Chaperone-mediated autophagy.
Chaperone-mediated autophagy (CMA) is a lysosomal pathway of proteolysis that is responsible for the degradation of 30% of cytosolic proteins under conditions of prolonged nutrient deprivation. Molecular chaperones in the cytosol and in the lysosomal lumen stimulate this proteolytic pathway. The molecular chaperones in the cytosol unfold substrate proteins prior to their translocation across the lysosomal membrane, while the chaperone in the lysosomal lumen is probably required to pull the substrate protein across the lysosomal membrane. A critical component for CMA is a receptor in the lysosomal membrane, the lysosome-associated membrane protein (LAMP) type 2A. LAMP-2A levels in the lysosomal membrane can be increased by reduced degradation and/or redistribution from the lysosomal lumen to the lysosomal membrane. Recent results show that CMA is also activated by oxidative stress, and in this case LAMP-2A is increased due to transcriptional regulation. CMA can be reduced by inhibitors of glucose-6-phosphate dehydrogenase and of the heat shock protein of 90 kDa. Reduction of levels of LAMP-2A using RNAi strategies reduces CMA activity, but macroautophagy is activated as a result. The decrease in CMA causes cells to be more susceptibile to oxidative and other stresses. LAMP-2A in the lysosomal membrane can be sequestered into cholesterol-rich microdomains where it is inactive. When CMA is activated, LAMP-2A moves out of these domains. The reduced CMA in aging is due to reduced LAMP-2A in the lysosomal membrane. This reduction is caused by an age-related increased degradation of LAMP-2A and an age-related reduced ability of LAMP-2A to reinsert into the lysosomal membrane. These findings reveal a rich complexity of mechanisms to control CMA activity.


Mechanical Cardiopulmonary Resuscitation In and On the Way to the Cardiac Catheterization Laboratory.
Cardiac arrest, though not common during coronary angiography, is increasingly occurring in the catheterization laboratory because of the expanding complexity of percutaneous interventions (PCI) and the patient population being treated. Manual chest compression in the cath lab is not easily performed, often interrupted, and can result in the provider experiencing excessive radiation exposure. Mechanical cardiopulmonary resuscitation (CPR) provides unique advantages over manual performance of chest compression for treating cardiac arrest in the cardiac cath lab. Such advantages include the potential for uninterrupted chest compressions, less radiation exposure, better quality chest compressions, and less crowded conditions around the catheterization table, allowing more attention to ongoing PCI efforts during CPR. Out-of-hospital cardiac arrest patients not responding to standard ACLS therapy can be transported to the hospital while mechanical CPR is being performed to provide safe and continuous chest compressions en route. Once at the hospital, advanced circulatory support can be instituted during ongoing mechanical CPR. This article summarizes the epidemiology, pathophysiology and nature of cardiac arrest in the cardiac cath lab and discusses the mechanics of CPR and defibrillation in that setting. It also reviews the various types of mechanical CPR and their potential roles in and on the way to the laboratory. (Circ J 2016; 80: 1292-1299).


Effects of Class IV laser therapy on fibromyalgia impact and function in women with fibromyalgia.
This study evaluated the effects of Class IV laser therapy on pain, Fibromyalgia (FM) impact, and physical function in women diagnosed with FM. The study was a double-blind, randomized control trial. Testing was completed at the university and Rheumatologist office and treatment was completed at a chiropractic clinic. Thirty-eight (38) women (52±11 years; mean±standard deviation) with FM were randomly assigned to one of two treatment groups, laser heat therapy (LHT; n=20) or sham heat therapy (SHT; n=18). Both groups received treatment twice a week for 4 weeks. Treatment consisted of application of LHT or SHT over seven tender points located across the neck, shoulders, and back. Treatment was blinded to women and was administered by a chiropractic physician for 7 minutes. Participants were evaluated before and after treatment for number and sensitivity of tender points, completed the FM Impact Questionnaire (FIQ) and the pain question of the FIQ, and were measured for function using the continuous scale physical functional performance (CS-PFP) test. Data were evaluated using repeated-measures analysis of variance with significance accepted at p≤0.05. There were significant interactions for pain measured by the FIQ (LHT: 7.1±2.3 to 6.2±2.1 units; SHT: 5.8±1.3 to 6.1±1.4 units) and for upper body flexibility measured by the CS-PFP (LHT: 71±17 to 78±12 units; SHT: 77±12 to 77±11 units) with the LHT improving significantly compared to SHT. There was a time effect for the measure of FM impact measured by the FIQ, indicating that FM impact significantly improved from pre- to post-treatment in LHT (63±20 to 57±18 units), while no change was observed in the SHT (57±11 to 55±12 units). This study provides evidence that LHT may be a beneficial modality for women with FM in order to improve pain and upper body range of motion, ultimately reducing the impact of FM.


Problematization of perspectives on health promotion and empowerment in mental health nursing--within the research network "MeHNuRse" and the Horatio conference, 2012.
Mental illness is increasing worldwide, while society's response seems to be a trend toward narrower and more specialized mental health care. This development is creating great demands on mental health nurses to include a health promotion perspective in care and support of persons with mental illness. A health promotion perspective emphasizes cooperation and communication with people who suffer from long-term mental illness, focusing on their independence and health. From a health perspective, every human being is an actor in his/her own life, with an inherent ability to make his/her own choices. However, persons who suffer from long-term mental illness are at risk of losing power and control over areas of their lives and their health. Mental health nurses are in a position to support these individuals in promoting health and in maintaining or regaining control over their lives. The emphasis of this paper is to problematize mental health nurses' responsibility to provide health-promoting nursing care in relation to empowerment by means of emancipation, self-efficacy, and self-management. We argue that mental health nurses can work from a health-promoting perspective by using these concepts and that this challenges some of the traditional ideas of health promotion in mental health nursing. The theoretical background discussions in this paper have their origin in the research network "Mental Health Nursing Research in Scandinavia" (MeHNuRse) and from the professional discussions developed during a 2012 workshop that included mental health nurses and researchers at the European Horatio Festival in Stockholm.


Heterogeneous photocatalysis in flow chemical reactors.
The synergy between photocatalysis and continuous flow chemical reactors has shifted the paradigms of photochemistry, opening new avenues of research with safer and scalable processes that can be readily implemented in academia and industry. Current state-of-the-art photocatalysts are homogeneous transition metal complexes that have favourable photophysical properties, wide electrochemical redox potentials, and photostability. However, these photocatalysts present serious drawbacks, such as toxicity, limited availability, and the overall cost of rare transition metal elements. This reduces their long-term viability, especially at an industrial scale. Heterogeneous photocatalysts (HPCats) are an attractive alternative, as the requirement for the separation and purification is largely removed, but typically at the cost of efficiency. Flow chemical reactors can, to a large extent, mitigate the loss in efficiency through reactor designs that enhance mass transport and irradiation. Herein, we review some important developments of heterogeneous photocatalytic materials and their application in flow reactors for sustainable organic synthesis. Further, the application of continuous flow heterogeneous photocatalysis in environmental remediation is briefly discussed to present some interesting reactor designs that could be implemented to enhance organic synthesis.


Isolation of time-dependent DNA damage induced by energetic carbon ions and their fragments using fluorescent nuclear track detectors.
High energetic carbon (C-) ion beams undergo nuclear interactions with tissue, producing secondary nuclear fragments. Thus, at depth, C-ion beams are composed of a mixture of different particles with different linear energy transfer (LET) values. We developed a technique to enable isolation of DNA damage response (DDR) in mixed radiation fields using beam line microscopy coupled with fluorescence nuclear track detectors (FNTDs). We imaged live cells on a coverslip made of FNTDs right after C-ion, proton or photon irradiation using an in-house built confocal microscope placed in the beam path. We used the FNTD to link track traversals with DNA damage and separated DNA damage induced by primary particles from fragments. We were able to spatially link physical parameters of radiation tracks to DDR in live cells to investigate spatiotemporal DDR in multi-ion radiation fields in real time, which was previously not possible. We demonstrated that the response of lesions produced by the high-LET primary particles associates most strongly with cell death in a multi-LET radiation field, and that this association is not seen when analyzing radiation induced foci in aggregate without primary/fragment classification. We report a new method that uses confocal microscopy in combination with FNTDs to provide submicrometer spatial-resolution measurements of radiation tracks in live cells. Our method facilitates expansion of the radiation-induced DDR research because it can be used in any particle beam line including particle therapy beam lines. Biological Physics and Response Prediction.


A corticotroph pituitary adenoma as the initial presentation of familial glucocorticoid deficiency.
CONTEXT; Familial glucocorticoid deficiency (FGD) is a rare autosomal recessive ACTH-resistance syndrome characterized by glucocorticoid deficiency in the absence of mineralocorticoid deficiency. Here, we report the case of a young woman with a corticotroph pituitary adenoma as the initial presentation of FGD. A 15-year-old girl was referred to our institution for a 16 mm pituitary adenoma associated with glucocorticoid deficiency. Clinical and biological features were evocative of FGD. DNA sequencing did not identify mutations in either the melanocortin 2 receptor (MC2R) or the MC2R accessory protein genes, indicating type 3 FGD. Despite adequate glucocorticoid replacement, plasma ACTH levels remained increased and pituitary magnetic resonance imaging (MRI) showed a progression of the tumour size resulting in optic chiasm compression with intra-tumoural haemorrhaging. When the patient was 26 years old, it was decided that she would undergo transsphenoidal surgery. The histomorphological analysis identified a well-individualized pituitary adenoma immunoreactive for ACTH. The proband's sister also exhibited type 3 FGD associated with pituitary hyperplasia upon MRI. This case highlights the relationship between FGD and hyperplasia of ACTH-producing cells, potentially leading to histologically proven pituitary corticotroph adenomas. This observation raises the question of the pituitary MRI's significance in the follow-up of FGD.


A probabilistic estimation of fluoride intake by infants up to the age of 4 months from infant formula reconstituted with tap water in the fluoridated regions of Ireland.
Two probabilistic models were developed to estimate the acute and chronic exposure to fluoride of exclusively formula-fed infants aged 0-4 months as a result of the consumption of infant formula reconstituted with fluoridated tap water in Ireland. The estimates were based on calculated infant formula consumption and accepted body weight standards, together with reported concentrations of fluoride in infant formula powder and measured values for the fluoride content of water in Ireland. The mean acute exposure of infants to fluoride on any single day in areas served by 387 fluoridated water supplies was estimated to be between 0.11 and 0.14 mg/kg body weight depending on age group (95th percentiles 0.2 and 0.26 mg/kg b.w., respectively). These predicted intakes were well below the intake of fluoride associated with acute toxic effects, which is considered to be 5 mg fluoride (F(-))/kg body weight. The mean chronic exposure of infants to fluoride was estimated to be between 0.106 and 0.170 mg/kg b.w./day depending on body weight (95th percentiles 0.108 and 0.172 mg/kg b.w./day, respectively). This estimate described the average daily fluoride intake of infants during the first 4 months of life residing in the areas served by 226 water supplies that achieved an average yearly fluoride concentration below 1.03 mg/l. Dental fluorosis may be considered to be the only risk at these low doses and from our work it is estimated that there is a very low risk of moderate dental fluorosis of the permanent dentition in infants exposed to fluoride at these levels.


B and T Cells Driving Multiple Sclerosis: Identity, Mechanisms and Potential Triggers.
Historically, multiple sclerosis (MS) has been viewed as being primarily driven by T cells. However, the effective use of anti-CD20 treatment now also reveals an important role for B cells in MS patients. The results from this treatment put forward T-cell activation rather than antibody production by B cells as a driving force behind MS. The main question of how their interaction provokes both B and T cells to infiltrate the CNS and cause local pathology remains to be answered. In this review, we highlight key pathogenic events involving B and T cells that most likely contribute to the pathogenesis of MS. These include (1) peripheral escape of B cells from T cell-mediated control, (2) interaction of pathogenic B and T cells in secondary lymph nodes, and (3) reactivation of B and T cells accumulating in the CNS. We will focus on the functional programs of CNS-infiltrating lymphocyte subsets in MS patients and discuss how these are defined by mechanisms such as antigen presentation, co-stimulation and cytokine production in the periphery. Furthermore, the potential impact of genetic variants and viral triggers on candidate subsets will be debated in the context of MS.


Low-back pain definitions in occupational studies were categorized for a meta-analysis using Delphi consensus methods.
To determine which literature-based definitions of low back pain (LBP) could be combined to produce sufficiently similar sets for use in a meta-analysis. A group of six international experts participated in an e-mail-administered Delphi process. Literature-based LBP definitions were preliminarily classified into 14 sets within four outcome types: pathology, symptoms and care-seeking, functional limitations, and participation. Experts independently rated their level of agreement that each outcome definition belonged in its assigned set using a seven-point Likert scale. After each round, results were synthesized and revised classifications were fed back to the experts who were asked to consider them before rerating the outcome definitions. The experts completed three Delphi rounds and reached consensus on the categorization of 115/119 (97%) of the outcome definitions. There were 20 final sets of outcomes identified: three sets of pathology outcomes, two sets each of functional limitation and participation outcomes, and 13 sets of symptom and care-seeking outcomes. In a research area that currently lacks uniformly accepted definitions of outcomes, we successfully used a Delphi consensus process to reach substantial agreement on combinable LBP outcomes that would be combinable for a meta-analysis.


Laparoscopically assisted anterior rectal wall resection and reanastomosis for deeply infiltrating endometriosis.
A 28-year-old woman, presented with a history of long-standing, severe pelvic and bowel endometriosis. Pronounced cul-de-sac tenderness and nodularity were noted on pelvic examination. Videolaseroscopy was undertaken, the rectum was mobilized, and the tumor was prolapsed to the level of the anus. Anterior rectal wall resection and reanastomosis were performed; the colon was returned to the pelvis under direct visualization via laparoscope.


Role of carglumic acid in the treatment of acute hyperammonemia due to N-acetylglutamate synthase deficiency.
N-acetylglutamate synthase (NAGS) deficiency is a rare inborn error of metabolism affecting ammonia detoxification in the urea cycle. The product of NAGS is N-acetylglutamate which is the absolutely required allosteric activator of the first urea cycle enzyme carbamoylphosphate synthetase 1. In defects of NAGS, the urea cycle function can be severely affected resulting in fatal hyperammonemia in neonatal patients or at any later stage in life. NAGS deficiency can be treated with a structural analog of N-acetylglutamate, N-carbamyl-L-glutamate, which is available for enteral use as a licensed drug. Since NAGS deficiency is an extremely rare disorder, reports on the use of N-carbamyl-L-glutamate are mainly based on single patients. According to these, the drug is very effective in treating acute hyperammonemia by avoiding the need for detoxification during the acute metabolic decompensation. Also during long-term treatment, N-carbamyl-L-glutamate is effective in maintaining normal plasma ammonia levels and avoiding the need for additional drug therapy or protein-restricted diet. Open questions remain which concern the optimal dosage in acute and long-term use of N-carbamyl-L-glutamate and potential additional disorders in which the drug might also be effective in treating acute hyperammonemia. This review focuses on the role of N-carbamyl-L-glutamate for the treatment of acute hyperammonemia due to primary NAGS deficiency but will briefly discuss the current knowledge on the role of N-carbamyl-L-glutamate for treatment of secondary NAGS deficiencies.


18F-FAZA PET imaging response tracks the reoxygenation of tumors in mice upon treatment with the mitochondrial complex I inhibitor BAY 87-2243.
We describe a noninvasive PET imaging method that monitors early therapeutic efficacy of BAY 87-2243, a novel small-molecule inhibitor of mitochondrial complex I as a function of hypoxia-inducible factor-1α (HIF1α) activity. Four PET tracers [(18)F-FDG, (18)F-Fpp(RGD)2, (18)F-FLT, and (18)F-FAZA] were assessed for uptake into tumor xenografts of drug-responsive (H460, PC3) or drug-resistant (786-0) carcinoma cells. Mice were treated with BAY 87-2243 or vehicle. At each point, RNA from treated and vehicle H460 tumor xenografts (n = 3 each) was isolated and analyzed for target genes. Significant changes in uptake of (18)F-FAZA, (18)F-FLT, and (18)F-Fpp(RGD)2 (P < 0.01) occurred with BAY 87-2243 treatment with (18)F-FAZA being the most prominent. (18)F-FDG uptake was unaffected. (18)F-FAZA tumor uptake declined by 55% to 70% (1.21% ± 0.10%ID/g to 0.35 ± 0.1%ID/g; n = 6, vehicle vs. treatment) in both H460 (P < 0.001) and PC3 (P < 0.05) xenografts 1 to 3 days after drug administration. (18)F-FAZA uptake in 786-0 xenografts was unaffected. Decline occurred before significant differences in tumor volume, thus suggesting (18)F-FAZA decrease reflected early changes in tumor metabolism. BAY 87-2243 reduced expression of hypoxia-regulated genes CA IX, ANGPTL4, and EGLN-3 by 99%, 93%, and 83%, respectively (P < 0.001 for all), which corresponds with reduced (18)F-FAZA uptake upon drug treatment. Heterogeneous expression of genes associated with glucose metabolism, vessel density, and proliferation was observed. Our studies suggest suitability of (18)F-FAZA-PET as an early pharmacodynamic monitor on the efficacy of anticancer agents that target the mitochondrial complex I and intratumor oxygen levels (e.g., BAY 87-2243).


Antithyroid drugs and their analogues: synthesis, structure, and mechanism of action.
Thyroid hormones are essential for the development and differentiation of all cells of the human body. They regulate protein, fat, and carbohydrate metabolism. In this Account, we discuss the synthesis, structure, and mechanism of action of thyroid hormones and their analogues. The prohormone thyroxine (T4) is synthesized on thyroglobulin by thyroid peroxidase (TPO), a heme enzyme that uses iodide and hydrogen peroxide to perform iodination and phenolic coupling reactions. The monodeiodination of T4 to 3,3',5-triiodothyronine (T3) by selenium-containing deiodinases (ID-1, ID-2) is a key step in the activation of thyroid hormones. The type 3 deiodinase (ID-3) catalyzes the deactivation of thyroid hormone in a process that removes iodine selectively from the tyrosyl ring of T4 to produce 3,3',5'-triiodothyronine (rT3). Several physiological and pathological stimuli influence thyroid hormone synthesis. The overproduction of thyroid hormones leads to hyperthyroidism, which is treated by antithyroid drugs that either inhibit the thyroid hormone biosynthesis and/or decrease the conversion of T4 to T3. Antithyroid drugs are thiourea-based compounds, which include propylthiouracil (PTU), methimazole (MMI), and carbimazole (CBZ). The thyroid gland actively concentrates these heterocyclic compounds against a concentration gradient. Recently, the selenium analogues of PTU, MMI, and CBZ attracted significant attention because the selenium moiety in these compounds has a higher nucleophilicity than that of the sulfur moiety. Researchers have developed new methods for the synthesis of the selenium compounds. Several experimental and theoretical investigations revealed that the selone (C═Se) in the selenium analogues is more polarized than the thione (C═S) in the sulfur compounds, and the selones exist predominantly in their zwitterionic forms. Although the thionamide-based antithyroid drugs have been used for almost 70 years, the mechanism of their action is not completely understood. Most investigations have revealed that MMI and PTU irreversibly inhibit TPO. PTU, MTU, and their selenium analogues also inhibit ID-1, most likely by reacting with the selenenyl iodide intermediate. The good ID-1 inhibitory activity of PTU and its analogues can be ascribed to the presence of the -N(H)-C(═O)- functionality that can form hydrogen bonds with nearby amino acid residues in the selenenyl sulfide state. In addition to the TPO and ID-1 inhibition, the selenium analogues are very good antioxidants. In the presence of cellular reducing agents such as GSH, these compounds catalytically reduce hydrogen peroxide. They can also efficiently scavenge peroxynitrite, a potent biological oxidant and nitrating agent.


An antigen-targeted approach to adoptive transfer therapy of cancer.
Previous attempts to treat human malignancies by adoptive transfer of tumor-specific CTLs have been limited by the difficulty of isolating T cells of defined antigen specificity. The recent development of MHC class I/antigenic peptide tetrameric complexes that allow direct identification of antigen-specific T cells has opened new possibilities for the isolation and in vitro expansion of tumor-specific T cells. In the present study, we have derived polyclonal monospecific cell lines from circulating Melan-A-specific CTL precursors of HLA-A*0201+ melanoma patients by combining stimulation with recently identified peptide analogues of the immunodominant epitope from the melanoma-associated antigen Melan-A with staining with fluorescent HLA-A*0201/Melan-A peptide tetramers. In vitro expansion of antigen-specific CD8+ T cells was monitored by flow cytometry with the fluorescent tetramers and anti-CD8 monoclonal antibody. This analysis revealed that Melan-A 26-35 peptide analogues were much more efficient than the parental peptides in stimulating a rapid in vitro expansion of antigen-specific CD8+ T cells. These cells were then isolated by tetramer-guided cell sorting and subsequently expanded in vitro by mitogen stimulation. The resulting polyclonal but monospecific CTLs fully cross-recognized the parental peptides and were able to efficiently lyse Melan-A-expressing tumor cells. Altogether, these results pave the way to a molecularly defined approach to antigen-specific adoptive transfer therapy of cancer.


From the archives of the AFIP: Gastrointestinal carcinoids: imaging features with clinicopathologic comparison.
Gastrointestinal carcinoids are well-differentiated endocrine neoplasms that belong to a diverse group of tumors that arise from cells of the diffuse endocrine system. A wide variety of specialized endocrine cells that populate the gastrointestinal mucosa and submucosa give rise to carcinoids. Consequently, carcinoids may occur throughout the gastrointestinal tract and produce a variety of hormones and protein products that are associated with specific clinical symptoms. Biologic behavior of carcinoids varies by site and cell type, but all gastrointestinal carcinoids are considered to have malignant potential. They may produce specific syndromes such as Zollinger-Ellison syndrome, or they may occur in association with inherited syndromes such as multiple endocrine neoplasia type 1 or neurofibromatosis type 1. Metastatic carcinoids may produce carcinoid syndrome. The small intestine is the most common location for gastrointestinal carcinoids. Most small intestinal carcinoids arise from enterochromaffin cells of the distal ileum that produce serotonin. Small intestinal carcinoids often have an aggressive biologic behavior and, as such, patients frequently have metastases to regional lymph nodes and the liver at initial presentation. Pathologic and radiologic manifestations of serotonin-producing small intestinal carcinoids are related to local and regional effects of serotonin and its metabolites. In contrast, carcinoids of the appendix and rectum are commonly discovered incidentally as small lesions that are unassociated with clinical evidence of hormone production and have a more indolent clinical course. Carcinoids of the stomach, duodenum, and colon are uncommon but have distinctive clinical, pathologic, and radiologic appearances. Knowledge of the diverse clinical, pathologic, and radiologic spectrum of gastrointestinal carcinoids is important in the imaging and management of patients with suspected carcinoids or focal gastrointestinal masses.


MicroRNA target site identification by integrating sequence and binding information.
High-throughput sequencing has opened numerous possibilities for the identification of regulatory RNA-binding events. Cross-linking and immunoprecipitation of Argonaute proteins can pinpoint a microRNA (miRNA) target site within tens of bases but leaves the identity of the miRNA unresolved. A flexible computational framework, microMUMMIE, integrates sequence with cross-linking features and reliably identifies the miRNA family involved in each binding event. It considerably outperforms sequence-only approaches and quantifies the prevalence of noncanonical binding modes.


Identifying and Validating Requirements of a Mobile-Based Self-Management System for People Living with HIV.
Due to the widespread use of mobile technology and the low cost of this technology, implementing a mobile-based self-management system can lead to adherence to the medication regimens and promotion of the health of people living with HIV (PLWH). We aimed to identify requirements of a mobile-based self-management system, and validate them from the perspective of infectious diseases specialists. This is a mixed-methods study that carried out in two main phases. In the first phase, we identified requirements of a mobile-based self-management system for PLWH. In the second phase, identified requirements were validated using a researcher made questionnaire. The statistical population was infectious diseases specialists affiliated to Tehran University of Medical Sciences. The collected data were analyzed using SPSS statistical software (version 19), and descriptive statistics. By full-text review of selected studies, we determined requirements of a mobile-based self-management system in four categories: demographic, clinical, strategically and technical capabilities. According to the findings, 6 data elements for demographic category, 11 data elements for clinical category, 10 items for self-management strategies, and 11 features for technical capabilities were selected. Using the identified preferences, it is possible to design and implement a mobile-based self-management system for HIV-positive people. Developing a mobile-based self-management system is expected to progress the skills of self-management PLWH, improve of medication regimen adherence, and facilitate communication with healthcare providers.


Electrochemically prepared composites of graphene oxide and conducting polymers: Cytocompatibility of cardiomyocytes and neural progenitors.
The cytocompatibility of cardiomyocytes derived from embryonic stem cells and neural progenitors, which were seeded on the surface of composite films made of graphene oxide (GO) and polypyrrole (PPy-GO) or poly(3,4-ethylenedioxythiophene) (PEDOT-GO) are reported. The GO incorporated in the composite matrix contributes to the patterning of the composite surface, while the electrically conducting PPy and PEDOT serve as ion-to-electron transducers facilitating electrical stimulation/sensing. The films were fabricated by a simple one-step electropolymerization procedure on electrically conducting indium tin oxide (ITO) and graphene paper (GP) substrates. Factors affecting the cell behaviour, i.e. the surface topography, wettability, and electrical surface conductivity, were studied. The PPy-GO and PEDOT-GO prepared on ITO exhibited high surface conductivity, especially in the case of the ITO/PPy-GO composite. We found that for cardiomyocytes, the PPy-GO and PEDOT-GO composites counteracted the negative effect of the GP substrate that inhibited their growth. Both the PPy-GO and PEDOT-GO composites prepared on ITO and GP significantly decreased the cytocompatibility of neural progenitors. The presented results enhance the knowledge about the biological properties of electroactive materials, which are critical for tissue engineering, especially in context stimuli-responsive scaffolds.


Thermodynamics of A:G mismatch poly(dG) synthesis by human immunodeficiency virus 1 reverse transcriptase.
Human immunodeficiency virus 1 (HIV-1) reverse transcriptase has been found to conduct error-prone synthesis on DNA and RNA templates. We find here that tolerance of an A:G mispair with poly(rA) as template is particularly strong, such that extensive poly(dG) synthesis is conducted. This type of extensive misincorporation is not observed with several reference DNA polymerases. Surprisingly, HIV reverse transcriptase processivity and kcat for dGMP misincorporation and normal dTMP incorporation are about the same. However, the Km value for dGTP in poly(dG) synthesis is approximately 1000-fold higher than the Km for dTTP in poly(dT) synthesis. Comparison of thermodynamic parameters for dGMP misincorporation and normal dNMP incorporation indicates a lower energy of activation for dGMP misincorporation than for normal dNMP incorporation. Entropy of activation (delta S*) for normal dTMP incorporation is positive (approximately 10 cal/kmol), whereas delta S* for dGMP misincorporation is negative (-36 cal/kmol). Since differences in delta S* are usually considered to reflect differences in solvation for the transition state complex, these results are consistent with the interpretation that the active site of HIV reverse transcriptase is flexible enough to misincorporate dGMP without the usual dispersion of water molecules.


The role of eye movements in motor sequence learning.
An experiment that utilized a 16-element movement sequence was designed to determine the impact of eye movements on sequence learning. The participants were randomly assigned to two experimental groups: a group that was permitted to use eye movements (FREE) and a second group (FIX) that was instructed to fixate on a marker during acquisition (ACQ). A retention test (RET) was designed to provide a measure of learning, and two transfer tests were designed to determine the extent to which eye movements influenced sequence learning. The results demonstrated that both groups decreased the response time to produce the sequence, but the participants in the FREE group performed the sequence more quickly than participants of the FIX group during the ACQ, RET and the two transfer tests. Furthermore, continuous visual control of response execution was reduced over the course of learning. The results of the transfer tests indicated that oculomotor information regarding the sequence can be stored in memory and enhances response production.


Epidemiologic features of the physical and sexual maltreatment of children in the Carolinas.
Child maltreatment remains a significant public health and social problem in the United States. Incidence data rely on substantiated reports of maltreatment known to official social service agencies. The objective of this study was to describe the epidemiologic features of child physical and sexual abuse, on the basis of maternal self-reports. Computer-assisted, anonymous, cross-sectional, telephone surveys (N = 1435) were conducted with mothers of children 0 to 17 years of age in North and South Carolina. Mothers were asked about potentially abusive behaviors used by either themselves or their husbands or partners in the context of other disciplinary practices. They were also asked about their knowledge of any sexual victimization their children might have experienced. The incidence of physical and sexual maltreatment determined through maternal reports. Use of harsh physical discipline, equivalent to physical abuse, occurred with an incidence of 4.3%. Shaking of very young children as a means of discipline occurred among 2.6% of children <2 years of age. Mothers reported more frequent physical discipline of their children, including shaking, for themselves than for fathers or father figures. Nearly 11 of 1000 children were reported by their mothers as having been sexually victimized within the past year. The incidence of physical abuse determined with maternal self-reports was 40 times greater than that of official child physical abuse reports, and the sexual abuse incidence was 15 times greater. For every 1 child who sustains a serious injury as a result of shaking, an estimated 150 children may be shaken and go undetected. There was no statistically significant difference in the overall rates of physical or sexual maltreatment between the 2 states. Official statistics underestimate the burden of child maltreatment. Supplemental data obtained with alternative strategies can assist policymakers and planners in addressing needs and services within communities and states. These data support the need for continued interventions to prevent maltreatment.


Occipital neuralgias: clinical recognition of a complicated headache. A case series and literature review.
The objective of this study was to assist clinicians in the diagnosis of the occipital neuralgia syndrome by describing its clinical characteristics. Bibliographies and clinical descriptions of occipital neuralgia syndrome were identified through a review of literature published between 1966 and 1993. A prospective case series was performed by the authors in a university emergency department during a 1-year period. Patients with unilateral aching pain of the head, coupled with pain in the distribution of the occipital nerve, Tinel's sign, and relief of pain after local anesthetic injection, were included. Patients rated pain relief on a 10-point scale. Twelve patients met the criteria for occipital neuralgia and were included in the study. All patients reported at least 80% decrease of pain after injection, and 42% had complete relief. Clinical features, other than headache, that were common in patients included tinnitus in 33%; scalp paresthesia, 33%; nausea, 42%; dizziness, 50%; and visual disturbances, 67%. Occipital neuralgia is a benign extracranial cause of headache, and it may be confused with other more serious headache syndromes. Recognition depends on an understanding of the symptoms along with a careful history and physical examination. Local anesthetic injections produce significant relief of the headaches and can aid in the diagnosis of the syndrome.


Additive approach for inactivation of Escherichia coli O157:H7, Salmonella, and Shigella spp. on contaminated fresh fruits and vegetables using bacteriophage cocktail and produce wash.
The incidence of foodborne outbreaks involving fresh produce is of worldwide concern. Lytic bacteriophage cocktails and a levulinic acid produce wash were investigated for their effectiveness against the foodborne pathogens Escherichia coli O157:H7, Shigella spp., and Salmonella on broccoli, cantaloupe, and strawberries. Inoculated samples were treated with bacteriophage cocktails (BC) before storage at 10°C for 24 h, a levulinic acid produce wash (PW) after storage at 10°C for 24 h, or a combination of the washes (BCPW) before and after storage. All three treatments were compared against a 200-ppm free available chlorine wash. Wash solutions were prepared using potable water and water with an increased organic content of 2.5 g/liter total dissolved solids and total organic carbon. BCPW was the most effective treatment, producing the highest log reductions in the pathogens. Produce treated with BCPW in potable water with a PW exposure time of 5 min resulted in the highest reduction of each pathogen for all samples tested. The type of produce and wash solution had significant effects on the efficacy of the individual treatments. The chlorine wash in water with higher organic content was the least effective treatment tested. An additive effect of BCPW was seen in water with higher organic content, resulting in greater than 4.0-log reductions in pathogens. Our findings indicate that the combination of antimicrobial BC with a commercial produce wash is a very effective method for treating produce contaminated with E. coli O157:H7, Shigella spp., and Salmonella even in the presence of high loads of organic matter.


Potentiation of oxyntic atrophy-induced gastric metaplasia in amphiregulin-deficient mice.
The loss of parietal cells from the gastric mucosa (oxyntic atrophy) is a critical step in the pathogenesis of chronic gastritis and gastric adenocarcinoma. Parietal cells are known to secrete epidermal growth factor receptor (EGFR) ligands, which are critical regulators of differentiation in the gastric mucosa. Although all of the actions of EGFR ligands are mediated through a common EGFR protein, individual ligands may produce different physiologic responses. Previous investigations have suggested that a deficit in EGFR signaling in waved-2 mice accelerates the emergence of metaplasia after induction of acute oxyntic atrophy. We sought to determine whether specific EGFR ligands regulate the metaplastic response to oxyntic atrophy. To induce spasmolytic polypeptide-expressing metaplasia (SPEM), amphiregulin (AR) and transforming growth factor-alpha-deficient mice and their wild-type littermates were treated with DMP-777 for 0-14 days and for 14 days followed by 14 days of recovery off drug. We evaluated the gastric mucosal response to oxyntic atrophy using cell lineage-specific markers. Although loss of transforming growth factor-alpha did not influence the induction of SPEM, loss of AR caused an acceleration and amplification in the induction of SPEM after acute oxyntic atrophy. Trefoil factor family 2/spasmolytic polypeptide and intrinsic factor dual-immunostaining cells significantly increased in the SPEM of AR-deficient mice. At the bases of glands, intrinsic factor immunoreactive cells also were costained for 5-bromo-2'-deoxyuridine, suggesting their re-entry into the cell cycle. The absence of AR promoted the rapid emergence of SPEM in response to oxyntic atrophy.


Clinical and epidemiological characteristics of patients with influenza A (H1N1) 2009 attended to at the emergency room of a children's hospital.
In June 2009, the first influenza pandemic of the twenty-first century, due to the swine origin influenza A (H1N1) 2009 virus, was declared. This study aimed to describe the epidemiological and clinical features, complications, lethality and risk factors for hospital admission of microbiologically confirmed cases of influenza A (H1N1) 2009 infection seen at the emergency department of a children's hospital. All cases of children with influenza A (H1N1) 2009 viral infection, confirmed microbiologically by real-time reverse transcription polymerase chain reactions and treated in the emergency room between July and December 2009, were prospectively included. Patients were compared according to admission requirement to study variables associated with the risk of hospitalisation. Oseltamivir was the antiviral used for the treatment and its safety was analysed. Four hundred and twelve patients with influenza A (H1N1) 2009 infection were included. The most frequent symptoms were: fever (96%), cough (95%) and coryza (90%). Eighty-five patients (20.6%) were admitted: three to the paediatric intensive care unit and two died. Hospitalised children were younger than those not admitted (median age 5 vs 8 years; p = 0.001). Age under 1 year (OR 6.01; CI 95% 2.77-13.05), pneumonia (OR 7.99; CI 95% 3.50-18.22) and haemoglobinopathy or underlying blood disorders (OR 5.99; CI 95% 1.32-27.30) were statistically significant risk factors for admission. No differences were observed regarding onset of antiviral treatment among admitted and non-admitted patients. Treatment with oseltamivir was well tolerated. In conclusion, the incidence of severe cases and lethality of influenza A (H1N1) 2009 infection were low in our setting, even in a population with risk factors for developing complications.


Neonatal hypothermia on admission to a special care unit in Dar-es-Salaam, Tanzania: a cause for concern.
To determine the prevalence and risk factors for hypothermia among neonates on admission to the Neonatal Care Unit. Hypothermia in newborn babies is a problem in tropical countries despite warm environmental conditions and it contributes to a high neonatal morbidity and mortality. A study was undertaken to determine the prevalence of hypothermia and its association with early neonatal outcome among neonates admitted to the Neonatal Care Unit of Muhimbili Medical Centre. At admission all neonates were examined and axillary temperature recorded using a low-reading thermometer. Six-hourly temperature was taken in all infants. Those with a temperature below 36.5 degrees C were recruited as cases and those with normal temperature served as controls. These neonates were followed up for early neonatal outcome. Hypothermia on admission was found in 366 out of 1,632 babies (22.4%). In none of these was hypothermia recorded or reported as a reason for admission. Thirteen percent of the hypothermic neonates had severe hypothermia, with body temperature below 32 degrees C on admission. Hypothermia was significantly associated with deliveries from outside hospitals and with those who had operative or instrumental delivery in the same hospital. It was also associated with prematurity, low birth weight babies, time taken to transfer the baby and inadequate clothing after delivery. It was found that hypothermic infants had a three fold higher mortality and morbidity. These infants had a longer stay in the unit and had a higher post natal weight loss. There was no low-reading thermometer in the unit. It is concluded that there is cause for concern about hypothermia in the neonates at Muhimbili Medical Centre. Efforts should be made to sensitize and educate all levels of staff dealing with neonates, and low-reading thermometers should be part of the essential kit in the unit.


Tricuspid Valve Repair for the Poor Right Ventricle: Tricuspid Valve Repair in Patients with Mild-to-Moderate Tricuspid Regurgitation Undergoing Mitral Valve Repair Improves In-Hospital Outcome.
Background Tricuspid regurgitation (TR) in patients undergoing surgery for mitral valve (MV) increases morbidity and mortality, especially in case of a poor right ventricle. Does repair of mild-to-moderate insufficiency of the tricuspid valve (TV) in patients undergoing MV surgery lead to a benefit in early postoperative outcome? Methods A total of 22 patients with mild-to-moderate TR underwent MV repair and concomitant TV repair with Tri-Ad (Medtronic ATS Medical Inc., Minneapolis, Minnesota, United States) and Edwards Cosgrove (Edwards Lifesciences Irvine, California, United States) rings. The severity of TR was assessed echocardiographically by using color-Doppler flow images. The tricuspid annular plane systolic excursion (TAPSE) was under 1.7 cm. Additional procedures included coronary artery bypass (n = 9) and maze procedure (n = 15). The following parameters were compared: postoperative and peak dose of noradrenaline (NA), pre/postoperative systolic pulmonary pressure (sPAP), extubation time, operation time, cross-clamp time, cardiopulmonary bypass (CPB) time, pre/postoperative ejection fraction (EF), intensive care unit (ICU)-stay, hospital stay, cell saver blood transfusion, intra/postoperative blood transfusion, and postoperative TR. Results The mean age was 67 ± 14.8 years, 45% were male. Mean EF was 47 ± 16.2%, postoperative 52 ± 12.4%. sPAP was 46 ± 20.1 mm Hg preoperatively, sPAP was 40.6 ± 9.4 mm Hg postoperatively, NA postoperatively was 12 ± 10 μg/min, NA peak was 18 ± 11 μg/min, operation time was 275 ± 92 minutes, CPB was 145 ± 49 minutes, ICU stay was 2.4 ± 2.4 days, hospital stay was 10.8 ± 3.5 days, cell saver blood transfusion was 736 ± 346 mL, intraoperative transfusions were 2.5 ± 1.6. Two patients needed postoperative transfusions. A total of 19 patients were extubated at the 1st postoperative day, 2 patients at the 2nd day, and 1 at the 4th postoperative day. Two patients required a pacemaker. No reintubation, no in-hospital mortality, and one reoperation because of bleeding complications. Conclusion Correction of mild-to-moderate TR at the time of MV repair does maintain TV function and avoid right ventricular dysfunction in the early postoperative period improving the clinical outcome.


Rab6 coordinates a novel Golgi to ER retrograde transport pathway in live cells.
We visualized a fluorescent-protein (FP) fusion to Rab6, a Golgi-associated GTPase, in conjunction with fluorescent secretory pathway markers. FP-Rab6 defined highly dynamic transport carriers (TCs) translocating from the Golgi to the cell periphery. FP-Rab6 TCs specifically accumulated a retrograde cargo, the wild-type Shiga toxin B-fragment (STB), during STB transport from the Golgi to the endoplasmic reticulum (ER). FP-Rab6 TCs associated intimately with the ER, and STB entered the ER via specialized peripheral regions that accumulated FP-Rab6. Microinjection of antibodies that block coatomer protein I (COPI) function inhibited trafficking of a KDEL-receptor FP-fusion, but not FP-Rab6. Additionally, markers of COPI-dependent recycling were excluded from FP-Rab6/STB TCs. Overexpression of Rab6:GDP (T27N mutant) using T7 vaccinia inhibited toxicity of Shiga holotoxin, but did not alter STB transport to the Golgi or Golgi morphology. Taken together, our results indicate Rab6 regulates a novel Golgi to ER transport pathway.


Reflections, resilience and recovery: a qualitative study of Covid-19's impact on an international adult population's mental health and priorities for support.
The impact of the coronavirus 2019 (Covid-19) pandemic on different countries and populations is well documented in quantitative studies, with some studies showing stable mental health symptoms and others showing fluctuating symptoms. However, the reasons behind why some symptoms are stable and others change are under-explored, which in turn makes identifying the types of support needed by participants themselves challenging. To address these gaps, this study thematically analysed 925 qualitative responses from five open-ended responses collected in the UCL-Penn Global COVID Study between 17 April and 31 July 2021 (Wave 3). Three key themes that comprised 13 codes were reported by participants across countries and ages regarding the impact of Covid-19 on their health, both mental and physical, and livelihoods. These include: (1) Outlook on self/life, (2) Self-improvement, and (3) Loved ones (friends and family). In terms of support, while 2.91% did not require additional support, 91% wanted support beyond financial support. Other unexpected new themes were also discussed regarding vulnerable populations suffering disproportionately. The pandemic has brought into sharp focus various changes in people's mental health, physical health and relationships. Greater policy considerations should be given to supporting citizens' continued access to mental health when considering pandemic recovery.


Characterization of a dipartite iron uptake system from uropathogenic Escherichia coli strain F11.
In the uropathogenic Escherichia coli strain F11, in silico genome analysis revealed the dicistronic iron uptake operon fetMP, which is under iron-regulated control mediated by the Fur regulator. The expression of fetMP in a mutant strain lacking known iron uptake systems improved growth under iron depletion and increased cellular iron accumulation. FetM is a member of the iron/lead transporter superfamily and is essential for iron uptake by the Fet system. FetP is a periplasmic protein that enhanced iron uptake by FetM. Recombinant FetP bound Cu(II) and the iron analog Mn(II) at distinct sites. The crystal structure of the FetP dimer reveals a copper site in each FetP subunit that adopts two conformations: CuA with a tetrahedral geometry composed of His(44), Met(90), His(97), and His(127), and CuB, a second degenerate octahedral geometry with the addition of Glu(46). The copper ions of each site occupy distinct positions and are separated by ∼1.3 Å. Nearby, a putative additional Cu(I) binding site is proposed as an electron source that may function with CuA/CuB displacement to reduce Fe(III) for transport by FetM. Together, these data indicate that FetMP is an additional iron uptake system composed of a putative iron permease and an iron-scavenging and potentially iron-reducing periplasmic protein.


DICER1 in the Pathogenesis of Age-related Macular Degeneration (AMD) - Alu RNA Accumulation versus miRNA Dysregulation.
DICER1 deficiency in the retinal pigment epithelium (RPE) was associated with the accumulation of Alu transcripts and implicated in geographic atrophy (GA), a form of age-related macular degeneration (AMD), an eye disease leading to blindness in millions of people. Although the exact mechanism of this association is not fully known, the activation of the NLRP3 inflammasome, maturation of caspase-1 and disruption in mitochondrial homeostasis in RPE cells were shown as critical for it. DICER1 deficiency results in dysregulation of miRNAs and changes in the expression of many genes important for RPE homeostasis, which may also contribute to AMD. DICER1 deficiency can change the functions of the miR-183/96/182 cluster that regulates photoreceptors and their synaptic transmission. Aging, the main AMD risk factor, is associated with decreased expression of DICER1 and changes in its diurnal pattern that are not synchronized with circadian regulation in the retina. The initial insult inducing DICER1 deficiency in AMD may be oxidative stress, another major risk factor of AMD, but further studies on the role of deficient DICER1 in AMD pathogenesis and its therapeutic potential are needed.


ICG-001 affects DRP1 activity and ER stress correlative with its anti-proliferative effect.
Mitochondria form a highly dynamic network driven by opposing scission and fusion events. DRP1 is an essential modulator of mitochondrial fission and dynamics within mammalian cells. Its fission activity is regulated by posttranslational modifications such as activating phosphorylation at serine 616. DRP1 activity has recently been implicated as being dysregulated in numerous human disorders such as cancer and neurodegenerative diseases. Here we describe the development of a cell-based screening assay to detect DRP1 activation. We utilized this to undertake focused compound library screening and identified potent modulators that affected DRP1 activity including ICG-001, which is described as WNT/β-catenin signaling inhibitor. Our findings elucidate novel details about ICG-001's mechanism of action (MOA) in mediating anti-proliferative activity. We show ICG-001 both inhibits mitochondrial fission and activates an early endoplasmic reticulum (ER) stress response to induce cell death in susceptible colorectal cancer cell lines.


Selected contribution: axial stretch increases spontaneous pacemaker activity in rabbit isolated sinoatrial node cells.
Isolated, spontaneously beating rabbit sinoatrial node cells were subjected to longitudinal stretch, using carbon fibers attached to both ends of the cell. Their electrical behavior was studied simultaneously in current-clamp or voltage-clamp mode using the perforated patch configuration. Moderate stretch ( approximately 7%) caused an increase in spontaneous beating rate (by approximately 5%) and a reduction in maximum diastolic and systolic potentials (by approximately 2.5%), as seen in multicellular preparations. Mathematical modeling of the stretch intervention showed the experimental results to be compatible with stretch activation of cation nonselective ion channels, similar to those found in other cardiac cell populations. Voltage-clamp experiments validated the presence of a stretch-induced current component with a reversal potential near -11 mV. These data confirm, for the first time, that the positive chronotropic response of the heart to stretch is, at least in part, encoded on the level of individual sinoatrial node pacemaker cells; all reported data are in agreement with a major contribution of stretch-activated cation nonselective channels to this response.


Clinician perspectives delivering telehealth interventions to children/families impacted by pediatric traumatic brain injury.
Technological advances have made the delivery of psychological interventions via web-based platforms increasingly feasible. In recent years, there has been growth in the delivery of psychological interventions through web-based modalities, that is, telepsychology. Although there is evidence supporting the usability and feasibility of telepsychology for a range of populations, there is limited literature on clinician perceptions delivering telepsychology, particularly to pediatric rehabilitation populations. In this mixed-methods study, we report on clinician perspectives and experiences delivering telepsychology to children/families impacted by pediatric traumatic brain injury. Seventeen clinicians (psychologists and advanced psychology doctoral students) who delivered telepsychology interventions to children/families impacted by pediatric brain injuries completed surveys and interviews. Overall, clinicians reported that telepsychology was equivalent to face-to-face treatment in many regards (e.g., therapeutic alliance, weekly progress, child/family engagement, and establishing rapport). Clinicians reported a number of advantages of telepsychology over face-to-face interventions for this pediatric population including greater ease of scheduling, increased understanding of the family and home environments, and less caregiver stigma of behavioral health care. Disadvantages of telepsychology included difficulties reading nonverbal cues, logistical/technological issues, and greater disruptions during sessions. Findings provide an important foundation for future investigations examining the merits of telepsychology versus traditional treatment for both pediatric rehabilitation populations. (PsycINFO Database Record (c) 2019 APA, all rights reserved).


Flexible Printed Monolithic-Structured Solid-State Dye Sensitized Solar Cells on Woven Glass Fibre Textile for Wearable Energy Harvesting Applications.
Previously, textile dye sensitised solar cells (DSSCs) woven using photovoltaic (PV) yarns have been demonstrated but there are challenges in their implementation arising from the mechanical forces in the weaving process, evaporation of the liquid electrolyte and partially shaded cells area, which all reduce the performance of the cell. To overcome these problems, this paper proposes a novel fabrication process for a monolithic-structured solid-state dye sensitized solar cell (ssDSSC) on textile using all solution based processes. A glass fibre textile substrate was used as the target substrate for the printed ssDSSC that contain multiple layers of electrodes and active materials. The printed ssDSSC on textile have been successfully demonstrated and compared with a reference device made with the same processes on a glass substrate. All PV textile devices were characterized under simulated AM 1.5 conditions and a peak efficiency of 0.4% was achieved. This approach is potentially suitable for the low cost integration of PV devices onto high temperature textiles, but to widen the range of applications future research is required to reduce the processing temperature to enable the device to be fabricated on the standard fabric substrates.


Open aortic arch reconstruction after previous cardiac surgery: outcomes of 168 consecutive operations.
Open arch interventions after previous cardiac surgery are considered high risk. We reviewed our outcomes in patients requiring aortic arch reconstruction after previous cardiovascular surgery. From March 2000 to March 2014, the data from 168 patients with previous sternotomy requiring aortic arch replacement were reviewed. The indications for surgery, perioperative data, and outcomes of reoperation were analyzed. The mean age was 61 ± 14 years, and 119 were men (70%). The indications for reoperation were aneurysm (57%), valvular disease (13%), impending rupture (12%), aortic dissection (9.0%), and endocarditis (7.7%). The median time from the previous operation to reoperation was 7 years. The mean aortic diameter was 55 mm. Total or partial arch replacement was performed in 38% and 62% of patients, respectively. Fifty-five patients (32.7%) had undergone previous ascending dissection repair and 45 (26.8%) had previous coronary bypass surgery. Deep hypothermic circulatory arrest was used in all. Selective cerebral perfusion was used in 39% and retrograde cerebral perfusion in 14%. The incidence of permanent stroke was 5.4%. Operative mortality (30-day) was 8.3%. Older age (odds ratio, 1.05; 95% confidence interval, 1.00-1.10; P = .04), New York Heart Association class III/IV (odds ratio, 3.15; 95% confidence interval, 1.01-9.86; P = .04), and extracorporeal circulation time (odds ratio, 1.01; 95% confidence interval, 1.00-1.02; P = .001) were predictors of perioperative death. The median follow-up was 3.0 years. Survival was 85%, 78%, and 68% at 1, 3, and 5 years, respectively. Reoperations to address the aortic arch have acceptable mortality and morbidity. Open repair under circulatory arrest is the benchmark to which endovascular therapies should be compared.


Sets2Networks: network inference from repeated observations of sets.
The skeleton of complex systems can be represented as networks where vertices represent entities, and edges represent the relations between these entities. Often it is impossible, or expensive, to determine the network structure by experimental validation of the binary interactions between every vertex pair. It is usually more practical to infer the network from surrogate observations. Network inference is the process by which an underlying network of relations between entities is determined from indirect evidence. While many algorithms have been developed to infer networks from quantitative data, less attention has been paid to methods which infer networks from repeated co-occurrence of entities in related sets. This type of data is ubiquitous in the field of systems biology and in other areas of complex systems research. Hence, such methods would be of great utility and value. Here we present a general method for network inference from repeated observations of sets of related entities. Given experimental observations of such sets, we infer the underlying network connecting these entities by generating an ensemble of networks consistent with the data. The frequency of occurrence of a given link throughout this ensemble is interpreted as the probability that the link is present in the underlying real network conditioned on the data. Exponential random graphs are used to generate and sample the ensemble of consistent networks, and we take an algorithmic approach to numerically execute the inference method. The effectiveness of the method is demonstrated on synthetic data before employing this inference approach to problems in systems biology and systems pharmacology, as well as to construct a co-authorship collaboration network. We predict direct protein-protein interactions from high-throughput mass-spectrometry proteomics, integrate data from Chip-seq and loss-of-function/gain-of-function followed by expression data to infer a network of associations between pluripotency regulators, extract a network that connects 53 cancer drugs to each other and to 34 severe adverse events by mining the FDA's Adverse Events Reporting Systems (AERS), and construct a co-authorship network that connects Mount Sinai School of Medicine investigators. The predicted networks and online software to create networks from entity-set libraries are provided online at http://www.maayanlab.net/S2N. The network inference method presented here can be applied to resolve different types of networks in current systems biology and systems pharmacology as well as in other fields of research.


The evolving impact of g protein-coupled receptor kinases in cardiac health and disease.
G protein-coupled receptors (GPCRs) are important regulators of various cellular functions via activation of intracellular signaling events. Active GPCR signaling is shut down by GPCR kinases (GRKs) and subsequent β-arrestin-mediated mechanisms including phosphorylation, internalization, and either receptor degradation or resensitization. The seven-member GRK family varies in their structural composition, cellular localization, function, and mechanism of action (see sect. II). Here, we focus our attention on GRKs in particular canonical and novel roles of the GRKs found in the cardiovascular system (see sects. III and IV). Paramount to overall cardiac function is GPCR-mediated signaling provided by the adrenergic system. Overstimulation of the adrenergic system has been highly implicated in various etiologies of cardiovascular disease including hypertension and heart failure. GRKs acting downstream of heightened adrenergic signaling appear to be key players in cardiac homeostasis and disease progression, and herein we review the current data on GRKs related to cardiac disease and discuss their potential in the development of novel therapeutic strategies in cardiac diseases including heart failure.


Impact of microbial Mn oxidation on the remobilization of bioreduced U(IV).
Effects of Mn redox cycling on the stability of bioreduced U(IV) are evaluated here. U(VI) can be biologically reduced to less soluble U(IV) species and the stimulation of biological activity to that end is a salient remediation strategy; however, the stability of these materials in the subsurface environments where they form remains unproven. Manganese oxides are capable of rapidly oxidizing U(IV) to U(VI) in mixed batch systems where the two solid phases are in direct contact. However, it is unknown whether the same oxidation would take place in a porous medium. To probe that question, U(IV) immobilized in agarose gels was exposed to conditions allowing biological Mn(II) oxidation (HEPES buffer, Mn(II), 5% O2 and Bacillus sp. SG-1 spores). Results show the oxidation of U(IV) to U(VI) is due primarily to O2 rather than to MnO2. U(VI) produced is retained within the gel to a greater extent when Mn oxides are present, suggesting the formation of strong surface complexes. The implication for the long-term stability of U in a bioremediated site is that, in the absence of competing ligands, biological Mn(II) oxidation may promote the immobilization of U(VI) produced by the oxidation of U(IV).


Isoliquiritigenin exhibits anti-proliferative properties in the pituitary independent of estrogen receptor function.
The plant flavonoid isoliquiritigenin (ISL) is a botanical estrogen widely taken as an herbal supplement to ease the symptoms of menopause. ISL has been also shown to have anti-tumor properties in a number of cancer cell backgrounds. However, the effects of ISL on normal cells are less well known and virtually unstudied in the context of the pituitary gland. We have established a pituitary explant culture model to screen chemical agents for gene expression changes within the pituitary gland during a period of active proliferation and differentiation. Using this whole-organ culture system we found ISL to be weakly estrogenic based on its ability to induce Cckar mRNA expression, an estrogen receptor (ER) mediated gene. Using a range of ISL from 200nM to 200μM, we discovered that ISL promoted cell proliferation at a low concentration, yet potently inhibited proliferation at the highest concentration. ICI 182,780 failed to antagonize ISL's repression of pituitary cell proliferation, indicating the effect is independent of ER signaling. Coincident with a decrease in proliferating cells, we observed down-regulation of transcript for cyclin D2 and E2 and a strong induction of mRNA and protein for the cyclin dependent kinase inhibitor Cdkn1a (p21). Importantly, high dose ISL did not alter the balance of progenitor vs. differentiated cell types within the pituitary explants and they seemed otherwise healthy; however, TUNEL staining revealed an increase in apoptotic cell death in ISL treated cultures. Our results merit further examination of ISL as an anti-tumor agent in the pituitary gland.


Effect of Acute Caffeine Intake on the Fat Oxidation Rate during Exercise: A Systematic Review and Meta-Analysis.
A number of previous investigations have been designed to determine the effect of acute caffeine intake on the rate of fat oxidation during exercise. However, these investigations have shown contradictory results due to the differences in the exercise protocols used or the co-ingestion of caffeine with other substances. Hence, to date, there is no consensus about the effect of caffeine on fat oxidation during exercise. The purpose of this study was to conduct a systematic review followed by a meta-analysis to establish the effect of acute intake of caffeine (ranging from 2 to 7 mg/kg of body mass) on the rate of fat oxidation during exercise. A total of 19 studies published between 1978 and 2020 were included, all of which employed crossover experimental designs in which the ingestion of caffeine was compared to a placebo. Studies were selected if the exercise intensity was consistent in the caffeine and placebo trials and if these were preceded by a fasting protocol. A subsequent meta-analysis was performed using the random effects model to calculate the standardized mean difference (SMD). The meta-analysis revealed that caffeine significantly (p = 0.008) increased the fat oxidation rate (SMD = 0.73; 95% CI = 0.19 to 1.27). This increment was consistent with a significant (p = 0.04) reduction of the respiratory exchange ratio (SMD = -0.33; 95% CI = -0.65 to -0.01) and a significant (p = 0.049) increase in the oxygen uptake (SMD = 0.23; 95% CI = 0.01 to 0.44). The results also showed that there was a dose-response effect of caffeine on the fat oxidation rate, indicating that more than 3.0 mg/kg is necessary to obtain a statistically significant effect of this stimulant on fat oxidation during exercise. Additionally, the ability of caffeine to enhance fat oxidation during exercise was higher in sedentary or untrained individuals than in trained and recreational athletes. In conclusion, pre-exercise intake of a moderate dose of caffeine may effectively increase fat utilization during aerobic exercise of submaximal intensity performed after a fasting period. However, the fitness level of the participant may modulate the magnitude of the effect of caffeine on fat oxidation during exercise.


Absence of effects of prolonged simvastatin therapy on nocturnal sleep in a large randomized placebo-controlled study. Oxford Cholesterol Study Group.
1. It has been suggested that lipophilic HMG CoA reductase inhibitors, like lovastatin and simvastatin, may cause sleep disturbance. 2. Six hundred and twenty-one patients at increased risk of coronary heart disease were randomized in a single centre to receive 40 mg daily simvastatin, 20 mg daily simvastatin or matching placebo. To assess the effects of prolonged use of simvastatin on nocturnal sleep quality and duration, a sleep questionnaire was administered to 567 patients (95% of 595 survivors) at an average of 88 weeks (range: 44-129 weeks) after randomization. 3. The main outcome measures were sleep-related problems and use of sleep-enhancing medications reported during routine study follow-up visits, and responses to the sleep questionnaire about changes in sleep duration and about various sleep events during the preceding month. 4. No differences were observed between the treatment groups in the frequency of sleep-related problems reported, in the proportion of follow-up visits at which such problems were reported, or in the use of sleep-enhancing medications. The numbers who stopped study treatment were similar in the different treatment groups, and no patient stopped principally because of insomnia. In response to the sleep questionnaire, there were no significant differences between the treatment groups in reports of various sleep events during the preceding month, except that slightly fewer patients allocated simvastatin reported waking often. No differences in sleep duration were observed. 5. This placebo-controlled trial does not indicate any adverse effects of prolonged treatment with simvastatin on systematically sought measures of sleep disturbance.


C9ORF72 intermediate repeat expansion in patients affected by atypical parkinsonian syndromes or Parkinson's disease complicated by psychosis or dementia in a Sardinian population.
The hexanucleotide repeat expansion GGGGCC in the C9ORF72 gene larger than 30 repeats has been identified as a major genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Recent papers investigated the possible pathogenic role and associated clinical phenotypes of intermediate C9ORF72 repeat expansion ranging between 20 and 30 repeats. Some studies suggested its pathogenicity for typical Parkinson's disease (PD), atypical parkinsonian syndromes, FTD with/without parkinsonism, and ALS with/without parkinsonism or with/without dementia. In our study, we aimed to screen patients affected by atypical parkinsonian syndromes or PD complicated by psychosis or dementia for the presence of C9ORF72 repeat expansions, and in unrelated age- and sex-matched healthy controls. Consecutive unrelated patients with atypical parkinsonian syndromes and patients with PD complicated by psychosis or dementia were included in this study. Atypical parkinsonian syndromes were further divided into two groups: one with patients who met the criteria for the classic forms of atypical parkinsonism [multiple system atrophy (MSA), Lewy body disease (LBD), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD)] ;and patients who did not meet the above criteria, named non-classical atypical parkinsonism with or without dementia. Ninety-two unrelated patients (48 men, 44 women) were enrolled. None of the patients was found to be carriers of C9ORF72 repeat expansions with more than 30 repeats. Intermediate 20-30 repeat expansions were detected in four female patients (4.3 %). Three of them presented clinical features of atypical parkinsonian syndromes, two with non-classical atypical parkinsonism and dementia FTD-like, and one with non-classical atypical parkinsonism without dementia. The other patient presented clinical features of typical PD complicated by psychosis. Among 121 control subjects, none presented long or short expansion for the C9ORF72 gene. Our findings seem to support the hypothesis that the hexanucleotide expansions of C9ORF72 gene with intermediate repetitions between 20 and 29 repetitions could be associated with typical PD with psychosis or dementia and atypical parkinsonisms with dementia (non-classical atypical parkinsonism with dementia FTD-like) or without dementia (non-classical atypical parkinsonism upper MND-like), although the causal relationship is still unclear. In these latter patients, parkinsonism, more or less levodopa responsive, constituted the symptomatological central core at onset.


Positron emission tomographic measurements of pulvinar activity during an attention task.
Positron emission tomographic scans were recorded from human subjects following an object-identification task, one version of which required attentional selection and the other version of which did not. In one experimental session, the attention-demanding displays were presented in the left visual field and the nonattention displays were presented in the right visual field. In a second session, the sides of the displays were reversed. Analysis of the scans indicated that, averaged across the 2 sessions, the pulvinar showed greater glucose uptake when it was contralateral to the display of the selective attention task than when it was contralateral to the display of the nonattention task. The pattern of the data indicated that the degree of the attention task effect on pulvinar glucose uptake may differ between the hemispheres. In view of known connections between the pulvinar and cortical areas that mediate object identification, the present finding suggests that the pulvinar operates interactively with these cortical structures when an identification process demands selective attention.


Bubblewrap: Online tiling and real-time flow prediction on neural manifolds.
While most classic studies of function in experimental neuroscience have focused on the coding properties of individual neurons, recent developments in recording technologies have resulted in an increasing emphasis on the dynamics of neural populations. This has given rise to a wide variety of models for analyzing population activity in relation to experimental variables, but direct testing of many neural population hypotheses requires intervening in the system based on current neural state, necessitating models capable of inferring neural state online. Existing approaches, primarily based on dynamical systems, require strong parametric assumptions that are easily violated in the noise-dominated regime and do not scale well to the thousands of data channels in modern experiments. To address this problem, we propose a method that combines fast, stable dimensionality reduction with a soft tiling of the resulting neural manifold, allowing dynamics to be approximated as a probability flow between tiles. This method can be fit efficiently using online expectation maximization, scales to tens of thousands of tiles, and outperforms existing methods when dynamics are noise-dominated or feature multi-modal transition probabilities. The resulting model can be trained at kiloHertz data rates, produces accurate approximations of neural dynamics within minutes, and generates predictions on submillisecond time scales. It retains predictive performance throughout many time steps into the future and is fast enough to serve as a component of closed-loop causal experiments.


EAP1/Daxx interacts with ETS1 and represses transcriptional activation of ETS1 target genes.
ETS1 is a member of the evolutionarily conserved family of ets genes, which are transcription factors that bind to unique DNA sequences, either alone or by association with other proteins. In this study, we have used the yeast two-hybrid system to identify an ETS1 interacting protein. The ETS1 N-terminal amino acid region was used as bait and an interaction was identified with the Daxx protein, referred to as EAP1 (ETS1 Associated Protein 1)/Daxx. This interactin has been shown to exist in yeast and in vitro. EAP1/Daxx and ETS1 are co-localized in the nucleus of mammalian cells. The region in EAP1/Daxx which specifically binds to ETS1 is located within its carboxy terminal 173 amino acid region. The ETS1 interaction region is located within its N-terminal 139 amino acids and is referred as the Daxx Interaction Domain (DID). The DID appears to be conserved in several other ets family members, as well as in other proteins known to interact with Daxx. The EAP1/Daxx interacts with both isoforms of ETS1, p51-ETS1 and p42-ETS1. Interaction of EAP1/Daxx with ETS1 causes the repression of transcriptional activation of the MMP1 and BCL2 genes. The interaction domains of both ETS1 and EAP1/Daxx are required for this repression and deletion of either domain abolishes this activity.


Detection and Quantification of β-Amyloid, Pyroglutamyl Aβ, and Tau in Aged Canines.
Canine cognitive dysfunction syndrome is an age-associated disorder that resembles many aspects of human Alzheimer disease. The characterization of canine cognitive dysfunction syndrome has been restricted to selected laboratory dogs and mongrels, thereby limiting our knowledge of potential breed-related and age-related differences. We examined the brains of 24 dogs from various breeds. The frontal cortex, hippocampus, and entorhinal cortex were investigated. Deposits of β-amyloid (Aβ) and tau were analyzed phenotypically and quantified stereologically. In all dogs aged 10 years or older, plaques containing pyroglutamyl Aβ and Aβ8-17 were detected. Within the ventral hippocampus, significantly more pyroglutamyl Aβ plaques were deposited in small and medium dogs than in large dogs. Hyperphosphorylated tau with formation of neurofibrillary tangles was observed in 3 animals aged 13 to 15 years. This study provides the first investigation of pyroglutamyl Aβ in comparison with total Aβ (as shown by Aβ8-17 immunoreactivity) in dogs of different breeds, sizes, and ages. Our results indicate that canine cognitive dysfunction syndrome is relatively common among aged canines, thereby emphasizing the relevance of such populations to translational Alzheimer disease research.


Internal models of limb geometry in the control of hand compliance.
The aim of this article is to describe the role of some neural mechanisms in the adaptive control of limb compliance during preplanned mechanical interaction with objects. We studied the EMG responses and the kinematic responses evoked by pseudorandom perturbations continuously applied by means of a torque motor before and during a catching task. The temporal changes of these responses were studied by means of an identification technique for time-varying systems. We found a transient reversal of EMG stretch reflex responses centered on the time of ball impact on the hand; this reversal results in a transient coactivation of antagonist muscles at both the elbow and the wrist. The kinematic responses describe the relation between torque input and position output. Thus, they provide a global measure of limb compliance. The changes in limb compliance during catching were quantified by computing error criteria either in the Cartesian coordinates of the hand or in the angular coordinates of the elbow and wrist joints. We found that only the hand compliance in Cartesian coordinates is consistently minimized around impact, in coincidence with the transient reversal of the stretch reflex responses. By contrast, the error criteria expressed in the angular coordinates of the joints have a variable time course and are not minimized around impact. It is known that hand compliance depends on both the pattern of muscle activities and the geometrical configuration of the limb. Therefore, the lack of consistent correlation between the changes in hand compliance and the changes in the geometrical configuration of the limb during catching indicates that the gating of the stretch reflex responses around impact time is based on an internal model of limb geometry.


Calcium/calmodulin-dependent protein kinase II links ER stress with Fas and mitochondrial apoptosis pathways.
ER stress-induced apoptosis is implicated in various pathological conditions, but the mechanisms linking ER stress-mediated signaling to downstream apoptotic pathways remain unclear. Using human and mouse cell culture and in vivo mouse models of ER stress-induced apoptosis, we have shown that cytosolic calcium resulting from ER stress induces expression of the Fas death receptor through a pathway involving calcium/calmodulin-dependent protein kinase IIgamma (CaMKIIgamma) and JNK. Remarkably, CaMKIIgamma was also responsible for processes involved in mitochondrial-dependent apoptosis, including release of mitochondrial cytochrome c and loss of mitochondrial membrane potential. CaMKII-dependent apoptosis was also observed in a number of cultured human and mouse cells relevant to ER stress-induced pathology, including cultured macrophages, endothelial cells, and neuronal cells subjected to proapoptotic ER stress. Moreover, WT mice subjected to systemic ER stress showed evidence of macrophage mitochondrial dysfunction and apoptosis, renal epithelial cell apoptosis, and renal dysfunction, and these effects were markedly reduced in CaMKIIgamma-deficient mice. These data support an integrated model in which CaMKII serves as a unifying link between ER stress and the Fas and mitochondrial apoptotic pathways. Our study also revealed what we believe to be a novel proapoptotic function for CaMKII, namely, promotion of mitochondrial calcium uptake. These findings raise the possibility that CaMKII inhibitors could be useful in preventing apoptosis in pathological settings involving ER stress-induced apoptosis.


Physical and economic consequences of climate change in Europe.
Quantitative estimates of the economic damages of climate change usually are based on aggregate relationships linking average temperature change to loss in gross domestic product (GDP). However, there is a clear need for further detail in the regional and sectoral dimensions of impact assessments to design and prioritize adaptation strategies. New developments in regional climate modeling and physical-impact modeling in Europe allow a better exploration of those dimensions. This article quantifies the potential consequences of climate change in Europe in four market impact categories (agriculture, river floods, coastal areas, and tourism) and one nonmarket impact (human health). The methodology integrates a set of coherent, high-resolution climate change projections and physical models into an economic modeling framework. We find that if the climate of the 2080s were to occur today, the annual loss in household welfare in the European Union (EU) resulting from the four market impacts would range between 0.2-1%. If the welfare loss is assumed to be constant over time, climate change may halve the EU's annual welfare growth. Scenarios with warmer temperatures and a higher rise in sea level result in more severe economic damage. However, the results show that there are large variations across European regions. Southern Europe, the British Isles, and Central Europe North appear most sensitive to climate change. Northern Europe, on the other hand, is the only region with net economic benefits, driven mainly by the positive effects on agriculture. Coastal systems, agriculture, and river flooding are the most important of the four market impacts assessed.


The intake of a high-fat diet and grape seed procyanidins induces gene expression changes in peripheral blood mononuclear cells of hamsters: capturing alterations in lipid and cholesterol metabolisms.
We previously demonstrated that hamsters that were fed either a standard diet (STD) or a high-fat diet (HFD) and treated with a grape seed procyanidin extract (GSPE) showed decreased adiposity and circulating levels of free fatty acids compared with hamsters treated with a vehicle (Caimari et al. in Int J Obes 37:576-83, 2013, doi: 10.1038/ijo.2012.75 ). Here, we tested whether the gene expression changes in peripheral blood mononuclear cells (PBMCs) can reflect these metabolic effects and the dyslipidaemia produced by the HFD feeding in the same cohort of animals. The mRNA levels of a subset of genes were also studied in the liver in order to evaluate the capacity of PBMCs to reflect the metabolic adaptations that occur in this organ. In PBMCs, we reported a simultaneous up-regulation of the lipid-related genes involved in both the anabolic (pparγ, acc1 and gpat) and the catabolic (pparα, ucp2, atgl and hsl) pathways in response to the GSPE treatment, similar but no identical to previous observations in retroperitoneal white adipose tissues of these animals. Furthermore, the key cholesterol metabolism genes srebp2 and ldlr were significantly down-regulated in PBMCs of both HFD-fed groups compared with the STD groups. Although the expression of srebp2 in the liver followed a similar pattern to that obtained in PBMCs, no comparable changes were found between the liver and PBMCs in the expression of most of the studied genes. In conclusion, our results highlight the potential of PBMCs as a high accessible tissue for the indirect study of cholesterol and adipose tissue metabolism dynamics.


Primordium of an artificial Bruch's membrane made of nanofibers for engineering of retinal pigment epithelium cell monolayers.
Transplanted retinal pigment epithelium (RPE) cells hold promise for treatment of age-related macular degeneration (AMD) and Stargardt disease (SD), but it is conceivable that the degenerated host Bruch's membrane (BM) as a natural substrate for RPE might not optimally support transplanted cell survival with correct cellular organization. We fabricated novel ultrathin three-dimensional (3-D) nanofibrous membranes from collagen type I and poly(lactic-co-glycolic acid) (PLGA) by an advanced clinical-grade needle-free electrospinning process. The nanofibrillar 3-D networks closely mimicked the fibrillar architecture of the native inner collagenous layer of human BM. Human RPE cells grown on our nanofibrous membranes bore a striking resemblance to native human RPE. They exhibited a correctly orientated monolayer with a polygonal cell shape and abundant sheet-like microvilli on their apical surfaces. RPE cells built tight junctions and expressed RPE65 protein. Flat 2-D PLGA film and cover glass as controls delivered inferior RPE layers. Our nanofibrous membranes may imitate the natural BM to such extent that they allow for the engineering of an in vivo-like human RPE monolayer that maintains the natural biofunctional characteristics. Such ultrathin membranes may provide a promising vehicle for a functional RPE cell monolayer implantation in the subretinal space in patients with AMD or SD.


Spectrum of Protein Location in Proteomes Captures Evolutionary Relationship Between Species.
The native subcellular location (also referred to as localization or cellular compartment) of a protein is the one in which it acts most frequently; it is one aspect of protein function. Do ten eukaryotic model organisms differ in their location spectrum, i.e., the fraction of its proteome in each of seven major cellular compartments? As experimental annotations of locations remain biased and incomplete, we need prediction methods to answer this question. After systematic bias corrections, the complete but faulty prediction methods appeared to be more appropriate to compare location spectra between species than the incomplete more accurate experimental data. This work compared the location spectra for ten eukaryotes: Homo sapiens (human), Gorilla gorilla (gorilla), Pan troglodytes (chimpanzee), Mus musculus (mouse), Rattus norvegicus (rat), Drosophila melanogaster (fruit/vinegar fly), Anopheles gambiae (African malaria mosquito), Caenorhabitis elegans (nematode), Saccharomyces cerevisiae (baker's yeast), and Schizosaccharomyces pombe (fission yeast). The two largest classes were predicted to be the nucleus and the cytoplasm together accounting for 47-62% of all proteins, while 7-21% of the proteins were predicted in the plasma membrane and 4-15% to be secreted. Overall, the predicted location spectra were largely similar. However, in detail, the differences sufficed to plot trees (UPGMA) and 2D (PCA) maps relating the ten organisms using a simple Euclidean distance in seven states (location classes). The relations based on the simple predicted location spectra captured aspects of cross-species comparisons usually revealed only by much more detailed evolutionary comparisons. Most interestingly, known phylogenetic relations were reproduced better by paralog-only than by ortholog-only trees.


Habituation of the cold shock response is inhibited by repeated anxiety: Implications for safety behaviour on accidental cold water immersions.
Accidental cold-water immersion (CWI) triggers the life-threatening cold shock response (CSR) which is a precursor to sudden death on immersion. One practical means of reducing the CSR is to induce an habituation by undergoing repeated short CWIs. Habituation of the CSR is known to be partially reversed by the concomitant experience of acute anxiety, raising the possibility that repeated anxiety could prevent CSR habituation; we tested this hypothesis. Sixteen participants (12 male, 4 female) completed seven, seven-minute immersions in to cold water (15°C). Immersion one acted as a control (CON1). During immersions two to five, which would ordinarily induce an habituation, anxiety levels were repeatedly increased (CWI-ANXrep) by deception and a demanding mathematical task. Immersions six and seven were counter-balanced with another high anxiety condition (CWI-ANXrep) or a further control (CON2). Anxiety (20cm visual analogue scale) and cardiorespiratory responses (cardiac frequency [fc], respiratory frequency [fR], tidal volume [VT], minute ventilation [V̇E]) were measured. Comparisons were made between experimental immersions (CON1, final CWI-ANXrep, CON2), across habituation immersions and with data from a previous study. Anxiety levels were sustained at a similar level throughout the experimental and habituation immersions (mean [SD] CON1: 7.0 [4.0] cm; CON2: 5.8 [5.2] cm cf CWI-ANXrep: 7.3 [5.5] cm; p>0.05). This culminated in failure of the CSR to habituate even when anxiety levels were not manipulated (i.e. CON2). These data were different (p<0.05) to previous studies where anxiety levels were allowed to fall across habituation immersions and the CSR consequently habituated. Repeated anxiety prevented CSR habituation. A protective strategy that includes inducing habituation for those at risk should include techniques to lower anxiety associated with the immersion event or habituation may not be beneficial in the emergency scenario.


Telomeric G-quadruplexes are a substrate and site of localization for human telomerase.
It has been hypothesized that G-quadruplexes can sequester the 3' end of the telomere and prevent it from being extended by telomerase. Here we purify and characterize stable, conformationally homogenous human telomeric G-quadruplexes, and demonstrate that human telomerase is able to extend parallel, intermolecular conformations in vitro. These G-quadruplexes align correctly with the RNA template of telomerase, demonstrating that at least partial G-quadruplex resolution is required. A highly purified preparation of human telomerase retains this extension ability, establishing that the core telomerase enzyme complex is sufficient for partial G-quadruplex resolution and extension. The parallel-specific G-quadruplex ligand N-methyl mesoporphyrin IX (NMM) causes an increase in telomeric G-quadruplexes, and we show that telomerase colocalizes with a subset of telomeric G-quadruplexes in vivo. The ability of telomerase to partially unwind, extend and localize to these structures implies that parallel telomeric G-quadruplexes may play an important biological role.


Moderate activation of IKK2-NF-kB in unstressed adult mouse liver induces cytoprotective genes and lipogenesis without apparent signs of inflammation or fibrosis.
The NF-kB signaling, regulated by IKK1-p52/RelB and IKK2-p65, is activated by various stresses to protect or damage the liver, in context-specific manners. Two previous studies of liver-specific expression of constitutive active IKK2 (IKK2ca) showed that strong activation of IKK2-NF-kB in mouse livers caused inflammation, insulin resistance, and/or fibrosis. The purpose of this study was to understand how moderate activation of IKK2-NF-kB in adult mouse livers alters hepatic gene expression and pathophysiology. We generated mice with adult hepatocyte-specific activation of Ikk2 (Liv-Ikk2ca) using Alb-cre mice and Ikk2ca Rosa26 knockin mice in which a moderate expression of Ikk2ca transgene was driven by the endogenous Rosa26 promoter. Surprisingly, compared to wild-type mice, adult male Liv-Ikk2ca mice had higher hepatic mRNA expression of Ikk2 and classical NF-kB targets (e.g. Lcn2 and A20), as well as IKK1, NIK, and RelB, but no changes in markers of inflammation or fibrosis. Blood levels of IL-6 and MCP-1 remained unchanged, and histology analysis showed a lack of injury or infiltration of inflammatory cells in livers of Liv-Ikk2ca mice. Moreover, Liv-Ikk2ca mice had lower mRNA expression of prooxidative enzymes Cyp2e1 and Cyp4a14, higher expression of antioxidative enzymes Sod2, Gpx1, and Nqo1, without changes in key enzymes for fatty acid oxidation, glucose utilization, or gluconeogenesis. In parallel, Liv-Ikk2ca mice and wild-type mice had similar levels of hepatic reduced glutathione, endogenous reactive oxygen species, and lipid peroxidation. Additionally, Liv-Ikk2ca mice had higher Cyp3a11 without down-regulation of most drug processing genes. Regarding nuclear proteins of NF-kB subunits, Liv-Ikk2ca mice had moderately higher p65 and p50 but much higher RelB. Results of ChIP-qPCR showed that the binding of p50 to multiple NF-kB-target genes was markedly increased in Liv-Ikk2ca mice. Additionally, Liv-Ikk2ca mice had moderate increase in triglycerides in liver, which was associated with higher lipogenic factors Pparγ, Lxr, Fasn, Scd1, and CD36. In summary, moderate activation of IKK2-NF-kB in unstressed adult mouse hepatocytes produces a cytoprotective gene expression profile and induces lipogenesis without apparent signs of inflammation or fibrosis, likely due to strong activation of the anti-inflammatory IKK1-RelB alternative NF-kB pathway as well as the Lxr.


Guidelines for the Acute Treatment of Cerebral Edema in Neurocritical Care Patients.
Acute treatment of cerebral edema and elevated intracranial pressure is a common issue in patients with neurological injury. Practical recommendations regarding selection and monitoring of therapies for initial management of cerebral edema for optimal efficacy and safety are generally lacking. This guideline evaluates the role of hyperosmolar agents (mannitol, HTS), corticosteroids, and selected non-pharmacologic therapies in the acute treatment of cerebral edema. Clinicians must be able to select appropriate therapies for initial cerebral edema management based on available evidence while balancing efficacy and safety. The Neurocritical Care Society recruited experts in neurocritical care, nursing, and pharmacy to create a panel in 2017. The group generated 16 clinical questions related to initial management of cerebral edema in various neurological insults using the PICO format. A research librarian executed a comprehensive literature search through July 2018. The panel screened the identified articles for inclusion related to each specific PICO question and abstracted necessary information for pertinent publications. The panel used GRADE methodology to categorize the quality of evidence as high, moderate, low, or very low based on their confidence that the findings of each publication approximate the true effect of the therapy. The panel generated recommendations regarding initial management of cerebral edema in neurocritical care patients with subarachnoid hemorrhage, traumatic brain injury, acute ischemic stroke, intracerebral hemorrhage, bacterial meningitis, and hepatic encephalopathy. The available evidence suggests hyperosmolar therapy may be helpful in reducing ICP elevations or cerebral edema in patients with SAH, TBI, AIS, ICH, and HE, although neurological outcomes do not appear to be affected. Corticosteroids appear to be helpful in reducing cerebral edema in patients with bacterial meningitis, but not ICH. Differences in therapeutic response and safety may exist between HTS and mannitol. The use of these agents in these critical clinical situations merits close monitoring for adverse effects. There is a dire need for high-quality research to better inform clinicians of the best options for individualized care of patients with cerebral edema.