembled those of the previously reported cholinergic and adrenergic nerves, respectively. Many blood vessels were found to be innervated by both types of immunoreactive nerves. Scattered substance P-immunoreactive fibers were occasionally seen, being present in the submucosa and around the detrusor muscles. The significance of these nerves remains to be elucidated.


Being "easy": Women's desire to accept help in later life.
This article examines the concept of being "easy" as a quality that female caregivers aspire to as they envision their potential care in later life. It draws on data from a qualitative study exploring the significance of caregiving to adult daughters' perspectives and plans for their own aging. Caregivers' experiences differ based upon their perception of how easy or difficult it was to care for their loved one. The former exemplifies a positive way to approach aging and the receiving of care. Being easy describes an attitude that these caregivers desired for their own later life.


Interpretation of the excess adsorption isotherms of organic eluent components on the surface of reversed-phase phenyl modified adsorbents.
The adsorption of three organic eluent components (acetonitrile, methanol, and tetrahydrofuran) from water were measured on four phenyl-type bonded phases using the minor disturbance method. The thicknesses of organic layer enriched above the phenyl-type bonded ligands were assessed and interpreted. Acetonitrile and tetrahydrofuran showed multilayer formation while methanol showed monomolecular adsorption. These results were compared to those obtained on alkyl bonded phases.


A comparison of canine normal hepatic alkaline phosphatase and variant alkaline phosphatase of serum and liver.
The isoenzyme of alkaline phosphatase from normal liver, the corticosteroid induced isoenzyme of alkaline phosphatase from serum and liver and a hepatocellular variant isoenzyme of alkaline phosphatase induced by lymphosarcoma have been partially purified and their the present modification incorporates Polybrene into buffer to eliminate this heparin interference. The proposed method shown excellent agreement with a reference procedure based on clottable protein, and excellent day-to-day precision (C.V.3.5%). The present method is easily adaptable to semi-automated measurements.


Psychiatric Morbidity Is Common Among Children With Juvenile Idiopathic Arthritis: A National Matched Cohort Study.
Juvenile idiopathic arthritis (JIA) is a chronic rheumatic disease that causes joint inflammation and pain. Previous studies have indicated affected mental health and increased risk of psychiatric conditions among patients with JIA. We aimed to explore differences in psychiatric morbidity between children with JIA and their peers. We further studied if parental socioeconomic status (SES) influences the association between JIA and the risk of psychiatric morbidity. We used a matched cohort design to estimate the association between JIA and psychiatric disease. Children with JIA, born between 1995 and 2014, were identified in Danish national registers. Based on birth registers, we randomly selected 100 age- and sex-matched children per index child. Index date was the date of the fifth JIA diagnosis code or the date of matching for reference children. End of follow-up was the date of psychiatric diagnosis, death, emigration, or December 31, 2018, whatever came first. Data were analyzed using a Cox proportional hazard model. We identified 2086 children with JIA with a mean age at diagnosis of 8.1 years. Children with JIA had a 17% higher instantaneous risk of a psychiatric diagnosis when compared with the reference group, with an adjusted hazard ratio of 1.17 (95% CI 1.02-1.34). Relevant associations were found only for depression and adjustment disorders. Stratifying our analysis for SES showed no modifying effect of SES. Children with JIA had a higher risk of psychiatric diagnoses compared to their peers, especially diagnoses of depression and adjustment disorders. The association between JIA and psychiatric disease did not depend on parental SES.


[An unusual case of elevated and persistent levels of LDH in the blood of a patient after acute myocardial infarct].
The Authors illustrate a peculiar case of elevated and persistent levels of LDH activity in serum of a patient after acute myocardial infarction. The patient was also affected by a testicular tumor. Surgical ablation of neoplasm normalized LDH activity serum.


Psychosocial factors associated with indices of cortisol production in women with breast cancer and controls.
The present study was designed to (i) explore which psychosocial factors were associated with indices representing the early morning peak, diurnal cortisol rhythm and area under the curve (AUC); (ii) examine whether the relationships between psychosocial functioning and these cortisol indices were consistent and (iii) explore whether these relationships were influenced by the clinical status of the participant. Newly diagnosed breast cancer patients (n = 85) and healthy control women (n = 59) were recruited. State and trait measures of psychosocial functioning (i.e. anxiety, depression, distress, neuroticism, extraversion, marital satisfaction and mastery) were undertaken. In addition, all participants provided four saliva samples (on waking, 30 min later, between 11 and 1 p.m., before lunch and between 8 and 10 p.m., at least 2h after evening meal) over two consecutive days to assess cortisol levels. The results highlighted the divergent nature of the four cortisol indices; revealed the presence of some significant relationships between the psychosocial measures and the cortisol indices; but highlighted inconsistencies in the relationships evident for patients and those observed for control women.


Diagnosis of Leishmania (Leishmania) chagasi infection in dogs and the relationship with environmental and sanitary aspects in the municipality of Palmas, State of Tocantins, Brazil.
The aim of the present study was to identify the presence of Leishmania (Leishmania) chagasi infection in dogs in the City of Palmas, Tocantins, Brazil, using the PCR technique to list the hot spots of infected dogs in the city and associate their occurrence to significant environmental changes at capture sites. DNA was extracted from blood of dogs, and the PCR were performed with primers RV1/RV2. After screening the population studied, the regions of the city that had the highest occurrence of canine infection were detected. These sites were visited, and ecological parameters denoting anthropogenic disturbance were evaluated. Some important features were listed in the regions visited, such as low urbanization, lack of public collection of sewage, limited garbage collection, vacant lots with tall vegetation, decaying organic matter, and, most importantly, the occurrence of stray dogs and poultry in homes. The methodology for screening the population was very efficient, especially in evaluating a large number of individuals in a short time, with a high degree of automation. The results indicate an association between the observed parameters and the occurrence of infection in dogs. The model presented in the city is ideal for studies of disease progression and expansion and for the evaluation of control measures adopted for canine VL.


Characterization of low-cost glycolipoprotein biosurfactant produced by Lactobacillus plantarum 60 FHE isolated from cheese samples using food wastes through response surface methodology and its potential as antimicrobial, antiviral, and anticancer activities.
Considering the need of new lactic acid bacteria (LAB) for the production of novel biosurfactant (BS) molecules, the current study brings out a new insight on the exploration of cheese samples for BS producers and process optimization for industrial applications. In view of this, Lactobacillus plantarum 60FHE, Lactobacillus paracasei 75FHE, and Lactobacillus paracasei 77FHE were selected as the most operative strains. The biosurfactants (BSs) described as glycolipoproteins via Fourier-transform infrared spectroscopy (FTIR) exhibited antimicrobial activity against the food-borne pathogens. L. plantarum 60FHE BS showed an anticancer activity against colon carcinoma cells and had a week antiviral activity against Hepatitis A virus. Furthermore, glycolipoprotein production was enhanced by 1.42-fold through the development of an optimized process using central composite design (CCD). Emulsifying activities were stable after 60-min incubation from 4 to 120 °C, at pH 2-12, and after the addition of NaCl (2-14%). Characterization by nuclear magnetic resonance spectroscopy (1H NMR) revealed that BS produced from strain 60FHE was glycolipoprotein. L. plantarum produced mixed BSs determined by Liquid Chromatography/Mass Spectrometry (LC-MS). Thus, indicating that BS was applied as a microbial food prevention and biomedical. Also, L. plantarum 60FHE BS was achieved with the use of statistical optimization on inexpensive food wastes.


Simultaneous determination of seven coumarins by UPLC-MS/MS: Application to a comparative pharmacokinetic study in normal and arthritic rats after oral administration of Huo Luo Xiao Ling Dan or single-herb extract.
A simple, sensitive and reliable ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for simultaneous quantitation of seven coumarins, the bio-active ingredients of Huo Luo Xiao Ling Dan (HLXLD), in rat plasma. The liquid-liquid extraction method with ether-dichloromethane (2:1, v/v) was used to prepare the plasma samples. Analytes and internal standard (IS) of bifendate were separated on a Shim-pack XR-ODS column (75mm×3.0mm, 2.2μm particles) using gradient elution with the mobile phase consisting of methanol and 0.05% formic acid in water at a flow rate of 0.4mL/min. Detection was performed on a triple quadrupole (TQ) tandem mass spectrometry equipped with an electrospray ionization source in the positive ionization and multiple reaction monitoring (MRM) mode. The lower limits of quantitation (LLOQ) were 0.03-0.25ng/mL for all the analytes. Intra- and inter-day precision and accuracy of the seven analytes were well within acceptance criteria (15%). The matrix effect and the mean extraction recoveries of the analytes and IS from rat plasma were all within satisfaction. The validated method has been successfully applied to compare pharmacokinetic profiles of the seven active ingredients in rat plasma between normal and arthritic rats after oral administration of HLXLD, Angelica pubescens extract and Notopterygium incisum extract, respectively. Results showed that there were remarkable differences in pharmacokinetic properties of the analytes among the different groups.


Regression of Acanthosis Nigricans with the Addition of Sitagliptin and Pioglitazone.
Acanthosis nigricans (AN) is a cutaneous disorder associated with various diseases. There are few documented cases of regression of AN. We discuss a case of a 48-year old diabetic woman with resolution of AN after treatment with sitagliptin and pioglitazone.


Insomnia as an expression of obstructive sleep apnea syndrome--the effect of treatment with nocturnal ventilatory support.
Obstructive sleep apnea syndrome (OSAS) and insomnia often coexist, and it is estimated that nearly half of those who suffer from the former report symptoms of the latter. The fact that these patients have no other causes of insomnia indicates that it is a sign of OSAS. The aim of the study is to evaluate the effectiveness of nocturnal ventilatory support (NVS) in the treatment of insomnia secondary to OSAS. In order to conduct the retrospective study, the authors reviewed the medical records of patients with insomnia and OSAS that had received NVS. Patients with psychiatric disorders, sleep movement disorders, psycho-physiological insomnia, circadian rhythm sleep disorders, inadequate sleep hygiene, use and abuse of hypnotic agents, stimulants, antidepressants, anxiolytics and alcohol, were excluded. For the selected patients, the effects of NVS in terms of clinical signs and symptoms of insomnia, apnea-hypopnea index (AHI), Epworth Sleepiness Scale (ESS) score, and number of sleep hours were analyzed, before and after treatment with NVS. After reviewing 1241 medical records, 56 patients were selected, with a mean age of 60.9±10.0 years. Twenty-two (39.3%) suffered from intermediate insomnia, 19 (33.9%) had initial insomnia, eight (14.3%) had the mixed type, and seven patients (12.5%) had terminal insomnia. The majority of patients (n=48; 85.7%) were treated with auto-titrating continuous positive airway pressure (APAP). Forty-four patients (78.6%) overcame insomnia; insomnia symptoms persisted in nine (16.1%), and three (5.4%) patients abandoned during the medical follow-up. There was an association between the type of insomnia and its resolution and, in percentage terms patients with the mixed type did not manage to overcome insomnia symptoms (75%). There was a statistically significant difference between patients that overcame insomnia and those who did not in terms of the average time which elapsed between the initiation of treatment with NVS and compliance with the adherence criteria: 161±61 days for the former, and 225±141 days for the latter (p=0.003). Before and after the NVS treatment, patients slept an average of 5.29±1.37 and 6.37±1.55h per night, respectively (p<0.001). Among the patients who overcame insomnia, six did not meet the treatment adherence criteria: five adhered more than 4h/night in less than 70% of all nights (60.6±3.2%), and one patient adhered less than 4h in all nights (3.5h/night). NVS has proved effective in treating insomnia secondary to OSAS, and favorable results could be observed even in patients that did not meet the criteria of NVS adherence. Results suggest that all insomnia subtypes, except the mixed subtype, may derive from OSAS.


Scanning electron microscope study on flat type membrane oxygenator after prolonged extracorporeal circulation.
Despite adequate anticoagulation, thrombi may develop onto any foreign surface subjected to circulating blood. Although the efficiency of membrane oxygenators can decrease, distal embolization can be more critical and dangerous. Scanning electron microscope observations carried out on the Travenol Modulung oxygenator used both on dogs and on patients for prolonged partial cardiopulmonary by-pass pointed out the following observations. Caution must be observed when translating the results from dogs to human beings. Heparin management can be delicate and critical. The problem of blood transfusion can be extremely important and must be extensively investigated.


A study using LC-MS/MS-based metabolomics to investigate the effects of iron oxide nanoparticles on rat liver.
Iron oxide nanoparticles (IONPs) are widely used in food additives, but their metabolic mechanism in the body is still unclear. In this study, male Sprague-Dawley rats were orally administered with IONPs for 28 days to investigate the adverse effect and metabolic mechanism on liver by the combination of traditional toxicology technology and liquid chromatography tandem-mass spectrometry (LC-MS/MS)-based metabolomics. The results showed that IONPs could increase the concentration of blood glucose and the metabolites in the liver of the control and IONPs-treated group were significantly changed. A total of 32 different metabolites were found, including choline, Phosphatidylcholine (PC), Phosphatidylethanolamine (PE), Phosphatidylserine (PS), etc. Pathway analysis based on KEGG database demonstrated that the glycerophospholipid metabolism pathway would be affected. And the expression of the key enzymes of altered metabolomics pathway was further verified at the transcription level. In short, our study clarified oral exposure to IONPs would induce lipid metabolism disorders in the liver of rats, which provided useful information about their safety and potential risks.


Recent Studies on Serotonin 5-HT2A Receptor Antagonists in Medicinal Chemistry: A Last Decades Survey.
In the last decades, much attention has been paid to the functioning of receptors to understand better how they work with various chemical motifs. Among different families, G-proteincoupled receptor (GPCR) families have drawn much attention in the twenty-first century. They are the most prominent signal transducer across the cell membrane, comprising thousand-odd proteins. One of the members of GPCRs is the serotonin 2A (5-HT2A) receptor, which has been associated with complex etiological mental illnesses. In this survey, we collected data on 5-HT2A, i.e., the role of 5- HT2A receptors in human and animal analogy, various binding site functionalities, advanced effects, and synthetic aspects.


No evidence for significant association between GABA receptor genes in chromosome 15q11-q13 and autism in a Japanese population.
The gamma-aminobutyric acid (GABA) receptor genes GABRB3, GABRA5, and GABRG3 located on chromosome 15q11-q13 have been major candidates for susceptibility genes for autism, a neurodevelopmental disorder with a complex genetic etiology. In this study, we first investigated the association between the GABA receptor genes and autism in a Japanese population by analyzing 11 single nucleotide polymorphisms (SNPs). Intron 3 of GABRB3 was densely mapped because the previous studies observed the association of the microsatellite 155CA-2 located in the region. We observed no significant difference in allelic frequencies or genotypic distributions of the 11 SNPs between patients and controls. A permutation test showed no significant global differences in estimated haplotype frequencies between patients and controls. Analysis after confining the subjects to males showed similar results. Thus, this study provides no positive evidence of an association between the GABA receptor genes and autism in a Japanese population. However, in a SNP (rs3212337) located near the microsatellite 155CA-2, a significant deviation from the Hardy-Weinberg equilibrium was observed in patients (p = 0.029, corrected for multiple testing). This finding may suggest further studies around the markers for more definitive conclusions.


The effects of an exercise training on steroid hormones in preadolescent children - a moderator for enhanced cognition?
Research demonstrates a variety of cognitive benefits induced by different chronic physiological stressors (e.g. motor vs. cardiovascular exercise training) and acute exercise to affect steroid hormone secretion, the present study aims to investigate 1.) motor vs. cardiovascular exercise training and their impact on salivary Testosterone, Estradiol and Progesterone and 2.) whether alterations in steroid hormones might moderate exercise-induced cognitive benefits. In a randomized control trial, 71 preadolescent children (39 girls) at the age of 9-10 years were randomly assigned to a cardiovascular exercise group (CE), a motor exercise group (ME) as well as a control group (CON) in which students were attending assisted homework. The participants attended their program three times per week, for a total of 10 weeks. Steroid hormones in the saliva, working memory performance and state-trait anxiety levels were determined before and after the intervention. Statistical analysis showed no chronic changes in hormones through the interventions. Stepwise hierarchical regression revealed the Testosterone*ME-interaction to be a significant moderator for working memory performance post-intervention (β = 0.48, p = 0.014). Within the ME, increases of Testosterone from t1 to t2 were predicting higher working memory performance post-intervention (β = 0.45, R2 = 0.16, p = 0.038). Our findings suggest 1.) that motor-exercise-induced cognitive benefits are moderated by changes in Testosterone and 2.) that a 10-week exercise training of moderate intensity and with a motor demanding vs a cardiovascular character does not affect the chronic concentration of salivary Testosterone, Estradiol and Progesterone nor anxiety levels in preadolescent children.


COVID-19, food insecurity and dietary diversity of households: Survey evidence from Nigeria.
The policy measures of the government of Nigeria to restrain the spread of COVID-19, particularly in the initial three months (April - June 2020) led to significant disruptions to household livelihoods and food security. We investigate the effects of COVID-19 on food security and dietary diversity of households; focusing on the pathways through which income loss, endowments of wealth, social capital, and safety net programs moderate the severity of households' food security and dietary diversity. Primary data obtained from a telephone survey of 1,031 Nigerian households were analyzed using ordered logit and negative binomial models. Our results show that income losses due to the COVID-19 restrictive measures had pushed households into a more severe food insecurity and less diverse nutritional outcomes. Regarding wealth effects, livestock ownership significantly cushioned households from falling into a more severe food insecurity amid the pandemic. We found that because of the pandemic's indiscriminate effect across communities, the potential of social capital as an informal support mechanism might have been eroded to enable households to cope with shocks. Furthermore, safety net programs by the government and NGOs did not provide significant protection to households from falling into severe food insecurity and malnutrition amid the pandemic. We suggest three policy propositions - prioritize investment in local job creation to curb income loss; build the wealth base of households (e.g., land tenure security or livestock) to enhance resilience to shocks; and target safety nets and other social support programs spatially, temporally, and across social groups to enhance the effectiveness of such programs amid shocks.


Constitutive expression of CYP3A mRNA in Bama miniature pig tissues.
The pig, particularly the miniature pig (minipig), is becoming an important animal model due to its physiological and anatomical similarities to humans. Bama minipigs (Sus scrofa domestica), a Chinese natural minipig breed, are a promising animal model. The pig is a useful model for drug metabolism and pharmacological studies due to the similar properties of cytochrome P450 (CYP)3A between pigs and humans. However, a detailed investigation regarding the abundance and expression of CYP3A in porcine tissues, particularly in minipig tissues, has not been performed. The present study investigated constitutive expression of CYP3A mRNA in Bama minipig tissues using real-time reverse transcriptase-polymerase chain reaction (RT-PCR). We found that the expression of CYP3A mRNAs relative to the endogenous control, β-actin (ACTB), was lower than when compared to the expression of the endogenous control, TATA-binding protein (TBP), except for the expression of CYP3A29 mRNA in the spleen, adrenal gland, testis, and epididymis, and CYP3A46 in the spleen. Expression levels of all three CYP3As were highest in the liver amongst all the tissues tested, and the order of relative mRNA expression level of the three CYP3As was different between other tissues. We also analyzed the relative expression of the three CYP3A mRNAs in each tissue. CYP3A46 had the highest expression in all extrahepatic tissues, whereas CYP3A22 had the highest expression in the liver, and CYP3A29 had the lowest expression in all tissues except in the duodenum, where it had higher expression than CYP3A22. Because CYP3A22 and CYP3A46 were the most highly expressed isoforms, it could follow that they are probably important functional CYP3A genes for Bama miniature pig. Our present work will broaden the understanding of the physiological functions of CYP3As in the Bama minipig and promote its application in drug metabolism and pharmacological studies. Our results also indicate that the breed, age, and castration status of the pig should be considered when using the pig as an animal model in pharmacological applications.


Quantitative determination of sodium monofluoroacetate "1080" in infant formulas and dairy products by isotope dilution LC-MS/MS.
A fast and easy-to-use confirmatory liquid-chromatography tandem mass-spectrometry (LC-MS/MS) based-method was developed for the analysis of the pesticide sodium monofluoroacetate (MFA, also called "1080") in infant formulas and related dairy products. Extraction of the compound encompassed sample reconstitution and liquid-liquid extraction under acidic conditions. Time-consuming solid-phase extraction steps for clean-up and enrichment and tedious derivatisation were thus avoided. Resulting sample extracts were analysed by electrospray ionisation (ESI) in negative mode. Quantification was performed by the isotopic dilution approach using (13)C-labelled MFA as internal standard. The procedure was validated according to the European document SANCO/12571/2013 and performance parameters such as linearity (r(2) > 0.99), precision (RSD(r) ≤ 9%, RSD(iR) ≤ 11%) and recovery (96-117%) fulfilled its requirements. Limit of quantifications (LOQ) was 1 µg kg(-1) for infant formulas and related dairy products except for whey proteins powders with a LOQ of 5 µg kg(-1). Method ruggedness was further assessed in another laboratory devoted to routine testing for quality control.


Binding of monoclonal anti-native DNA autoantibodies to DNA of varying size and conformation.
A microchemical assay for phosphorus was applied to the measurement of DNA in immune complexes formed with monoclonal or serum anti-DNA autoantibodies and DNA of varying size and conformation. Two monoclonal antibodies were produced by hybridomas derived from spleen cells of autoimmune MRL-lpr/lpr mice and were purified from culture fluid by affinity chromatography on columns of goat anti-mouse Ig-Sepharose. Double-helical DNA fragments were prepared by brief digestion of calf thymus DNA with micrococcal and S1 nucleases and fractionation on Sepharose 4B; their double-stranded structures was confirmed by measurement of thermal denaturation. Immune complexes were formed with monoclonal or serum antibodies and native DNA or DNA fragments or denatured DNA; the complexes were precipitated with goat anti-mouse IgG and washed, and DNA phosphorus content of the precipitates was measured. With one monoclonal autoantibody (H241), there were discontinuous increases in the amount of DNA that could be bound (and decreases in the antigen concn required for half-maximal binding) as the DNA size increased. There were especially marked increases in binding efficiency as fragment size increased from an average of 100 (range 85-105) to an average of 150 (range 105-170) base pairs, and again between 450 (range 360-620) and 600 (range 425-825) base pairs. A second monoclonal antibody (H143) did not show significant variation in binding with DNA fragments larger than 300 base pairs. With smaller fragments, the amount of DNA bound by H143 was reduced, but the DNA concn required for half-maximal binding was not. Affinities of these monoclonal antibodies were within the spectrum of human systemic lupus erythematosus serum IgG anti-DNA autoantibodies. The dependence of binding on mol. wt is important in the evaluation of these monoclonal antibodies as biochemical reagents and as potential participants in formation of immune complexes in vivo.


Single tryptophan of disordered loop from Plasmodium falciparum purine nucleoside phosphorylase: involvement in catalysis and microenvironment.
Among various tropical diseases, malaria is a major life-threatening disease caused by Plasmodium parasite. Plasmodium falciparum is responsible for the deadliest form of malaria, so-called cerebral malaria. Purine nucleoside phosphorylase from P. falciparum is a homohexamer containing single tryptophan residue per subunit that accepts inosine and guanosine but not adenosine for its activity. This enzyme has been exploited as drug target against malaria disease. It is important to draw together significant knowledge about inherent properties of this enzyme which will be helpful in better understanding of this drug target. The enzyme shows disorder to order transition during catalysis. The single tryptophan residue residing in conserved region of transition loop is present in purine nucleoside phosphorylases throughout the Plasmodium genus. This active site loop motif is conserved among nucleoside phosphorylases from apicomplexan parasites. Modification of tryptophan residue by N-bromosuccinamide resulted in complete loss of activity showing its importance in catalysis. Inosine was not able to protect enzyme against N-bromosuccinamide modification. Extrinsic fluorescence studies revealed that tryptophan might not be involved in substrate binding. The tryptophan residue localised in electronegative environment showed collisional and static quenching in the presence of quenchers of different polarities.


Modeling insufficient effort responses in mixed-worded scales.
Surveys often add reverse-coded questions to monitor respondents with insufficient effort responses (IERs) but often wrongly assume that all respondents consistently answer all questions with full effort. By contrast, this study expanded the mixture model for IERs and ran a simulation via LatentGOLD to show the harmful consequences of ignoring IERs to positively and negatively worded questions: less test reliability, bias and less accuracy in slope and intercept parameters. We showed its practical application to two public data sets: Machiavellianism (five-point scale) and self-reported depression (four-point scale).


Cellular mechanisms of action of mineralocorticoid hormones.
Mineralocorticoid hormones are a subset of steroid hormones that act primarily in epithelial tissues to regulate ion transport of Na+, K+ and H+. Cellular specificity is conferred by receptors which act in the nucleus to stimulate gene expression. Transcription and subsequent translation result in the production of new proteins which mediate the physiologic effects. The mechanisms involved in receptor specificity and localization, in regulation of gene activation, and in expression of transport effects are reviewed. The cellular actions of mineralocorticoids fit well with the general model of steroid hormone action but considerable questions remain at each step in the process.


Identification of domains of the human A1 adenosine receptor that are important for binding receptor subtype-selective ligands using chimeric A1/A2a adenosine receptors.
To provide new insights into the regions of the human A1 adenosine receptor (A1AR) involved in ligand binding, a series of chimeric human A1 and rat A2a adenosine receptors (A1/A2a) were studied. Binding studies were initially performed on acutely transfected COS cells using fixed doses of the A2aAR agonist [3H]CGS-21680, the A1AR agonist [3H]2-chloro-N6-cyclopentyladenosine (CCPA), and the A1AR antagonist [3H]8-cyclopentyl-1,3-dipropylxanthine (DPCPX). When the region of the A2aAR from the amino terminus to the end of transmembrane (TM) 1 was replaced by the corresponding region of the A1AR (A1TM1/A2a), [3H]CGS-21680 and [3H]CCPA binding was detectable. When an A1TM1-2/A2a construct was studied, [3H]CGS-21680 binding was lost and [3H] DPCPX binding appeared. Saturation studies using [3H]CCPA revealed that the A1TM1/A2a construct had low affinity. However, with the subsequent addition of A1AR TMs 2-4 receptor affinity improved markedly. Saturation studies using [3H]DPCPX also revealed that the TMs 1-4 of the A1AR conferred wild-type receptor affinity. When the ligand binding properties of A1TM1-4/A2a, A1TM1-6/A2a, and wild type A1AR constructs were directly compared, no differences were found using 10 different compounds. When truncated A1ARs that extended from the amino terminus to shortly after TM4 were examined, no binding was detectable suggesting that the amino half of the receptor alone is not sufficient for ligand binding. Collectively, these data suggest that the important determinants for A1AR agonist and antagonist binding and ligand specificity are present in TMs 1-4.


Solvent Effect on the Solvothermal Synthesis of Mesoporous NiO Catalysts for Activation of Peroxymonosulfate to Degrade Organic Dyes.
In this work, we successfully prepared three different mesoporous NiO nanostructures with preferential (111) planes using three different solvents-water, a water-ethanol mixture, and a water-ethylene glycol mixture. The NiO nanosheets prepared from the water-ethylene glycol mixture and denoted as NiO-EG showed a nanosheet morphology thinner than 10 nm, whereas the water-ethanol and water samples were 30-40 nm and above 100 nm thick, respectively. The NiO-EG catalyst was found to exhibit a high catalyzing ability to activate peroxymonosulfate (PMS) for decoloring dyes, by which 94.4% of acid orange 7 (AO7) was degraded under the following reaction conditions: AO7 = 50 mg/L, catalyst = 0.2 g/L, PMS = 0.8 g/L, pH = 7, and 30 min reaction time. The dye degradation rate was investigated as a function of the catalyst dosage, pH, and dye concentration. According to quenching experiments, it was found that SO4 •-, HO•, and O2 •- were the dominant radicals for AO7 degradation, and oxygen vacancies played a significant role in the generation of radicals. High surface area, thin flaky structure, rich oxygen vacancies, fast charge transport, and low diffusion impedance all enhanced the catalytic activity of NiO-EG, which exhibited the highest degradation ability due to its abundant accessible active sites for both adsorption and catalysis.


A patient presenting with acute abdomen due to metastatic small bowel melanoma: a case report.
Malignant tumors of the small bowel are rare. Melanoma of the small intestine is in most cases metastatic from a primary skin lesion. Perforation of small bowel melanoma is an extremely rare entity. To the best of our knowledge this is the fifth case published to date. We report a rare case of acute abdomen due to perforated metastatic small bowel melanoma in a 38-year-old Caucasian man. In the majority of cases small bowel melanoma represents metastasis from cutaneous sites. Although rare, the possibility of abdominal metastatic melanoma presenting with the clinical picture of acute abdomen must be always considered by the operating surgeon in patients with a history of primary cutaneous malignant lesion.


Arterial vasodilator, systemic and coronary hemodynamic effects of nisoldipine in congestive heart failure secondary to ischemic or dilated cardiomyopathy.
The systemic and coronary hemodynamic and neurohumoral effects of nisoldipine, a calcium antagonist drug with high vascular specificity, were investigated in 17 patients with chronic congestive heart failure (CHF). Brachial artery infusions (n = 9) decreased forearm vascular resistance in a dose-dependent manner, attesting to its powerful arterial vasodilator properties. A dose of 3 micrograms/kg intravenously decreased mean blood pressure 16% and systemic vascular resistance 33%, while increasing stroke index 19% and ejection fraction 21% at rest and similarly during exercise. Pulmonary capillary wedge pressure decreased significantly during exercise. Intravenous infusion of nisoldipine increased rest coronary sinus flow 10% (p less than 0.05, n = 7), decreased rest and exercise coronary vascular resistance 28% and 19% (p less than 0.01) and rest myocardial oxygen consumption 14% (p less than 0.05). In 13 patients similar systemic hemodynamic results were found after treatment with oral nisoldipine, 2 X 20 mg for 4 weeks. Stroke index and stroke work index increased long-term more than acutely (31% and 12.4% vs 19% and 4.5% at rest, both p less than 0.05), which may indicate a compensated mild cardiodepressant effect of intravenous nisoldipine. Changes in forearm vascular resistance after intra-arterial administration did not correlate with changes of systemic vascular resistance after intravenous administration, suggesting that factors other than vascular calcium entry blockade importantly influence hemodynamic responses. Elevated control plasmas norepinephrine and renin levels, on average, did not change during chronic therapy but individual changes were compatible with a reduction of sympathetic activity in patients with hemodynamic improvement.(ABSTRACT TRUNCATED AT 250 WORDS)


Age related changes in the rate of stress relaxation within the rat respiratory system.
Stress relaxation within the respiratory system was examined in young rats at different ages, 0-1 day old (GPI), 4-5 days old (GPII) and 30-40 days old (GPIII). Each rat was anesthetized, tracheostomized and placed inside a saline filled plethysmograph with the tracheal cannula projecting through the wall of the plethysmograph. Volume history was standardized by three inflations to a mean transrespiratory system pressure (P) of 20-25 cm H2O, then P was set to zero and lung volume was abruptly increased by rapidly withdrawing fluid from the plethysmograph. Following the volume step, lung volume was maintained constant and changes in P due to stress relaxation were recorded for 30 sec. The rate of stress relaxation was obtained by calculating the slope (R) of the normalized change in P per unit of time on a semi-log plot. GPIII rats exhibited the slowest rate (R = 0.068 +/- 0.004 SD) whereas GPII rats demonstrated the fastest rate (R = 0.092 +/- 0.011 SD). Stress relaxation in GPI was intermediate (R = 0.076 +/- 0.005 SD). Values of dynamic (Cdyn) and static (Cstat) compliance were determined for rats similar in age to the three age groups used to determine R. The difference between Cstat and Cdyn increased with R and was greatest in rats between 0 and 6 days old. We conclude that: (1) stress relaxation within the rat respiratory system at 30-40 days is less than that found in rats during the first week after birth, and (2) changes in the viscoelastic properties of the respiratory system may contribute to age related variation in the difference between Cstat and Cdyn.


Implantation of artificial sphincter for anal incontinence. Report of five cases.
The implantation of an artificial anal sphincter in five patients with anal incontinence of neuromuscular origin is described. The prosthesis, an AMS 800 artificial urinary sphincter (American Medical Systems), worked well with solid or semisolid stool, but less satisfactorily when diarrhea was present. Manometric and defecographic studies seem to indicate that the effect of the prosthesis may be ascribed partly to maintenance of an acute anorectal angle even during straining.


The effectiveness and safety of direct-acting antivirals for hepatitis C virus treatment: A single-center experience in Saudi Arabia.
The introduction of direct-acting antivirals (DAA) to treat the hepatitis C virus (HCV) overcame many drawbacks of interferon-based therapy. DAA achieved sustained viral response (SVR) rates above 90% and overcame many drawbacks of pegylated interferon regimens.The HCV genotype (GT) distribution varies by geographical area, with GT-4 being most prevalent in the Middle East region, including Saudi Arabia. Yet, the real-world evidence about using DAAs in the Saudi population is limited.Thus, the aim of this study to investigate the effectiveness and safety of DAAs in Saudi patients with HCV infection. A retrospective cohort study included patients treated with DAAs from 2015 to 2017 at a tertiary care hospital in Riyadh, Saudi Arabia. All patients with HCV treated with either ledipasvir plus sofosbuvir (LDS/SOF) ± ribavarin (RBV) or ombitasvir-paritaprevir-ritonavir (OBV/PTV/r) ± dasabuvir (DSV) ± RBV were included. Using a per-protocol analysis, the effectiveness outcome was the end-of-treatment response (EOTr) and Sustained virologic reponce12 weeks after competing the regimen (SVR12). The secondary safety outcome was the adverse event related to the therapy reported by the patients. A total of 97 patients were included; with the majority infected with GT-4 (64 %), followed by GT-1 (18 %), in addition to 8 % having a mixed GT (1 + 4). The EOTr and SVR12 rates were 98 % and 96 %, respectively. SVR12 was 94.4 % within the LDS/SOF ± RBV group and 97.7 % within the OBV/PTV/r ± DSV ± RBV group. Only 4 % had a response failure due to relapse or breakthrough, and all were infected with mixed GT1 + 4. Medications were well tolerated with minimal side effects, including vomiting, nausea, and weakness. DAAs regimens are associated with high rates of SVR12 and are well tolerated with a good safety profile in Saudi HCV-infected patients.


Rhizophagus proliferus genome sequence reiterates conservation of genetic traits in AM fungi, but predicts higher saprotrophic activity.
Arbuscular mycorrhizal (AM) fungi are ubiquitous endosymbionts of terrestrial plants. It helps plants to extract more nutrients from the soil and enhances the plant tolerance to various ecological stress factors. The AM fungal genome sequence helps to identify the gene repertoires that are crucial for adaptation to different habitat and mechanisms for interaction with host plant. The present work comprises the first draft of the genome sequence of Rhizophagus proliferus, which is an important AM species present in biofertilizer consortia for agricultural purpose. The estimated genome size of R. proliferus is ~ 110 Mbps and its genomic assembly is 94.35% complete. Genome mining was carried out to identify putative gene families important for biological functions. A total of 22,526 protein-coding genes were estimated in the genome, with an abundance of kinases and reduced number of glycoside hydrolases as compared to other fungal classes. A striking finding in the R. proliferus genome was higher number of carbohydrate esterases (CE), which may suggest towards presence of higher saprotrophic activity in this species as compared to the previously reported AM fungi, which may indicate towards its role as a link between plants and soil mineral nutrients. The genome sequence and annotation of R. proliferus presented here would serve as an important reference for functional genomics studies required for developing biofertilizer formulations in future. In addition, the findings from this work may also prove important in deciphering molecular mechanisms in AM fungi that govern the host-specific interaction and associated agriculture benefits.


Flexing and stretching in nonribosomal Peptide synthetases.
Re-engineering of nonribosomal peptide synthetase molecular assembly lines has been hampered by a lack of detailed knowledge concerning inter-domain substrate transfer. Recent structural studies of catalytically relevant domain-domain interactions provide valuable insights into this problem (Liu et al., 2011; Sundlov et al., 2012 [in this issue of Chemistry & Biology]).


The generation of the flower by self-organisation.
The essence of the Turing-Child theory (Schiffmann, 1991, 2017) is the direct and spontaneous conversion of chemical energy into simultaneous differentiation and morphogenesis, and all localised biological work and localised entropy-reducing processes. This is done via the identification of the Turing instability with cAMP and ATP being the Turing morphogens that mutually fulfil the five Turing inequalities. A flower model like the ABC model is derived from experiments with mutations. But what actually generates the model in real development? That is, how do genes of class A come to be expressed in the sepal and petal whorls, genes of class B in the petal and stamen whorls, and genes of class C in the stamen and carpel whorls. We suggest that the generation of the ABC model occurs via sequential compartmentalisation by Turing-Child eigenfunction patterns similar to the one occurring in Drosophila (Schiffmann, 2012). We also suggest a similar mechanism for the generation of the dorso-lateral-ventral polarity and bilateral symmetry. A mechanism for the generation of the regular location of the floral organs is also suggested. The symmetry and regularity of flowers, which are the source of their attraction and beauty, stem from the symmetry and regularity of the Turing-Child eigenfunctions. The central problem in developmental biology is the endless regress. This endless regress is halted by the Turing-Child pre-patterns and this is illustrated on a central example in flower generation. Both the shape and the chemistry - the steady-state rate of ATP synthesis and hydrolysis - of the Turing-Child pre-patterns are exactly what is required. Art and science meet in flower formation.


An autoradiographic study of 3H-thymidine incorporation and distribution in the dermis of healing skin incisions in the pig.
The dermal response to injury in healing incisional wounds aged from 1 to 24 days was studied by in vitro and in vivo 3H-thymidine (3HT) labelling and the use of colcemid to induce metaphase arrest. In vitro 3HT labelling provided mean numbers (+/- s.e. mean) of labelled dermal cells/section within the incision and in 3 zones in the dermis up to 0.875 mm lateral to the incision. There was a progressive increase in the numbers of DNA-synthesizing cells in the incisions as granulation tissue developed from Day 1 to Day 7, and a sharp decline by Day 9. Immediately adjacent to the incision there was a dramatic increase in the number of labelled cells on Day 2, with reduced numbers laterally. This response, which was largely maintained up to 7 days, paralleled the cellular reaction around blood vessels and skin appendages but also included labelled dermal fibroblasts. The in vivo 3HT study, taken in conjunction with the colcemid and in vitro 3HT studies, supported the idea that during the development of granulation tissue there is a continual recruitment of paraincisional cells, including dermal fibroblasts, either by direct migration or by division and then migration, as well as by continued mitosis of cells already within the incision.


Macrocolonies (Granules) Formation as a Cause of False-Negative Results in the MGIT 960 System: Cause Analysis and Correlation with Mycobacterial Species.
The viable mycobacterial bacilli can sometimes form granules in the Mycobacterium Growth Indicator Tube (MGIT) to produce instrument-negative outcomes when BACTEC MGIT 960 culture is performed. The cause of this phenomenon has never been analyzed. Thirty-one instrument-negative, granule presenting MGIT vials were collected for conducting acid-fast staining and also liquid and solid subculture. Species identification and drug susceptibility test were performed with the recovered strains. Cultivation test was done by inoculating small amount of bacilli into the MGIT vials. Twenty-four and twenty-nine of the tested MGIT vials were smear and culture positive, respectively. In total, 18, 4, and 7 of the cultivated strains were identified as Mycobacterium tuberculosis complex, M. intracellulare, and M. xenopi, respectively. When a limited amount of bacilli was inoculated, the granule formation was observed for M. xenopi strains in the MGIT system. The granules observed in the instrument-negative MGIT vials consisted of viable bacilli, which emphasized the need of visual inspection to increase recovery rate. Limited bacterial load and specific species might be the cause of granule forming.


Northern blot and in situ hybridization analyses for the neurogranin mRNA in the developing monkey cerebral cortex.
Neurogranin is a postsynaptic substrate for protein kinase C, and its expression is related to dendritic spine development and postsynaptic plasticity. Using both Northern blot analysis and in situ hybridization techniques, we investigated the developmental changes of neurogranin expression in the monkey cerebral cortex. In each of four neocortical areas examined, i.e., the prefrontal area (area FD of von Bonin and Bailey), the temporal association area (TE), the primary somatosensory area (PB), and the primary visual area (OC), the Northern blot analysis showed that the amount of neurogranin mRNA was low during the prenatal and perinatal periods until postnatal day 8. It increased during postnatal development and reached its peak value at postnatal day 70 (in area OC) or postnatal month 6 (in area FD, TE, and PB). After that, the amount of neurogranin mRNA in the cerebral neocortex decreased gradually until postnatal years 2-3. The in situ hybridization experiments also showed a transient increase of neurogranin mRNA in the neocortex during postnatal day 70 to postnatal month 6. The transient increase was prominent in layers II and III of areas FD and TE; deep in layer III of area PB; and in layers II, III, and IV of area OC. In the hippocampus, in contrast to the results in the neocortex, the expression of neurogranin mRNA was decreased almost continuously during the postnatal period. The transiently increased expression of neurogranin in the postnatal neocortex may be a molecular basis for the postsynaptic modification of afferent inputs possibly from subcortical structures.


Changes in airway hyperresponsiveness following smoking cessation: comparisons between Mch and AMP.
Given the observed association between smoking, inflammation and airway hyperresponsiveness (AHR) one may predict that smoking cessation may improve AHR. However, only a few studies have investigated the effect of smoking on AHR and their results appear to be conflicting depending on the stimulus used in their bronchoprovocation protocol. The aim of the current study was to compare changes in AHR between direct (methacholine (Mch)) and indirect (adenosine 5'monophosphate (AMP)) stimuli before and at different time points after smoking cessation in smokers with allergic rhinitis (+/-asthma). We have prospectively studied changes in AHR to inhaled Mch and AMP in smokers with allergic rhinitis (+/-asthma), before and at 6 and 12 months after smoking cessation. It was found that 28% (16/57) of the participants had quit smoking by the end of the study. No significant change in AHR was observed in smoking cessation failures. A significant improvement in AHR to AMP but not Mch was observed 6 months after smoking cessation in quitters; a 1.2 doubling concentrations change in PC20 AMP was measured whereas only a 0.4 doubling concentrations change was observed for PC20 Mch. However, after 12 months smoking cessation the improvement in AHR became significant for both AMP and Mch, their dose-response curves being displaced to the right to a similar extent (1.4 and 1.1 doubling concentrations for AMP and Mch, respectively). Smoking cessation can improve AHR in smokers who quit with a 6 months improvement being reported for the airways response to AMP but not Mch. AMP challenge may detect earlier changes in AHR in smokers during smoking cessation.


A latent variable approach in simultaneous modeling of longitudinal and dropout data in schizophrenia trials.
Dropouts impact clinical trial outcome analyses. Ignoring missing data is not an acceptable option when planning, conducting or interpreting the analysis of a clinical trial. Treatment related efficacy and safety data observed in the trial may not always be sufficient in explaining the dropouts' mechanism. Nevertheless, these dropout data may carry important treatment-related information and present as an outcome by itself. Traditional analyses involve the use of the time-to-event approach assuming that the dropouts' hazard is solely related to the efficacy or safety profiles observed in a study. A latent variable approach was developed to generalize this approach and to implement a more flexible dropout hazard function in a schizophrenia trial. This unobserved latent variable was used to jointly model the longitudinal efficacy data and dropout profiles across treatments. The analysis provides a framework to model informative dropouts simultaneously with primary efficacy outcomes and make intelligent decisions in drug development.


Enhancing insulin sensitivity by dual PPARγ partial agonist, β-catenin inhibitor: Design, synthesis of new αphthalimido-o-toluoyl2-aminothiazole hybrids.
Partial PPARγ agonists attracted substantially heightened interest as safer thiazolidinediones alternatives. On the other hand, Wnt/β-catenin antagonists have been highlighted as promising strategy for type 2 diabetes management via up-regulating PPARγ gene expression. We aimed at synthesizing novel partial PPARγ agonists with β-catenin inhibitory activity which could enhance insulin sensitivity and avoid the side effects of full PPARγ agonists. We synthesized novel series of α-phthlimido-o-toluoyl-2-aminothiazoles hybrids for evaluating their antidiabetic activity and discovering its mechanistic pathway. We assessed effect of the new hybrids on PPARγ activation using a luciferase reporter assay system. Moreover, intracellular triglyceride levels, gene levels of c/EBPα, PPARγ and PPARγ targets including GLUT4, adiponectin, aP2 were measured in 3T3-L1 cells. Uptake of 2-DOG together with PPARγ and β-catenin protein levels were evaluated in 3T3-L1cells. In addition, molecular docking studies with PPARγ LBD, physicochemical properties and structure activity relationship of the novel hybrids were also studied. Three of the synthesized hybrids showed partial PPARγ agonistic activity and distinct PPARγ binding pattern. These compounds modulated PPARγ gene expression and PPARγ target genes; and increased glucose uptake in 3T3-L1 and slightly induced adipogenesis compared to rosiglitazone. Moreover, these compounds reduced β-catenin protein level which reflected in increased both PPARγ gene and protein levels that leads to improved insulin sensitivity and increased GLUT4 and adiponectin gene expression. Our synthesized compounds act as novel partial PPARγ agonists and β-catenin inhibitors that have potent insulin sensitizing activity and mitigate the lipogenic side effects of TZDs.


Integrative detection and analysis of structural variation in cancer genomes.
Structural variants (SVs) can contribute to oncogenesis through a variety of mechanisms. Despite their importance, the identification of SVs in cancer genomes remains challenging. Here, we present a framework that integrates optical mapping, high-throughput chromosome conformation capture (Hi-C), and whole-genome sequencing to systematically detect SVs in a variety of normal or cancer samples and cell lines. We identify the unique strengths of each method and demonstrate that only integrative approaches can comprehensively identify SVs in the genome. By combining Hi-C and optical mapping, we resolve complex SVs and phase multiple SV events to a single haplotype. Furthermore, we observe widespread structural variation events affecting the functions of noncoding sequences, including the deletion of distal regulatory sequences, alteration of DNA replication timing, and the creation of novel three-dimensional chromatin structural domains. Our results indicate that noncoding SVs may be underappreciated mutational drivers in cancer genomes.


Development of resistance to FGFR inhibition in urothelial carcinoma via multiple pathways in vitro.
Alterations of fibroblast growth factor receptors (FGFRs) are common in bladder and other cancers and result in disrupted signalling via several pathways. Therapeutics that target FGFRs have now entered the clinic, but, in common with many cancer therapies, resistance develops in most cases. To model this, we derived resistant sublines of two FGFR-driven bladder cancer cell lines by long-term culture with the FGFR inhibitor PD173074 and explored mechanisms using expression profiling and whole-exome sequencing. We identified several resistance-associated molecular profiles. These included HRAS mutation in one case and reversible mechanisms resembling a drug-tolerant persister phenotype in others. Upregulated IGF1R expression in one resistant derivative was associated with sensitivity to linsitinib and a profile with upregulation of a YAP/TAZ signature to sensitivity to the YAP inhibitor CA3 in another. However, upregulation of other potential therapeutic targets was not indicative of sensitivity. Overall, the heterogeneity in resistance mechanisms and commonality of the persister state present a considerable challenge for personalised therapy. Nevertheless, the reversibility of resistance may indicate a benefit from treatment interruptions or retreatment following disease relapse in some patients. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.


Injections of deuterated tryptamine into the nucleus accumbens of the rat: effects on locomotor activity and monoamine metabolism.
Previous studies have shown that the systemic injection of tryptamine stimulates locomotion in rats and that the nucleus accumbens, a region involved in locomotion, contains the largest concentrations of binding sites for tryptamine in the brain of the rat. The present study examined the behavioral and neurochemical effects of bilateral injections into the accumbens of a deuterated analog of tryptamine, a,a-[2H]tryptamine. Injections of 25 micrograms a,a-[2H]tryptamine increased movements in rats at 25-70 min after injection and increased vertical (rearing) activity at 25-40 min. Injections of 50 micrograms of a,a-[2H]tryptamine produced a transient suppression of movement and vertical activity at 5-15 min, followed by increases in these activities at 40-65 min after injection that were comparable to the increases elicited by 10 micrograms of d-amphetamine. At 30 min after the injection of 50 micrograms a,a-[2H]tryptamine the concentration of dopamine in the nucleus accumbens was increased by 87%, and was preceded by a transient decrease in the level of the metabolite of dopamine homovanillic acid. The levels of 5-hydroxytryptamine and its major metabolite, 5-hydroxyindoleacetic acid in the nucleus accumbens were not changed. Thus, a,a-[2H]tryptamine may interact with tryptamine receptors in the nucleus accumbens to modulate locomotor behavior through mesolimbic dopamine neurons.


Postnatal clinical and imaging follow-up of infants with prenatal isolated mild ventriculomegaly: a series of 101 cases.
Postnatal imaging and clinical outcome of fetuses with isolated mild ventriculomegaly (IMV) have never been systematically analysed. To evaluate the postnatal clinical outcomes of a large cohort of fetuses with IMV and to correlate them with pre- and postnatal imaging. We report a prospective study of 101 fetuses with IMV (10-15 mm ventriculomegaly with otherwise normal US, MRI, karyotype and TORCH screening). IMV was divided into minor (10-11.9 mm) and moderate (12-15 mm) ventriculomegaly. Ventriculomegaly was considered uni- or bilateral, stable, progressive, regressive or resolved according to the prenatal US follow-up. Clinical follow-up was performed by a neuropaediatrician. Postnatal imaging included cranial US (n = 71) and MRI (n = 76). The outcome of minor and moderate IMV was excellent in 94% and 85% of infants, respectively. It was not different between uni- and bilateral IMV, and between stable, regressive and resolved IMV, and was independent of gestational age at diagnosis and gender. Fixed neurological abnormalities were observed in nine infants. Postnatal MRI showed white-matter abnormalities in 14 infants, including 6 of the 9 infants with a poor outcome. The prognosis was slightly better in minor IMV than in moderate IMV. Postnatal MRI showed white-matter abnormalities in two-thirds of the infants with a poor outcome.


Gait deficits and dynamic stability in children and adolescents with cerebral palsy: A systematic review and meta-analysis.
Studies have demonstrated that ambulatory children and adolescents with cerebral palsy demonstrate atypical gait patterns. Out of numerous gait variables, identification of the most deteriorated gait parameters is important for targeted and effective gait rehabilitation. Therefore, this study aimed to identify the gait parameters with the most discriminating nature to distinguish cerebral palsy gait from normal gait. Multiple databases were searched to include studies on ambulatory children and adolescents with cerebral palsy that included gait (spatio-temporal, kinematic, and kinetic) and dynamic stability variables. Of 68 studies that met the inclusion criteria, 35 studies were included in the meta analysis. Effect size was used to assess the discriminative strength of each variable. A large effect (≥ 0.8) of cerebral palsy on double limb support time (Standardized Mean Difference = 0.98), step length (Standardized Mean Difference = 1.65), step width (Standardized Mean Difference = 1.21), stride length (Standardized Mean Difference = 1.75), and velocity (Standardized Mean Difference = 1.42) was observed at preferred-walking speed. At fast-walking speed, some gait variables (i.e. velocity and stride length) exhibited larger effect size compared to preferred-walking speed. For some kinematic variables (e.g. range of motion of pelvis), the effect size varied across the body planes. Our systematic review detects the most discriminative features of cerebral palsy gait. Non-uniform effects on joint kinematics across the anatomical planes support the importance of 3D gait analysis. Differential effects at fast versus preferred speeds emphasize the importance of measuring gait at a range of speeds.


Ultrasonic Cole-Cole diagram for solutions and application to alpha-chymotrypsin.
Deconvolution of ultrasonic data into single relaxations is rarely feasible when only the absorption or the velocity of the waves is measured. Here we use both series of data to construct a Cole-Cole diagram for a solution. When applied to alpha-chymotrypsin, this method shows two relaxations that are well separated on the time scale, a result that will help simplify analyses of the ultrasonic data for this enzyme.


Metabolic Regulation of Mammalian Stem Cell Differentiation.
Formation of normal tissue structure, homeostasis maintenance, and tissue damage repair require proliferation and differentiation of stem cells. A distinctive feature of these cells is a unique organization of metabolic pathways, in which contribution of energy production mechanisms to the general cellular metabolism is principally different from that in differentiated cells. Moreover, metabolic changes during differentiation of embryonic and postnatal stem cells have several specific features. The alterations in the stem cell metabolism are not simply consequences of cell differentiation, but also active regulators of this process. Metabolic enzymes and intermediates control and guide the maintenance of stemness, self-renewal, and differentiation of stem cells. The review discusses the patterns and molecular mechanisms of the switch in the metabolism of stem cells during their transition from the pluripotent to differentiated state with the special emphasis on how metabolic processes occurring in the stem cells regulate their functions, ability to differentiate, and the choice of the direction for development.


Computer programs for modeling mammalian cell batch and fed-batch cultures using logistic equations.
A MATLAB(®) toolbox was developed for applying the logistic modeling approach to mammalian cell batch and fed-batch cultures. The programs in the toolbox encompass sensitivity analyses and simulations of the logistic equations in addition to cell specific rate estimation. The toolbox was first used to generate time courses of the sensitivity equations for characterizing the relationship between the logistic variable and the model parameters. Subsequently, the toolbox was used to describe CHO cell data from batch and fed-batch mammalian cell cultures. Cell density, product, glucose, lactate, glutamine, and ammonia data were analyzed for the batch culture while fed-batch analysis included cell density and product concentration. In all instances, experimental data were well described by the logistic equations and the resulting specific rate profiles were representative of the underlying cell physiology. The 6-variable batch culture data set was also used to compare the logistic specific rates with those from polynomial fitting and discrete derivative methods. The polynomial specific rates grossly misrepresented cell behavior in the initial and final stages of culture while those based on discrete derivatives had high variability due to computational artifacts. The utility of logistic specific rates to guide process development activities was demonstrated using specific protein productivity versus growth rate trajectories for the 3 cultures examined in this study. Overall, the computer programs developed in this study enable rapid and robust analysis of data from mammalian cell batch and fed-batch cultures which can help process development and metabolic flux estimation.


Preventive Effects of Different Drugs on Asymptomatic Lower Extremities Deep Venous Thrombosis After Artificial Joint Replacement: A Mixed Treatment Comparison.
This network meta-analysis aims to compare the preventive effects of 8 drugs (edoxaban, dabigatran, apixaban, rivaroxaban, warfarin, bemiparin, ximelagatran, and enoxaparin) on asymptomatic deep venous thrombosis (DVT) of lower extremities after artificial joint replacement. PubMed, Cochrane Library, and Embase were searched from their inception through October 2015 for randomized controlled trials comparing 8 drugs for the prevention of asymptomatic DVT of lower extremities after artificial joint replacement. Network meta-analysis combined the direct and indirect evidence to evaluate odd ratios (ORs) and surface under the cumulative ranking curves values. A total of 15 randomized controlled trials satisfying the inclusion criteria were enrolled. Edoxaban, apixaban, and rivaroxaban had poorer preventive effects on asymptomatic DVT of lower extremities after undergoing artificial joint replacement when compared with warfarin [OR = 0.16, 95% confidence interval (CI), 0.04-0.60; OR = 0.22, 95% CI, 0.07-0.64; OR = 0.16, 95% CI, 0.05-0.49, respectively]. When compared with enoxaparin, the preventive effects of edoxaban and rivaroxaban were poorer (OR = 0.37, 95% CI, 0.15-0.85; OR = 0.37, 95% CI, 0.21-0.59, respectively). The preventive effects of edoxaban and rivaroxaban were poorer than dabigatran (OR = 0.38, 95% CI, 0.14-0.99; OR = 0.38, 95% CI, 0.18-0.73, respectively). The surface under the cumulative ranking curves values showed that warfarin had better preventive effects on asymptomatic DVT of lower extremities after undergoing artificial joint replacement. Among the 8 drugs (edoxaban, dabigatran, apixaban, rivaroxaban, warfarin, bemiparin, ximelagatran, and enoxaparin), warfarin had better preventive effects on asymptomatic DVT of lower extremities after undergoing artificial joint replacement.


Infections and allergy - helminths, hygiene and host immune regulation.
There is increasing evidence that helminth infections can protect the host against Th2-mediated allergic pathologies, even though helminths themselves are strong Th2 inducers. In murine model systems, alleviation of allergy is not achieved through immune deviation to Th1, but is linked to expansion of regulatory T cell activity. Parasite infection does not prevent allergen sensitisation, but restricts the Th2 effector phase responsible for inflammation. Suppression of allergic inflammation can be transferred by Treg phenotype cells from an infected, allergen-naïve animal to an uninfected, sensitized recipient. Patent allergy in humans is also known to be modulated by helminth infections, suggesting that a similar regulatory network may be controlling immunopathologic disease in man.


Stabilization of metals in acidic mine spoil with amendments and red fescue (Festuca rubra L.) growth.
Stabilization of metals with amendments and red fescue (Festuca rubra, cv. Keszthelyi 2) growth was studied on an acidic and phytotoxic mine spoil (pH(KCl) 3.20-3.26; Cd 7.1 mg kg(-1), Cu 120 mg kg(-1), Pb 2154 mg kg(-1) and Zn 605 mg kg(-1)) from Gyöngyösoroszi, Hungary in a pot experiment. Raising the pH above 5.0 by lime (CaCO(3)), and supplementing with 40 mg kg(-1)nitrogen (NH(4)NO(3)) made this material suitable for plant growth. All cultures were limed with 0.5% (m/m) CaCO(3) (treatment 1), which was combined with 5% (m/m) municipal sewage sludge compost (treatment 2), 5% (m/m) peat (treatment 3), 7.5% (m/m) natural zeolite (clinoptilolite) (treatment 4), and 0.5 (m/m) KH(2)PO(4) (treatment 5). Treatments 1-5 were combined with each other (treatment 6). After 60 days of red fescue growth, pH of the limed mine spoil decreased in all cultures units. Application of peat caused the highest pH decrease (1.15), while decrease of pH was less than 0.23 in treatments 2, 5 or 6. Application of lime significantly reduced concentrations of metals in the 'plant available' fraction of mine spoil compared to non-limed mine spoil. Amendments added to limed mine spoil changed variously the ratio of Cd, Cu, Pb and Zn in exchangeable or 'plant available' fractions, differently influencing the phytoavailability of these metals. Most of the metals were captured in the roots of test plants. Treatment 2 caused the appearance of less Cd in shoots (<0.1 microg g(-1)) or roots (3.11 microg g(-1)), while treatment 5 resulted in the highest Cd concentration (2.13 microg g(-1)) in shoots. Treatments did not influence significantly the Cu accumulation in shoots. The Pb accumulation of roots (44.7 microg g(-1)) was most effectively inhibited by combined treatment, while the highest value (136 microg g(-1)) was found in the culture treated with potassium phosphate. Pb concentration in shoots was below the detection limit, except for treatments 5 and 6. Peat application resulted in higher Zn concentration (448 microg g(-1)) in shoots than other amendments, where these values were around 100 microg g(-1). All amendments influenced positively the dry matter yield of red fescue grown in limed mine spoil, however the application of 0.5 phosphate was less favourable. Liming, application of amendments and growth of red fescue can stabilize metals in acidic and phytotoxic mine spoil, and by phytostabilization they can reduce the risk of metal contamination of the food chain.


Modulation of cellular phosphoprotein profiles in transformation and redifferentiation of murine AKR embryonic fibroblastic cells.
Cellular phosphoprotein profiles from normal mouse embryonic fibroblast AKR-2B cells were compared to those of their permanently, chemically transformed malignant counterparts AKR-MCA cells, and AKR-2B cells reversibly transformed by transforming growth factor (AKR-TGF). Similar 32P-phosphorylation profiles were observed for both the AKR-TGF and AKR-MCA cells which were distinct from that of the normal AKR-2B cells. Dimethylformamide (DMF)-induced differentiation of the AKR-MCA cells resulted in a restoration of the normal AKR-2B phosphorylation profile to the malignant AKR-MCA cells.


Environmental quality as an issue in the legislative history of the National Health Planning and Resources Development Act of 1974 (PL93-641).
It is the purpose of this brief communication to examine the history of the most recently enacted health planning legislation (PL 93-641) to assess how environmental health issues were addressed and resolved during the legislative process.


Rapid divergence and gene flow at high latitudes shape the history of Holarctic ground squirrels (Urocitellus).
Across the animal tree of life, the prevalence and evolutionary role(s) of hybridization remain incompletely understood. Rapidly radiating clades can serve as important systems for investigating these issues; however, such groups are often characterized by additional, widespread sources of gene tree discordance (e.g., incomplete lineage sorting). In this paper, we employed a multilocus dataset, Bayesian gene tree inference, and multiple species tree reconstruction methods to infer phylogeny of Holarctic ground squirrels (Urocitellus). We tested phylogenetic hypotheses based on previous morphological, cytological and single-locus datasets, and began to parse the causes of pervasive gene tree discordance that was observed. There is widespread incomplete lineage sorting in Urocitellus, consistent with rapid diversification embedded within the larger radiation of marmotine ground squirrels. We also recovered strong support for 2 instances of mitonuclear discord due to ancient hybridization among members of the high-latitude parryii-richardsonii-elegans clade. These results add to a growing number of documented hybridization events in ground squirrels, suggesting their radiation is a fertile system for understanding the interplay of diversification and hybridization in animal evolution.


Effect of isoleucine deprivation of rabbitpox virus DNA synthesis in mouse L cells.
Synthesis of rabbitpox DNA was inhibited in mouse L cells deprived of isoleucine. Time-course patterns of incorporation of radiolabelled precursors into viral DNA revealed that synthesis of viral DNA began about 6 h after reversal of the isoleucine-deficient state.


Multimodal wavelet embedding representation for data combination (MaWERiC): integrating magnetic resonance imaging and spectroscopy for prostate cancer detection.
Recently, both Magnetic Resonance (MR) Imaging (MRI) and Spectroscopy (MRS) have emerged as promising tools for detection of prostate cancer (CaP). However, due to the inherent dimensionality differences in MR imaging and spectral information, quantitative integration of T(2) weighted MRI (T(2)w MRI) and MRS for improved CaP detection has been a major challenge. In this paper, we present a novel computerized decision support system called multimodal wavelet embedding representation for data combination (MaWERiC) that employs, (i) wavelet theory to extract 171 Haar wavelet features from MRS and 54 Gabor features from T(2)w MRI, (ii) dimensionality reduction to individually project wavelet features from MRS and T(2)w MRI into a common reduced Eigen vector space, and (iii), a random forest classifier for automated prostate cancer detection on a per voxel basis from combined 1.5 T in vivo MRI and MRS. A total of 36 1.5 T endorectal in vivo T(2)w MRI and MRS patient studies were evaluated per voxel by MaWERiC using a three-fold cross validation approach over 25 iterations. Ground truth for evaluation of results was obtained by an expert radiologist annotations of prostate cancer on a per voxel basis who compared each MRI section with corresponding ex vivo wholemount histology sections with the disease extent mapped out on histology. Results suggest that MaWERiC based MRS T(2)w meta-classifier (mean AUC, μ = 0.89 ± 0.02) significantly outperformed (i) a T(2)w MRI (using wavelet texture features) classifier (μ = 0.55 ± 0.02), (ii) a MRS (using metabolite ratios) classifier (μ = 0.77 ± 0.03), (iii) a decision fusion classifier obtained by combining individual T(2)w MRI and MRS classifier outputs (μ = 0.85 ± 0.03), and (iv) a data combination method involving a combination of metabolic MRS and MR signal intensity features (μ = 0.66 ± 0.02).


Atypical tibial fracture in breast cancer patient with bone metastasis receiving denosumab therapy: a case report and review of the literature.
Denosumab therapy is often used to reduce skeletal-related events in metastatic bone disease. On the other hand, there have been some instances of atypical femoral fracture in patients with metastatic bone disease treated with denosumab. In this case report, we describe a patient with metastatic bone disease due to breast cancer who had been using denosumab for 4 years to prevent skeletal-related events and suffered an atypical tibial fracture. We report here the case of an 82-year-old Japanese woman who had received yearly intravenous denosumab for 4 years and presented with a fracture fulfilling the criteria for an atypical fracture, except for being located at the tibial diaphysis. She was found to have stage 4 breast cancer with multiple bone metastases 4 years prior. She had difficulty walking due to her tibial pain and underwent surgical treatment. Four months after surgery, the tibial fracture site exhibited bone fusion. In patients with long-term use of denosumab to prevent skeletal-related events in metastatic bone disease, it is important to be aware of shin and thigh pain and to examine for signs of atypical tibial fractures to pay attention to atypical femoral fractures.


Colour flow duplex scanning in suspected acute deep vein thrombosis; experience with routine use.
To determine the accuracy of colour flow Duplex scanning (CFDS) in the diagnosis of deep vein thrombosis (DVT) and subsequently to investigate its diagnostic value in patients who have normal deep veins despite symptoms. Prospective open clinical study. Vascular laboratory and radiology departments of University Hospital. In the first part 112 limbs in 103 patients, 94 with symptoms of acute DVT and nine with pulmonary embolism (PE) were examined prospectively with CFDS and venography. Subsequently, in the second part, 328 legs in 304 patients were examined by CFDS alone for acute symptoms of DVT or PE. DVT was detected in 55 limbs by venography: proximal DVT was seen in 23 limbs, distal DVT in 25 limbs and both proximal and distal in seven limbs. CFDS was 100% sensitive and 98.8% specific in detecting proximal DVT whereas its sensitivity and specificity was 87.5% and 98.7% for distal DVT. Positive and negative predictive values were over 95% for both limb segments. The overall accuracy for the proximal DVT was 99.4% and for the distal 93.1%. In the second part, CFDS alone detected DVT in 156 limbs (47.6%); DVT was in the proximal segment in 82, distal segment in 61 and both in 13. In 172 limbs other causes of symptoms were identified in 34 (20%). We have demonstrated that CFDS is as accurate as venography when used by experienced operators. The average time of examination is 15-20 minutes and compares favourably with venography. Other causes of leg symptoms can also be diagnosed by CFDS in around 20% of patients who are found to have normal veins.


Age-Dependent Neurochemical Remodeling of Hypothalamic Astrocytes.
The hypothalamus is a crucial integrative center in the central nervous system, responsible for the regulation of homeostatic activities, including systemic energy balance. Increasing evidence has highlighted a critical role of astrocytes in orchestrating hypothalamic functions; they participate in the modulation of synaptic transmission, metabolic and trophic support to neurons, immune defense, and nutrient sensing. In this context, disturbance of systemic energy homeostasis, which is a common feature of obesity and the aging process, involves inflammatory responses. This may be related to dysfunction of hypothalamic astrocytes. In this regard, the aim of this study was to evaluate the neurochemical properties of hypothalamic astrocyte cultures from newborn, adult, and aged Wistar rats. Age-dependent changes in the regulation of glutamatergic homeostasis, glutathione biosynthesis, amino acid profile, glucose metabolism, trophic support, and inflammatory response were observed. Additionally, signaling pathways including nuclear factor erythroid-derived 2-like 2/heme oxygenase-1 p38 mitogen-activated protein kinase, nuclear factor kappa B, phosphatidylinositide 3-kinase/Akt, and leptin receptor expression may represent putative mechanisms associated with the cellular alterations. In summary, our findings indicate that as age increases, hypothalamic astrocytes remodel and exhibit changes in their neurochemical properties. This process may play a role in the onset and/or progression of metabolic disorders.


Changes of Antioxidant Function and the mRNA Expression Levels of Apoptosis Genes in Duck Ovaries Caused by Molybdenum or/and Cadmium.
To investigate the effects of molybdenum (Mo) combined with cadmium (Cd) on the antioxidant function and the mRNA expression levels of apoptosis-related genes in duck ovaries, 60 healthy 11-old-day female ducks were treated with hexaammonium molybdate ([(NH4)6Mo7O24·4H2O]) or/and cadmium sulfate (3CdSO4·8H2O) at different doses on a daily basis for 120 days. On the 120th day, ten female birds in each group were euthanized, and the ovaries and blood were collected to determine the antioxidant indexes and the mRNA expression levels of Bak-1, Bcl-2, and caspase-3 in ovaries. In addition, ovary tissues were subjected to histopathological analysis with optical microscope. The total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity decreased significantly (P < 0.01) in treated groups comparing with control while the nitric oxide synthase (NOS) activity increased (P < 0.01) both in ovary tissue and serum. The Bak-1 and caspase-3 expressions were upregulated while the Bcl-2 was downgraded by Mo or/and Cd. Biomolecules were affected in all metal-treated groups, whereas combined-treated animals showed greater effects. What is more, pathological damage in Mo and Cd combination treated groups was more severe. The results from the present study indicated that Mo or/and Cd caused oxidative stress and apoptosis in duck ovaries. Combination of Mo and Cd showed additive or synergistic effect leading to apoptosis and oxidative stress, and the pathway might be the mitochondrial pathway.


Human plasma lipid transfer protein catalyzes the speciation of high density lipoproteins.
The role of purified plasma lipid transfer protein complexes in determining the particle size distribution of human plasma high density lipoproteins (HDL) was examined in vitro. Incubation of HDL2 or HDL3, isolated from normolipemic subjects with very low density lipoproteins (VLDL) or VLDL-remnants and lipid transfer protein complex had little or no effect on HDL particle size. In contrast, HDL isolated from patients with hypertriglyceridemia, designated HDL3D, showed speciation of particle size distribution when incubated with VLDL-remnants and the transfer protein. Incubation of HDL3D with VLDL-remnants and lipid transfer complex resulted in the production of two particles of radius 4.3 and 3.7 nm; incubation with VLDL or in the absence of the transfer protein did not result in a redistribution of particle size. We suggest that the action of lipid transfer protein complex on triacylglycerol-rich lipoprotein remnants and HDL accounts for the low levels of HDL-cholesterol observed in subjects with severe hypertriglyceridemia.


Prospective use of molecular minimal residual disease for risk stratification in children and adolescents with acute lymphoblastic leukemia : Long-term results of the AIEOP-BFM ALL 2000 trial in Austria.
Since 1979 Austrian children and adolescents with acute lymphoblastic leukemia (ALL) have been treated according to protocols of the Berlin-Frankfurt-Münster (BFM) study group. The Associazione Italiana di Ematologia e Oncologia Pediatrica and BFM (AIEOP-BFM) ALL 2000 study was designed to prospectively study patient stratification into three risk groups using minimal residual disease (MRD) on two time points during the patient's early disease course. The MRD levels were monitored by detection of clone-specific rearrangements of the immunoglobulin and T‑cell receptor genes applying a quantitative polymerase chain reaction-based technique. The 7‑year event-free survival (EFS) and overall survival rates for all 608 Austrian patients treated between June 1999 and December 2009 within the AIEOP-BFM 2000 study were 84 ± 2% and 91 ± 1%, respectively, with a median observation time of 6.58 years. Event-free survival for patients with precursor B‑cell and T‑cell ALL were 84 ± 2% (n = 521) and 84 ± 4% (n = 87; p = 0.460), respectively. The MRD assessment was feasible in 94% of the patients and allowed the definition of precursor B‑cell ALL patients with a low, intermediate or high risk of relapse even on top of clinically relevant subgroups. A similar finding with respect to MRD relevance in T‑ALL patients was not possible due to the small number of patients and events. Since this pivotal international AIEOP-BFM ALL 2000 trial, molecular response to treatment has been continuously used with additional refinements to stratify patients into different risk groups in all successive trials of the AIEOP-BFM ALL study group.


[Combined injuries of the pelvic bones and bladder in children].
On the basis of observations of 15 children with associated injuries of the pelvic bones and urinary bladder the author describes the clinical picture, diagnosis and treatment depending on the kind of injuries of the urinary bladder.


Top-income adjustments and official statistics on income distribution: the case of the UK.
UK official statistics on income distribution have incorporated top-income adjustments to household survey data since 1992. This article reviews the work undertaken by the Department for Work and Pensions and the Office for National Statistics, and the academic research that influenced them, and reflects on the lessons to learn from the UK experience.


Mid-term Outcomes in Nonelderly Adults Undergoing Surgery for Isolated Aortic Valve Infective Endocarditis: Results From Two Canadian Centers.
Little is known about the mid-term prognosis of nonelderly patients (≤60 years) after the surgical treatment of isolated aortic valve infective endocarditis (IE). Better characterization of these outcomes could help in tailoring the surgical management in these patients. From 2000 to 2015, 164 adult patients ≤60 years of age (mean 46 ± 11 years, 81% male) underwent surgical treatment for isolated aortic valve IE in 2 high-volume Canadian centers. Twenty-three patients (14%) were intravenous drug users (IVDUs). Patients with recurrent IE or concomitant endocarditis on other valves were excluded. The aortic valve was replaced with a mechanical prosthesis (44%), a tissue valve (30%), a homograft (18%), or a Ross procedure (9%). Mean follow-up was 6.2 ± 4.6 years (92% complete). Thirty-day mortality was 7%. Actuarial survival rates at 5 and 10 years were 80 ± 3% and 71 ± 4%, respectively. IVDU (hazard ratio [HR] 3.8, 95% CI 1.4-10.1; P = 0.01) and prosthetic valve endocarditis (HR 2.6, 95% CI 1.1-6.4; P = 0.04) were associated with increased mid-term mortality. Mid-term survival was best in non-IVDU patients with native valve endocarditis, yet lower than a matched elective aortic valve replacement (AVR) population. Overall, freedom from recurrence of IE at 1, 5, and 10 years was 94 ± 2%, 91 ± 3%, and 89 ± 3%, respectively. IVDU was associated with higher rates of recurrence, especially in the first year after surgery. In nonelderly adults undergoing surgery for aortic valve IE, mid-term survival is suboptimal. Although non-IVDU patients with native valve endocarditis have better mid-term outcomes, survival remains lower than a matched population of elective AVR in nonelderly patients.