Alcohol and Puberty: Mechanisms of Delayed Development
Adolescence represents a vulnerable period for developing youth. Alcohol use and misuse are especially problematic behaviors during this time. Adolescents are more sensitive to alcohol and less tolerant of its detrimental effects than are adults. Research in humans and animals has revealed that early alcohol consumption can result in delayed pubertal development. Animal studies have shown that alcohol detrimentally affects neuroendocrine systems within the hypothalamic region of the brain that are associated with the normal, timely onset of the pubertal process. To effectively restore development and shorten recovery time associated with the adverse effects of alcohol on puberty, researchers must first understand the molecular and physiological mechanisms by which alcohol interferes with critical hypothalamic functions.


Antiproliferative activity of Vietnamese medicinal plants.
Methanol, methanol-water (1:1) and water extracts were prepared from seventy-seven Vietnamese medicinal plants and tested for their antiproliferative activities against human HT-1080 fibrosarcoma cells. Among them, fifteen extracts including seven methanol extracts of Caesalpinia sappan, Catharanthus roseus, Coscinium fenestratum, Eurycoma longifolia, Hydnophytum formicarum and Streptocaulon juventas (collected at two areas), six methanol-water (1:1) extracts of Cae. sappan, Cat. roseus, Co. fenestratum, H. formicarum and S. juventas (at two areas), and two water extracts of Cae. sappan and S. juventas exhibited antiproliferative activities in a concentration-dependent manner. Their antiproliferative activities against human cervix HeLa adenocarcinoma, human lung A549 adenocarcinoma, murine colon 26-L5 carcinoma, murine Lewis lung carcinoma (LLC) and murine B16-BL6 melanoma cells were then examined. Co. fenestratum showed selective activity against lung carcinoma and/or lung metastatic cell lines, A549, LLC and B16-BL6, while H. formicarum and S. juventas showed selective activity against human tumor cell lines, HeLa and A549. Characteristic morphological change and DNA fragmentation indicated the antiproliferative activity to be due to the induction of apoptosis.


Interdisciplinary Team Meetings in Practice: an Observational Study of IDTs, Sensemaking Around Care Transitions, and Readmission Rates.
Interdisciplinary teams (IDTs) have been implemented to improve collaboration in hospital care, but their impact on patient outcomes, including readmissions, has been mixed. These mixed results might be rooted in differences in organization of IDT meetings between hospitals, as well as variation in IDT characteristics and function. We hypothesize that relationships between IDT members are an important team characteristic, influencing IDT function in terms of how members make sense of what is happening with patients, a process called sensemaking OBJECTIVE: (1) To describe how IDT meetings are organized in practice, (2) assess differences in IDT member relationships and sensemaking during patient discussions, and (3) explore their potential association with risk-stratified readmission rates (RSRRs). Observational, explanatory convergent mixed-methods case-comparison study of IDT meetings in 10 Veterans Affairs hospitals. Clinicians participating in IDTs and facility leadership. Three-person teams observed and recorded IDT meetings during week-long visits. We used observational data to characterize relationships and sensemaking during IDT patient discussions. To assess sensemaking, we used 2 frameworks that reflected sensemaking around each patient's situation generally, and around care transitions specifically. We examined the association between IDT relationships and sensemaking, and RSRRs. We observed variability in IDT organization, characteristics, and function across 10 hospitals. This variability was greater between hospitals than between teams at the same hospital. Relationship characteristics and both types of sensemaking were all significantly, positively correlated. General sensemaking regarding each patient was significantly negatively associated with RSRR (- 0.65, p = 0.044). IDTs vary not only in how they are organized, but also in team relationships and sensemaking. Though our design does not allow for inferences of causation, these differences may be associated with hospital readmission rates.


Preoperative sizing of grafts for transfemoral endovascular aneurysm management: a prospective comparative study of spiral CT angiography, arteriography, and conventional CT imaging.
To define the impact of spiral computed tomographic angiography (CTA) with image reconstruction on graft selection for Transfemoral Endovascular Aneurysm Management (TEAM) by comparing it to conventional computed tomography (CT) and contrast arteriography. Twenty-one candidates for TEAM were included. The diameters of the superior and inferior aneurysm necks and lengths between the graft attachment sites were measured using the three imaging techniques. These measurements and their consequences on graft selection were studied. The difference in length sizing between spiral CTA and arteriography never exceeded 1 cm; however, lengths measured by conventional CT scanning resulted in underestimation of graft length in 91% of patients. Graft diameters were chosen too small in 62% of the patients when based on arteriographic diameter measurements. A graft of similar diameter was selected by spiral CTA and conventional CT scanning in 81% of the patients, while minor oversizing by conventional CT scanning was found in 14%. Neither conventional CT scanning nor arteriography is adequate as a sole preoperative radiological investigation for TEAM graft sizing. Spiral CTA with image processing produces all information required for selection of the optimal graft size and should be regarded the method of first choice for this purpose.


[Treatment of Mallory-Weiss syndrome by endoscopic diathermocoagulation].
The work analyses the results of treatment of 111 patients with the Mallory-Weiss syndrome by endoscopic diathermocoagulation. It proved ineffective from the very beginning in 11 (9.9%) cases, in another 23 (20.7%) patients hemorrhage recurred in the immediate 4 days but in one third of them its source was another disease. The follow-up of 77 patients revealed recurrence of the disease in the late-term period (from 12 months to 9 years) after diathermocoagulation in 12 (15.6%), but only 2 of them had to be operated on. The operative mortality was 17%, but when surgery was combined with diathermocoagulation the mortality rate was 4.5%.


Design, syntheses, and studies of supramolecular porphyrin-fullerene conjugates, using bis-18-crown-6 appended porphyrins and pyridine or alkyl ammonium functionalized fullerenes.
Photoinduced electron-transfer processes in cis and trans functionalized bis-18-crown-6 porphyrin self-assembled with fullerene functionalized with pyridine or alkylammonium cation entities are reported. The structural integrity of the newly formed supramolecular conjugates was accomplished by optical absorption and emission, electron spray ionization mass, electrochemistry, and semiempirical PM3 calculations. A 1:2 stoichiometry of the supramolecular porphyrin:fullerene conjugates was deduced from these studies. The conjugates revealed stable "two-point"' binding involving metal-ligand coordination and alkylammonium cation-crown ether binding or only the latter type of binding depending upon the functionality of the fullerene and metal ion in the porphyrin cavity. The effect of the variation on free energy changes of charge separation and the charge recombination was achieved by varying the metal ion in the porphyrin cavity. The charge-separation rates (k(CS)) determined from the picosecond time-resolved emission studies were generally higher for the cis bis-crown functionalized porphyrins than those of the corresponding trans ones. A comparison of the k(CS) values reported earlier for 1:1 porphyrin-fullerene conjugates with a similar self-assembly mechanism suggested that employing a higher number of acceptor entities improves the electron-transfer rates. The calculated charge-recombination rates (k(CR)) were 2-3 orders of magnitude smaller than the k(CS) values, suggesting the occurrence of the charge recombination process in the Marcus inverted region. The lifetimes of the radical ion pair (tau(RIP)) ranged between 46 and 233 ns indicating charge stabilization in the studied conjugates.


Fluid resuscitation following injury: rationale for the use of balanced salt solutions.
Initial fluid management of the injured patient involves replacement of fluid lost incident to the trauma as well as prompt recognition and treatment of shock. Prompt fluid replacement based on these concepts will result in a reduction in mortality and prevention of many complications resulting from prolonged inadequate tissue perfusion. In this discussion, an attempt will be made to facilitate this approach by: (1) discussing these fluid changes; (2) presenting a clinically applicable classification of shock; and (3) outlining a rational approach to the early treatment of hemorrhagic shock, the most frequent form of shock occurring early after injury.


Evidence for the involvement of the glucocorticoid receptor gene in bipolar disorder in an isolated northern Swedish population.
Dysfunction of the hypothalamus-pituitary-adrenal (HPA) axis is one of the most consistent findings in the pathophysiology of mood disorders. The potential role of genes related to HPA axis function has been investigated extensively in major depression. However, in bipolar disorder (BPD) such studies are scarce. We performed a systematic HapMap-based association study of six genes crucial for HPA axis function in relation to BPD. Haplotype tagging single nucleotide polymorphisms (htSNPs) were selected in order to identify all haplotypes with a frequency of more than 1% in the genes encoding the glucocorticoid receptor (GR), mineralocorticoid receptor (MR), corticotrophin releasing hormone receptor 1 (CRH-R1) and 2 (CRH-R2), CRH binding protein (CRH-BP), and FK binding protein 5 (FKBP5). This resulted in a total selection of 225 SNPs that were genotyped and analyzed in 309 BPD patients and 364 matched control individuals all originating from an isolated northern Swedish population. Consistent evidence for an association with BPD was found for NR3C1, the gene encoding GR. Almost all SNPs in two adjacent haplotype blocks contributed to the positive signal, comprised of significant single marker, sliding window, and haplotype-specific p-values. All these results point to a moderately frequent (10-15%) susceptibility haplotype covering the entire coding region and 3' untranslated region (UTR) of NR3C1. This study contributes to the growing evidence for a role of the glucocorticoid receptor gene (NR3C1) in vulnerability to mood disorders, and BPD in particular, and warrants further in vitro investigation of the at-risk haplotypes with respect to disease etiology. However, this association might be restricted to this specific population, as it is observed in a rather small sample from an isolated population without replication, and data from large meta-analyses for genome-wide association studies in BPD do not show the GR as a very strong candidate.


Phosphorylation of Eukaryotic Initiation Factor 4G1 (eIF4G1) at Ser1147 Is Specific for eIF4G1 Bound to eIF4E in Delayed Neuronal Death after Ischemia.
Ischemic strokes are caused by a reduction in cerebral blood flow and both the ischemic period and subsequent reperfusion induce brain injury, with different tissue damage depending on the severity of the ischemic insult, its duration, and the particular areas of the brain affected. In those areas vulnerable to cerebral ischemia, the inhibition of protein translation is an essential process of the cellular response leading to delayed neuronal death. In particular, translation initiation is rate-limiting for protein synthesis and the eukaryotic initiation factor (eIF) 4F complex is indispensable for cap-dependent protein translation. In the eIF4F complex, eIF4G is a scaffolding protein that provides docking sites for the assembly of eIF4A and eIF4E, binding to the cap structure of the mRNA and stabilizing all proteins of the complex. The eIF4F complex constituents, eIF4A, eIF4E, and eIF4G, participate in translation regulation by their phosphorylation at specific sites under cellular stress conditions, modulating the activity of the cap-binding complex and protein translation. This work investigates the phosphorylation of eIF4G1 involved in the eIF4E/eIF4G1 association complex, and their regulation in ischemia-reperfusion (IR) as a stress-inducing condition. IR was induced in an animal model of transient cerebral ischemia and the results were studied in the resistant cortical region and in the vulnerable hippocampal CA1 region. The presented data demonstrate the phosphorylation of eIF4G1 at Ser1147, Ser1185, and Ser1231 in both brain regions and in control and ischemic conditions, being the phosphorylation of eIF4G1 at Ser1147 the only one found in the eIF4E/eIF4G association complex from the cap-containing matrix (m7GTP-Sepharose). In addition, our work reveals the specific modulation of the phosphorylation of eIF4G1 at Ser1147 in the vulnerable region, with increased levels and colocalization with eIF4E in response to IR. These findings contribute to elucidate the molecular mechanism of protein translation regulation that underlies in the balance of cell survival/death during pathophysiological stress, such as cerebral ischemia.


[Glutamic acid oxidation and its regulation in purified mitochondria and nerve endings of rat cerebral cortex].
A comparative study of respiration of mitochondrial and synaptosomal fractions of adult rat brain cerebral cortex has shown that glutamic acid is oxidized in both fractions and that its oxidation is potentiated by ADP. Addition of ADP has, however, a different effect on the endogenous respiration of these two fractions; while it has no effect on the endogenous respiration of the synaptosomal fraction, it increases considerably the weak endogenous respiration of the mitochondrial fraction.


Soluble BTN2A1 Is a Potential Prognosis Biomarker in Pre-Treated Advanced Renal Cell Carcinoma.
The development of immune checkpoint inhibitors (ICI) has dramatically changed the landscape of therapies for metastatic renal cell carcinoma. However, many patients do not benefit from such therapy and prognostic or predictive validated biomarker validated for ICI are still needed to better select and treat patient. Plasmatic soluble immune checkpoints have been described as potential immune biomarkers in hematological malignancies and solids tumors, then, we would like to explore the prognostic value of different soluble immune checkpoints in patients with mRCC treated with nivolumab after TKI. We prospectively collected plasma samples before nivolumab infusion from 38 patients previously treated for mRCC with TKI at Paoli-Calmettes Institute, from the NIVOREN GETUG-AFU 26 study (NCT03013335). Enzyme-linked immunosorbent assays (ELISA) were performed for soluble forms of PD-1, PD-L1, global BTN3, BTLA, BTN3A1 and BTN2A1. Among the different soluble checkpoints analyzed, only high baseline plasmatic level of BTN2A1 was significantly associated with shorter PFS: median PFS was 3.95 months for sBTN2A1high vs 14.30 months for sBTN2A1low (sBTN2A1 cut-off: 6.7ng/mL; HR = 2.26, 95%CI [0.68 - 4.60], p = 0.0307). There was no statistical difference in OS between sBTN2A1high and sBTN2A1low. Our results suggest that the baseline level of plasmatic BTN2A1 could be an independent prognosis factor of PFS after nivolumab for pre-treated patient with mRCC. However, these results need to be validated in a larger prospective cohort and the biological role of BTN subfamily and γδ T cell immunity in mRCC must be elucidated.


A Study on the Characteristics of Healthy Volunteers who Participate in Phase I Clinical Trials in Korea.
The phase I trial is the first step in administering a drug to humans, but it has no therapeutic purpose. Under the absence of therapeutic purpose, healthy volunteers demonstrated different motivations, unlike the actual patients participating in trials. There were many reported motivations, such as financial motivation, contributing to the health science, accessing ancillary health care benefits, scientific interest or interest in the goals of the study, meeting people, and general curiosity. The aim of this study was to identify the motivation and characteristics of healthy volunteers participating in phase I trials in the Republic of Korea. We gave surveys to 121 healthy volunteers to study their demographic characteristics and the reasons of participation. We identified whether the decision to participate in the research was influenced by demographic factors and whether the perception and attitudes toward the research were influenced by the characteristics of the healthy volunteers. After completion of the first survey, 12 healthy volunteers who had participated in a phase I clinical trial were selected to answer the second interview. According to our survey, most healthy volunteers were unmarried men and economically dependent. Most of them participated in the study because of financial reward. The most important factor to measure financial reward was the research period. Also, 43% of the volunteers were university students, 42% answered "university graduation" and 55% were residing in family-owned houses. Many healthy volunteers were found to be living in family homes and to have a student status or lack of economic independence. Results of the survey showed that 64% of respondents indicated having more than one clinical trial participation. In-depth interviews showed that healthy volunteers had diverse motivation to participate in research and that healthy volunteer perceive the clinical trial positively. The main motivation for healthy volunteers' participation in research was "financial reward." Healthy volunteers also considered research schedules, processes, and safety, and had a positive perception of clinical trials, but they thought that the public has a negative perception.


Applying Machine Learning Approaches to Suicide Prediction Using Healthcare Data: Overview and Future Directions.
Objective: Early identification of individuals who are at risk for suicide is crucial in supporting suicide prevention. Machine learning is emerging as a promising approach to support this objective. Machine learning is broadly defined as a set of mathematical models and computational algorithms designed to automatically learn complex patterns between predictors and outcomes from example data, without being explicitly programmed to do so. The model's performance continuously improves over time by learning from newly available data. Method: This concept paper explores how machine learning approaches applied to healthcare data obtained from electronic health records, including billing and claims data, can advance our ability to accurately predict future suicidal behavior. Results: We provide a general overview of machine learning concepts, summarize exemplar studies, describe continued challenges, and propose innovative research directions. Conclusion: Machine learning has potential for improving estimation of suicide risk, yet important challenges and opportunities remain. Further research can focus on incorporating evolving methods for addressing data imbalances, understanding factors that affect generalizability across samples and healthcare systems, expanding the richness of the data, leveraging newer machine learning approaches, and developing automatic learning systems.


Hyaluronidase Hyal1 Increases Tumor Cell Proliferation and Motility through Accelerated Vesicle Trafficking.
Hyaluronan (HA) turnover accelerates metastatic progression of prostate cancer in part by increasing rates of tumor cell proliferation and motility. To determine the mechanism, we overexpressed hyaluronidase 1 (Hyal1) as a fluorescent fusion protein and examined its impact on endocytosis and vesicular trafficking. Overexpression of Hyal1 led to increased rates of internalization of HA and the endocytic recycling marker transferrin. Live imaging of Hyal1, sucrose gradient centrifugation, and specific colocalization of Rab GTPases defined the subcellular distribution of Hyal1 as early and late endosomes, lysosomes, and recycling vesicles. Manipulation of vesicular trafficking by chemical inhibitors or with constitutively active and dominant negative Rab expression constructs caused atypical localization of Hyal1. Using the catalytically inactive point mutant Hyal1-E131Q, we found that enzymatic activity of Hyal1 was necessary for normal localization within the cell as Hyal1-E131Q was mainly detected within the endoplasmic reticulum. Expression of a HA-binding point mutant, Hyal1-Y202F, revealed that secretion of Hyal1 and concurrent reuptake from the extracellular space are critical for rapid HA internalization and cell proliferation. Overall, excess Hyal1 secretion accelerates endocytic vesicle trafficking in a substrate-dependent manner, promoting aggressive tumor cell behavior.


[Hypomania/mania induced by cessation of antidepressant drugs].
Although rarely reported, the induction of hypomanic/manic episodes due to sudden or gradual cessation of antidepressant drugs is a phenomenon observed in clinical settings. Herein we present 2 patients that had manic episodes induced by gradual cessation of antidepressant drugs. Common features of both cases were as follows: patients were female; a major depressive episode was the reason for starting treatment; familial loading for unipolar depressive disorder; venlafaxine was administered for treatment of the episode; mood elevation symptoms while gradually decreasing the medication dose; absence of physical symptoms related to withdrawal; antipsychotic and mood stabilizing drugs were required for the treatment of the episode. In both cases 1) a hypomanic/manic episode induced by the use of antidepressants, 2) agitated depression, 3) physical withdrawal syndrome, and 4) spontaneous episodes in the natural course of the illness were the 4 different states that were taken into consideration for differential diagnosis. Hypomanic/manic episodes induced by cessation of antidepressant drugs are thought to shed light on the etiology of bipolar disorder, which this report discusses with reference to the case reports.


Ovine fetal metabolism during norepinephrine infusion.
Although stress in fetal life not only increases fetal catecholamine concentration but also decreases fetal growth, there have been few studies that define the specific role of catecholamines in mediating the fetal response to stress. None, however, have investigated effects on fetal amino acid or protein metabolism, processes that should be affected during aberrant fetal growth. Therefore, hormone concentrations as well as oxygen, glucose, lactate, and amino nitrogen, leucine, and protein metabolism were measured with and without norepinephrine infusion in fetuses of eight pregnant ewes (118-125 days of gestation). Transumbilical uptake of oxygen increased during norepinephrine infusion, whereas uptake of glucose remained constant and that of lactate and amino acids fell. The proportion of fetal oxidative metabolism that could be supported by transplacental uptake of exogenous substrates was < 1, indicating that endogenous substrates were used to maintain fetal oxidative metabolism and therefore that fetal growth was diminished. Both fetal leucine uptake and oxidation decreased during norepinephrine infusion, as did fetal protein synthesis and proteolysis. Fetal protein synthesis fell more than proteolysis, however. Consequently, fetal protein accretion, a variable closely related to fetal growth, also fell. Thus the effects of norepinephrine infusion in fetuses suggest that fetal catecholamines play an important role not only in altering fetal metabolism but also in regulating fetal growth.


Comparative Evaluation of Antibacterial Effect of GIC Containing Chlorhexidine and Miswak on Streptococcus mutans and Streptococcus sobrinus in Early Childhood Caries Children: A PCR Study.
The therapeutic procedures used in the treatment of caries do not always eliminate all the microorganisms. Persisting cariogenic bacteria can cause recurrent caries and failure of restoration. Incorporation of an antimicrobial agent in the restorative material may be of paramount significance. The purpose of this study was to evaluate and compare the antibacterial effect of glass ionomer cement (GIC) containing CHX and miswak extract on Streptococcus mutans and Streptococcus sobrinus in ECC children using polymerase chain reaction (PCR). Forty-five children with ECC in the age-group 3-6 years were selected. The children were randomly allocated into three groups. Supragingival plaque samples (S1) were collected from sound buccal or labial surfaces of primary teeth. Cavity preparation was done and the teeth were restored according to the group to which the child had been allotted. The second plaque sample (S2) was collected 1 month and the final sample after 3 months of restoring all the decayed teeth. All the samples were sent for PCR analysis. Intergroup analysis was done using Kruskal-Wallis test followed by Mann-Whitney post hoc test showed statistically significant difference in S. mutans and S. sobrinus count between group I (CHX) and group III (control) and group II (miswak) and group III (control) but no statistically significant difference between group I (CHX) and group II (miswak) in S. mutans and S. sobrinus count. 1% chlorhexidine digluconate and aqueous extract of miswak are equally effective against S. mutans and S. sobrinus. Miswak can be used as an alternative herbal antimicrobial that can be incorporated in anhydrous GIC. Kalpavriksha AJ, Siddaiah SB, Bilichodmath S, et al. Comparative Evaluation of Antibacterial Effect of GIC Containing Chlorhexidine and Miswak on Streptococcus mutans and Streptococcus sobrinus in Early Childhood Caries Children: A PCR Study. Int J Clin Pediatr Dent 2021;14(2):229-234.


Association Between Use of Oral Contraceptives and Folate Status: A Systematic Review and Meta-Analysis.
To conduct a systematic review and meta-analysis of the effect of oral contraceptive use on plasma and red blood cell (RBC) folate concentrations. We searched Medline, EMBASE, Web of Science, and the Cochrane library for human studies published from inception to June 2013 evaluating oral contraceptive use and folate status. Case-control studies, cohort studies, and clinical trials were included. A random-effects model of outcomes was used for the meta-analysis. A total of 2831 women in 17 studies were included in the analysis. In those whose plasma folate concentrations were available, there was a significant folate-lowering effect of oral contraceptives observed (mean reduction 1.27 µg/L; 95% CI 1.85 to 0.69, P < 0.001). Similarly, after analyzing data from 1389 women in 12 studies whose RBC folate concentrations were available, significantly lower folate status was observed among oral contraceptive users (mean reduction 59.32 µg/L; 95% CI 58.03 to 23.04, P < 0.001). Because of the reduction in blood folate concentrations associated with the use of oral contraceptives, it is critical for women of childbearing age to continue folate supplementation during oral contraceptive use.


Association between MTHFR C677T polymorphism and venous thromboembolism risk in the Chinese population: a meta-analysis of 24 case-controlled studies.
The association between methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms and venous thromboembolism (VTE) risk in the Chinese population has been widely reported, but results were inconsistent and underpowered. To elucidate the variable results, a meta-analysis and systematic review were performed from all case-controlled studies relating MTHFR C677T polymorphism by pooling data on them. We estimated the pooled odds ratio with its 95% confidence intervals to assess this possible association. Finally, a total of 24 studies with 2339 cases and 4048 controls were included in the current meta-analysis. Significant association was found with VTE risk for all genetic models. Subgroup analyses by type of VTE further identified the above-mentioned association in deep vein thrombosis/pulmonary embolism and splanchnic vein thrombosis. The findings from our meta-analysis support the associations of MTHFR C677T polymorphism with VTE risk in the Chinese population.


Comprehensive Genomic Discovery of Non-Coding Transcriptional Enhancers in the African Malaria Vector Anopheles coluzzii.
Almost all regulation of gene expression in eukaryotic genomes is mediated by the action of distant non-coding transcriptional enhancers upon proximal gene promoters. Enhancer locations cannot be accurately predicted bioinformatically because of the absence of a defined sequence code, and thus functional assays are required for their direct detection. Here we used a massively parallel reporter assay, Self-Transcribing Active Regulatory Region sequencing (STARR-seq), to generate the first comprehensive genome-wide map of enhancers in Anopheles coluzzii, a major African malaria vector in the Gambiae species complex. The screen was carried out by transfecting reporter libraries created from the genomic DNA of 60 wild A. coluzzii from Burkina Faso into A. coluzzii 4a3A cells, in order to functionally query enhancer activity of the natural population within the homologous cellular context. We report a catalog of 3,288 active genomic enhancers that were significant across three biological replicates, 74% of them located in intergenic and intronic regions. The STARR-seq enhancer screen is chromatin-free and thus detects inherent activity of a comprehensive catalog of enhancers that may be restricted in vivo to specific cell types or developmental stages. Testing of a validation panel of enhancer candidates using manual luciferase assays confirmed enhancer function in 26 of 28 (93%) of the candidates over a wide dynamic range of activity from two to at least 16-fold activity above baseline. The enhancers occupy only 0.7% of the genome, and display distinct composition features. The enhancer compartment is significantly enriched for 15 transcription factor binding site signatures, and displays divergence for specific dinucleotide repeats, as compared to matched non-enhancer genomic controls. The genome-wide catalog of A. coluzzii enhancers is publicly available in a simple searchable graphic format. This enhancer catalogue will be valuable in linking genetic and phenotypic variation, in identifying regulatory elements that could be employed in vector manipulation, and in better targeting of chromosome editing to minimize extraneous regulation influences on the introduced sequences. Importance: Understanding the role of the non-coding regulatory genome in complex disease phenotypes is essential, but even in well-characterized model organisms, identification of regulatory regions within the vast non-coding genome remains a challenge. We used a large-scale assay to generate a genome wide map of transcriptional enhancers. Such a catalogue for the important malaria vector, Anopheles coluzzii, will be an important research tool as the role of non-coding regulatory variation in differential susceptibility to malaria infection is explored and as a public resource for research on this important insect vector of disease.


OmpR coordinates the expression of virulence factors of Enterohemorrhagic Escherichia coli in the alimentary tract of Caenorhabditis elegans.
Enterohemorrhagic Escherichia coli (EHEC), an enteropathogen that colonizes in the intestine, causes severe diarrhea and hemorrhagic colitis in humans by the expression of the type III secretion system (T3SS) and Shiga-like toxins (Stxs). However, how EHEC can sense and respond to the changes in the alimentary tract and coordinate the expression of these virulence genes remains elusive. The T3SS-related genes are known to be regulated by the locus of enterocyte effacement (LEE)-encoded regulators, such as Ler, as well as non-LEE-encoded regulators in response to different environmental cues. Herein, we report that OmpR, which participates in the adaptation of E. coli to osmolarity and pH alterations, is required for EHEC infection in Caenorhabditis elegans. OmpR protein was able to directly bind to the promoters of ler and stx1 (Shiga-like toxin 1) and regulate the expression of T3SS and Stx1, respectively, at the transcriptional level. Moreover, we demonstrated that the expression of ler in EHEC is in response to the intestinal environment and is regulated by OmpR in C. elegans. Taken together, we reveal that OmpR is an important regulator of EHEC which coordinates the expression of virulence factors during gastrointestinal infection in vivo.


Tislelizumab in advanced/metastatic esophageal squamous cell carcinoma: health-related quality of life in Asian patients.
Post-hoc analysis examined health-related quality of life and esophageal squamous cell carcinoma (ESCC) symptoms in the Asian subgroup of patients in RATIONALE-302 (NCT03430843). Patients were randomized 1:1 to either tislelizumab or investigator-chosen chemotherapy (paclitaxel, docetaxel, or irinotecan). Health-related quality of life was measured using the EORTC QLQ-C30 and the QLQ-OES18. Least-squares mean score changes from baseline to weeks 12 and 18 in health-related quality-of-life scores were assessed using a mixed model for repeated measurements. Reported nominal p-values are for descriptive purposes only. Of the 512 patients, this analysis was conducted in 392 Asian patients (tislelizumab, n = 192; investigator-chosen chemotherapy, n = 200). The tislelizumab arm had stable global health status/quality of life, but fatigue scores worsened in both arms. The change from baseline was similar for physical functioning in both arms at weeks 12 and 18. Eating and dysphagia scores remained stable in the tislelizumab arm. Reflux improved at week 12 in the tislelizumab arm and worsened in the investigator-chosen chemotherapy arm. Overall, the health-related quality of life and ESCC-related symptoms of patients receiving tislelizumab in the Asian subgroup remained stable or improved, while patients receiving investigator-chosen chemotherapy experienced worsening. These results in Asian patients corroborate the findings in the intent-to-treat population, suggesting tislelizumab is a potential new second-line treatment option for patients with advanced or metastatic ESCC. The RATIONALE-302 study is registered on clinicaltrials.gov as NCT03430843.


Impact of type, size and shape of plates on hospital patients' perceptions of the quality of meals and satisfaction with foodservices.
Hospital meals are important for supporting nutrition goals and the overall hospital experience. The aim of this study was to assess if patients' perceptions of quality attributes of hot meals were impacted by the type of plate on which the meal was served. Eligible patients selected from medical and/or surgical wards at a 929 bed acute care hospital were randomised to receive hot meals served on either ceramic or disposable plates at eight evening meals. Patients from a multi-centre trial of pre-packaged meals where re-thermalised meals were presented on a variety of colour and shaped disposable plates were included in a second phase. Patient assessment of meal quality was undertaken using the Meal Assessment Tool (MAT)(Hannan-Jones & Capra, 2017) with data collected by interview. In phase one, 137 eligible patients competed the MAT (response 46.8%), with results showing no significant difference in the rating of appearance, quality, expectations or satisfaction when meals were served on disposable or ceramic plates. In phase two data were collected from 6189 patients (response 47.4%). No practical difference in rating of meals was found in relation to colour and sectioning of plates, however a statistical but non-practical difference was found regarding plate shape, with meals served on oblong/rectangular plates rated more positively. Of 3078 comments made concerning the meals, 99 (3.2%) concerned the plates on which the meals were served. This study has shown that quality, appearance and taste of meals, rather than the serving ware are important to patients. Findings suggest that attending to other quality aspects of meals may be far more important than the plate itself, and deserve attention.


Synchronous growth of Rhodopseudomonas palustris from the swarmer phase.
Rhodopseudomonas palustris was chosen as a model organism for studying bacterial differentiation. Synchronous populations selected by sucrose gradient centrifugation yielded more than 95% swarmer cells. The appearance and disappearance of cell morphological groupings and the doubling of cell numbers in cultures of such swarmer populations were very well defined. Cells were only motile for the first half of the division cycle, but motility was regained before division. Development gave rise to a distinct and characteristic pattern of extinction increase and particle volume distribution. The development of swarmers into mother cells and the dimorphic division of R. palustris are discussed as simple examples of differentiation.


Longitudinal analysis of minority women's perceptions of cohesion: the role of cooperation, communication, and competition.
Interaction in the form of cooperation, communication, and friendly competition theoretically precede the development of group cohesion, which often precedes adherence to health promotion programs. The purpose of this manuscript was to explore longitudinal relationships among dimensions of group cohesion and group-interaction variables to inform and improve group-based strategies within programs aimed at promoting physical activity. Ethnic minority women completed a group dynamics-based physical activity promotion intervention (N = 103; 73% African American; 27% Hispanic/Latina; mage = 47.89 + 8.17 years; mBMI = 34.43+ 8.07 kg/m2) and assessments of group cohesion and group-interaction variables at baseline, 6 months (post-program), and 12 months (follow-up). All four dimensions of group cohesion had significant (ps < 0.01) relationships with the group-interaction variables. Competition was a consistently strong predictor of cohesion, while cooperation did not demonstrate consistent patterns of prediction. Facilitating a sense of friendly competition may increase engagement in physical activity programs by bolstering group cohesion.


Unmet Need for Social and Emotional Support and Lack of Recalled Screening Is Associated with Depression in the Long-Term Course After Stroke.
Details on adequate care and prevalence of depression in long-term stroke aftercare are limited. We aimed to determine long-term depression rates after stroke and to test for an association between depression and inadequate screening, socio-economic complications and lack of sub-optimal care. In this cross-sectional study, 57 patients were re-invited into the clinic 2-3 years after stroke. Patients were interviewed about recalled screening concerning depression and unmet needs. Depression, the patient's social situation, and confounders were assessed by standardized scores. In our study, 20% (n = 11) of patients were classified as depressed by the HDRS-17 score result. However, only 36% of all patients recalled to have been previously screened for depression and only 43% of those patients also recalled out-patient screening. Patients classified as depressed reported significantly lower recalled screening rates (9% vs 43%; p = 0.036) and higher rates of self-reported unmet need with emotional problems (72% vs 18%; p < 0.001). Depression in our study was further associated with a worse socio-economic situation, fewer social contacts, unmet needs with regard to emotional problems and higher rates of recommendations to apply for additional social support. Our data suggest that systematic out-patient screening for depression is lacking in stroke aftercare. Furthermore, the high rate of unmet emotional needs, the poor socio-economic situation and the higher rates of recommendations for social counselling and application for benefits suggest an undersupply of care in the out-patient setting that is more prominent in patients with depression and warrants further studies to investigate the underlying causes.


Quantitative assessment of the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis against the green peach aphid Myzus persicae.
A two-step method was developed to quantitatively assess the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis (Li) Humber, on the green peach aphid, Myzus persicae (Sulzer). Firstly, a standard time-dose-mortality relationship, established by modeling data from bioassay 1 at varying conidial dosages (0.4 - 10.4 conidia/mm2) of Z. anhuiensis F97028, was used to yield an estimate of expected mortality probability at a given dosage. Secondly, bioassay 2 was conducted by simultaneously exposing six < or = 4-day-old nymphal colonies to a shower of Z. anhuiensis conidia at each of four dosages (resulting from exposures of 0.3 - 8.0 min). Subsequently, the colonies were separately immersed in a 0.1% chlorothalonil solution for 0.5 min to disinfect all surviving conidia on the host integument from 1 - 12 h after exposure under temperature treatments of 15 and 20 degrees C, respectively. The infection rate during a specific period from the end of the exposure to the immersion was then estimated as the ratio of the observed mortality over the expected mortality probability at a particular dosage. The results showed that the infection of M. persicae from Z. anhuiensis was highly rapid with little difference between aphid colonies maintained at 15 and 20 degrees C before being immersed in the fungicidal solution after exposure. The first 6-hour period after exposure was most crucial to successful infection of the fungus with the infection rate greatly depending on conidial dosages. It took < or = 1 h to infect > 50% of the aphids at a dosage of > 1.5 conida/mm2 and > 90% at > 50 conidia/mm2.


Selective alterations in prefrontal cortical GABA neurotransmission in schizophrenia: a novel target for the treatment of working memory dysfunction.
Disturbances in critical cognitive processes, such as working memory, are now regarded as core features of schizophrenia, but available pharmacological treatments produce little or no improvement in these cognitive deficits. Although other explanations are possible, these cognitive deficits appear to reflect a disturbance in executive control, the processes that facilitate complex information processing and behavior and that include context representation and maintenance, functions dependent on the dorsolateral prefrontal cortex (DLPFC). Studies in non-human primates indicate that normal working memory function depends upon appropriate GABA neurotransmission in the DLPFC, and alterations in markers of GABA neurotransmission are well documented in the DLPFC of subjects with schizophrenia. Thus, the purpose of this paper is to review the nature of the altered GABA neurotransmission in the DLPFC in schizophrenia, and to consider how these findings might inform the search for new treatments for cognitive dysfunction in this illness. Postmortem studies suggest that markers of reduced GABA neurotransmission in schizophrenia may be selective for, or at least particularly prominent in, the subclass of GABA neurons, chandelier cells, that provide inhibitory input to the axon initial segment of populations of pyramidal neurons. Given the critical role that chandelier cells play in synchronizing the activity of pyramidal neurons, the pharmacological amelioration of this deficit may be particularly effective in normalizing the neural network activity required for working memory function. Because GABA(A) receptors containing the a(2) subunit are selectively localized to the axon initial segment of pyramidal cells, and appear to be markedly up-regulated in schizophrenia, treatment with novel benzodiazepine-like agents with selective activity at GABA(A) receptors containing the a(2) subunit may be effective adjuvant agents for improving working memory function in schizophrenia.


The influence of different drying methods on constituents and antioxidant activity of saffron from china.
More and more saffron has been cultivated in China because of the increasing saffron demand, but no paper has studied the influence of drying methods on the quality of Chinese saffron. In this paper, three different dehydration treatments applied in actual production were evaluated: dehydration with electric oven, vacuum oven, and microwave. We determined that the highest quality of saffron will be obtained when fresh saffron is treated at higher temperatures (no more than 70°C) for a long time by electric oven drying and vacuum oven drying. In microwave drying, treatments at lower microwave power and longer time benefit the quality of saffron. In addition, the influence of the drying method on antioxidants in saffron is discussed. The correlation between individual saffron profiles and the antioxidant value was estimated by spectrum-effect relationships analysis.


EEG signatures change during unilateral Yogi nasal breathing.
Airflow through the left-and-right nostrils is said to be entrained by an endogenous nasal cycle paced by both poles of the hypothalamus. Yogic practices suggest, and scientific evidence demonstrates, that right-nostril breathing is involved with relatively higher sympathetic activity (arousal states), while left-nostril breathing is associated with a relatively more parasympathetic activity (stress alleviating state). The objective of this study was to further explore this laterality by controlling nasal airflow and observing patterns of cortical activity through encephalographic (EEG) recordings. Thirty subjects participated in this crossover study. The experimental session consisted of a resting phase (baseline), then a period of unilateral nostril breathing (UNB) using the dominant nasal airway, followed by UNB using the non-dominant nasal airway. A 64-channel EEG was recorded throughout the whole session. The effects of nostril-dominance, and nostril-lateralization were assessed using the power spectral density of the neural activity. The differences in power-spectra and source localization were calculated between EEG recorded during UNB and baseline for delta, theta, alpha, beta and gamma bands. Cluster-based permutation tests showed that compared to baseline, EEG spectral power was significantly (1) decreased in all frequency bands for non-dominant nostril UNB, (2) decreased in alpha, beta and gamma bands for dominant nostril UNB, (3) decreased in all bands for left nostril UNB, and (4) decreased in all bands except delta for right nostril UNB. The beta band showed the most widely distributed changes across the scalp. our source localisation results show that breathing with the dominant nostril breathing increases EEG power in the left inferior frontal (alpha band) and left parietal lobule (beta band), whereas non-dominant nostril breathing is related to more diffuse and bilateral effects in posterior areas of the brain.These preliminary findings may stimulate further research in the area, with potential applications to tailored treatment of brain disorders associated with disruption of sympathetic and parasympathetic activity.


Evaluation of ultrasound Tissue Velocity Imaging: a phantom study of velocity estimation in skeletal muscle low-level contractions.
Tissue Velocity Imaging (TVI) is an ultrasound based technique used for quantitative analysis of the cardiac function and has earlier been evaluated according to myocardial velocities. Recent years several studies have reported applying TVI in the analysis of skeletal muscles. Skeletal tissue velocities can be very low. In particular, when performing isometric contractions or contractions of low force level the velocities may be much lower compared to the myocardial tissue velocities. In this study TVI was evaluated for estimation of tissue velocities below the typical myocardial velocities. An in-house phantom was used to see how different PRF-settings affected the accuracy of the velocity estimations. With phantom peak velocity at 0.03 cm/s the error ranged from 31% up to 313% with the different PRF-settings in this study. For the peak velocities at 0.17 cm/s and 0.26 cm/s there was no difference in error with tested PFR settings, it is kept approximately around 20%. The results from the present study showed that the PRF setting did not seem to affect the accuracy of the velocity estimation at tissue velocities above 0.17 cm/s. However at lower velocities (0.03 cm/s) the setting was crucial for the accuracy. The PRF should therefore preferable be reduced when the method is applied in low-level muscle contraction.


[Case Report of Four Patients Received Ramucirumab Monotherapy as Second-Line Chemotherapy for Advanced Gastric Cancer].
Ramucirumab monotherapy is one of the conditionally recommended regimens in second-line chemotherapy for advanced gastric cancer. However, there are few clinical data on ramucirumab monotherapy in Japanese patients. Herein, we present 4 case reports of advanced gastric cancer patients who received ramucirumab monotherapy. The 4 patients' age ranged from 65-81 years old(median: 70 years old), with a 3:1 male to female ratio. Since all cases were in poor performance status, administration of cell-killing anticancer drugs such as paclitaxel was contraindicated, and ramucirumab monotherapy was selected as an alternative. Ramucirumab was administrated 2-8 times(median: 3 times), resulting to a stable disease in 1 patient, and progression-free survival was noted to be 3-16 weeks(median: 5 weeks). Regarding complications, Grade 2 hypertension occurred in 1 patient, and no serious adverse events were observed. Ramucirumab monotherapy is a well-tolerated second-line chemotherapy for patients with advanced gastric cancer in poor performance status, and it is expected to have some disease control effect.


Population correlates of circulating mercury levels in Korean adults: the Korea National Health and Nutrition Examination Survey IV.
Prior studies focused on bioaccumulation of mercury (Hg) and on large, long-lived fish species as the major environmental source of Hg, but little is known about consumption of small-sized fish or about non-dietary determinants of circulating Hg levels. The purpose of this study was to evaluate whole blood mercury concentration (WBHg) and its major dietary and non-dietary correlates in Korean adults. We analyzed cross-sectional data from 3,972 (male = 1,994; female = 1,978) participants who completed the Korean National Health and Nutrition Examination Survey IV, 2008 to 2009. Relevant factors included diet, geographic location of residence, demographics, and lifestyle. WBHg concentration was measured using cold-vapor atomic absorption spectrometry. Multivariable linear models assessed independent correlates of dietary and non-dietary factors for WBHg levels. Median levels of WBHg were 5.1 μg/L in men and 3.7 μg/L in women. Higher levels of fish/shellfish intake were associated with higher levels of WBHg. Higher consumption of small-sized fish was linked to higher levels of WBHg. Non-dietary predictors of higher WBHg were being male, greater alcohol consumption, higher income and education, overweight/obesity, increasing age, and living in the southeast region. Both dietary and non-dietary factors were associated with WBHg levels in the Korean population. There is significant geographic variation in WBHg levels; residents living in the mid-south have higher WBHg levels. We speculate that uncontrolled geographic characteristics, such as local soil/water content and specific dietary habits are involved.


Fukuoka and AGA Criteria Have Superior Diagnostic Accuracy for Advanced Cystic Neoplasms than Sendai Criteria.
The aim of this study was to compare the American Gastroenterological Association guidelines (AGA criteria), the 2012 (Fukuoka criteria), and 2006 (Sendai criteria) International Consensus Guidelines for the diagnosis of advanced pancreatic cystic neoplasms. All patients who underwent surgical resection of a pancreatic cyst from August 2007 through January 2016 were retrospectively analyzed at a single tertiary academic center. Relevant clinical and imaging variables along with pathology results were collected to determine appropriate classification for each guideline. Advanced pancreatic cystic neoplasms were defined by the presence of either high-grade dysplasia or cystic adenocarcinoma. Diagnostic accuracy was measured by ROC analysis. A total of 209 patients were included. Both the AGA and Fukuoka criteria had a higher diagnostic accuracy for advanced neoplastic cysts than the Sendai criteria: AGA ROC 0.76 (95% CI 0.69-0.81), Fukuoka ROC 0.78 (95% CI 0.74-0.82), and Sendai ROC 0.65 (95% CI 0.61-0.69) (p < 0.0001). There was no difference between the Fukuoka and the AGA criteria. While the sensitivity was higher in the Fukuoka criteria compared to the AGA criteria (97.7 vs. 88.6%), the specificity was higher in the AGA criteria compared to the Fukuoka criteria (62.4 vs. 58.2%). In a surgical series of patients with pancreatic cysts, the AGA and Fukuoka criteria had superior diagnostic accuracy for advanced neoplastic cysts compared to the original Sendai criteria.


[Graded physical loading in assessing the efficacy of hypotensive pharmacotherapy].
Standardized graded exercise was used to examine 76 patients suffering from stage I and II essential hypertension, in whom the treatment with hypothiazid, hemiton, adelphanesidrex, propranolol and metoprolol resulted in arterial pressure normalization at rest. Hypothiazid did not protect the patients from the development of pathological (excessive) hypertensive response to graded exercise. Hemiton and adelphanesidrex allowed the attainment of normal (physiological) hypertensive response but in a small part of patients. The treatment with beta-adrenoblockers alone and combined with hypothiazid provided physiological hypertensive response to graded exercise in the absolute majority of the patients.


Clinical and genetic investigation of amantadine-associated corneal edema.
Amantadine use has been temporally associated with bilateral corneal edema in a series of cases; however, its pathophysiological mechanisms have yet to be elucidated. We sought to rule out subclinical Fuchs dystrophy as a contributor, characterize its pattern of corneal edema, and describe the long-term outcome of concurrent topical steroids while resuming amantadine. After a 44-year-old woman presented with new acute onset bilateral corneal edema, amantadine was discontinued, with clinical improvement. However, neurological decompensation required restarting amantadine, which she did concurrently with topical loteprednol. To determine whether subclinical Fuchs dystrophy might be present, triplet-primed polymerase chain reaction was conducted to measure copy number of the CTG18.1 trinucleotide repeat in TCF4. Specular microscopy and Scheimpflug imaging were conducted and followed for 32 months to assess for resolution and stability. Literature review was conducted to assess for consistency of the clinical phenotype. Corneal edema resolved clinically 4 weeks after discontinuation of amantadine. Serial Scheimpflug imaging demonstrated resolution of posterior and central corneal edema and specular microscopy revealed intracellular opacities with loss of endothelial cell density. Despite resuming amantadine, Scheimpflug imaging and specular microscopy measurements remained stable at 32 months. Triplet-primed PCR of CTG18.1 in TCF4 revealed no trinucleotide repeat expansion. Amantadine-associated corneal edema is characteristically posterior and central and appears unlikely to represent early or subclinical decompensation of Fuchs dystrophy. We describe the unique outcome of continued corneal clearance after restarting amantadine concurrently with steroids, a pattern that has persisted over 32 months to date.


Effects of bilateral ibotenate-induced lesions of the nucleus basalis magnocellularis upon selective cholinergic biochemical markers in the rat anterior cerebral cortex.
The relationship of choline acetyltransferase (ChAT) activity and high affinity binding of the potent and selective sodium-dependent choline uptake inhibitor [3H]hemicholinium-3 ([3H]HC-3) to high-affinity binding of the muscarinic agonist [3H](+)-cis-methyldioxolane ([3H](+)CD), the putative M1 selective antagonist [3H]pirenzepine ([3H]PZ) and the classical antagonist [3H](-)-quinuclidinyl benzilate ([3H](-)QNB) in homogenates of the rat neocortex was studied. ChAT activity was 42% lower in rats with ibotenate-induced lesions of the nucleus basalis magnocellularis (nbm) when compared to controls, and [3H]HC-3 binding was similarly reduced by 44%. However, equilibrium dissociation constants (Kd values) for [3H]HC-3 (0.8-1.0 nM), [3H](-)QNB (11-24 pM), [3H]PZ (4.0-4.3 nM) and [3H](+)CD (2.1-2.9 nM) were each unchanged. Mean Bmax values (total binding site densities) for [3H](+)CD were significantly altered in both hemispheres of the anterior cerebral cortex, showing a 25% reduction in the number of sites which display the highest affinity conformation for this potent muscarinic agonist. The decreased ChAT activity and [3H]HC-3 binding after nbm lesions were associated with only slight reductions in putative M1 muscarinic site density (14%) and [3H](-)QNB binding site density (13%). Thus, it appears that while [3H]PZ and [3H](-)QNB label predominantly postsynaptic muscarinic binding sites, a significant number of sites labeled by [3H](+)CD may be associated with presynaptic cholinergic nerve terminals. These data suggest that cholinergic input differentially regulates the drug binding sites of anterior cerebral cortical muscarinic receptors, exerting a substantial effect upon the highest affinity conformational state for agonists.


Computational imaging of sperm locomotion.
Not only essential for scientific research, but also in the analysis of male fertility and for animal husbandry, sperm tracking and characterization techniques have been greatly benefiting from computational imaging. Digital image sensors, in combination with optical microscopy tools and powerful computers, have enabled the use of advanced detection and tracking algorithms that automatically map sperm trajectories and calculate various motility parameters across large data sets. Computational techniques are driving the field even further, facilitating the development of unconventional sperm imaging and tracking methods that do not rely on standard optical microscopes and objective lenses, which limit the field of view and volume of the semen sample that can be imaged. As an example, a holographic on-chip sperm imaging platform, only composed of a light-emitting diode and an opto-electronic image sensor, has emerged as a high-throughput, low-cost and portable alternative to lens-based traditional sperm imaging and tracking methods. In this approach, the sample is placed very close to the image sensor chip, which captures lensfree holograms generated by the interference of the background illumination with the light scattered from sperm cells. These holographic patterns are then digitally processed to extract both the amplitude and phase information of the spermatozoa, effectively replacing the microscope objective lens with computation. This platform has further enabled high-throughput 3D imaging of spermatozoa with submicron 3D positioning accuracy in large sample volumes, revealing various rare locomotion patterns. We believe that computational chip-scale sperm imaging and 3D tracking techniques will find numerous opportunities in both sperm related research and commercial applications.


Knowledge and attitude towards zoonoses among animal health workers and livestock keepers in Arusha and Tanga, Tanzania.
Zoonoses are infections naturally transmitted between vertebrate animals and humans. An exploratory questionnaire-based survey of animal health workers(n=36) and livestock keepers(n=43) was carried out from April 2001 to March 2002 in Tanga and Arusha regions, northern Tanzania, to assess local knowledge, attitudes and public awareness for animal zoonoses. A combination of closed and open-ended questions, focus group discussions and ranking techniques were employed to gather information on perceptions concerning the type of zoonotic diseases prevalent in the study area, level of risk, mode of transmission and methods of preventing disease transmission from animals to humans. The results demonstrated that rabies, tuberculosis and anthrax were considered the three most common zoonotic diseases. Sharing living accommodation with animals, consumption of un-treated livestock products (i.e. milk, meat or eggs) and attending to parturition were perceived as routes of transmission. Knowledge about zoonosis was higher in smallholder dairy (92%; 33/36) than traditional livestock keepers (P<0.05). On the contrary, the perceived risk of contracting a zoonosis was significantly higher in traditional livestock (86%; 6/7) than smallholder dairy keepers (P<0.05). Stratification of the risk of zoonosis by farm location revealed that rural farms (85%; 7/8) were considered significantly at a higher risk when compared to peri or urban located farms (P<0.05). Most of the respondents stated cooking of meat or boiling of milk as a way to prevent transmission. However, there was a significant difference in the perception of the risk posed by contact with potentially infected animals /or animal products with animal health workers having a much higher level of perception compared to livestock keepers. These results suggest that in the Tanga and Arusha, Tanzania, patchy awareness and knowledge of zoonoses, combined with food consumption habits and poor animal husbandry are likely to expose respondents to an increased risk of contracting zoonoses. Public health promotion on education and inter-disciplinary one-health collaboration between vets, public health practitioners and policy makers should result in a more efficient and effective joint approach to the diagnosis and control of zoonoses in Tanzania.


Periphytic biofilm: An innovative approach for biodegradation of microplastics.
Microplastics (MPs) have been gaining the attention of environmental researchers since the 1960s anecdotal reports of plastic entanglement and ingestion by marine creatures. Due to their increasing accretion in aquatic environments, as well as resistance towards degradation, marine litter research has focused on microplastics more recently. In the present study, a relatively new method of biodegradation was implemented for the biodegradation of three structurally different MPs i.e. polypropylene (PP), polyethylene (PE) and polyethylene terephthalate (PET). Periphytic biofilm was used for this purpose in various backgrounds of carbon sources (glucose, peptone, and glucose and peptone). Biodegradation of MPs was estimated in terms of weight loss. It was observed that the addition of glucose enhanced the biodegradation of MPs by periphyton biofilm for all MPs (from 9.52%-18.02%, 5.95%-14.02% and 13.24-19.72% for PP, PE and PET respectively) after 60 days compared to natural biofilm alone. To the contrary, peptone, and glucose and peptone together, were inhibitory. Biodegradation was further confirmed by morphological changes observed using SEM, FTIR spectra and GPC lent further support to the results whereby new peaks appeared along with reduction in old peaks and decrease in peak intensities. MiSeq sequencing shows that Deinococcus-thermus > Proteobacteria > Cyanobacteria are the dominant phyla in natural biofilms, and their relative abundances increase after the addition of glucose. However, the abundances shifted to Deinococcus-thermus > Cyanobacteria > Firmicutes > Bacteroidetes, when the biofilms were treated with either peptone alone, or with glucose and peptone together. Therefore, the change in biodegradation capability might also be due to the change in the microbial community structures after addition of the C-sources. These experiments provide an innovative approach towards effective biodegradation of MPs using a relatively new environment-friendly method.


Development and initial validation of a theory of planned behaviour questionnaire to assess the nursing students' Intention to be self-employed.
Self-employment represents a new work opportunity for new nursing graduates; however, little is known about nursing students' Intention to meet this new demand from the labour market. To develop a theory-driven questionnaire to assess students' Intentions for self-employment and psychometrically test this scale. A cross-sectional study was conducted investigating 344 nursing students in their final year in an Italian university. The content, face, construct, convergent and discriminant validity, and reliability were evaluated. We also tested the theory of planned behaviour. The Planned Self-Employment Scale demonstrated satisfactory validity and good reliability. A moderate Intention to be self-employed emerged among nurses (3.6 ± 1.5 out of 7). This level of Intention was predicted by the Attitudes and Perceived Behaviour Control dimensions. The Planned Self-Employment Scale exhibited good validity and reliability and can thus be used with nursing students. Educators need to design a curriculum to improve students' decision-making and autonomy. At the central level, there is a need to produce guidelines that offer guidance to both nurses and patients, as well as the system as a whole with regard to this new form of nursing service.


Substance use disorder and the baby boom generation: Does Berlin outpatient addiction care face a sustained change?
The ageing of baby boomers is expected to confront addiction care with new challenges. This cohort had greater exposure to psychoactive substances in youth than earlier cohorts. In this study, we aimed to investigate whether Berlin addiction care is confronted with a sustained change in its clientele initiated by the baby boomers. Using data from Berlin outpatient addiction care facilities, we contrasted type of primary substance use disorder and number of comorbid substance use disorders in baby boomers with an earlier and a later cohort. To isolate cohort effects, two-level random intercept regression models were applied in the overlapping age groups of the baby boomer cohort with each of the other cohorts. Compared with the earlier cohort, alcohol use disorder lost importance whereas illicit substance use disorder gained importance in the baby boomers. Baby boomers presented a higher number of comorbid substance use disorders than the earlier cohort. Comparing baby boomers with the later cohort, these relationships pointed in the opposite direction. Outpatient addiction care faces a sustained change to more illicit and comorbid substance use disorders. With increasing life expectancy and the ageing of baby boomers marked by higher substance use than previous cohorts, older clients, who had been under-represented in outpatient addiction care, will gain relevance. Hence, addiction care has to adapt its offers to appropriately meet the changing needs of its clientele.


Antibody response to Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan in patients after solid-organ transplantation.
Cryptococcosis is an important complication of solid-organ transplantation, but the risk factors for disease are poorly understood. The goal of this study was to investigate whether specific or nonspecific serum immunoglobulin levels determined in samples obtained before and after solid-organ transplantation differed in patients who did or did not develop cryptococcosis after transplantation. We analyzed pretransplantation sera from 25 subjects, 15 who subsequently developed cryptococcosis and 10 who did not, and posttransplantation sera from 24 subjects, 13 who developed cryptococcosis and 11 who did not. All subjects received a tacrolimus-based immunosuppressive regimen. Total immunoglobulin levels were measured by immunodiffusion, and Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan (GXM)-specific serum antibody levels were determined by enzyme-linked immunosorbent assays. The results showed that solid-organ transplantation had a significant effect on total immunoglobulin and GXM-reactive antibody levels. GXM-reactive antibody levels differed in subjects who did and did not develop cryptococcosis. In pretransplant serum samples, the levels of GXM-reactive immunoglobulin M (IgM) were significantly lower in subjects who developed cryptococcosis after transplantation than in those who did not. For posttransplant serum samples, the levels of GXM-reactive IgM and IgG were significantly higher among the subjects who developed cryptococcosis than among those who did not. These findings suggest that perturbations in the preexisting antibody or B-cell repertoire and/or related to treatment of rejection, transplantation, or immunosuppressive therapy could translate into an increased risk for transplant-associated cryptococcosis.


Nitrosyl hydride (HNO) replaces dioxygen in nitroxygenase activity of manganese quercetin dioxygenase.
Quercetin dioxygenase (QDO) catalyzes the oxidation of the flavonol quercetin with dioxygen, cleaving the central heterocyclic ring and releasing CO. The QDO from Bacillus subtilis is unusual in that it has been shown to be active with several divalent metal cofactors such as Fe, Mn, and Co. Previous comparison of the catalytic activities suggest that Mn(II) is the preferred cofactor for this enzyme. We herein report the unprecedented substitution of nitrosyl hydride (HNO) for dioxygen in the activity of Mn-QDO, resulting in the incorporation of both N and O atoms into the product. Turnover is demonstrated by consumption of quercetin and other related substrates under anaerobic conditions in the presence of HNO-releasing compounds and the enzyme. As with dioxygenase activity, a nonenzymatic base-catalyzed reaction of quercetin with HNO is observed above pH 7, but no enhancement of this basal reactivity is found upon addition of divalent metal salts. Unique and regioselective N-containing products ((14)N/(15)N) have been characterized by MS analysis for both the enzymatic and nonenzymatic reactions. Of the several metallo-QDO enzymes examined for nitroxygenase activity under anaerobic condition, only the Mn(II) is active; the Fe(II) and Co(II) substituted enzymes show little or no activity. This result represents an enzymatic catalysis which we denote nitroxygenase activity; the unique reactivity of the Mn-QDO suggests a metal-mediated electron transfer mechanism rather than metal activation of the substrate's inherent base-catalyzed reactivity.


Effectiveness of Vestibular Rehabilitation Therapy for Treatment of Concussed Adolescents With Persistent Symptoms of Dizziness and Imbalance.
Adolescents who suffer sport concussion typically respond to a prescription of cognitive and physical rest in the acute phases of healing; however, some adolescents do not respond to rest alone. Dizziness, unsteadiness, and imbalance are impairments, which may linger longer than 30 days, leading to a diagnosis of postconcussion syndrome (PCS). Vestibular assessment and therapy may benefit adolescents suffering from these persistent symptoms. Does vestibular rehabilitation therapy (VRT) rather than continued prescription of rest (cognitive and physical) reduce recovery time and persistent symptoms of dizziness, unsteadiness, and imbalance in adolescents (12-18 y) who suffer PCS following a sports-related concussion? Summary of Key Findings: All 4 studies selected included adolescents suffering from PCS, specifically continued dizziness, unsteadiness, and imbalance. VRT was an effective intervention for this population. Adolescents presenting with this cluster of symptoms may also demonstrate verbal and visual memory loss linked to changes in the vestibular system postconcussion. Improved screening tools can help better understand vestibular system changes, identify adolescents who may benefit from VRT sooner, and decrease long-term impairments. Clinical Bottom Line: Moderate evidence supports that adolescents who suffer from persistent symptoms of dizziness, unsteadiness, and imbalance following sport concussion should be evaluated more specifically and earlier for vestibular dysfunction and can benefit from participation in individualized VRT. Early evaluation and treatment may result in a reduction of time lost from sport as well as a return to their premorbid condition. For these adolescents, VRT may be more beneficial than continued physical and cognitive rest when an adolescent's symptoms last longer than 30 days. Strength of Recommendation: Grade B evidence exists to support that VRT is more effective than continued cognitive and physical rest in reducing persistent symptoms of dizziness, unsteadiness, and imbalance in adolescents who suffer PCS.


Metabolomic biosignature differentiates melancholic depressive patients from healthy controls.
Major depressive disorder (MDD) is a heterogeneous disease at the level of clinical symptoms, and this heterogeneity is likely reflected at the level of biology. Two clinical subtypes within MDD that have garnered interest are "melancholic depression" and "anxious depression". Metabolomics enables us to characterize hundreds of small molecules that comprise the metabolome, and recent work suggests the blood metabolome may be able to inform treatment decisions for MDD, however work is at an early stage. Here we examine a metabolomics data set to (1) test whether clinically homogenous MDD subtypes are also more biologically homogeneous, and hence more predictiable, (2) devise a robust machine learning framework that preserves biological meaning, and (3) describe the metabolomic biosignature for melancholic depression. With the proposed computational system we achieves around 80 % classification accuracy, sensitivity and specificity for melancholic depression, but only ~72 % for anxious depression or MDD, suggesting the blood metabolome contains more information about melancholic depression.. We develop an ensemble feature selection framework (EFSF) in which features are first clustered, and learning then takes place on the cluster centroids, retaining information about correlated features during the feature selection process rather than discarding them as most machine learning methods will do. Analysis of the most discriminative feature clusters revealed differences in metabolic classes such as amino acids and lipids as well as pathways studied extensively in MDD such as the activation of cortisol in chronic stress. We find the greater clinical homogeneity does indeed lead to better prediction based on biological measurements in the case of melancholic depression. Melancholic depression is shown to be associated with changes in amino acids, catecholamines, lipids, stress hormones, and immune-related metabolites. The proposed computational framework can be adapted to analyze data from many other biomedical applications where the data has similar characteristics.


Normal characteristics of the ocular vestibular evoked myogenic potential.
Stimulus-evoked electromyographic changes can be recorded from the extraocular muscles. These short-latency negative-polarity evoked myogenic potentials are called ocular vestibular evoked myogenic potentials (oVEMPs). To date there has not yet been a large-scale study examining the effects of age on the amplitude, latency, threshold, and interaural differences of the oVEMP to air-conducted stimuli. Further, before the oVEMP can become a useful clinical tool, the test-retest reliability of the response must be established. The oVEMP response, once more completely understood, may provide diagnostic information that is complementary to the cervical vestibular evoked myogenic potential (cVEMP; i.e., sternocleidomastoid muscle). To describe the normal characteristics of oVEMP in a cohort of age-stratified subjects, to assess the test-retest reliability of the oVEMP, and to determine if reference contamination occurs using a common recommended infraorbital reference electrode derivation. A prospective, descriptive study design was used for an investigation with a threefold purpose in which oVEMP recordings were made from the extraocular muscles (e.g., inferior oblique muscle). Fifty otologically and neurologically normal adults and children served as subjects. Subjects ranged in age from 8 to 88 yr. In Investigation 1, oVEMPs were recorded from the ipsilateral and contralateral inferior oblique muscles for all subjects. The stimulus was a 95 dB nHL 500 Hz tone burst. Next, oVEMP thresholds were obtained. Amplitude, latency, and thresholds were tabulated, and descriptive statistics were used to calculate normative values. Age-related differences in oVEMP component latencies, amplitudes, interaural amplitude asymmetries (IAAs), and thresholds were determined using an analysis of variance. In Investigation 2, oVEMPs were recorded twice in 10 subjects, once (test) and once approximately 10 weeks later (retest). Test-retest reliability for the oVEMP peak-to-peak amplitude, n1 latency, p1 latency, n1 threshold, and IAA were assessed with intraclass correlation coefficients (ICCs) calculated using a two-way random-effects, absolute-agreement model. In Investigation 3, a four-channel oVEMP recording was conducted in 10 subjects. Both observational methods and paired-sample t-tests were used to evaluate the effect that reference electrode location had on the oVEMP. oVEMP responses were present bilaterally in 90% of our subjects. The upper limit of oVEMP amplitude asymmetry, defined as the mean plus two standard deviations, was 34% (mean = 14%, SD 10), and the mean n1 latency was 12.5 (SD 1.0) msec. The amplitude of the response significantly decreased and the threshold significantly increased with increasing age, with the greatest age effects occurring in subjects 50 yr and older. Test-retest reliability was acceptable (ICCs for the measurement variables ranged from .53 to .87). Using conventional recommended recording techniques, evidence of reference contamination occurred for all subjects, resulting in a mean amplitude reduction of 30% (range = 18%-43%). Age results in systematic changes in oVEMP measurement parameters. The test-retest reliability is acceptable, and reference contamination averaging 30% is guaranteed using a second infraorbital electrode as the inverting input (i.e., reference electrode) for bipolar recordings. The oVEMP can be used as a complementary diagnostic tool to the cVEMP in evaluating subjects with suspected peripheral vestibular disorders.


Neuromodulatory and anti-inflammatory ingredient for sensitive skin: in vitro assessment.
The manifestation of sensitive skin occurs as a consequence of increased permeability of the Stratum corneum, besides the involvement of neuro-immune-endocrine system. In this study, we evaluated the effects of an active ingredient SensC on the production of neuropeptides substance P (SP), enkephalin and β-endorphin; eicosanoids prostaglandin E2 (PGE2) and leukotriene B4 (LTB4); histamine, transient receptor potential vanilloid subfamily member 1 (TRPV1), and envelope proteins filaggrin and involucrin, using an in vitro model of human cell culture. Our results demonstrated that treatment of keratinocyte cultures with SensC prevented the increase of all evaluated inflammatory mediators induced by interleukin-1 alpha (IL-1α). As the same way, SensC provides decrease in the synthesis of TRPV1. Regarding the synthesis of envelope proteins, SensC promoted increases for filaggrin and involucrin levels, when compared to control group. Considering the absence of appropriate treatment, the availability of ingredients, such as SensC, with antiinflammatory and protective barrier properties can be a significant tool for preventing neurosensorial symptoms associated with sensitive skin.


Determination of cannabinoids from a surrogate hops matrix using multiple reaction monitoring gas chromatography with triple quadrupole mass spectrometry.
Cannabinoids are the primary bioactive constituents of Cannabis sativa and Cannabis indica plants. In this work, gas chromatography in conjunction with triple quadrupole mass spectrometry in multiple reaction monitoring mode was explored for determination of cannabinoids from a surrogate hops matrix. Gas chromatography with mass spectrometry is a reasonable choice for the analysis of these compounds; however, such methods are susceptible to false positives for Δ9-tetrahydrocannabinol, due to decarboxylation of Δ9-tetrahydrocannabinolic acid, its acid precursor, in the hot injection port. To avoid this transformation, the carboxyl group of Δ9-tetrahydrocannabinolic acid was protected through a silylation reaction. Multiple reaction monitoring transitions for both unmodified and silylated cannabinoids were developed and the fragmentation pathways for the different species were assigned. Precision and accuracy were evaluated for cannabinoids spiked into hops at different levels. The developed methods provided good linearity (R2 > 0.99) for all the cannabinoids with a linear range from 0.15 to 20 mg/L, and with limits of detection in the orders of low- to mid-picogram on column. The recoveries for the cannabinoids were generally between 75 and 120%. Precisions (<6% coefficient of variation) were within acceptable ranges.


Child outcomes after placement of a cervical pessary in women with a multiple pregnancy: A 4-year follow-up of the ProTWIN trial.
The ProTWIN trial previously showed no beneficial effect of treatment with a cervical pessary vs usual care to prevent preterm birth in women with a multiple pregnancy. However, in women with a midtrimester short cervix (<38 mm), pessary did reduce the composite outcome of neonatal morbidity and mortality. This follow-up study evaluates the long-term outcomes of all children born to mothers who participated in the ProTWIN trial at 4 years of age. Parents received the Ages and Stages Questionnaire, Strength and Difficulties Questionnaire and a health questionnaire. All questionnaires were reported separately and as a combined outcome (abnormal child outcome). A linear mixed effects model was used to adjust for correlated data in twins and correction for confounders was performed. In exploratory analysis, a composite outcome of death or survival with abnormal child outcome was used by combining extrapolated data on child outcome with survival data. All data were analyzed for the total group and the subgroup of women with midtrimester short cervix. Of the original 813 women of the ProTWIN trial, we approached 579, of whom 258 participated (45%) in follow-up. We received questionnaires of 514 children (281 pessary vs 233 control), with 119 children in the subgroup of women with midtrimester short cervix. An abnormal child outcome was found in 23% in the pessary group vs 16% in the control group (odds ratio 1.58; 95% confidence interval 0.94-2.65). In exploratory analysis with extrapolated data on child outcome (n = 815), no difference in abnormal child outcome was seen between the pessary and control group. In the subgroup of women with a short cervix (n = 268), this composite outcome indicated a favorable outcome for children born to mothers with pessary. In women with a multiple pregnancy, the use of a cervical pessary did not improve development, behavior or physical outcomes of the surviving children at age 4.


Colutellin A, an immunosuppressive peptide from Colletotrichum dematium.
Colletotrichum dematium is an endophytic fungus recovered from a Pteromischum sp. growing in a tropical forest in Costa Rica. This fungus makes a novel peptide antimycotic, colutellin A, with a MIC of 3.6 microg ml(-1) (48 h) against Botrytis cinerea and Sclerotinia sclerotiorum. Collutellin A has a mass of 1127.7 Da and contains residues of Ile, Val, Ser, N-methyl-Val and beta-aminoisobutryic acid in nominal molar ratios of 3 : 2 : 1 : 1 : 1, respectively. Independent lines of evidence suggest that the peptide is cyclic and sequences of Val-Ile-Ser-Ile and Ile-Pro-Val have been deduced by MS/MS as well as Edman degradation methods. Colutellin A inhibited CD4(+) T-cell activation of interleukin 2 (IL-2) production with an IC(50) of 167.3+/-0.38 nM, whereas cyclosporin A in the same test yielded a value of 61.8 nM. Inhibition of IL-2 production by collutellin A at such a low concentration indicates the potential immunosuppressive activity of this compound. In repeated experiments, cyclosporin A at or above 8 microg ml(-1) exhibited high levels of cytotoxicity on human peripheral blood mononuclear cells, whereas collutellin A or DMSO (carrier) alone, after 24 and 48 h of culture, exhibited no toxicity. Because of these properties collutellin A has potential as a novel immunosuppressive drug.


Biodiversity mediates the effects of stressors but not nutrients on litter decomposition.
Understanding the consequences of ongoing biodiversity changes for ecosystems is a pressing challenge. Controlled biodiversity-ecosystem function experiments with random biodiversity loss scenarios have demonstrated that more diverse communities usually provide higher levels of ecosystem functioning. However, it is not clear if these results predict the ecosystem consequences of environmental changes that cause non-random alterations in biodiversity and community composition. We synthesized 69 independent studies reporting 660 observations of the impacts of two pervasive drivers of global change (chemical stressors and nutrient enrichment) on animal and microbial decomposer diversity and litter decomposition. Using meta-analysis and structural equation modeling, we show that declines in decomposer diversity and abundance explain reduced litter decomposition in response to stressors but not to nutrients. While chemical stressors generally reduced biodiversity and ecosystem functioning, detrimental effects of nutrients occurred only at high levels of nutrient inputs. Thus, more intense environmental change does not always result in stronger responses, illustrating the complexity of ecosystem consequences of biodiversity change. Overall, these findings provide strong evidence that the consequences of observed biodiversity change for ecosystems depend on the kind of environmental change, and are especially significant when human activities decrease biodiversity.


Postural stability of elderly submmitted to multisensorial physical therapy intervention.
The present study investigated the influence of a regular Physical Therapy program with emphasis on proprioceptive stimulation, on the center pressure trajectory and muscle adjustments necessary to maintain balance on orthostatic position. After medical assessment, 29 subjects (63.06 +/- 2.84 years) were submitted to electromyographic and baropodometric evaluation and, subsequently, to a program of 12 weeks of physical therapy intervention focusing on proprioceptive stimulation. The oscillation of the pressure center (Pressure platform-MatScanTekscan) was evaluated concomitantly to electromyographic signal envelopment and median frequency assessment of anterior tibial and gastrocnemius muscles (signal conditioner module--MCS 1000-V2-LYNX ). The Wilcoxon test was used for comparison of means, with the significance level of 5%. The pressure center trajectory unipodal position with open eyes (UO) decreased significantly after physical therapy intervention. In this position, there was a reduction of the electromyographic signal envelopment and an increase on median frequency of the gastrocnemius muscle, while the anterior tibial muscle showed increased median frequency after intervention. For the unipodal position with closed eyes (UC), the anterior tibial muscle showed significant electromyographic signal envelopment decrease and increase of median frequency, which also increased for the gastrocnemius muscle. It is concluded that the physical therapy intervention promoted decrease of body oscillation, recruitment improvement and adjustment of the gastrocnemius and anterior tibial muscles, contributing to the postural balance improvement in orthostatic position on senior population.


Test Bench for Evaluation of a Soft Robotic Link.
In this paper we describe the control approaches tested in the improved version of an existing soft robotic neck with two Degrees Of Freedom (DOF), able to achieve flexion, extension, and lateral bending movements similar to those of a human neck. The design is based on a cable-driven mechanism consisting of a spring acting as a cervical spine and three servomotor actuated tendons that let the neck to reach all desired postures. The prototype was manufactured using a 3D printer. Two control approaches are proposed and tested experimentally: a motor position approach using encoder feedback and a tip position approach using Inertial Measurement Unit (IMU) feedback, both applying fractional-order controllers. The platform operation is tested for different load configurations so that the robustness of the system can be checked.


Hydroxyzine Use and Mortality in Patients Hospitalized for COVID-19: A Multicenter Observational Study.
(1) Background: Based on its antiviral activity, anti-inflammatory properties, and functional inhibition effects on the acid sphingomyelinase/ceramide system (FIASMA), we sought to examine the potential usefulness of the H1 antihistamine hydroxyzine in patients hospitalized for COVID-19. (2) Methods: In a multicenter observational study, we included 15,103 adults hospitalized for COVID-19, of which 164 (1.1%) received hydroxyzine within the first 48 h of hospitalization, administered orally at a median daily dose of 25.0 mg (SD = 29.5). We compared mortality rates between patients who received hydroxyzine at hospital admission and those who did not, using a multivariable logistic regression model adjusting for patients' characteristics, medical conditions, and use of other medications. (3) Results: This analysis showed a significant association between hydroxyzine use and reduced mortality (AOR, 0.51; 95%CI, 0.29-0.88, p = 0.016). This association was similar in multiple sensitivity analyses. (4) Conclusions: In this retrospective observational multicenter study, the use of the FIASMA hydroxyzine was associated with reduced mortality in patients hospitalized for COVID-19. Double-blind placebo-controlled randomized clinical trials of hydroxyzine for COVID-19 are needed to confirm these results, as are studies to examine the potential usefulness of this medication for outpatients and as post-exposure prophylaxis for individuals at high risk for severe COVID-19.


Activating mineralocorticoid receptor mutation in hypertension exacerbated by pregnancy.
Hypertension and pregnancy-related hypertension are major public health problems of largely unknown causes. We describe a mutation in the mineralocorticoid receptor (MR), S810L, that causes early-onset hypertension that is markedly exacerbated in pregnancy. This mutation results in constitutive MR activity and alters receptor specificity, with progesterone and other steroids lacking 21-hydroxyl groups, normally MR antagonists, becoming potent agonists. Structural and biochemical studies indicate that the mutation results in the gain of a van der Waals interaction between helix 5 and helix 3 that substitutes for interaction of the steroid 21-hydroxyl group with helix 3 in the wild-type receptor. This helix 5-helix 3 interaction is highly conserved among diverse nuclear hormone receptors, suggesting its general role in receptor activation.


Functional consequences of mutations in the smooth muscle myosin heavy chain at sites implicated in familial hypertrophic cardiomyopathy.
Familial hypertrophic cardiomyopathy (FHC) is frequently associated with mutations in the beta-cardiac myosin heavy chain. Many of the implicated residues are located in highly conserved regions of the myosin II class, suggesting that these mutations may impair the basic functions of the molecular motor. To test this hypothesis, we have prepared recombinant smooth muscle heavy meromyosin with mutations at sites homologous to those associated with FHC by using a baculovirus/insect cell expression system. Several of the heavy meromyosin mutants, in particular R403Q, showed an increase in actin filament velocity in a motility assay and an enhanced actin-activated ATPase activity. Single molecule mechanics, using a laser trap, gave unitary displacements and forces for the mutants that were similar to wild type, but the attachment times to actin following a unitary displacement were markedly reduced. These results suggest that the increases in activity are due to a change in kinetics and not due to a change in the intrinsic mechanical properties of the motor. In contrast to earlier reports, we find that mutations in residues implicated in FHC affect motor function by enhancing myosin activity rather than by a loss of function.


GBM-associated mutations and altered protein expression are more common in young patients.
Geriatric glioblastoma (GBM) patients have a poorer prognosis than younger patients, but IDH1/2 mutations (more common in younger patients) confer a favorable prognosis. We compared key GBM molecular alterations between an elderly (age ≥ 70) and younger (18 < = age < = 45) cohort to explore potential therapeutic opportunities. Alterations more prevalent in the young GBM cohort compared to the older cohort (P < 0.05) were: overexpression of ALK, RRM1, TUBB3 and mutation of ATRX, BRAF, IDH1, and TP53. However, PTEN mutation was significantly more frequent in older patients. Among patients with wild-type IDH1/2 status, TOPO1 expression was higher in younger patients, whereas MGMT methylation was more frequent in older patients. Within the molecularly-defined IDH wild-type GBM cohort, younger patients had significantly more mutations in PDGFRA, PTPN11, SMARCA4, BRAF and TP53. GBMs from 178 elderly patients and 197 young patients were analyzed using DNA sequencing, immunohistochemistry, in situ hybridization, and MGMT-methylation assay to ascertain mutational and amplification/expressional status. Significant molecular differences occurred in GBMs from elderly and young patients. Except for the older cohort's more frequent PTEN mutation and MGMT methylation, younger patients had a higher frequency of potential therapeutic targets.


Adrenal and thyroid axis function in preterm ventilated baboons.
Cortisol and thyroid hormones are critical to normal fetal development and neonatal transition, and baseline values and stimulation tests are abnormal after preterm birth. To evaluate cortisol and thyroxine (T4) responses that are not influenced by uncontrolled antenatal events associated with human preterm labor, we measured cortisol and T4 after standard-dose adrenocorticotropin (ACTH) and corticotropin-releasing hormone (CRH) stimulation tests, as well as high-dose CRH and thyrotropin-releasing hormone stimulation tests in baboons that were delivered for 3 separate protocols at 125 days of gestation (term is 186 days). The animals were surfactant treated and ventilated for up to 14 days. Some fetuses were exposed to fetal or maternal betamethasone, and some newborns were treated with 10 microg/kg T4 for 9 days after birth. Baseline cortisol levels were in a stress range of 30-60 microg/dl by day 5. Cortisol did not increase consistently until day 11 in response to a high CRH dose or ACTH. T4 treatment for 9 days after birth suppressed the cortisol responses and subsequent baseline T4 levels. The hypothalamic-pituitary-adrenal (HPA) axis was unresponsive to standard dose stimulation tests until 11 days of age in preterm baboons, indicating HPA immaturity.