Detoxification of the phytoalexin pisatin by a fungal cytochrome P-450.
The fungus Nectria haematococca, a pathogen of garden pea (Pisum sativum), can demethylate pisatin, an antimicrobial compound synthesized by infected pea tissue. The phenolic product is less toxic than pisatin to many microorganisms. Cell extracts catalyzing pisatin demethylation were obtained from N. haematococca, and the properties of the reaction were examined. The enzyme activity was greatest in the high-speed pellet fraction, in which rates up to 20 nmol/min/mg protein were observed. The Km for pisatin was relatively low, less than 5 microM. The reaction was dependent on NADPH, which could not be replaced by any other cofactor tested. However, in the presence of NADPH, NADH increased the rate of demethylation. Oxygen uptake by the enzyme was stimulated by addition of pisatin, the increment of oxygen utilization being approximately equimolar with pisatin added. Formaldehyde was a product of the reaction. The effects of various inhibitors were tested to determine whether this reaction is mediated by cytochrome P-450. The respiratory inhibitors KCN (1 mM) and antimycin A strongly inhibited the demethylation of pisatin by intact cells of the fungus, but not by the NADPH-supplemented enzyme. The cytochrome P-450 inhibitors SKF 525-A and 1-(2-isopropylphenyl)imidazole inhibited demethylation both in whole cells and in the enzyme preparation, though the latter compound was effective only at high concentrations. Most other cytochrome P-450 inhibitors tested had little effect. However the reaction was quite sensitive to CO, and this inhibition was readily reversed by light at wavelengths near 450 nm. It is concluded that pisatin demethylase is a cytochrome P-450 monooxygenase.


Combined analysis of the ORCHID and OCAY studies: does eradication of Helicobacter pylori lead to sustained improvement in functional dyspepsia symptoms?
Currently, one of the most important unresolved questions concerning Helicobacter pylori is whether eradication of the organism leads to a sustained improvement in symptoms in patients diagnosed with functional (non-ulcer) dyspepsia. Recently, two very similar studies, the ORCHID and OCAY studies, have been completed and the combined results of these two multicentre, multinational, randomised, double blind, controlled clinical trials are reviewed.


Bienzymatic synergism of vanadium oxide nanodots to efficiently eradicate drug-resistant bacteria during wound healing in vivo.
Antibiotic resistance is a common phenomenon observed during treatment with antibacterials. Use of nanozymes, especially those with synergistic enzyme-like activities, as antibacterials could overcome this problem, but their synthesis is limited by their high cost and/or complex production process. Herein, vanadium oxide nanodots (VOxNDs) were prepared via a one-step bottom-up ethanol-thermal method using vanadium trichloride as the precursor. VOxNDs alone possess bienzyme mimics of peroxidase and oxidase. Accordingly, highly efficient antibacterials against drug-resistant bacteria can be obtained through synergistic catalysis; the oxidase-like activity decomposes O2 to generate superoxide anion radical (O2-) and hydroxyl radicals (OH), and the intrinsic peroxidase-like activity can further induce the production of OH from external H2O2. Consequently, H2O2 concentration could decrease up to four magnitude orders with VOxNDs to achieve an antibacterial efficacy similar to that of H2O2 alone. Wound healing in vivo further confirms the high antibacterial efficiency, good biocompatibility, and application potential of the synergistic antibacterial system due to the "nano" structure of VOxNDs. The method of synthesis of nanodot antibacterials described in this paper is inexpensive, and the results of this study reveal the multi-enzymatic synergism of nanozymes.


Accelerated DNA methylation age: Associations with PTSD and neural integrity.
Accumulating evidence suggests that posttraumatic stress disorder (PTSD) may accelerate cellular aging and lead to premature morbidity and neurocognitive decline. This study evaluated associations between PTSD and DNA methylation (DNAm) age using recently developed algorithms of cellular age by Horvath (2013) and Hannum et al. (2013). These estimates reflect accelerated aging when they exceed chronological age. We also examined if accelerated cellular age manifested in degraded neural integrity, indexed via diffusion tensor imaging. Among 281 male and female veterans of the conflicts in Iraq and Afghanistan, DNAm age was strongly related to chronological age (rs ∼.88). Lifetime PTSD severity was associated with Hannum DNAm age estimates residualized for chronological age (β=.13, p=.032). Advanced DNAm age was associated with reduced integrity in the genu of the corpus callosum (β=-.17, p=.009) and indirectly linked to poorer working memory performance via this region (indirect β=-.05, p=.029). Horvath DNAm age estimates were not associated with PTSD or neural integrity. Results provide novel support for PTSD-related accelerated aging in DNAm and extend the evidence base of known DNAm age correlates to the domains of neural integrity and cognition.


Assessment of Pericardial Disease with Cardiovascular MRI.
Disorders of the pericardium are common and can result in significant morbidity and mortality. Advances in multimodality imaging have enhanced our ability to diagnose and stage pericardial disease and improve our understanding of the pathophysiology of the disease. Cardiovascular MRI (CMR) can be used to define pericardial anatomy, identify the presence and extent of active pericardial inflammation, and assess the hemodynamic consequences of pericardial disease. In this way, CMR can guide the judicial use of antiinflammatory and immune modulatory medications and help with timing of pericardiectomy. CMR can also be used to diagnose congenital disorders of the pericardium. Furthermore, CMR can be used to define pericardial masses and understand their malignant potential.


Optimization of cultural conditions for growth associated chromate reduction by Arthrobacter sp. SUK 1201 isolated from chromite mine overburden.
Arthrobacter sp. SUK 1201, a chromium resistant and reducing bacterium having 99% sequence homology of 16S rDNA with Arthrobacter sp. GZK-1 was isolated from chromite mine overburden dumps of Orissa, India. The objective of the present study was to optimize the cultural conditions for chromate reduction by Arthrobacter sp. SUK 1201. The strain showed 67% reduction of 2mM chromate in 7 days and was associated with the formation of green insoluble precipitate, which showed characteristic peak of chromium in to energy dispersive X-ray analysis. However, Fourier transform infrared spectra have failed to detect any complexation of end products of Cr(VI) reduction with the cell mass. Reduction of chromate increased with increased cell density and was maximum at 10(10)cells/ml, but the reduction potential decreased with increase in Cr(VI) concentration. Chromate reducing efficiency was promoted when glycerol and glucose was used as electron donors. Optimum pH and temperature of Cr(VI) reduction was 7.0 and 35 °C respectively. The reduction process was inhibited by several metal ions and metabolic inhibitors but not by Cu(II) and DNP. These findings suggest that Arthrobacter sp. SUK 1201 has great promise for use in Cr(VI) detoxification under a wide range of environmental conditions.


A Pilot Study of Self-Rated and Psychophysical Olfactory Dysfunction in Men Living with HIV.
Olfactory loss is associated with poor quality of life, malnutrition, and increased risk of depression, yet few studies have examined unawareness of olfactory dysfunction in men living with HIV (MLWH). MLWH (n = 51) completed olfaction self-ratings, psychophysical odor identification testing, cognitive measures, and questionnaires assessing smell habits, mood, cognitive failures, and quality of life. The sensitivity and specificity of olfactory self-ratings was calculated, and t-tests were used to examine factors contributing to discordance between self-rated and psychophysical olfaction dysfunction. We found that 33.3% (17 of 51 MLWH) of our sample demonstrated discordance between self-reported and psychophysical olfactory scores. Those unaware of olfaction dysfunction reported using less scented products in daily life but showed no other differences across demographic, clinical, or cognitive indices. Our results cohere with prior studies of cognitively normal older adults, traumatic brain injury, and Parkinson's disease, which found that olfactory self-ratings may inadequately capture the full range of a person's olfactory status. Our work extends these findings to MLWH, with discordance rates ranging from 35 to 61% for self-rated and psychophysical olfactory dysfunction. Given the differing rates of self-rated and psychophysical olfaction in our sample, psychophysical olfactory measures may be useful to consider in the neuropsychological assessment and clinical care of PLWH. The online version contains supplementary material available at 10.1007/s12078-022-09305-x.


Effects of the built environment and human factors on the spread of COVID-19: A systematic literature review.
Soon after its emergence, COVID-19 became a global problem. While different types of vaccines and treatments are now available, still non-pharmacological policies play a critical role in managing the pandemic. The literature is enriched enough to provide comprehensive, practical, and scientific insights to better deal with the pandemic. This research aims to find out how the built environment and human factors have affected the transmission of COVID-19 on different scales, including country, state, county, city, and urban district. This is done through a systematic literature review of papers indexed on the Web of Science and Scopus. Initially, these databases returned 4264 papers, and after different stages of screening, we found 166 relevant papers and reviewed them. The empirical papers that had at least one case study and analyzed the effects of at least one built environment factor on the spread of COVID-19 were selected. Results showed that the driving forces can be divided into seven main categories: density, land use, transportation and mobility, housing conditions, demographic factors, socio-economic factors, and health-related factors. We found that among other things, overcrowding, public transport use, proximity to public spaces, the share of health and services workers, levels of poverty, and the share of minorities and vulnerable populations are major predictors of the spread of the pandemic. As the most studied factor, density was associated with mixed results on different scales, but about 58 % of the papers reported that it is linked with a higher number of cases. This study provides insights for policymakers and academics to better understand the dynamic roles of the non-pharmacological driving forces of COVID-19 at different levels.


Evidence for a role of brain thyrotropin-releasing hormone (TRH) on stress gastric lesion formation in rats.
Specific polyclonal antibodies raised against synthetic thyrotropin-releasing hormone (TRH) infused intracerebroventricularly (ICV) significantly decreased gastric lesions induced by cold restraint stress. The antiulcer effect of immunologic blockade of brain TRH was specific. Normal rabbit serum or antibodies raised against somatostatin, alpha-MSH, Leu-enkephalin, gonadotropin-releasing hormone and atrial natriuretic factor were ineffective. These findings suggest that brain TRH may play an important role in experimental stress ulcer formation.


Assessment of hypnotic effects and patient satisfaction in empirical use of sleep medicines.
We assessed the hypnotic effects of and patient satisfaction with three types of hypnotics prescribed empirically: ultra-short-acting (US-a), short-acting (S-a), and intermediate- and long-acting (IL-a) agents. We studied 310 insomniac patients (age 60.5 +/- 15.0 years) treated with US-a (n = 124), S-a (n = 149) or IL-a (n = 37) agents. Patients were interviewed to evaluate individual satisfaction and drug efficacy. Efficacy, as assessed by total sleep time (TST) and sleep latency time (SLT), was compared between satisfied and dissatisfied patient groups. Nocturnal awaking curve for each hypnotic was used for comparing the effects between satisfied and dissatisfied patient groups in each type of hypnotics. Thirty-two patients (25.8%) were dissatisfied with US-a, 35 (23.5%) with S-a and 11 (29.7%) with IL-a. TST differed significantly between satisfied and dissatisfied groups: 424 +/- 88 vs. 345 +/- 101 min for US-a (P < 0.001), 440 +/- 84 vs. 359 +/- 111 min for S-a (P < 0.001) and 453 +/- 96 vs. 345 +/- 125 min for IL-a (P < 0.01), respectively. With IL-a agents, the SLT of dissatisfied patients was longer than in satisfied ones (81 +/- 52 vs. 33 +/- 22 min, P < 0.05). Twenty (62.5%) dissatisfied patients taking US-a agents awoke before 05:00 hours - a rate significantly higher than satisfied patients (n = 23, 25.0%, P < 0.001). These characteristics of dissatisfied patients were reflected by the patterns of nocturnal awaking curves, although the patterns for satisfied patients were similar among the three types of hypnotics. Between 24% and 30% of patients were dissatisfied with their hypnotics. Shorter TST was common in dissatisfied patients receiving any agent, for reasons differing among hypnotics (longer SLT with IL-a agents and early awakening with US-a). Drug efficacy and patient satisfaction in empirical use of hypnotics can be assessed by nocturnal awaking curves for each hypnotic.


Leptospirosis diagnosed by molecular DNA detection on skin biopsy.
Leptospirosis is one of the most common anthropozoonoses in the world. Humans are accidental hosts who get infected through damaged skin or conjunctiva from environmental sources such as soil and water contaminated by urine or tissues from infected animals. We report the case of a young man for whom the diagnosis was obtained by polymerase chain reaction (PCR) detection from a skin biopsy.


A regulatory role for cortisol in muscle glycogen metabolism in rainbow trout Oncorhynchus mykiss Walbaum.
To test the hypothesis that cortisol has a regulatory role in fish muscle glycogenesis post-exercise, rainbow trout were treated 1 h prior to exercise with either saline (control) or metyrapone (2-methyl-1, 2-di-3-pyridyl-1-propanone) to block cortisol synthesis. Following exercise (time 0), half of the metyrapone-treated fish received a single injection of cortisol, to mimic the post-exercise rise usually observed. Muscle glycogen and the relative activities of glycogen phosphorylase a (Phos a) and glycogen synthase I (GSase I), regulatory enzymes for glycogen resynthesis, were monitored 4 h post-exercise. Metyrapone treatment succeeded in blocking the post-exercise rise in plasma cortisol (17+/-2 vs 118+/-13 ng ml(-1) in controls at time 0), and cortisol injection resulted in a larger and more prolonged cortisol increase than in controls (159+/-22 vs 121+/-14 ng ml(-1) in controls at 1 h). Muscle glycogen was completely restored in the metyrapone-treated fish within 2 h after exercise (8.3+/-0.6 vs 8+/-0.7 micromol g(-1) pre-exercise), only partially restored in control fish at 4 h (5.4+/-01.4 vs 8.8+/-1.3 micromol g(-1) pre-exercise), and not at all in cortisol-treated fish (1.0+/-0.5 micromol g(-1) at 4 h). The rapid glycogen resynthesis in the metyrapone-treated fish was associated with a more rapid inactivation of Phos a and stimulation of GSase I compared to controls. In cortisol-treated fish, Phos a activity persisted throughout 4 h post-exercise; there was also a significant stimulation of GSase I activity. As a consequence of dual activation of Phos a and GSase I, glycogen cycling probably occurred, thus preventing net synthesis. This explains why the post-exercise elevation of cortisol inhibits net glycogen synthesis in trout muscle.


Worldwide Burden, Risk Factors, and Temporal Trends of Ovarian Cancer: A Global Study.
This study aimed to investigate the most updated worldwide incidence and mortality, risk factors, and epidemiologic trend of ovarian cancer in different countries, regions, and age groups. The Global Cancer Observatory database was used for incidence and mortality rates of ovarian cancer in 2020. Data from Cancer Incidence in Five Continents and the WHO mortality database was accessed for trend analysis. Age-standardized rates (ASRs, per 100,000 persons) were calculated for incidence and mortality. The 10-year annual average percent change (AAPC) was estimated by Joinpoint regression analysis. There was an overall decreasing trend of ovarian cancer, yet its burden has been increasing in lower-income countries and among younger females in some countries. Intensive lifestyle modifications are warranted, especially for the populations at high risk for ovarian cancer, including smoking cessation, alcohol use reduction, physical activity, weight control, and treatment of metabolic diseases.


Short-lived success: assessment of an intervention to improve pregnancy weight gain in Colorado.
Inadequate weight gain in pregnancy is a major contributor to low birth weight in Colorado, where the low birth weight rate is among the highest in the nation. In 2004, the Colorado Department of Public Health and Environment implemented a population-based intervention in 9 counties, including provider training and a public media campaign, to encourage pregnant women to gain an adequate amount of weight in pregnancy as defined by the 1990 Institute of Medicine guidelines. Pregnancy Risk Assessment Monitoring System survey data were used to track weight gain in pregnancy in 1997 through 2004 (baseline), 2005 (post-intervention), and 2006 and 2007 (after the intervention had concluded). During the period immediately after the implementation of the intervention, the percentage of women delivering in the 9 study counties who gained an inadequate amount of weight during pregnancy dropped from 18.4 at baseline to 12.8 in 2005. However, this progress was reversed in 2006, when the percentage of women with inadequate weight gain rose to 19.7. Training providers to educate women about the importance of adequate weight gain in pregnancy, in conjunction with a social marketing campaign, appears to be a promising approach to addressing a major contributor to low birth weight. However, a time-limited intervention is likely to have temporary results.


Sequence of a RNA templated by the 3'-OH RNA terminus of defective interfering particles of vesicular stomatitis virus.
We have sequenced the endogenous RNA polymerase product produced by disrupted purified virions of vesicular stomatitis virus defective interfering particles by using the newer one-dimensional rapid gel sequencing techniques and confirming this with a modified two-dimensional gel vectoring technique. The sequence of this 46-nucleotide RNA is: 5'(pp)pACGAAGACCACAAAACCA-GAUAAAAAAUAAAAACCACAAGAGGG(U)COH3'. We infer that this sequence is identical to the sequence at the 5' end of infectious vesicular stomatitis virus RNA and is complementary to the sequence of the 3'-OH terminus of this defective interfering particle genome RNA.


[Diseases associated with bloodstream infections caused by the new species included in the old Streptococcus bovis group].
We sought to identify possible diseases associated with bloodstream infections caused by new species of S. bovis group isolated in blood cultures and by studying patient records Forty-four consecutive blood culture isolates initially designated S. bovis were further characterised using phenotypic methods Patient records were examined. We identified 15 Streptococcus gallolyticus subsp. gallolyticus, 24 Streptococcus gallolyticus subsp. pasteurianus, and 5 Streptococcus infantarius isolates in 44 BSI episodes. The association between S. bovis bacteraemia and endocarditis and/or colon carcinoma is highly dependent on the causative species. Streptococcus gallolyticus subsp. gallolyticus is a surrogate for endocarditis and/or bowel disease, whereas Streptococcus gallolyticus subsp. pasteurianus is a surrogate for hepato-biliary disease.


Which Probiotic Is the Most Effective for Treating Acute Diarrhea in Children? A Bayesian Network Meta-Analysis of Randomized Controlled Trials.
Acute diarrhea is a major cause of morbidity and mortality in children under five. Probiotics are beneficial for treating acute diarrhea in children, but unclear which specific probiotic is the most effective. We performed a Bayesian network meta-analysis to examine the comparative effectiveness of probiotics. By searching EMBASE, PubMed, and the Cochrane Library up to 31 March 2021, randomized clinical trials (RCTs) on probiotics for treating acute diarrhea in children were included. Primary outcomes included the duration of diarrhea and diarrhea lasting ≥2 days, and secondary outcomes included the mean stool frequency on day 2 and duration of hospitalization, fever, and vomiting. We assessed the certainty of the evidence of outcomes according to Grading of Recommendations Assessment, Development, and Evaluation (GRADE) guideline. Eighty-four studies with twenty-one different interventions in 13,443 children were included. For the primary outcomes, moderate evidence indicated that, Lactobacillus reuteri [mean difference (MD) = -0.84 day; 95% confidence interval (CI), -1.39, -0.29], Bifidobacterium lactis (MD = -0.98 day; 95%CI, -1.82, -0.14), Saccharomyces boulardii (MD = -1.25 day; 95%CI, -1.59, -0.91), Lactobacillus species (spp.) plus Bifidobacterium spp. plus Saccharomyces spp. (MD = -1.19 day; 95%CI, -1.81, -0.58), and Bacillus spp. plus Enterococcus spp. plus Clostridium spp. (MD = -1.1 day; 95%CI, -1.84, -0.35) significantly reduced the duration of diarrhea when compared with placebo. Saccharomyces boulardii [Odds ratio (OR) = 0.22; 95%CI, 0.11, 0.41] and Lactobacillus reuteri (OR = 0.23; 95%CI, 0.090, 0.60) significantly reduced the risk of diarrhea lasting ≥2 days when compared with placebo or no treatment, with moderate evidence. Among all probiotics, Saccharomyces boulardii may be the most effective in reducing both duration of diarrhea (compared with placebo) and risk of diarrhea lasting ≥2 days (compared with placebo or no treatment), with moderate evidence. To be conclusive, Saccharomyces boulardii may be the most effective probiotic for treating acute diarrhea in children, followed by several other single-strain and multi-strain probiotics.


The Hypothalamic Paraventricular Nucleus Is the Center of the Hypothalamic-Pituitary-Thyroid Axis for Regulating Thyroid Hormone Levels.
Background: Thyrotropin-releasing hormone (TRH) was the first hypothalamic hormone isolated that stimulates pituitary thyrotropin (TSH) secretion. TRH was also later found to be a stimulator of pituitary prolactin and distributed throughout the brain, gastrointestinal tract, and pancreatic β cells. We previously reported the development of TRH null mice (conventional TRHKO), which exhibit characteristic tertiary hypothyroidism and impaired glucose tolerance due to insufficient insulin secretion. Although in the past five decades many investigators, us included, have attempted to determine the hypothalamic nucleus responsible for the hypothalamic-pituitary-thyroid (HPT) axis, it remained obscure because of the broad expression of TRH. Methods: To determine the hypothalamic region functionally responsible for the HPT axis, we established paraventricular nucleus (PVN)-specific TRH knockout (PVN-TRHKO) mice by mating Trh floxed mice and single-minded homolog 1 (Sim1)-Cre transgenic mice. We originally confirmed that most Sim1 was expressed in the PVN using Sim1-Cre/tdTomato mice. Results: These PVN-TRHKO mice exhibited tertiary hypothyroidism similar to conventional TRHKO mice; however, they did not show the impaired glucose tolerance observed in the latter, suggesting that TRH from non-PVN sources is essential for glucose regulation. In addition, a severe reduction in prolactin expression was observed in the pituitary of PVN-TRHKO mice compared with that in TRHKO mice. Conclusions: These findings are conclusive evidence that the PVN is the center of the HPT axis for regulation of serum levels of thyroid hormones and that the serum TSH levels are not decreased in tertiary hypothyroidism. We also noted that TRH from the PVN regulated prolactin, whereas TRH from non-PVN sources regulated glucose metabolism.


High expression of Talin-1 is associated with poor prognosis in patients with nasopharyngeal carcinoma.
Talin-1 is a cytoskeletal protein that plays an important role in tumourgenesis, migration and metastasis in several malignant tumors. The aim of this study was to evaluate the expression and prognostic value of Talin-1 in nasopharyngeal carcinoma (NPC). Talin-1 mRNA and protein expression were examined in NPC cell lines and clinical nasopharyngeal tissues by quantitative RT-PCR, agarose gel electrophoresis and western blotting. The expression of Talin-1 was analyzed by immunohistochemical staining in 233 paraffin-embedded NPC specimens with clinical follow-up data and cox regression analysis was used to identify independent prognostic factors. The functional role of Talin-1 in NPC cell lines was evaluated by small interfering RNA-mediated depletion of the protein followed by the wound healing and transwell invasion assays. The expression of Talin-1 was significantly upregulated in most NPC cell lines and clinical tissues at both the mRNA and protein levels. High expression of Talin-1 was significantly associated with distant metastasis (P = 0.001) and patient death (P = 0.001). In addition, high expression of Talin-1 was associated with significantly poorer overall survival (OS: HR, 2.15; 95% CI, 1.28-3.63; P = 0.003) and poorer distant metastasis-free survival (DMFS: HR, 2.39; 95% CI, 1.38-4.15; P = 0.001). Cox regression analysis indicated that high expression of Talin-1 and TNM stage were independent prognostic indicators (both P < 0.05). Stratified analysis demonstrated that high expression of Talin-1 was associated with significantly poorer survival in patients with advanced stage disease (stage III-IV, HR, 1.91; 95% CI, 1.09-3.35; P = 0.02 for OS and HR, 2.22; 95% CI, 1.24-3.99; P = 0.006 for DMFS). Furthermore, the depletion of Talin-1 suppressed the migratory and invasive ability of NPC cells in vitro. Our data demonstrate that high expression of Talin-1 is associated with significantly poorer OS and poorer DMFS in NPC and depletion of Talin-1 expression inhibited NPC cell migration and invasion. Talin-1 may serve as novel prognostic biomarker in NPC.


Systemic corticosteroids improve tendon healing when given after the early inflammatory phase.
Inflammation initiates tendon healing and then normally resolves more or less completely. Unresolved inflammation might disturb the remodeling process. We hypothesized that suppression of inflammation during the early remodeling phase by systemic dexamethasone treatment can improve healing. 36 rats underwent Achilles tendon transection and were randomized to dexamethasone or saline on days 0-4 after surgery (early inflammatory phase), and euthanasia day 7. Another 54 rats received injections days 5-9 (early remodeling phase) and were euthanized day 12 for mechanical, histological and flow cytometric evaluation. Dexamethasone treatment days 0-4 reduced the cross-sectional area, peak force and stiffness by day 7 to less than half (p < 0.001 for all), while material properties (peak stress and elastic modulus) were not significantly affected. In contrast, dexamethasone treatment days 5-9 increased peak force by 39% (p = 0.002) and stiffness by 58% (p < 0.001). The cross-sectional area was reduced by 42% (p < 0.001). Peak stress and elastic modulus were more than doubled (p < 0.001 for both). Semi-quantitative histology at day 12 showed that late dexamethasone treatment improved collagen alignment, and flow cytometry revealed reduced numbers of CD8a+ cytotoxic T cells in the tendon callus. These results suggest that downregulation of lingering inflammation during the early remodeling phase can improve healing.


Expression of a synthetic cry1EC gene for resistance against Spodoptera litura in transgenic peanut (Arachis hypogaea L.).
The tobacco cutworm (Spodoptera litura) is a polyphagous foliage insect and a major pest on peanut (Arachis hypogaea L.). S. litura is susceptible to the chimeric delta-endotoxin Cry1EC reported earlier. De-embryonated cotyledon explants of peanut were transformed using Agrobacterium tumefaciens strain EHA101 harboring a synthetic cry1EC gene driven by the CaMV 35S promoter. Transgenic plants of peanut with a single copy insertion of cry1EC were selected in the T(0) generation by Southern blot hybridization. Real-time PCR, Western blot and ELISA analysis indicated that expression of the cry1EC gene was higher in single copy T(1) plants. Immunoassay showed expression of Cry1EC up to 0.13% of total soluble protein in T(1) plants. Leaf feeding bioassay on highly expressing transgenic lines showed 100% killing of larvae at the 2(nd) instar stage of S. litura. This is the first report of transgenic peanut plants with resistance to S. litura.


Folliculin controls lung alveolar enlargement and epithelial cell survival through E-cadherin, LKB1, and AMPK.
Spontaneous pneumothoraces due to lung cyst rupture afflict patients with the rare disease Birt-Hogg-Dubé (BHD) syndrome, which is caused by mutations of the tumor suppressor gene folliculin (FLCN). The underlying mechanism of the lung manifestations in BHD is unclear. We show that BHD lungs exhibit increased alveolar epithelial cell apoptosis and that Flcn deletion in mouse lung epithelium leads to cell apoptosis, alveolar enlargement, and an impairment of both epithelial barrier and overall lung function. We find that Flcn-null epithelial cell apoptosis is the result of impaired AMPK activation and increased cleaved caspase-3. AMPK activator LKB1 and E-cadherin are downregulated by Flcn loss and restored by its expression. Correspondingly, Flcn-null cell survival is rescued by the AMPK activator AICAR or constitutively active AMPK. AICAR also improves lung condition of Flcn(f/f):SP-C-Cre mice. Our data suggest that lung cysts in BHD may result from an underlying defect in alveolar epithelial cell survival, attributable to FLCN regulation of the E-cadherin-LKB1-AMPK axis.


Is Donated Breast Milk Better Than Formula for Feeding Very Low Birth Weight Infants? A Systematic Review and Meta-Analysis.
The role of donated breast milk and formula feeding in very low birth weight infants (VLBWIs) remains unclear. The objective for this study was to evaluate the efficacy and safety of breast milk and formula for feeding VLBWIs. A comprehensive search of PubMed databases was conducted on November 30, 2018, to identify related randomized controlled trials (RCTs). Pooled odds ratio (OR) and mean differences (MD) with 95% confidence intervals (CI) were calculated using either a fixed or random-effects model. Seven RCTs with a total of 876 infants were included. The formula showed more advantages in increasing infant weight (MD = -6.58, 95% CI: -11.19 to -1.98) and length (MD = -0.30, 95% CI: -0.41 to -0.20) than donated breast milk, but formula could also significantly increase the risks of necrotizing enterocolitis (OR = 0.33, 95% CI: 0.18 to 0.59) and length of hospital stay (MD = -3.78, 95% CI: -6.84 to -0.72) when compared with donated breast milk. No significant differences on the head circumference gain (MD = -0.16, 95% CI: -0.33 to 0.01), sepsis (OR = 1.21, 95% CI: 0.84 to 1.75), retinopathy of prematurity (OR = 1.22, 95% CI: 0.73 to 2.05) and mortality (OR = 0.84, 95% CI: 0.47 to 1.52) were found. The donated breast milk shows more advantages in reducing the incidence of NEC and length of hospital stay, but also more disadvantages in increasing the weight and length of VLBWIs and ELBWIs when compared with preterm formula feeding. Currently, there is no solid evidence demonstrating a significant advantage of donated breast milk over formula in the feeding of VLBWIs. Larger-scale RCTs with rigorous designs are needed to elucidate the feeding plans of VLBWIs.


Carbon nanotubes-ionic liquid nanocomposites sensing platform for NADH oxidation and oxygen, glucose detection in blood.
An excellent electrochemical sensing platform has been designed by combining the huge specific surface area of carbon nanotubes (CNTs) and the remarkable conductivity of ionic liquid (IL). IL can easily untangle CNTs bundles and disperse CNTs by itself under grinding condition due to the π-π interaction between CNTs and IL. The resulting nanocomposites showed an augmentation on the voltammetric and amperometric behaviors of electrocatalytic activity toward O(2) and NADH. Therefore, such an efficient platform was developed to fabricate mediator-free oxygen sensor and glucose biosensor based on glucose dehydrogenase (GDH). O(2) could be determined in the range of zero to one hundred percent of O(2) content with the detection limit of 126 μg L(-1) (S/N=3). The glucose biosensor which was constructed by entrapping GDH into chitosan on the nanocomposites modified glassy carbon electrode surface, exhibited good electrocatalytic oxidation toward glucose with a detection limit of 9 μM in the linear range of 0.02-1mM. We also applied the as-prepared sensors to detect oxygen and glucose in real blood samples and acquired satisfied results.


Central precocious puberty may be a manifestation of endocrine dysfunction in pediatric patients with mitochondrial disease.
We retrospectively reviewed the data of 140 female pediatric patients with rare mitochondrial diseases (MDs) confirmed using muscle biopsy. We evaluated patients who were diagnosed with central precocious puberty (PP) with early pubertal development to determine whether PP is a clinical manifestation of MDs. We also examined the clinical, auxiological, laboratory, and radiological parameters after 1 year of gonadotropin-releasing hormone treatment for central PP. Among the 140 girls with MDs, 29 had early pubertal development and underwent endocrine evaluation. Ten (7.1%) patients were diagnosed with central PP; the prevalence of central PP was higher than was that previously thought. Patients with central PP exhibited bone age advancement over 1 year and increased sex hormone levels despite their young age at diagnosis. Serum estradiol levels were significantly higher in younger patients than in older patients (P = 0.004). Patients with central PP treated with gonadotropin-releasing hormone had favorable outcomes, and their pubertal development was suppressed for 1 year.Conclusion: Central PP may be a manifestation of endocrine dysfunction in young girls with MDs. What is Known: • The general characteristics of mitochondrial diseases include developmental delays and retarded growth. • Precocious puberty has rarely been suggested as a clinical manifestation of mitochondrial diseases. What is New: • Among the 140 girls with mitochondrial diseases, 10 (7.1%) were diagnosed with central precocious puberty. • Serum estradiol levels were significantly higher in younger patients than in older patients.


[Postural effect of vestibular galvanic stimulation in patients with Parkinson's disease and camptocormia: Case series].
The objective of this paper is to explore the postural effect of galvanic vestibular stimulation (GVS) in camptocormia. It is a retrospective case series of 7 Parkinson disease and camptocormia patients. Binaural monopolar GVS was administered. Main outcome variable was trunk forward flexion (TFF), assessed pre and immediately post GVS, and one month after. The TFF was assessed in the standing position, with open and closed eyes. The Berg balance scale was assessed before and 1 month after GVS. The TFF immediately after the GVS showed a reduction of 2.3 cm (p = 0.091) in the first reading, and after a 1-min stance (p = 0.025). Four of 5 patients maintained or even improved this effect a month after (p = 0.082). No changes in Berg balance scale were observed. Our results suggest an acute improvement in posture after one session of GVS, which significantly decreased TFF. Most of the results had a marginal significance due to small sample size.


Sleep deprivation aggravated lipopolysaccharide/D-galactosamine-induced acute liver injury by suppressing melatonin production.
Sleep loss is common in patients with liver injury, but the effects of sleep deprivation (SD) on liver injury remain unclear. In the present study, the potential effects of SD on acute liver injury and the underlying mechanisms have been investigated. The sleep of male BALB/c mice has been deprived by using a modified multiple platform water bath for 3 days and acute liver injury was induced by intraperitoneal injection of lipopolysaccharide (LPS) and D-galactosamine (D-Gal). The degree of liver injury was detected by aminotransferase determination, histopathology and survival rate analysis. Inflammatory response and melatonin (MT) were measured by enzyme-linked immunosorbent assay (ELISA). In addition, hepatocyte apoptosis was determined by caspase activity measurement and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. We observed that SD increased plasma aminotransferases, TUNEL-positive hepatocytes, histological abnormalities and mortality rates in mice with LPS/D-Gal treatment. SD also promoted LPS/D-Gal-induced production of TNF-α and upregulated hepatic caspase-8, caspase-9, and caspase-3 activities in LPS/D-Gal-exposed mice. In addition, SD significantly decreased MT contents in plasma of mice with acute liver injury, but supplementation with MT reversed these SD-promoted changes. Our data suggested that SD exacerbated LPS/D-Gal-induced liver injury via decreasing melatonin production.


Non-invasive estimation of hepatic blood perfusion from H2 15O PET images using tissue-derived arterial and portal input functions.
The liver is perfused through the portal vein and the hepatic artery. When its perfusion is assessed using positron emission tomography (PET) and (15)O-labeled water (H(2) (15)O), calculations require a dual blood input function (DIF), i.e., arterial and portal blood activity curves. The former can be generally obtained invasively, but blood withdrawal from the portal vein is not feasible in humans. The aim of the present study was to develop a new technique to estimate quantitative liver perfusion from H(2) (15)O PET images with a completely non-invasive approach. We studied normal pigs (n=14) in which arterial and portal blood tracer concentrations and Doppler ultrasonography flow rates were determined invasively to serve as reference measurements. Our technique consisted of using model DIF to create tissue model function and the latter method to simultaneously fit multiple liver time-activity curves from images. The parameters obtained reproduced the DIF. Simulation studies were performed to examine the magnitude of potential biases in the flow values and to optimize the extraction of multiple tissue curves from the image. The simulation showed that the error associated with assumed parameters was <10%, and the optimal number of tissue curves was between 10 and 20. The estimated DIFs were well reproduced against the measured ones. In addition, the calculated liver perfusion values were not different between the methods and showed a tight correlation (r=0.90). In conclusion, our results demonstrate that DIF can be estimated directly from tissue curves obtained through H(2) (15)O PET imaging. This suggests the possibility to enable completely non-invasive technique to assess liver perfusion in patho-physiological studies.


Exploring the transition of undergraduate medical students into a clinical clerkship using organizational socialization theory.
Transitions in medical education are emotionally and socially dynamic; this may affect learning. Students transitioning from preclinical to clinical training may experience negative consequences. Less is understood about students' experiences during transitions within clinical training and influential factors. The authors used organizational socialization theory to explore a transition within the clinical years. Final-year medical students experienced a nine-week internal medicine clerkship; willing students participated. Students (n = 101; 97 %) completed a questionnaire with open-ended questions at the beginning and end of the clerkship and participated in six consecutive focus groups, until data saturation occurred (n = 37). Data were thematically analyzed. Socialization was challenging. Many students experienced difficulty developing relationships with team members. Students with a positive attitude experienced a smoother transition. Many students were uncertain of their roles, concerned about the workload and desired guidance to meet clerkship demands. This transition resulted in varied outcomes from enjoyment, increased confidence and student development through to disinterest. Transitions within clinical training are complex. Faculty should focus on adequate socialization in a new clerkship as this may facilitate a smoother transition. This may necessitate orientations, staff training, and formal student support. Further research is needed on the impact of these recommendations on learning and well-being.


MicroRNA-7 promotes neural differentiation of trabecular meshwork mesenchymal stem cell on nanofibrous scaffold.
The purpose of this study was to investigate miR-7 overexpression effects on neural differentiation of mesenchymal stem cells (MSCs) on both two-dimensional (2D) and three-dimensional (3D) culture systems. We upregulated miR-7 through lentiviral vector in trabecular meshwork MSCs (TMMSCs) and polymers of poly l-lactic acid/polycaprolactone fibrous scaffold were fabricated by electrospinning and characterized using scanning electron microscopy (SEM) and Fourier transform infrared (FTIR). Neural markers expression was evaluated through quantitative-polymerase chain reaction (q-PCR) and immunostaining. The results showed that the high percentage of cell transduction (84.9%) and miR-7 expression (folds: 677.93 and 556.4) was detected in TMMSCs-miR-7(+). SEM and FTIR established the fabrication of the hybrid scaffold. q-PCR analysis showed that on days 14 and 21 of transduction, the expression level of Nestin and glial fibrillary acidic protein (GFAP) genes were significantly higher in the scaffold (3D) compared with tissue culture polystyrene (2D) culture. The expression of microtubule-associated protein-2 (MAP-2) and GFAP genes in TMMSCs-miR-7(+) cells were significantly higher than those miR-7(-) cells after 21 days of cell culture. Also, MAP-2 and Nestin proteins were detected in TMMSCs-miR-7(+) cells. Our results demonstrate that miR-7 is involved in neural differentiation of TMMSCs and scaffold can improve differentiate into glial and neural progenitor cells. These findings provided some information for future use of microRNAs and scaffold in tissue engineering and cell therapy for neurological diseases.


Audit of investigation and follow-up of patients with raised monoclonal proteins.
To assess the adequacy of investigation and later review of patients with serum monoclonal proteins detected in our laboratory. To review all monoclonal proteins detected and to search through the laboratory computerized records for evidence of further investigation and later review. Nine per cent of monoclonal proteins detected were not followed up at all. A urine examination to detect Bence-Jones protein was not requested in 12% of those regularly followed up. The mean age of the patients studied was 69 years, but there were eight patients under 60 years who had multiple myeloma.


[Contribution of contrast enhanced ultrasonography in the characterization of carotid lesions].
Harmonic mode ultrasound with injection of a contrast enhancement agent allows visualization of mobile microbubbles in the carotid plaque corresponding to neovessels secondary to an inflammation or hypoxia. These neovessels could be considered "precursor" markers of the vulnerable plaque. The aim of this work was to give an update on ultrasound contrast imaging acquisition in the exploration of carotid artery both for atheromatous lesions and for large vessel vasculitis. A precise description of the material to be used, the image acquisition methodology and the environmental conditions is discussed, emphasizing the pitfalls to be avoided as well as proper image interpretation. Microbubbles in a plaque are significantly associated with an increase in cardiovascular events (infarction and acute coronary syndrome) and ipsilateral cerebral ischemic events. Wall irregularities, microfissures and ulcer plaque detection are facilitated by the use of contrast compared to the CT scan. No studies have yet validated contrast enhanced ultrasound in the exploration of asymptomatic carotid stenosis. Contrast enhanced ultrasound also allows to detect vasculitis of the large vessels active phases by the presence of microbubbles in the carotid wall thickening and to monitor the regression under appropriate medical treatment. Future validation studies or even registries are needed to allow better use of this tool in everyday clinical practice.


Markers of Hippo-Pathway Activity in Tumor Forming Liver Lesions.
Hepatocellular Carcinoma (HCC) is a lethal cancer worldwide. Recently, the hippo signaling pathway has been implicated in tumorigenesis of HCC and other malignant tumors. Aim of the study was therefore to evaluate the hippo signaling pathway activity and its clinico-pathological associations and crosstalk in different tumor forming hepatocellular lesions (HCC, hepatocellular adenoma (HCA), focal nodular hyperplasia (FNH) and cirrhosis). A tissue micro array (TMA) from paired human tumorous and non-tumorous (NT) tissue samples of HCC (n = 92), HCA (n = 25), FNH (n = 28) and cirrhosis (n = 28; no NT) was constructed. The hippo-pathway related proteins of MST1/2, (nuclear(n)/cytoplasmic(c)) YAP and (phospho(p)) TAZ and interactors as Glypican3, RASSF1a, pAKT, pERK and pP70S6K were evaluated by immunohistochemistry (IHC). Proliferation was assessed by Ki67-IHC and apoptosis by TUNEL-technique. MST1/2- and nYAP-immunoreactivity was associated with lymph node status (p = 0.048, p = 0.001), higher grading (p = 0.012, p = 0.24) and unfavorable relapse-free survival (p = 0.004, p = 0.003). MST1/2, c/nYAP and pTAZ were significantly different between HCC/NT (p < 0.001, p = 0.029, p < 0.001, p < 0.001) and mono-/polyclonal hepatocellular lesions (HCC/HCA vs. FNH/cirrhosis; all p ≤ 0.001). Phospho-TAZ-negativity and nYAP-positivity were almost exclusively and MST1/2 exclusively detected in HCC. MST1/2 correlated with pP70S6K (p = 0.002), pERK (p = 0.042), RASSF1a-IRS (p = 0.002) and GPC3 (p < 0.001) and nYAP with GPC3 (p = 0.025), higher Ki67-indices (p = 0.016) and lower apoptosis rate (p = 0.078). MST1/2 and nYAP are unfavorable prognostic markers associated with an aggressive tumor-phenotype in HCC. Positive nYAP- and negative pTAZ-immunostaining were strong indicators of a monoclonal hepatocellular lesion. The unexpected findings for MST1/2 remain to be elucidated.


Symphysitis following transrectal biopsy of the prostate.
Transrectal ultrasound-guided needle biopsy of the prostate is a widely accepted technique to obtain prostatic tissue for histological examination. Severe complications are rarely seen. We report a case of symphysitis causing hospitalization and severe pain and discomfort of the patient. Possible etiologic factors are traumatic osseous lesions and transport of rectal bacteria to the periosseous region. Especially in small prostates, care should be taken to avoid this condition. Prolonged perioperative antibiotic prophylaxis is mandatory.


A rapid method for the determination of radioactive caesium in live animals and carcasses, and its practical application in Norway after the Chernobyl nuclear reactor accident.
A technique was developed for the measurement of levels of caesium radionuclides (137Cs + 134Cs) in live reindeer, cattle, and sheep and in carcasses from these species. The instrument used was a sodium iodide scintillation detector coupled to a portable multi-channel analyser. Based on a combination of background measurements and measurements of impulses from animals with the detector in different anatomical positions we recommend the following procedures: LAMB: The detector placed on os sacrum (standing animal). REINDEER: The detector placed between the hind legs (animal lying on its side). CATTLE: The detector placed on the back of the standing animal, midway between os sacrum and trochanter major. Average geometrical factors for live animals were estimated. It was a linear correlation between measured activity levels in meat samples and counted impulses per sec in live animals. Geometrical factors were estimated at 95% confidence level with uncertainty between 6-14%. The detection limits varied between 50-200 Bq (becquerel)/kg in areas with ground depositions between 5-200 kBq/m2. Since the winter 1986/87 the technique has been the standard procedure for monitoring slaughter animals and carcasses for radiocaesium activity concentrations.


Intensive Care Unit Capacity and Its Associated Risk Factors During the COVID-19 Surge in the Republic of Korea: Analysis Using Nationwide Health Claims Data.
To identify risk factors for intensive care unit (ICU) admission and mechanical ventilator usage among confirmed coronavirus disease (COVID-19) patients and estimate the effects of mitigation efforts on ICU capacity in Korea. Data on profiles and medical history of all confirmed COVID-19 patients in the past 1 year were extracted from the Korean National Health Insurance System's claims database to assess risk factors for ICU admission and ventilator use. We used a time-series epidemic model to estimate the ICU census in Daegu from the reported hospital data. Multivariate regression analysis revealed male sex, old age, and residing in Daegu city as significant risk factors for ICU admission. The number of patients requiring ICU admission exceeded the bed capacity across all Daegu hospitals before March 9, 2020, and therefore, critically ill patients were transferred to nearby hospitals outside Daegu. This finding was consistent with our prediction that the ICU census in Daegu would peak on March 16, 2020, at 160 through mitigation efforts, without which it would have reached 300 by late March 2020. Older age and male sex were risk factors for ICU admission. In addition, the geographic location of the hospital seems to contribute to the severity of the COVID-19 patients admitted to the ICU and to the ICU capacity.


The incidence of stressful life events of elementary school-aged children.
Certain events, commonly found in the life histories of children, have been characterized as potentially stressful (Rutter, 1981). There is some evidence to support the hypothesis that these events play a role in the etiology of various disorders, although the nature of that relationship is not clear (Dohrenwend & Dohrenwend, 1974). Yet that relationship remains the subject of considerable interest, as can be seen by the number of studies on the influence of stressful life events on children's emotional adjustment and school performance (Boike, Gesten, Cowen, Felner, & Francis, 1978; Felner, Ginter, Boike, & Cowen, 1981; Felner, Stolberg, & Cowen, 1975; Gesten, Scher, & Cowen, 1981; Felner, Stolberg, & Cowen, 1975; Gesten, Scher, & Cowen, 1978; Rolf & Garmezy, 1974; Sandler & Block, 1979; Siegel, 1981). Sandler and Ramsey (1980) have suggested that the role of life events as stressors can be understood to be a function of, among other things, the frequency of occurrence of events. For research and clinical assessment purposes, baseline data on the incidence of stressful life events of children seem to be necessary. In order to collect and organize baseline data, the relevant demographic variables must be identified. It was hypothesized that there might be two relevant demographic variables: age and socioeconomic status (SES). The purposes of this study were to (a) conduct a survey to gather baseline data on the incidence of stressful life events of children, and (b) examine the effects of age and SES on the number of events reported.


Identification and reconstitution of the nucleoside transporter of CEM human leukemia cells.
The major nucleoside transporter of the human T leukemia cell line CEM has been identified by photoaffinity labeling with the transport inhibitor nitrobenzylmercaptopurine riboside (NBMPR). The photolabeled protein migrates on SDS-PAGE gels as a broad band with a mean apparent molecular weight (75,000 +/- 3000) significantly higher than that reported for the nucleoside transporter in human erythrocytes (55,000) (Young et al. (1983) J. Biol. Chem. 258, 2202-2208). However, after treatment with endoglycosidase F to remove carbohydrate, the NBMPR-binding protein in CEM cells migrates as a sharp peak with an apparent molecular weight (47,000 +/- 3000) identical to that reported for the deglycosylated protein in human erythrocytes (Kwong et al. (1986) Biochem. J. 240, 349-356). It therefore appears that the difference in the apparent molecular weight of the NBMPR-sensitive nucleoside transporter between the CEM cell line and human erythrocytes is a result of differences in glycosylation. The NBMPR-binding protein from CEM cells has been solubilized with 1% octyl glucoside and reconstituted into phospholipid vesicles by a freeze-thaw sonication technique. Optimal reconstitution of uridine transport activity was achieved using a sonication interval of 5 to 10 s and lipid to protein ratios of 60:1 or greater. Under these conditions transport activity in the reconstituted vesicles was proportional to the protein concentration and was inhibited by NBMPR. Omission of lipid or protein, or substitution of a protein extract prepared from a nucleoside transport deficient mutant of the CEM cell line resulted in vesicles with no uridine transport activity. The initial rate of uridine transport, in the vesicles prepared with CEM protein, was saturable with a Km of 103 +/- 11 microM and was inhibited by adenosine, thymidine and cytidine. The Km for uridine and the potency of the other nucleosides as inhibitors of uridine transport (adenosine greater than thymidine greater than cytidine) were similar to intact cells. Thus, although the nucleoside transporter of CEM cells has a higher molecular weight than the human erythrocyte transporter, it exhibits typical NBMPR-sensitive nucleoside transport activity both in the intact cell and when reconstituted into phospholipid vesicles.


De novo transcriptome analysis and identification of defensive genes in garlic (Allium sativum L.) using high-throughput sequencing.
Garlic (Allium sativum L.) is the second most widely cultivated Allium which is mainly grown in temperate regions and used as a flavoring agent in a wide variety of foods. Garlic contains various bioactive compounds whose metabolic pathways, plant-pathogen interactions, defensive genes, identify interaction networks, and functional genomics were not previously predicted in the garlic at the genomic level. To address this issue, we constructed two garlic Illumina 2000 libraries from tissues of garlic clove and leaf. Approximately 43 million 125 bp paired-end reads were obtained in the two libraries. A total of 239,973 contigs were generated by de novo assembly of both samples and were compared with the sequences in the NCBI non-redundant protein database (Nr). In total, 42% of contigs were matched to known proteins in public databases including Nr, Gene Ontology (GO), and Cluster Orthologous Gene Database (COG), and then, contigs were mapped to 138 via functional annotation against the Kyoto Encyclopedia of Genes and Genomes pathway database (KEGG). In addition, a number of regulatory genes including the CCHC (Zn) family, followed by WD40, bromodomain, bZIP, AP2-EREBP, BED-type (Zn) proteins, and defense response proteins related to different conserved domains, such as RGA3, NBS-LRR, TIR-NBS-LRR, LRR, NBS-ARC, and CC-NBS-LRR were discovered based on the transcriptome dataset. We compared the ortholog gene family of the A. sativum transcriptome to A. thaliana, O. sativa, and Z. mays and found that 12,077 orthologous gene families are specific to A. sativum L. Furthermore, we identified genes involved in plant defense mechanisms, their protein-protein interaction network, and plant-pathogen interaction pathways. Our study contains an extensive sequencing and functional gene-annotation analysis of A. sativum L. The findings provide insights into the molecular basis of TFs, defensive genes, and a reference for future studies on the genetics and breeding of A. sativum L.


Effect of arginine on oligomerization and stability of N-acetylglutamate synthase.
N-acetylglutamate synthase (NAGS; E.C.126.96.36.199) catalyzes the formation of N-acetylglutamate (NAG) from acetyl coenzyme A and glutamate. In microorganisms and plants, NAG is the first intermediate of the L-arginine biosynthesis; in animals, NAG is an allosteric activator of carbamylphosphate synthetase I and III. In some bacteria bifunctional N-acetylglutamate synthase-kinase (NAGS-K) catalyzes the first two steps of L-arginine biosynthesis. L-arginine inhibits NAGS in bacteria, fungi, and plants and activates NAGS in mammals. L-arginine increased thermal stability of the NAGS-K from Maricaulis maris (MmNAGS-K) while it destabilized the NAGS-K from Xanthomonas campestris (XcNAGS-K). Analytical gel chromatography and ultracentrifugation indicated tetrameric structure of the MmMNAGS-K in the presence and absence of L-arginine and a tetramer-octamer equilibrium that shifted towards tetramers upon binding of L-arginine for the XcNAGS-K. Analytical gel chromatography of mouse NAGS (mNAGS) indicated either different oligomerization states that are in moderate to slow exchange with each other or deviation from the spherical shape of the mNAGS protein. The partition coefficient of the mNAGS increased in the presence of L-arginine suggesting smaller hydrodynamic radius due to change in either conformation or oligomerization. Different effects of L-arginine on oligomerization of NAGS may have implications for efforts to determine the three-dimensional structure of mammalian NAGS.


Differentiated thyroid cancer is associated with less aggressive disease and better outcome in patients with coexisting Hashimotos thyroiditis.
Evaluation of surgical specimens suggests that patients with Hashimoto thyroiditis (HT) have a higher prevalence of differentiated thyroid cancer. Although patients with HT are reported to present with earlier stage disease, there is controversy as to whether these patients have better prognosis when adjusted for histology and stage at presentation. To investigate differences between patients with differentiated thyroid cancer patients and without HT for aggressiveness of disease and clinical outcome, and the decline rate of antithyroglobulin antibodies titers over time. A retrospective study using the Rabin Medical Center Thyroid Cancer Registry. Seven hundred fifty-three patients were included and divided into 2 groups of patients with and without HT at diagnosis. Disease severity at presentation was evaluated using the entire cohort, whereas a control group matched for age, gender, histology, and stage was used to evaluate disease course and outcome. HT was present in 14.2% (n = 107) of included patients and was associated with smaller primary tumor (17.9 vs 21.2 mm, P = .01) and less lymph node involvement (23% vs 34%, P = .02) at presentation. When matched groups were compared, patients with HT received less additional radioactive iodine (RAI) treatments (1.24 vs 1.45, P = .03) and showed lower rates of persistence at 1 year (13% vs 26%, P = .04) and higher rates of disease remission at the end of follow-up (90% vs 79%, P = .05). On multivariate analysis HT was predictive of a lower rate of lymph nodes involvement (odds ratio 0.34, 95% confidence interval 0.17-0.66) and persistent disease at the end of follow-up (odds ratio 0.48, 95% confidence interval 0.24-0.93). Antithyroglobulin antibodies slowly disappeared in most patients with no evidence of disease. Our study demonstrates that HT is associated with a less aggressive form of differentiated thyroid cancer and a better long-term outcome.


One-step amino acid selective isotope labeling of proteins in prototrophic Escherichia coli strains.
Amino acid selective isotope labeling is a useful approach to simplification of nuclear magnetic resonance (NMR) spectra of large proteins. Cell-free protein synthesis offers essentially unlimited flexibility of labeling patterns but is labor-intensive and expensive. In vivo labeling is simple in principle but generally requires auxotrophic strains, inhibitors of amino acid synthesis, or complex media formulations. We describe a simple procedure for amino acid selective labeling of proteins expressed in prototrophic Escherichia coli strains. Excellent labeling selectivity was achieved for histidine, lysine, methionine, and alanine. Simplicity and robustness of this protocol make it a useful tool for protein NMR.


Laminin G1 residues of protein S mediate its TFPI cofactor function and are competitively regulated by C4BP.
Protein S is a cofactor in the tissue factor pathway inhibitor (TFPI) anticoagulant pathway. It enhances TFPIα-mediated inhibition of factor (F)Xa activity and generation. The enhancement is dependent on a TFPIα-protein S interaction involving TFPIα Kunitz 3 and protein S laminin G-type (LG)-1. C4b binding protein (C4BP), which binds to protein S LG1, almost completely abolishes its TFPI cofactor function. However, neither the amino acids involved in TFPIα enhancement nor the mechanisms underlying the reduced TFPI cofactor function of C4BP-bound protein S are known. To screen for functionally important regions within protein S LG1, we generated 7 variants with inserted N-linked glycosylation attachment sites. Protein S D253T and Q427N/K429T displayed severely reduced TFPI cofactor function while showing normal activated protein C (APC) cofactor function and C4BP binding. Based on these results, we designed 4 protein S variants in which 4 to 6 surface-exposed charged residues were substituted for alanine. One variant, protein S K255A/E257A/D287A/R410A/K423A/E424A, exhibited either abolished or severely reduced TFPI cofactor function in plasma and FXa inhibition assays, both in the presence or absence of FV-short, but retained normal APC cofactor function and high-affinity C4BP binding. The C4BP β-chain was expressed to determine the mechanisms behind the reduced TFPI cofactor function of C4BP-bound protein S. Like C4BP-bound protein S, C4BP β-chain-bound protein S had severely reduced TFPI cofactor function. These results show that protein S Lys255, Glu257, Asp287, Arg410, Lys423, and Glu424 are critical for protein S-mediated enhancement of TFPIα and that binding of the C4BP β-chain blocks this function.


A two-stage design for randomized trials with binary outcomes.
A two-stage rule for a randomized clinical trial with a dichotomous outcome is presented. It is based on a rule discussed by Ellenberg and Eisenberger (Cancer Treat Rep 69:1147-1154, 1985). The rule is shown to lead to a reduction in expected sample size if the test treatment is ineffective, with almost no loss in power. All the mathematical formulas required to design such a rule are given, as well as some time-saving approximations. The accuracy of the approximations is shown.


An environmentally-relevant mixture of organochlorines and its vehicle control, dimethylsulfoxide, induce ultrastructural alterations in porcine oocytes.
Organochlorine chemicals accumulate in the environment, particularly in the Arctic, and constitute potential developmental hazards to wildlife and human health. Although some of their harmful effects are recognized, their mechanisms of action within the target cells need to be better understood. This study was designed to test the hypothesis that an environmentally-relevant organochlorine mixture alters oocyte ultrastructure in the porcine model. Immature cumulus-oocyte complexes (COCs), partially cultured (18 hr) COCs without treatment or exposed to the organochlorine mixture or its vehicle (0.1% dimethysulfoxide; DMSO) during culture were processed for light and transmission electronic microscopy (TEM). The organochlorines induced major ultrastructural changes in the COCs: decreased density of the lipid droplets, increased smooth endoplasmic reticulum (SER) volume and increased interactions among SER, mitochondria, lipid droplets and vesicles. We suggest that these ultrastructural changes facilitate energy formation necessary to produce metabolizing enzymes. Other ultrastructural changes may reflect some degree of organochlorine toxicity: fewer gap junctions and decreased electron density of the cortical granules. Unexpectedly, the DMSO control treatment also induced similar ultrastructural changes, but to a lesser degree than the organochlorine mixture. This study is the first to demonstrate the effect of environmental contaminants on mammalian oocyte ultrastructure.


Prevalence and Trends in Management of Idiopathic Normal Pressure Hydrocephalus in the United States: Insights from the National Inpatient Sample.
Over the past 2 decades, management of idiopathic normal pressure hydrocephalus (iNPH) has evolved significantly. In the current study, we sought to evaluate the national prevalence and management trends of iNPH in the United States using a national database. The National Inpatient Sample was queried for patients with an International Classification of Diseases diagnosis code for iNPH from 2007 to 2017. Trends in prevalence and procedure type were evaluated per 100,000 discharges and as a percentage of discharges, using weighted discharges. Utilization of procedure type across U.S. regions and hospital types was also compared. From 2007 to 2017, 302,460 weighted discharges with any diagnosis code for iNPH, aged ≥60 years, were identified. Prevalence ranged from 0.04% to 0.20% (41/100,000 to 202/100,000) among admitted patients ≥60 years old, giving an average prevalence during the study duration of 0.18% (179/100,000). Of 66,759 weighted discharges with a primary diagnosis code of iNPH undergoing surgical management, ventriculoperitoneal shunt (72.0% of discharges, n = 48,977) was most commonly used; of these, 9.3% (n = 4567) were performed laparoscopically. This result was followed by lumbar peritoneal shunt (15.1% of discharges, n = 10,441). Up to 15.1% (n = 9990) of discharges reported only a lumbar puncture, assumed to be only diagnostic, for screening, or part of serial cerebrospinal fluid removal procedures. Significant discrepancies in procedure utilization were also identified among hospitals in the Western, Southern, Northeast and Midwest regions, as well as between urban and rural hospitals (P < 0.05). We have summarized the national prevalence of iNPH, trends in its management over the previous decade and trends by region and hospital type.


A new model for Schizosaccharomyces pombe telomere recognition: the telomeric single-stranded DNA-binding activity of Pot11-389.
The protection of telomeres 1 (Pot1) proteins specifically recognize the single-stranded 3' end of the telomere, an activity essential for sustained cellular viability and proliferation. The current model for the telomeric single-stranded DNA (ssDNA) binding activity of Schizosaccharomyces pombe Pot1 is based on a 20 kDa fragment, Pot1pN. Recent biochemical studies suggest that SpPot1 contains a larger ssDNA-binding domain and we have identified a novel ssDNA-binding domain similar in size to the human Pot1 domain. This domain, Pot1(1-389), binds extremely tightly to an oligonucleotide consisting of two conserved hexameric S. pombe telomere repeats, d(GGTTACGGTTAC), with an affinity approximately 4000-fold tighter than Pot1pN binds its cognate ssDNA. The Pot1(1-389)/ssDNA complex exhibits a half-life of 53 min, consistent with that estimated for full-length SpPot1 and significantly longer than that of Pot1pN. Single nucleotide substitutions reveal that, in contrast to Pot1pN, tandem trinucleotide repeats (GTT) within d(GGTTACGGTTAC) are specifically recognized by Pot1(1-389). Interestingly, certain single nucleotide substitutions that impacted Pot1pN binding exhibited no effect on binding affinity by Pot1(1-389). However, these substitutions reduced binding affinity when simultaneously substituted in each hexameric repeat. The non-additive nature of these substitutions suggests that certain nucleotides are coupled through the ability of the flexible ssDNA oligonucleotide to adopt alternate, thermodynamically equivalent conformations. The biochemical behavior of Pot1(1-389) is more similar to that of the full-length SpPot1 protein than to that of Pot1pN, making Pot1(1-389) a valuable domain for the future study of how full-length SpPot1 interacts with telomeric ssDNA.


Death after surgery among patients with chronic disease: prospective study of routinely collected data in the English NHS.
Five million surgeries take place in the NHS each year. Little is known about the prevalence of chronic diseases among these patients, and the association with postoperative outcomes. Analysis of routine data from all NHS hospitals in England including patients aged ≥18 yr undergoing non-obstetric surgery between January 1, 2010 and December 31, 2015. The primary outcome was death within 90 days after surgery. For each chronic disease, we adjusted for age, sex, presence of other diseases, emergency surgery, and year using logistic regression models. We defined high-risk diseases as those with an adjusted odds ratio (OR) for death ≥2 and report associated 2-yr survival. We included 8 624 611 patients (median age, 53 [36-68] yr), of whom 6 913 451 (80.2%) underwent elective surgery and 1 711 160 (19.8%) emergency surgery. Overall, 2 311 600 (26.8%) patients had a chronic disease, of whom 109 686 (4.7%) died within 90 days compared with 24 136 (0.4%) of 6 313 011 without chronic disease. Respiratory disease (1 002 281 [11.6%]), diabetes mellitus (662 706 [7.7%]), and cancer (310 363; 3.6%) were the most common. Four chronic diseases accounted for 7.7% of patients but 59.0% of deaths: cancer (37 693 deaths [12.1%]; OR=8.3 [8.2-8.5]), liver disease (8638 deaths [10.3%]; OR=4.5 [4.4-4.7]), cardiac failure (26 604 deaths [12.6%]; OR=2.4 [2.4-2.5]), and dementia (19 912 deaths [17.9%]; OR=2.0 [1.9-2.0]). Two-year survival was 67.7% among patients with high-risk chronic disease, compared with 97.1% without. One in four surgical patients has a chronic disease with an associated 10-fold increase in risk of postoperative death. Two-thirds of all deaths after surgery occur among patients with high-risk diseases (cancer, cardiac failure, liver disease, dementia).


Xenobiotica-metabolizing enzymes in the lung of experimental animals, man and in human lung models.
The xenobiotic metabolism in the lung, an organ of first entry of xenobiotics into the organism, is crucial for inhaled compounds entering this organ intentionally (e.g. drugs) and unintentionally (e.g. work place and environmental compounds). Additionally, local metabolism by enzymes preferentially or exclusively occurring in the lung is important for favorable or toxic effects of xenobiotics entering the organism also by routes other than by inhalation. The data collected in this review show that generally activities of cytochromes P450 are low in the lung of all investigated species and in vitro models. Other oxidoreductases may turn out to be more important, but are largely not investigated. Phase II enzymes are generally much higher with the exception of UGT glucuronosyltransferases which are generally very low. Insofar as data are available the xenobiotic metabolism in the lung of monkeys comes closed to that in the human lung; however, very few data are available for this comparison. Second best rate the mouse and rat lung, followed by the rabbit. Of the human in vitro model primary cells in culture, such as alveolar macrophages and alveolar type II cells as well as the A549 cell line appear quite acceptable. However, (1) this generalization represents a temporary oversimplification born from the lack of more comparable data; (2) the relative suitability of individual species/models is different for different enzymes; (3) when more data become available, the conclusions derived from these comparisons quite possibly may change.


Quantitative Assay of SARS-CoV-2 RNA and Level of Proinflammatory Protein Gene Transcripts in Peripheral Blood Leukocytes after a Novel Coronavirus Infection.
The possibility of finding persistent SARS-CoV-2 viral particles in human peripheral blood leukocytes after a novel coronavirus infection was shown. The results of droplet digital PCR showed that 19 of 24 examined subjects had from 4 to 555 copies of the Nsp4 SARS-CoV-2 gene in 5-6 months after infection. The presence of this transcript in peripheral blood leukocytes was associated with reduced expression of FOXP3 gene and increased level of RORγ gene mRNA. The copy number of the Nsp4 gene negatively correlated with the level of FOXP3 gene mRNA (r=-0.45; p=0.028), but showed a positive correlation with the DANCR long non-coding RNA (r=0.94; p<0.001). In SARS-CoV-2-positive healthy individuals, the level of TLR2, NLRP3, and IL1B gene transcripts was higher than in SARS-CoV-2-negative donors. The presence of SARS-CoV-2 in a persistent form is probably associated with impaired immunosuppression and the development of chronic inflammation in apparently healthy volunteers after a new coronavirus infection.


Xanthogranulomatous pyelonephritis in children: diagnostic and therapeutic aspects.
Xanthogranulomatous pyelonephritis (XGP) is a rare, chronic inflammatory disease of the kidneys. It is characterized by destruction of renal parenchyma and accumulation of lipid-laden foamy macrophages. Diffuse and focal forms are known. The condition is mainly observed in middle-aged women, and it is very rare in childhood. Of 32 nephrectomies carried out in children for various diseases in our hospital over the course of 2 years, there were two cases of diffuse XGP. In both cases, the preoperative diagnosis based on ultrasound findings was highly suggestive of XGP. We present the two cases and define the typical ultrasonographic signs for distinguishing XGP from other renal masses. The diagnostic and therapeutic management is discussed and an overview of the literature is given.


Asymmetric segregation of mitochondria and mortalin correlates with the multi-lineage potential of inner ear sensory cell progenitors in vitro.
The sensory epithelia of the inner ear include hair cells and supporting cells that share a common precursor. One possible mechanism involved in the genesis of these cell types is through asymmetric cell division. In this work we have studied asymmetric division of inner ear sensory cell progenitors in vitro in an attempt to understand how the different cell phenotypes are generated. In the search for molecules that will segregate asymmetrically we have found that mitochondria in general, and a mitochondrial protein named mortalin in particular, are asymmetrically segregated during certain cell divisions. In one conditionally immortal cell line (UB/OC-1), which represents a population of committed hair cell precursors, mortalin is uniformly distributed in the cytoplasm and shared equally between sibling cells during division. In another cell line (UB/UE-1), which represents a bipotent, vestibular supporting cell that can produce both neonatal hair cells as well as supporting cells, mortalin segregates asymmetrically. In UB/UE-1, approximately 12% of the cells display an asymmetric distribution of mortalin and mitochondria. The proportion of asymmetric cells increases immediately after the release of the immortalizing gene and before the onset of differentiation. The asymmetric segregation of mortalin in the bipotent cell line and its uniform distribution in a committed, lineage-restricted cell line raises the possibility that it may play a role in cell fate determination.


Activity of fosmidomycin in an in vitro model of the treatment of bacterial cystitis.
The response to fosmidomycin of four strains of Escherichia coli was studied in an in vitro model of the treatment of bacterial cystitis. Three susceptible strains of E. coli responded well to relatively low concentrations of fosmidomycin: doses achieving peak concentrations of 50 or 250 mg/l suppressed bacterial growth for 13 h or more; however, when the surviving bacteria were challenged with a second dose, a reduced response was observed. When a fully resistant strain was exposed to fosmidomycin, bacterial growth was also suppressed for 13 h or more, even when the peak concentration achieved was below the conventionally determined minimum inhibitory concentration. Resistant variants which emerged after exposure to fosmidomycin were also resistant to fosfomycin in the absence of the potentiating agent, glucose-6-phosphate. In the presence of glucose-6-phosphate, complete (or partial) susceptibility to fosmidomycin and fosfomycin was retained by three of the four strains. These results suggest that fosmidomycin and fosfomycin are transported into E. coli by a similar mechanism, and that deletion of the hexose phosphate transport system does not occur following exposure to fosmidomycin in the absence of glucose-6-phosphate.


Efficient in vitro direct shoot organogenesis and regeneration of fertile plants from embryo explants of Bambara groundnut ( Vigna subterranea L. Verdc.).
An efficient protocol has been developed for direct shoot organogenesis from embryo axes derived from mature seeds of two different landraces of Bambara groundnut. Multiple shoots were initiated on several media containing different concentrations and combinations of benzylaminopurine (BAP) or thidiazuron (TDZ). Efficient regeneration occurred when the embryo axes were first plated for 6 days on a medium containing high concentrations of BAP (1 mg/l) and alpha-naphthaleneacetic acid (NAA, 1 mg/l) and then cut transversely and transferred onto a medium containing 1.5 mg/l BAP. Shoot regeneration frequency was 100% and from five to eight shoots per explant were obtained. The importance of using embryo explants and cytokinins in the culture media, with respect to controlling the development of a highly organogenic system, was demonstrated. Histological studies revealed that proliferating buds originated directly from the superficial layers of the explants without an intermediate callus phase. The regenerated shoots were rooted on a medium containing 1 mg/l NAA and then transferred to the greenhouse. Flow cytometric analyses and chloroplast counts of guard cells suggested that the regenerants were diploid. All were morphologically normal and fertile. The short duration, high efficiency and low frequency of somaclonal variation of this system make it well suited for wider biotechnological applications of Bambara groundnut-a neglected and under-utilized crop.


Type VII Collagen Deficiency in the Oncogenesis of Cutaneous Squamous Cell Carcinoma in Dystrophic Epidermolysis Bullosa.
Dystrophic epidermolysis bullosa is a rare genetic skin disorder caused by COL7A1 sequence variations that result in type VII collagen deficits and cutaneous and extracutaneous manifestations. One serious complication of dystrophic epidermolysis bullosa is cutaneous squamous cell carcinoma, a leading driver of morbidity and mortality, especially among patients with recessive dystrophic epidermolysis bullosa. Type VII collagen deficits alter TGFβ signaling and evoke multiple other cutaneous squamous cell carcinoma progression-promoting activities within epidermal microenvironments. This review examines cutaneous squamous cell carcinoma pathophysiology in dystrophic epidermolysis bullosa with a focus on known oncogenesis pathways at play and explores the idea that therapeutic type VII collagen replacement may reduce cutaneous squamous cell carcinoma risk.


Asian Corn Borer (Ostrinia furnacalis) Infestation Increases Fusarium verticillioides Infection and Fumonisin Contamination in Maize and Reduces the Yield.
Field trials based on manual infestation of the Asian corn borer (ACB) (Ostrinia furnacalis [Guenée]) and Fusarium verticillioides (Nirenberg) atomization were conducted on four maize hybrids to investigate the relationship between ACB infestation and F. verticillioides infection, yield loss, and fumonisin contamination in maize. Analysis of fumonisins B1 and B2 was carried out using an LC-MS/MS system. In this study, manual ACB infestation significantly promoted F. verticillioides infection (both symptomatic and symptomless) and grain fumonisin levels. Ear rot incidence and severity, symptomless kernel infection, and fumonisin contamination were significantly correlated to each other and to ACB damage severity. Manual ACB infestation increased fumonisin levels from 580 to 4,418 µg/kg in 2018; 6,059 to 10,681 µg/kg in 2019 spring-sown maize (2019A); and 2,042 to 5,060 µg/kg in 2019 summer-sown maize (2019B), with the threshold of the European Union (EU) being 4,000 µg/kg. The threshold was exceeded in spring of 2019 in untreated controls. Regarding yield, significant negative correlation between ACB damage and ear weight was observed in three seasons. These results indicated that ACB infestation can lead to severe quality degradation and yield loss of maize. Kernel fumonisin levels may exceed the concentration threshold of the EU in certain conditions, threatening the health of livestock and humans. Measures should be taken to reduce ACB infestation to ensure food and feed security.


Spontaneous fistula formation in renal pelvis of transplanted kidney.
A case of spontaneous fistula formation in renal pelvis of transplanted kidney is reported. Three attempts at surgical correction of the fistula failed, and cure was achieved by longstanding nephrostomy drainage.


Associations between exposure to brominated flame retardants and metabolic syndrome and its components in U.S. adults.
Humans are simultaneously exposed to numerous of environmental brominated flame retardants (BFRs). We aim to explore the overall associations of BFRs mixture on metabolic syndrome (MetS) and its components and further identify significant chemicals. This study included 4641 adults from the National Health and Nutrition Examination Survey (NHANES) in 2007-2016. The weighted logistic regression was conducted to estimate the association of a single BFR exposure with MetS and its components. Meanwhile, the weighted quantile sum (WQS) regression and Bayesian kernel machine regression (BKMR) were adopted to evaluate the overall associations of BFRs mixture on MetS and its components, and to identify significant chemicals. We also evaluated potential associations modified by sex. In the weighted logistic regression model, PBB153 were positively associated with MetS in a dose-dependent manner (Ptrend < 0.05). For its components, increasing quartiles of most BFRs were positively associated with abdominal obesity, hypertriglyceridemia, and low HDL. However, we found no statistically significant associations between BFRs and hypertension and hyperglycemia. WQS analyses found that BFRs mixture was positively associated with MetS (OR: 1.30; 95%CI:1.14, 1.46), abdominal obesity (OR: 1.15; 95%CI:1.03, 1.27), hypertriglyceridemia (OR:1.43; 95%CI:1.19, 1.67), and low HDL (OR: 1.15; 95%CI:1.01, 1.29). BKMR showed associations in a similar direction as WQS for BFRs mixture. For MetS, hypertriglyceridemia and Low HDL, PBB153, PBDE28 and PBDE209 were the most heavily weighting chemicals and had the highest the posterior inclusion probabilities in the WQS and BKMR, respectively. BFRs showed stronger associations of MetS and its components in males than in females. The present study suggested exposure to BFRs mixture was positively associated with MetS and its components in adults, and PBB153, PBDE28 and PBDE209 were the significant chemicals. However, prospective cohort studies are still needed to confirm the causal effect between BFRs mixture and MetS.


Simultaneous determination of macitentan and its active metabolite in human plasma by liquid chromatography-tandem mass spectrometry.
Macitentan is a newly approved endothelin receptor antagonist (ERA) for the long-term treatment of PAH with superior receptor-binding properties and a longer duration of action compared to other available ERAs. However, analytical methods for simultaneous determination of macitentan and its active metabolite, ACT-132577, in human plasma have not been fully reported in the literature. In this work, a fast, sensitive, and reliable high-performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) was firstly developed and completely validated for simultaneous determination of macitentan and its active metabolite in human plasma. Plasma samples were processed with a protein precipitation using acetonitrile, followed by chromatographic separation using an Inertsil ODS-SP column (100×2.1mm, 3.5μm) under isocratic elution with a mobile phase consisting of acetonitrile and 0.2% formic acid at a flow rate of 0.3mL/min. Quantification was operated in multiple reaction monitoring (MRM) mode using the transitions m/z 547.1→201.0 for macitentan, m/z 589.0→203.0 for ACT-132577, and m/z 380.5→243.3 for the IS (donepezil). The assay exhibited a linear range of 1-500ng/mL for both macitentan and ACT-132577. The accuracy and the intra- and inter-precisions were within acceptable ranges and no significant matrix effect was observed during the method validation. The developed method was successfully utilized to a human pharmacokinetic study of macitentan as well as ACT-132577 after oral administration of 10mg macitentan tablet in healthy Chinese volunteers.


Type I, II, and III Interferons: Regulating Immunity to Mycobacterium tuberculosis Infection.
Interferons (IFNs) are cytokines released by host cells in response to the presence of pathogens or tumor cells. The aim of this review was to present the previously known and new findings about the role of interferons type I and II, and recently discovered type III in Mycobacterium tuberculosis (M. tuberculosis) infection control. Infection of various cell types with M. tuberculosis induce both IFN-α and IFN-β synthesis. The majority of the studies support the findings that IFN type I actually promotes infection with M. tuberculosis. It has been well establish that IFN-γ has protective function against M. tuberculosis and the other mycobacteria and that the primary source of this cytokine are CD4(+) and CD8(+) T cells. Recently, it has been shown that also the innate lymphocytes, γδ T cells, natural killer (NK) T cells, and NK cells can also be the source of IFN-γ in response to mycobacterial infection. Several studies have shown that CD4(+) T cells protect mice against M. tuberculosis independently of IFN-γ. The balance between IFN-γ and different cytokines such as IL-10 and other Th2 cell cytokines is likely to influence disease outcome. Type I IFN appears to be detrimental through at least three separate, but overlapping, type I IFN-mediated mechanisms: induction of excessive apoptosis, specific suppression of Th1 and IFN-γ responses, and dampening of the immune response by strong IL-10 induction. Recently it has been found that M. tuberculosis infection in A549 lung epithelial cells stimulate up-regulation of IFN-λ genes in vitro. IFN-λs also have a role in modulation of Th1/Th2 response. IFN-λs are not essential for M. tuberculosis infection control, but can give some contribution in immune response to this pathogen.


Isolation and characterization of a new dimeric esterase from Lactobacillus sakei 23K.
The M(w) of a Lactobacillus sakei intracellular esterase, determined by gel filtration, was compared to those obtained from SDS-PAGE or MALDI-TOF, pointing to a dimeric structure. Its N-terminal sequence and peptide mass fingerprint suggest that it is the putative LSA044 protein from L. sakei 23K genome.


Supplementation of Korean fermented soy paste doenjang reduces visceral fat in overweight subjects with mutant uncoupling protein-1 allele.
The purpose of this study was to examine the hypothesis that the antiobesity effect of doenjang, a Korean fermented soy paste is different between the mutant and the wild-type alleles of a polymorphism upstream of the uncoupling protein-1 (UCP-1) gene in overweight subjects. In our randomized, double-blind, placebo-controlled trial, a total of 51 subjects with a body mass index of 23 kg/m(2) or greater and a waist-to-hip ratio of 0.90 or greater for men or 0.85 or greater for women were randomly assigned to take 9.9 g/d of either a placebo or doenjang for 12 weeks. The relative frequency of the mutant G allele of the UCP-1 polymorphism was 0.60 in the placebo group and 0.62 in the doenjang group. Supplementation of doenjang had no significant effect on the visceral fat area compared with that of the placebo group, but there was a significantly reduced amount of visceral fat in subjects with the G allele of UCP-1 polymorphism. Doenjang supplementation was found to significantly increase the free fatty acid concentration in subjects with both the A allele and the G allele. There was a significant association between visceral fat and age in study subjects with both the wild-type and mutant alleles of the UCP-1 gene. Doenjang supplementation significantly reduced visceral fat and increased the free fatty acid concentrations in subjects with the G allele of the UCP-1 polymorphism, which suggests that doenjang may be related to increased free fatty acid levels caused by elevated lipolysis in these subjects.


Human health and ecological risk assessment of trace elements in urban soils of 101 cities in China: A meta-analysis.
Urban soils pollution by trace elements arouses the growing interest in China. The aim of this study was to assess urban soil pollution by As, Cd, Cr, Cu, Hg, Ni, Pb and Zn in China and the possible impacts on urban inhabitants and urban green spaces (UGS). Data from more than 17,000 samples were applied to characterize the status of 101 cities. The pollution assessment proved that 11% of the cities are heavily polluted. According to the Hazard Index, the value of risk for the infant population in 15 cities exceed the standardly accepted tolerable levels. The carcinogenic risk assessment demonstrated the potential threat in the cities with the total population approximately 20,566,900. Cr and As were detected to be the most hazardous elements. UGS may be seriously threatened by trace elements toxicity in 38 cities. Cd was found to be the riskiest element for UGS. Ecosystem services of UGS can be significantly disrupted under the current situation in China and their status is expected to deteriorate in the future. For this reason, it is essential to alter the policy of the urbanization process and develop functional concepts of urban green infrastructures adapted to the high level of contamination which shall improve human well-being in China.


Benefit of combined therapy with nicorandil and colchicine in preventing monocrotaline-induced rat pulmonary arterial hypertension.
This study tested the hypothesis that combined therapy with nicorandil and colchicine is superior to either alone in attenuating monocrotaline (MCT)-induced rat pulmonary arterial hypertension (PAH). Adult male Sprague-Dawley rats (n=50) were equally randomized into group 1 (sham control), group 2 [MCT (60 mg/kg i.p.)], group 3 [MCT-Nicorandil (5.0 mg/kg/day)], group 4 [MCT-Colchicine (1.0 mg/kg/day)], and group 5 (MCT-Nicorandil-Colchicine). Drugs were given on day 5. All animals were sacrificed on day 90 after MCT administration. Right ventricular systolic blood pressure (RVSBP) and RV weight were increased in group 2 compared to group 1, reduced in groups 3 and 4 compared to group 2, and further reduced in group 5, whereas arterial-oxygen saturation showed an opposite pattern (all p<0.001). Pulmonary damage severity (thickened alveolar septum and pulmonary arteriolar wall, decreased alveolar-sac numbers), number of CD3+ cells, and protein expressions of inflammatory (MMP-9, NF-κB, VCAM-1, angiotensin II-receptor), apoptotic (Bax, caspase 3, cleaved PARP), and fibrotic (TGF-β, Smad3) biomarkers showed an identical pattern compared to that of RVSBP, whereas pulmonary expressions of anti-apoptotic (Bcl-2) and anti-fibrotic (BMP-2, Smad1/5) biomarkers displayed a reverse pattern (all p<0.01). The protein expressions of RV damage markers (BNP, caspase 3) were increased, whereas expression of biomarker for RV functional preservation (Cx43) was reduced in group 2 compared with group 1, elevated in groups 3 and 4 compared to group 2, and further increased in group 5 (all p<0.01). Combined therapy with nicorandil and colchicine is superior to either alone in attenuating MCT-induced PAH in rats.


Does Religion Prevent LGBTQ Acceptance? A Case Study with Queer and Trans Muslims in Toronto, Canada.
This paper looks at the terms and conditions of acceptance for queer and trans Muslims in dominant Muslim groups. The social tension between LGBTQ Identity and religion is explored through ethnographic research with "hybrid" LGBTQ Muslim organizations and "non-hybrid" Muslim ones in Toronto, Canada. The study draws on forty-seven in-depth interviews conducted with leaders of both hybrid groups and non-hybrid ones, in order to explore (1) what the acceptance of LGBTQ Muslims constitutes, (2) whether acceptance is acquired/conferred, and (3) whether religion prevents acceptance. I measure degrees of acceptance by comparing non-hybrid interviewee responses to characteristics that constitute ideal types of "acceptance" and "non-acceptance." Responses are weighed against interviews with hybrid group leaders as well as participant-observation data. Results show a range of four acceptance levels for non-hybrid Muslim groups. The data also show that religious belief alone does not explain or account for acceptance.


Three cases of Charles Bonnet Syndrome in patients with advanced glaucomatous visual field loss but preserved visual acuity.
To describe three cases of Charles Bonnet syndrome (CBS) in glaucoma patients with preserved visual acuity. Three glaucoma patients who had taken part in a recent CBS study were interviewed about their hallucinations. The patients underwent macular optical coherence tomography (OCT) of both eyes. The visual function was evaluated with visual field measurement (Humphrey visual field analyser) and visual acuity testing (Snellen scale). All three patients had preserved visual acuity (≥0.5 in both eyes) and at least one eye with advanced visual field defect (Mean Deviation worse than -12.00 decibel). They all reported vivid visual hallucinations with insight into the unreal nature of the hallucinations. Charles Bonnet syndrome can occur in glaucoma despite preserved visual acuity. Awareness of this relation is desirable among clinicians, as it will improve communication with patients.


An alternate delivery system improves vaccine performance against foot-and-mouth disease virus (FMDV).
Foot-and-mouth disease virus (FMDV) causes vesicular disease of cloven-hoofed animals with severe agricultural and economic implications. One of the most highly infectious and contagious livestock pathogens known, the disease spreads rapidly in naïve populations making it critical to have rapidly acting vaccines. Needle inoculation of killed virus vaccine is an efficient method of swiftly vaccinating large numbers of animals, either in eradication efforts or in outbreak situations in disease free countries, although, to be efficient, this requires utilizing the same needle with multiple animals. Here we present studies using a needle free system for vaccination with killed virus vaccine, FMDV strain O1 Manisa, as a rapid and consistent delivery platform. Cattle were vaccinated using a commercially available vaccine formulation at the manufacturer's recommended dose as well as four and sixteen fold less antigen load per dose. Animals were challenged intradermalingually (IDL) with live, virulent virus, homologous strain O1 Manisa, at various times following vaccination. All non-vaccinated control cattle exhibited clinical disease, including fever, viremia and lesions, specifically vesicle formation. Cattle vaccinated with the 1/16× and 1/4× doses using the needle free device were protected when challenged at both 7 and 28 days after vaccination. These data suggest that effective protection against disease can be achieved with 1/16 of the recommended vaccine dose when delivered using the needle free, intradermal delivery system, indicating the current vaccine stockpile that can be extended by many fold using this system.