Infectivity in chimpanzees (Pan troglodytes) of plasma collected before HCV RNA detectability by FDA-licensed assays: implications for transfusion safety and HCV infection outcomes.
Serial plasma aliquots (50 mL) obtained from 10 commercial donors who converted from hepatitis C virus (HCV) RNA negative to positive were transfused into 2 chimpanzees to assess infectivity during early HCV infection. Plasma, obtained 4 days before HCV RNA detectability by licensed assays, transmitted HCV infection to chimpanzee X355. The infectious PCR-negative plasma was subsequently shown to be positive in 2 of 23 replicates using a sensitive transcription-mediated amplification (TMA) assay, and estimated to contain 1.2 HCV RNA copies/mL (60 copies/50 mL transfused). Plasma units obtained up to 8 weeks earlier were not infectious in a second susceptible chimp, even when from donors with low-level, intermittent HCV RNA detection. Chimp x355 developed acute viremia with subsequent seroconversion, but cleared both virus and Ab in 17 weeks. When rechallenged 38 months later with 6000 RNA copies/mL from the same donor, X355 was transiently reinfected and again rapidly lost all HCV markers. We conclude that: (1) transfusions can transmit HCV infection before RNA detection, but the interval of test-negative infectivity is very brief; (2) early "blips" of HCV RNA appear noninfectious and can be ignored when calculating residual transfusion risk; and (3) markers of HCV infection can be lost rapidly after exposure to low-dose inocula.


Eight New Species of Charinus Simon, 1892 (Arachnida: Amblypygi: Charinidae) Endemic for the Brazilian Amazon, with Notes on Their Conservational Status.
Eight new species of Charinus Simon, 1892 are described for the Brazilian Amazon, from the states of Pará (C. bichuetteae sp. n., C. bonaldoi sp. n., C. carajas sp. n., C. ferreus sp. n., C. guto sp. n. and C. orientalis sp. n.) and Amazonas (Charinus brescoviti sp. n. and C. ricardoi sp. n.). All new species can be differentiated from the other species of the genus by the number of pseudo-articles in basitibia IV, the presence/absence of median eyes, and the shape of the female gonopod. Brazil now becomes the country with the largest diversity of Amblypygi in the world, with 25 known species. Half of the new species described here have a high degree of endangerment: C. bichuetteae sp. n. is threatened by the flood caused by the hydroelectric dam of Belo Monte, and C. carajas sp. n., C. ferreus sp. n. and C. orientalis sp. n. are endangered by the iron mining in Carajás municipality and surroundings. The Charinus species here described are endemic to the Amazon Region, so in order to assure their preservation, it is strongly recommended a special care with their habitats (type localities) which are facing increasing rates of destruction and deforestation.


Expression of the RNA-binding protein RBM3 is associated with a favourable prognosis and cisplatin sensitivity in epithelial ovarian cancer.
We recently demonstrated that increased expression of the RNA-binding protein RBM3 is associated with a favourable prognosis in breast cancer. The aim of this study was to examine the prognostic value of RBM3 mRNA and protein expression in epithelial ovarian cancer (EOC) and the cisplatin response upon RBM3 depletion in a cisplatin-sensitive ovarian cancer cell line. RBM3 mRNA expression was analysed in tumors from a cohort of 267 EOC cases (Cohort I) and RBM3 protein expression was analysed using immunohistochemistry (IHC) in an independent cohort of 154 prospectively collected EOC cases (Cohort II). Kaplan Meier analysis and Cox proportional hazards modelling were applied to assess the relationship between RBM3 and recurrence free survival (RFS) and overall survival (OS). Immunoblotting and IHC were used to examine the expression of RBM3 in a cisplatin-resistant ovarian cancer cell line A2780-Cp70 and its cisplatin-responsive parental cell line A2780. The impact of RBM3 on cisplatin response in EOC was assessed using siRNA-mediated silencing of RBM3 in A2780 cells followed by cell viability assay and cell cycle analysis. Increased RBM3 mRNA expression was associated with a prolonged RFS (HR = 0.64, 95% CI = 0.47-0.86, p = 0.003) and OS (HR = 0.64, 95% CI = 0.44-0.95, p = 0.024) in Cohort I. Multivariate analysis confirmed that RBM3 mRNA expression was an independent predictor of a prolonged RFS, (HR = 0.61, 95% CI = 0.44-0.84, p = 0.003) and OS (HR = 0.62, 95% CI = 0.41-0.95; p = 0.028) in Cohort I. In Cohort II, RBM3 protein expression was associated with a prolonged OS (HR = 0.53, 95% CI = 0.35-0.79, p = 0.002) confirmed by multivariate analysis (HR = 0.61, 95% CI = 0.40-0.92, p = 0.017). RBM3 mRNA and protein expression levels were significantly higher in the cisplatin sensitive A2780 cell line compared to the cisplatin resistant A2780-Cp70 derivative. siRNA-mediated silencing of RBM3 expression in the A2780 cells resulted in a decreased sensitivity to cisplatin as demonstrated by increased cell viability and reduced proportion of cells arrested in the G2/M-phase. These data demonstrate that RBM3 expression is associated with cisplatin sensitivity in vitro and with a good prognosis in EOC. Taken together these findings suggest that RBM3 may be a useful prognostic and treatment predictive marker in EOC.


Predicting the release profile of small molecules from within the ordered nanostructured lipidic bicontinuous cubic phase using translational diffusion coefficients determined by PFG-NMR.
The ordered nanostructured lipidic bicontinuous cubic phase has demonstrated potential as a drug release material, due to its ability to encapsulate a wide variety of compounds, which may undergo sustained, diffusion controlled release over time. Control of drug release has been shown to depend on the nanostructural parameters of the lipid mesophase. Herein, the diffusion and release of two amino acids, encapsulated within a range of different lipidic cubic mesophases are investigated. Pulsed-field gradient NMR was used to determine the diffusion coefficient of the encapsulated amino acid, which was found to be correlated with the nanoscale diameter of the water channels within the cubic mesophase. This information was used to predict the release profiles of encapsulated compounds from within the cubic mesophase, which was verified by directly measuring the release of each amino acid in vitro. Predicted release profiles tracked reasonably close to the measured release profiles, indicating that NMR determined diffusion measurements can be used to predict release profiles.


Oxygen sensing, hypoxia-inducible factor-1 and the regulation of mammalian gene expression.
A great many aspects of the anatomy and physiology of large animals are constrained by the need to match oxygen supply to cellular metabolism and appear likely to involve the regulation of gene expression by oxygen. Some insight into possible underlying mechanisms has been provided by studies of erythropoietin, a haemopoietic growth factor which stimulates red cell production in response to hypoxia. Studies of hypoxia-inducible cis-acting sequences from the erythropoietin gene have led to the recognition of a widespread transcriptional response to hypoxia based on the activation of a DNA-binding complex termed hypoxia-inducible factor-1 (HIF-1). Perturbation of the transcriptional response by particular transition metal ions, iron chelators and certain redox-active agents have suggested a specific oxygen sensing mechanism, perhaps involving a haem protein in a flavoprotein/cytochrome system. In addition to erythropoietin, HIF-1-responsive genes include examples with functions in cellular energy metabolism, iron metabolism, catecholamine metabolism, vasomotor control and angiogenesis, suggesting an important role in the coordination of oxygen supply and cellular metabolism. In support of this, we have demonstrated an important role for HIF-1 in tumour angiogenesis. HIF-1 itself consists of a heterodimer of two basic-helix-loop-helix proteins of the PAS family, termed HIF-1alpha and HIF-1beta, although other closely related members of this family may also contribute to the response to hypoxia. We have fused domains of HIF-1 genes to heterologous transcription factors to assay for regulatory function. These experiments have defined several domains in HIF-1alpha which can independently confer the hypoxia-inducible property, and they suggest a mechanism of HIF-1 activation in which post-translational activation/derepression of HIF-1alpha is amplified by changes in HIF-1alpha abundance most probably arising from suppression of proteolytic breakdown. Pursuit of the mechanism(s) underlying these processes should ultimately lead to better definition of the oxygen-sensing process.


Advancing vector biology research: a community survey for future directions, research applications and infrastructure requirements.
Vector-borne pathogens impact public health, animal production, and animal welfare. Research on arthropod vectors such as mosquitoes, ticks, sandflies, and midges which transmit pathogens to humans and economically important animals is crucial for development of new control measures that target transmission by the vector. While insecticides are an important part of this arsenal, appearance of resistance mechanisms is increasingly common. Novel tools for genetic manipulation of vectors, use of Wolbachia endosymbiotic bacteria, and other biological control mechanisms to prevent pathogen transmission have led to promising new intervention strategies, adding to strong interest in vector biology and genetics as well as vector-pathogen interactions. Vector research is therefore at a crucial juncture, and strategic decisions on future research directions and research infrastructure investment should be informed by the research community. A survey initiated by the European Horizon 2020 INFRAVEC-2 consortium set out to canvass priorities in the vector biology research community and to determine key activities that are needed for researchers to efficiently study vectors, vector-pathogen interactions, as well as access the structures and services that allow such activities to be carried out. We summarize the most important findings of the survey which in particular reflect the priorities of researchers in European countries, and which will be of use to stakeholders that include researchers, government, and research organizations.


An E-Learning Adaptation of an Evidence-Based Media Literacy Curriculum to Prevent Youth Substance Use in Community Groups: Development and Feasibility of REAL Media.
There is a need for evidence-based substance use prevention efforts that target high school-aged youth that are easy to implement and suitable for dissemination in school and community groups. The Youth Message Development (YMD) program is a brief, four-lesson, in-person curriculum that aims to prevent youth substance use through the development of youth media literacy. Specifically, YMD aims to increase understanding of advertising reach and costs, along with the techniques used to sell products; develop counterarguing and critical thinking skills in response to advertisements; and facilitate application of these skills to the development of youth-generated antisubstance messages. Although YMD has demonstrated evidence of success, it is limited by its delivery method and focus on alcohol and smoking. Study objectives were two-fold: (1) to adapt the YMD curriculum to a self-paced, interactive, electronic-learning (e-learning) format and expand its content to cover alcohol, combustible cigarettes, e-cigarettes, smokeless tobacco, marijuana, and prescription drugs, and (2) to test the feasibility of the adapted curriculum in partnership with a national youth organization. An iterative process was employed in partnership with the 4-H youth development organization and a technology developer and consisted of six phases: (1) focus groups to guide adaptation, (2) adaptation to an e-learning format renamed REAL media, (3) pilot-testing of the REAL media prototype to determine feasibility and acceptability, (4) program revisions, (5) usability testing of the revised prototype, and (6) final revisions. Focus groups and pilot and usability testing were conducted with 4-H youth club members and adult club leaders. Focus group feedback guided the build of an e-learning prototype of REAL media, which consisted of five online levels and interactive content guided by a mix of narration and on-screen text. Results of a pilot test of the prototype were neutral to positive, and the program was refined based on end-user feedback. An independent usability test indicated that youth 4-H members felt favorably about navigating REAL media, and they reported high self-efficacy in applying skills learned in the program. Additional refinements to the program were made based on their feedback. The iterative build process involving the end user from the outset yielded an overall successful technology-driven adaptation of an evidence-based curriculum. This should increase the likelihood of effectively impacting behavioral outcomes as well as uptake within community organizations.


Distinct profiles of brain atrophy in frontotemporal lobar degeneration caused by progranulin and tau mutations.
Neural network breakdown is a key issue in neurodegenerative disease, but the mechanisms are poorly understood. Here we investigated patterns of brain atrophy produced by defined molecular lesions in the two common forms of genetically mediated frontotemporal lobar degeneration (FTLD). Nine patients with progranulin (GRN) mutations and eleven patients with microtubule-associated protein tau (MAPT) mutations had T1 MR brain imaging. Brain volumetry and grey and white matter voxel-based morphometry (VBM) were used to assess patterns of cross-sectional atrophy in the two groups. In a subset of patients with longitudinal MRI rates of whole-brain atrophy were derived using the brain-boundary-shift integral and a VBM-like analysis of voxel-wise longitudinal volume change was performed. The GRN mutation group showed asymmetrical atrophy whereas the MAPT group showed symmetrical atrophy. Brain volumes were smaller in the GRN group with a faster rate of whole-brain atrophy. VBM delineated a common anterior cingulate-prefrontal-insular pattern of atrophy in both disease groups. Additional disease-specific profiles of grey and white matter loss were identified on both cross-sectional and longitudinal imaging: GRN mutations were associated with asymmetrical inferior frontal, temporal and inferior parietal lobe grey matter atrophy and involvement of long intrahemispheric association white matter tracts, whereas MAPT mutations were associated with symmetrical anteromedial temporal lobe and orbitofrontal grey matter atrophy and fornix involvement. The findings suggest that the effects of GRN and MAPT mutations are expressed in partly overlapping but distinct anatomical networks that link specific molecular dysfunction with clinical phenotype.


The Neuroprotective Effect of Glycyrrhizic Acid on an Experimental Model of Focal Cerebral Ischemia in Rats.
Cerebral ischemia is still one of the most important topics in neurosciences. Our study aimed to investigate the neuroprotective and anti-oxidant effects of glycyrrhizic acid on focal cerebral ischemia in rats. Twenty-four rats were divided equally into three groups. A middle cerebral artery occlusion model was performed in this study where sham and glycyrrhizic acid were administered intraperitoneally following middle cerebral artery occlusion. Group I was evaluated as control. Malondialdehyde (MDA), superoxide dismutase (SOD), and nuclear respiratory factor-1 (NRF1) levels were analyzed biochemically on the right cerebral hemisphere, while ischemic histopathological studies were completed to investigate the anti-oxidant status. Biochemical results showed that SOD and NRF1 levels were significantly increased in the glycyrrhizic acid group compared with the sham group while MDA levels were significantly decreased. On histopathological examination, cerebral edema, vacuolization, degeneration, and destruction of neurons were decreased in the glycyrrhizic acid group compared with the sham group. Cerebral ischemia was attenuated by glycyrrhizic acid administration. These observations indicate that glycyrrhizic acid may have potential as a therapeutic agent in cerebral ischemia by preventing oxidative stress.


Stress-induced changes in spatial memory are sexually differentiated and vary across the lifespan.
Stress exposure, depending on intensity and duration, elicits adaptive or maladaptive physiological changes. The same general pattern of advantageous versus deleterious stress effects appears to exist for some cognitive functions, particularly spatial learning and memory performance. This article reviews sex differences in response to stress on a variety of spatial tasks. In general, females are more resistant than males to stress-induced impairments on spatial tasks, including the radial arm maze and object placement. In young adulthood, chronic stress (restraint, 6 h per day for 21 days) impairs male performance on both tasks but leads to behavioural enhancements in females. Furthermore, these sex-dependent stress effects are influenced by both organisational and activational oestrogenic effects. Additionally, sex-specific stress responses vary depending on developmental age at the time of stress exposure. Male behavioural stress responses appear fixed across the lifespan (i.e. stress-induced cognitive impairments) whereas female stress responses appear more variable (i.e. stress-induced enhancements observed in young adulthood are different in response to prenatal stress and diminished following stress exposure at old age). These findings underscore the point that many effects obtained in males cannot be generalised to females and highlight the need to investigate the stress response at different ages and in both sexes.


Testing Wound-healing Activity in T15 Fibroblast Cultures: A Morphometric Analysis.
The purpose was to evaluate the use of mouse T15 fibroblast cell cultures for the investigation of wound-healing activity. In order to investigate their mechanisms of action, the effects of drugs with wound-healing activities were compared by using morphometric analyses by microscopy after cell staining. A number of parameters were used to evaluate the effects of titrated extracts from Centella asiatica and dexpanthenol (drugs that have been used in medical practice for their wound-healing activities) on cultured mouse T15 fibroblasts. These parameters were : the total number of cells ; the number of T15 cells in mitosis ; the percentages of fusiform, polygonal, round and vacuole-containing cells ; and the number of intracellular collagen granules. The results indicate that these two drugs exhibit wound-healing activities by activating fibroblast cells, and have cytoprotective effects, although their mechanisms of action on mouse T15 fibroblasts were different. On the basis of our findings, mouse T15 fibroblast cell cultures seem to be useful for the pharmacological screening of compounds with wound-healing activity.


Human adipose-derived stem cells ameliorate repetitive behavior, social deficit and anxiety in a VPA-induced autism mouse model.
Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder characterized by impairments in social interaction and communication, and patients often display co-occurring repetitive behaviors. Although the global prevalence of ASD has increased over time, the etiology and treatments for ASD are poorly understood. Recently, some researchers have suggested that stem cells have therapeutic potential for ASD. Thus, in the present study, we investigated the therapeutic effects of human adipose-derived stem cells (hASCs), a kind of autologous mesenchymal stem cells (MSCs) isolated from adipose tissue, on valproic acid (VPA)-induced autism model mice. Human ASCs were injected into the neonatal pups (P2 or P3) intraventricularly and then we evaluated major behavior symptoms of ASD. VPA-treated mice showed increased repetitive behaviors, decreased social interactions and increased anxiety but these autistic behaviors were ameliorated through transplantation of hASCs. In addition, hASCs transplantation restored the alteration of phosphatase and tensin homolog (PTEN) expression and p-AKT/AKT ratio in the brains of VPA-induced ASD model mice. The decreased level of vascular endothelial growth factor (VEGF) and interleukin 10 (IL-10) by VPA were rescued in the brains of the hASC-injected VPA mice. With these results, we experimentally found hASCs' therapeutic effects on autistic phenotypes in a ASD model mice for the first time. This animal model system can be used to elucidate further mechanisms of therapeutic effects of hASCs in ASD.


Visible quantum cutting in LiGdF4:Eu3+ through downconversion
For mercury-free fluorescent lamps and plasma display panels, alternative luminescent materials are required for the efficient conversion of vacuum ultraviolet radiation to visible light. Quantum cutting involving the emission of two visible photons for each vacuum ultraviolet photon absorbed is demonstrated in Eu3+-doped LiGdF4 with the concept of downconversion. Upon excitation of Gd3+ with a high-energy photon, two visible photons can be emitted by Eu3+ through an efficient two-step energy transfer from Gd3+ to Eu3+, with a quantum efficiency that approaches 200 percent.


Experiences with a vaccine being developed for the control of swine dysentery.
A prototype vaccine that is being developed for the control of swine dysentery (SD) was tested in two groups of experimental pigs. Vaccination induced high circulating antibody titres against the aetiological agent, Serpulina (Treponema) hyodysenteriae. Pigs in the first trial were vaccinated twice before being challenged orally with the bacteria. Five of 6 unvaccinated animals developed dysentery within a fortnight of challenge, but only 1 of 6 vaccinated pigs showed signs of disease at this time. Unexpectedly, 1 mo after challenge, the surviving unvaccinated pig and 2 remaining healthy vaccinated animals succumbed to the disease. The reason for the development of this late-onset form of dysentery was not clear. In the second trial, 8 pigs were vaccinated 3 times. Only 2 of these animals (25%) developed severe dysentery after being mixed with infected pigs, whereas 7 of 8 (88%) unvaccinated control pigs in the same pen became diseased. The late-onset form of dysentery was not observed. The prototype vaccine for SD provided a useful level of protection, and could be used in programs to control the disease in Australia.


Rockefeller braces for Baltimore.
In the article "Rockefeller braces for Baltimore" by Barbara J. Culliton (News & Comment, 12 Jan., p. 148), the pictures of David Baltimore and Joshua Lederberg were incorrectly credited. The credit for the Baltimore picture should have read, "M. Lampert, Boton." The credit for the Lederberg picture should have read, "Rockefeller University."


2.45 GHz microwave radiation impairs learning, memory, and hippocampal synaptic plasticity in the rat.
Microwave (MW) radiation has a close relationship with neurobehavioral disorders. Due to the widespread usage of MW radiation, especially in our homes, it is essential to investigate the direct effect of MW radiation on the central nervous system. Therefore, this study was carried out to determine the effect of MW radiation on memory and hippocampal synaptic plasticity. The rats were exposed to 2.45 GHz MW radiation (continuous wave with overall average power density of 0.016 mW/cm2 and overall average whole-body specific absorption rate value of 0.017 W/kg) for 2 h/day over a period of 40 days. Spatial learning and memory were tested by radial maze and passive avoidance tests. We evaluated the synaptic plasticity and hippocampal neuronal cells number by field potential recording and Giemsa staining, respectively. Our results showed that MW radiation exposure decreased the learning and memory performance that was associated with decrement of long-term potentiation induction and excitability of CA1 neurons. However, MW radiation did not have any effects on short-term plasticity and paired-pulse ratio as a good indirect index for measurement of glutamate release probability. The evaluation of hippocampal morphology indicated that the neuronal density in the hippocampal CA1 area was significantly decreased by MW.


Enantioselective gold(i)-catalyzed rearrangement of cyclopropyl-substituted 1,6-enynes into 2-oxocyclobutyl-cyclopentanes.
A gold(i)-catalyzed cycloisomerization/ring expansion sequence allows the highly enantioselective synthesis of 2-oxocyclobutylcyclopentane derivatives from cyclopropyl-substituted enynes. The bimetallic [(R)-MeO-DTBM-BIPHEP-(AuCl)2] complex was found to be the best precatalyst, affording the desired cyclobutanones in high yields and enantioselectivities (up to 99% ee). The usefulness of the method was further demonstrated by preparing the tricyclic core scaffold of russujaponol D.


Operative management of typhoid ileal perforation in children.
Intestinal perforation resulting from complicated typhoid fever still causes high morbidity and mortality. The purpose of the present study is to evaluate the outcome of its surgical management in Nigerian children. Emergency laparotomy and repair of the ileum was performed on 46 children with typhoid ileal perforation at the Federal Medical Centre (FMC), Azare, Nigeria, between January 2004-December 2008. This was followed by copious peritoneal lavage with warm normal saline and mass closure of the abdomen. There were 28 (60.86%) boys and 18 (39.13%) girls, with a mean age of 9.5 +/- 3.22 (range, 15 months-15 years). Abdominal pain (45), fever (44), and abdominal distention (36) were the most common presenting symptoms and majority of the patients (36) perforated within 14 days of illness. Solitary ileal perforations were the most common pathology, found in 31 (67.4%) cases. Simple closure of the perforations after debridement of the edges was the most frequent operative procedure performed. A total of 21 patients had one or more complications which included wound infection (21), postoperative fever (16), and wound dehiscence (6). Postoperative anaemia was a problem in 23 (50%) patients. The mortality rate was (13) 28.3%. The mean duration of hospital stay for survivors was 22.9 +/- 12.3 (range, 6-46 days). This was not significantly affected by the location or number of perforations on the ileum. The clinical course of typhoid ileal perforation may be different for the very young. The typically high rate of complications can be reduced if operation is undertaken earlier. Solitary ileal perforations can be managed safely with simple closure.


Prenatal folic acid treatment suppresses acrania and meroanencephaly in mice mutant for the Cart1 homeobox gene.
The paired-class homeobox-containing gene, Cart1, is expressed in forebrain mesenchyme, branchial arches, limb buds and cartilages during embryogenesis. Here, we show that Cart1-homozygous mutant mice are born alive with acrania and meroanencephaly but die soon after birth-a phenotype that strikingly resembles a corresponding human syndrome caused by a neural tube closure defect. Developmental studies suggest that Cart1 is required for forebrain mesenchyme survival and that its absence disrupts cranial neural tube morphogenesis by blocking the initiation of closure in the midbrain region that ultimately leads to the generation of lethal craniofacial defects. Prenatal treatment of Cart1 homozygous mutants with folic acid suppresses the development of the acrania/meroanencephaly phenotype.


What do patients think about during their consultations? A qualitative study.
The consultation has been widely examined, but chiefly from the doctor's standpoint. Comparatively little is known about how patients view the event even though shared understanding is an accepted goal of each consultation. The aim of the study was to describe the range and types of thoughts which patients have during their consultations. In semistructured interviews, prompted by video playback and transcript, 18 patients gave detailed accounts of their thoughts and feelings during a recent consultation. The study confirmed patients' central desire for understanding but also revealed a complex mix of other thoughts. Multiple problems and aims were usual. Patients routinely considered their relationship with the doctor, the doctor's willingness, ability and available time, and altered their behaviour accordingly. Patients and doctors have different models of the consultation-better understanding of how patients consult should lead to better training for doctors and improve shared understanding.


NTP Toxicology and Carcinogenesis Studies of 8-Hydroxyquinoline (CAS No. 148-24-3) in F344/N Rats and B6C3F1 Mice (Feed Studies).
Carcinogenesis studies of 8-hydroxyquinoline (99% pure), a metal chelator and antimicrobial agent, were conducted by administering the test chemical in feed to groups of 50 male and 50 female F344/N rats and B6C3F1 mice at concentrations of 0, 1,500, or 3,000 ppm for 103 weeks. These concentrations were selected because the chemical at higher concentrations resulted in reduced feed consumption, decreases in mean body weights, and deaths in the 15-day and 13-week studies. The average daily doses were estimated to be 73 and 143 mg/kg for male rats, 89 and 166 mg/kg for female rats, 217 and 396 mg/kg for male mice, and 349 and 619 mg/kg for female mice. Survival of dosed male and female rats and mice in the 2-year studies was comparable to that of the corresponding controls. The high dose rats and mice of each sex exhibited slight decreases in mean body weights and decreased feed consumption. Compound-related gross or microscopic pathologic effects were not observed in either species in the 15-day or 13-week studies. In the 2-year studies, C-cell adenomas/carcinomas of the thyroid gland showed a positive trend (P=0.03) for male rats (control, 1/50; low dose, 1/49; high dose, 6/47). The incidence of C-cell neoplasms in the high dose was not significantly increased compared with the controls, and the occurrence of C-cell hyperplasia was not elevated (4/50; 3/49; 1/47). The incidence of alveolar/bronchiolar adenomas or carcinomas (combined) in male rats with a positive trend, and the incidence in the high dose group was greater than that in the controls (0/50; 3/50; 4/50). This marginal effect was not supported by an increase in epithelial hyperplasia (5/50; 5/50; 3/50). These marginal increases in male rats were not regarded as being related to the administration of 8-hydroxyquinoline. In in vitro tests, 8-hydroxyquinoline did not induce either unscheduled DNA synthesis in rat hepatocytes or transformation of BALB/c-3T3 cells. An audit of the experimental data for these carcinogenesis studies on 8-hydroxyquinoline was conducted. No data discrepancies were found that significantly influenced the final interpretations. Under the conditions of these studies, there was no evidence of carcinogenicity for male and female F344/N rats or for male and female B6C3F1 mice given 8-hydroxyquinoline in feed at concentrations of 1,500 or 3,000 ppm for 103 weeks. Synonyms: 8-quinolinol; oxine; hydroxybenzopyridine


NMR spectroscopy and DFT calculations of a self-assembled arene ruthenium rectangle obtained from a combination of coordination and hydrogen bonds.
The hydrogen-bonded arene ruthenium metalla-rectangle, [(p-cymene)2Ru2(OO∩OO)(UPy)2]2(4+), obtained from 1-(4-oxo-6-undecyl-1,4-dihydropyrimidin-2-yl)-3-(pyridin-4-ylmethyl)urea (UPy) and the dinuclear arene ruthenium clip (p-cymene)2Ru2(OO∩OO)Cl2 (OO∩OO = 2,5-dioxido-1,4-benzoquinonato), is investigated by means of solution-phase NMR spectroscopy. Rotating frame nuclear Overhauser effect measurements are used to probe the H-bond network that drives the UPy self-assembly as well as the full rectangular supramolecular system. An effective distance that takes into account both intra- and intermolecular polarization-transfer pathways is utilised for data analysis. The experimental findings are corroborated by DFT calculations of NMR parameters and internuclear distances, thus confirming the formation of a very stable tetranuclear metalla-assembly.


A single chromosome assembly of Bacteroides fragilis strain BE1 from Illumina and MinION nanopore sequencing data.
Second and third generation sequencing technologies have revolutionised bacterial genomics. Short-read Illumina reads result in cheap but fragmented assemblies, whereas longer reads are more expensive but result in more complete genomes. The Oxford Nanopore MinION device is a revolutionary mobile sequencer that can produce thousands of long, single molecule reads. We sequenced Bacteroides fragilis strain BE1 using both the Illumina MiSeq and Oxford Nanopore MinION platforms. We were able to assemble a single chromosome of 5.18 Mb, with no gaps, using publicly available software and commodity computing hardware. We identified gene rearrangements and the state of invertible promoters in the strain. The single chromosome assembly of Bacteroides fragilis strain BE1 was achieved using only modest amounts of data, publicly available software and commodity computing hardware. This combination of technologies offers the possibility of ultra-cheap, high quality, finished bacterial genomes.


Isolation and characterization of an estrogen-inhibited variant derived from the MCF-7 breast cancer cell line.
The mechanism by which pharmacological concentrations of estrogen can paradoxically inhibit the growth of human breast cancer is unknown. We have selected for a variant line of MCF-7 which may help to understand this process. The variant was selected by exposing MCF-7 cells to high-specific-activity 16 alpha-[125I]iodoestradiol. These cells were viably frozen for two isotopic half-lives, defrosted once, then reexposed to 16 alpha-[125I]iodoestradiol to allow maximal radiation damage mediated by isotope associated with binding sites. This cell line (113) is one of 55 lines cloned from MCF-7 cells that survived this treatment. The growth response to estradiol of the 113 breast cancer cells grown in monolayer is normal for 4 to 6 days, and then the cell number plateaus as the cells appear to round up and detach. Concomitantly, a decrease in [3H]thymidine incorporation occurs. The cells cannot be rescued by removing estradiol from the medium. The inhibition is dose dependent and can be seen in concentrations of estradiol as low as 10(-10) M. The 113 cells are also inhibited by antiestrogens. They have normal levels of estrogen receptors which bind to DNA cellulose with activation. Progesterone receptors are estrogen inducible, although the levels are one-third that of wild-type MCF-7 cells. The morphological changes determined by electron microscopy of estrogen-treated cells are typical of degenerative cells. To investigate the possibility that inhibitory factors are secreted into the medium by 113 cells, conditioned medium from estrogen-exposed 113 cells is added to normal MCF-7 and 113 cells. No decrease in [3H]thymidine incorporation compared to controls is observed. When the secreted proteins are labeled with [35S]methionine and analyzed by sodium dodecyl sulfate-acrylamide gels, no major differences are apparent in the 113 and MCF-7 cells. Thus, the source of the defect is still unknown. It remains to be seen if the growth-inhibitory effects of 17 beta-estradiol on this cell line are receptor mediated or related to specific gene products which can be identified.


The RNA helicase/transcriptional co-regulator, p68 (DDX5), stimulates expression of oncogenic protein kinase, Polo-like kinase-1 (PLK1), and is associated with elevated PLK1 levels in human breast cancers.
p68 (DDX5) acts both as an ATP-dependent RNA helicase and as a transcriptional co-activator of several cancer-associated transcription factors, including the p53 tumor suppressor. p68 is aberrantly expressed in a high proportion of cancers, but the oncogenic drive for, or the consequences of, these expression changes remain unclear. Here we show that elevated p68 expression in a cohort of human breast cancers is associated significantly with elevated levels of the oncogenic protein kinase, Polo-like kinase-1 (PLK1). Patients expressing detectable levels of both p68 and PLK1 have a poor prognosis, but only if they also have mutation in the TP53 gene (encoding p53), suggesting that p68 can regulate PLK1 levels in a manner that is suppressed by p53. In support of this hypothesis, we show that p68 stimulates expression from the PLK1 promoter, and that silencing of endogenous p68 expression downregulates endogenous PLK1 gene expression. In the absence of functional p53, p68 stimulates the expression of PLK1 both at basal levels and in response to the clinically relevant drug, etoposide. In keeping with a role as a transcriptional activator/co-activator, chromatin immuno-precipitation analysis shows that p68 is associated with the PLK1 promoter, irrespective of the p53 status. However, its recruitment is stimulated by etoposide in cells lacking p53, suggesting that p53 can oppose association of p68 with the PLK1 promoter. These data provide a model in which p68 and p53 interplay regulates PLK1 expression, and which describes the behavior of these molecules, and the outcome of their interaction, in human breast cancer.


Combined inhibition of 20-hydroxyeicosatetraenoic acid formation and of epoxyeicosatrienoic acids degradation attenuates hypertension and hypertension-induced end-organ damage in Ren-2 transgenic rats.
Recent studies have shown that the renal CYP450 (cytochrome P450) metabolites of AA (arachidonic acid), the vasoconstrictor 20-HETE (20-hydroxyeicosatetraenoic acid) and the vasodilator EETs (epoxyeicosatrienoic acids), play an important role in the pathophysiology of AngII (angiotensin II)-dependent forms of hypertension and the associated target organ damage. The present studies were performed in Ren-2 renin transgenic rats (TGR) to evaluate the effects of chronic selective inhibition of 20-HETE formation or elevation of the level of EETs, alone or in combination, on the course of hypertension and hypertension-associated end-organ damage. Both young (30 days of age) prehypertensive TGR and adult (190 days of age) TGR with established hypertension were examined. Normotensive HanSD (Hannover Sprague-Dawley) rats served as controls. The rats were treated with N-methylsulfonyl-12,12-dibromododec-11-enamide to inhibit 20-HETE formation and/or with N-cyclohexyl-N-dodecyl urea to inhibit soluble epoxide hydrolase and prevent degradation of EETs. Inhibition in TGR of 20-HETE formation combined with enhanced bioavailability of EETs attenuated the development of hypertension, cardiac hypertrophy, proteinuria, glomerular hypertrophy and sclerosis as well as renal tubulointerstitial injury. This was also associated with attenuation of the responsiveness of the systemic and renal vascular beds to AngII without modifying their responses to noradrenaline (norepinephrine). Our findings suggest that altered production and/or action of 20-HETE and EETs plays a permissive role in the development of hypertension and hypertension-associated end-organ damage in this model of AngII-dependent hypertension. This information provides a basis for a search for new therapeutic approaches for the treatment of hypertension.


Parental perception of family functioning in everyday life with a child with ADHD.
The aim of this study was to describe and investigate family characteristics in relation to support, behaviour of the child, family functioning and sense of coherence from the parents' perspective in families with a child with attention deficit hyperactivity disorder (ADHD). A further aim was to explore predictors of family functioning. The study population consisted of 1964 parents of children with ADHD aged 15 years old and younger. In all, 265 parents responded to a questionnaire (response rate 48.2%; 217 mothers and 48 fathers). In addition to questions about the parents, children, family characteristics and support from health services, the questionnaire included the Family Assessment Device (FAD), the Strengths and Difficulties Questionnaire (SDQ), Sense of Coherence (SOC) and Social Cohesion and Support Index (SCS). Data were analysed with descriptive, comparative and standard multiple regression analyses. Parents with ADHD reported a weaker SOC and poorer FAD in the family than parents without ADHD. Parents with children medicated for ADHD were significantly more satisfied with social support (SCS), rated their children's behaviour as less problematic and reported better family functioning. Parents' age, SOC, SCS, SDQ and support from the community health services explained 46.2% of the variation in FAD. A strong SOC, social support and support from the community health services were strongly associated with a positive effect on family functioning. Parents with ADHD reported a weaker SOC and poorer family functioning than parents without ADHD. This knowledge may be useful and should be taken into account when planning support for such families.


How to ablate long-standing persistent atrial fibrillation?
Long-standing persistent (LSP) atrial fibrillation is the most challenging arrhythmia to treat. Catheter ablation of atrial fibrillation has reached satisfactory results for the long-term treatment of paroxysmal atrial fibrillation, but not for the treatment of LSP atrial fibrillation. Several approaches with various outcomes have been described in the literature. The purpose of this review is to summarize the ablation approach that we developed at our institution. During ablation of LSP atrial fibrillation, in addition to pulmonary vein antrum and posterior wall isolation, ablation of nonpulmonary vein triggers disclosed by high dosage of isoproterenol seems to be of utmost importance to achieve long-term success after a single procedure. The location of the nonpulmonary vein triggers includes the coronary sinus, the anterior part of the septum, the left atrial appendage and the superior vena cava. Termination of atrial fibrillation during ablation does not seem to influence the outcome. Increasing radiofrequency power from 30 up to 45 W seems an important factor to favour durable lesions. The approach described in this review will guide the reader to what we believe is the best approach for the ablation of patients with LSP atrial fibrillation.


Is RuAs2 a candidate for high temperature thermoelectric applications?
The mineral inspired material RuAs2 shows promise as a thermoelectric material with its high stability and attractive band structure. In order to validate these expectations phase-pure polycrystalline ruthenium arsenide was synthesized and densified using Spark Plasma Sintering. RuAs2 is an n-type semiconductor with an indirect band gap 0.69 eV as estimated from temperature dependent resistivity data, while the band gap calculated with DFT is 0.64 eV. The thermal conductivity and electrical resistivity are both high with room temperature values of 16 W m-1 K-1 and 170 mΩ cm respectively, leading to modest thermoelectric properties for the intrinsic system. Band structure calculations suggest that chemical modification should preferably be done at the As site to improve the intrinsic properties. Synchrotron powder X-ray diffraction and Rietveld structural refinements show RuAs2 to be a stable line phase up to 1000 K in both in air and in vacuum, and both as a powder and as a dense pellet. No indication of preferential orientation or material gradients are observed.


A spoonful of sugar helps the medicine go down: a novel technique to improve oral gavage in mice.
Oral gavage is a common route of precise oral dosing for studies in rodents. Complications including tracheal administration, esophageal trauma, and aspiration are common and usually related to animal resistance to the procedure, and the stress induced by oral gavage can be a confounding variable in many studies. The taste of sucrose conveys a pacifying and analgesic effect in newborns, whereas sour solutions can induce the swallow reflex in humans that are dysphagic. We hypothesized that precoating a gavage needle with sucrose or citrate (or both) would pacify mice and induce them to swallow, reducing the stress and complications associated with the technique. To validate this hypothesis, we quantitated time to passage, stress-related behavioral reactions to the procedure, and plasma corticosterone levels in mice after precoating gavage needles with water, sucrose, citrate, sucrose and citrate, or sodium chloride prior to oral gavage. Precoating needles with sucrose reduced the time to passage, decreased observable stress-related reactions to the procedure, and maintained plasma corticosterone levels similar to those in ungavaged control mice. Coating needles with water, sucrose and citrate, or citrate had no beneficial effects on these parameters. Our findings describe a novel, validated technique that measurably decreases signs of stress and thereby improves animal welfare during oral gavage. Furthermore, the use of sucrose may be a valuable tool to refine other minor or nonsurgical procedures in the field of laboratory animal research.


Coexistence of electrical conductivity and ferromagnetism in a hybrid material formed from reduced graphene oxide and manganese oxide.
The coexistence of electrical conductivity and ferromagnetism has been achieved in a reduced graphene oxide/manganese oxide hybrid (rGO-Mn) synthesized by chemical reduction of a graphene oxide and Mn(2+) (as its GO-Mn(2+) complex) using hydrazine. The rGO-Mn and GO-Mn(2+) complexes were characterized by Fourier transform infrared spectroscopy (FT-IR), Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and transmission electron microscopy (TEM). In rGO-Mn the Mn was present as manganese oxide nanoparticles located on the rGO nanosheets. This rGO-Mn exhibits both electrical conductivity and ferromagnetism. The synthesis of hybrids incorporating rGO and metal oxides is proposed as a useful strategy for generation of new multifunctional nano-composite materials.


Bottom-up determination of air-sea momentum exchange under a major tropical cyclone.
As a result of increasing frequency and intensity of tropical cyclones, an accurate forecasting of cyclone evolution and ocean response is becoming even more important to reduce threats to lives and property in coastal regions. To improve predictions, accurate evaluation of the air-sea momentum exchange is required. Using current observations recorded during a major tropical cyclone, we have estimated this momentum transfer from the ocean side of the air-sea interface, and we discuss it in terms of the drag coefficient. For winds between 20 and 48 meters per second, this coefficient initially increases and peaks at winds of about 32 meters per second before decreasing.


Vestibular development in marsupials and monotremes.
The young of marsupials and monotremes are all born in an immature state, followed by prolonged nurturing by maternal lactation in either a pouch or nest. Nevertheless, the level of locomotor ability required for newborn marsupials and monotremes to reach the safety of the pouch or nest varies considerably: some are transferred to the pouch or nest in an egg (monotremes); others are transferred passively by gravity (e.g. dasyurid marsupials); some have only a horizontal wriggle to make (e.g. peramelid and didelphid marsupials); and others must climb vertically for a long distance to reach the maternal pouch (e.g. diprotodontid marsupials). In the present study, archived sections of the inner ear and hindbrain held in the Bolk, Hill and Hubrecht collections at the Museum für Naturkunde, Berlin, were used to test the relationship between structural maturity of the vestibular apparatus and the locomotor challenges that face the young of these different mammalian groups. A system for staging different levels of structural maturity of the vestibular apparatus was applied to the embryos, pouch young and hatchlings, and correlated with somatic size as indicated by greatest body length. Dasyurids are born at the most immature state, with the vestibular apparatus at little more than the otocyst stage. Peramelids are born with the vestibular apparatus at a more mature state (fully developed semicircular ducts and a ductus reuniens forming between the cochlear duct and saccule, but no semicircular canals). Diprotodontids and monotremes are born with the vestibular apparatus at the most mature state for the non-eutherians (semicircular canals formed, maculae present, but vestibular nuclei in the brainstem not yet differentiated). Monotremes and marsupials reach the later stages of vestibular apparatus development at mean body lengths that lie within the range of those found for laboratory rodents (mouse and rat) reaching the same vestibular stage.


Expression of retinoid receptor genes and proteins in non-small-cell lung cancer.
Retinoids can suppress carcinogenesis in high-risk non-neoplastic bronchial lesions and can reduce the risk of second primary non-small-cell lung cancer (NSCLC). The effects of retinoids are mediated by nuclear receptors, i.e., the retinoic acid receptors (RARalpha, RARbeta, and RARgamma) and the retinoid X receptors (RXRalpha, RXRbeta, and RXRgamma). We investigated whether abnormalities in the in vivo expression of retinoid receptors are observed in NSCLC. Expression of retinoid receptors in paired specimens of normal and cancerous tissues from the lungs of 76 patients with NSCLC was studied by use of antiretinoid receptor antibodies (except those against RXRgamma) and immunohistochemistry. RAR messenger RNAs were analyzed by use of in situ hybridization and by reverse transcription-polymerase chain reaction (RT-PCR). Samples were also studied for loss of heterozygosity (LOH) at chromosome 3p24. All P values are two-sided. All studied receptors were expressed in normal lung cells and in high- risk non-neoplastic lesions. In tumor cells, overexpression of RXRalpha and RARalpha was frequently observed. In contrast, RXRbeta expression decreased in 18% of the tumor specimens. Furthermore, there was a marked decrease in the expression of RARbeta in 63% of the tumors (P<.0001). Decreased expression of RARgamma was observed by RT-PCR in 41% of the tumors (P<.0001). LOH at 3p24 was observed in 41% of the tumor specimens from informative patients and in 20% of the non-neoplastic lesions. Expression of RARalpha and RXRalpha is either normal or elevated in NSCLC. In contrast, a large percentage of tumors show a marked decrease in the expression of RARbeta, RARgamma, and RXRbeta as well as a high frequency of LOH at 3p24, which was also observed in non-neoplastic lesions. These data suggest that altered retinoid receptor expression may play a role in lung carcinogenesis.


Physical association of base excision repair enzymes with parental and replicating DNA in BHK-21 cells.
The physical association of mammalian excision repair enzymes with DNA was examined as a function of cell proliferation. The molecular weight distribution of two nuclear base excision repair enzymes, the uracil DNA glycosylase and the hypoxanthine DNA glycosylase, were examined by sucrose step gradient analysis. The sedimentation of DNA polymerase activity as well as the distribution of parental and replicating DNA were determined simultaneously. In confluent BHK-21 fibroblasts, basal levels of both DNA glycosylases, DNA polymerase beta, and parental DNA sedimented to the 20%/40% sucrose border. In proliferating BHK-21 cells, induced levels of both DNA glycosylases, DNA polymerase alpha, and replicating DNA sedimented to the 40%/50% sucrose border. The physical association of the repair enzymes with DNA was demonstrated by detergent treatment and by DNase digestion. As defined by [35S] methionine pulse labeling analysis, newly synthesized DNA repair enzymes were localized with either parental or replicating DNA. These results suggested that the physical association of mammalian DNA repair enzymes with nuclear DNA was dependent on the proliferative state of the cell.


Multisensory contributions to the 3-D representation of visuotactile peripersonal space in humans: evidence from the crossmodal congruency task.
In order to determine precisely the location of a tactile stimulus presented to the hand it is necessary to know not only which part of the body has been stimulated, but also where that part of the body lies in space. This involves the multisensory integration of visual, tactile, proprioceptive, and even auditory cues regarding limb position. In recent years, researchers have become increasingly interested in the question of how these various sensory cues are weighted and integrated in order to enable people to localize tactile stimuli, as well as to give rise to the 'felt' position of our limbs, and ultimately the multisensory representation of 3-D peripersonal space. We highlight recent research on this topic using the crossmodal congruency task, in which participants make speeded elevation discrimination responses to vibrotactile targets presented to the thumb or index finger, while simultaneously trying to ignore irrelevant visual distractors presented from either the same (i.e., congruent) or a different (i.e., incongruent) elevation. Crossmodal congruency effects (calculated as performance on incongruent-congruent trials) are greatest when visual and vibrotactile stimuli are presented from the same azimuthal location, thus providing an index of common position across different sensory modalities. The crossmodal congruency task has been used to investigate a number of questions related to the representation of space in both normal participants and brain-damaged patients. In this review, we detail the major findings from this research, and highlight areas of convergence with other cognitive neuroscience disciplines.


Effect and cost of an after-school dance programme on the physical activity of 11-12 year old girls: The Bristol Girls Dance Project, a school-based cluster randomised controlled trial.
The aim of this study was to examine the effectiveness and cost of an after-school dance intervention at increasing the physical activity levels of Year 7 girls (age 11-12). A cluster randomised controlled trial was conducted in 18 secondary schools. Participants were Year 7 girls attending a study school. The Bristol Girls Dance Project (BGDP) intervention consisted of up to forty, 75-minute dance sessions delivered in the period immediately after school by experienced dance instructors over 20-weeks. The pre-specified primary outcome was accelerometer assessed mean minutes of weekday moderate to vigorous physical activity (MVPA) at time 2 (52 weeks are T0 baseline assessments). Secondary outcomes included accelerometer assessed mean minutes of weekday MVPA at time 1 (while the intervention was still running) and psychosocial outcomes. Intervention costs were assessed. 571 girls participated. Valid accelerometer data were collected from 549 girls at baseline with 508 girls providing valid accelerometer data at baseline and time 2. There were no differences between the intervention and control group for accelerometer assessed physical activity at either time 1 or time 2. Only one third of the girls in the intervention arm met the pre-set adherence criteria of attending two thirds of the dance sessions that were available to them. Instrumental variable regression analyses using complier average causal effects provided no evidence of a difference between girls who attended the sessions and the control group. The average cost of the intervention was £73 per girl, which was reduced to £63 when dance instructor travel expenses were excluded. This trial showed no evidence that an after-school dance programme can increase the physical activity of Year 7 girls. The trial highlighted the difficulty encountered in maintaining attendance in physical activity programmes delivered in secondary schools. There is a need to find new ways to help adolescent girls to be physically active via identifying ways to support and encourage sustained engagement in physical activity over the life course. ISRCTN52882523.


Translocon component Sec62 acts in endoplasmic reticulum turnover during stress recovery.
The endoplasmic reticulum (ER) is a site of protein biogenesis in eukaryotic cells. Perturbing ER homeostasis activates stress programs collectively called the unfolded protein response (UPR). The UPR enhances production of ER-resident chaperones and enzymes to reduce the burden of misfolded proteins. On resolution of ER stress, ill-defined, selective autophagic programs remove excess ER components. Here we identify Sec62, a constituent of the translocon complex regulating protein import in the mammalian ER, as an ER-resident autophagy receptor. Sec62 intervenes during recovery from ER stress to selectively deliver ER components to the autolysosomal system for clearance in a series of events that we name recovER-phagy. Sec62 contains a conserved LC3-interacting region in the C-terminal cytosolic domain that is required for its function in recovER-phagy, but is dispensable for its function in the protein translocation machinery. Our results identify Sec62 as a critical molecular component in maintenance and recovery of ER homeostasis.


Implementation and Extended Evaluation of the Euroimmun Anti-SARS-CoV-2 IgG Assay and Its Contribution to the United Kingdom's COVID-19 Public Health Response.
In March 2020, the Rare and Imported Pathogens Laboratory at the UK Health Security Agency (UKHSA) (formerly Public Health England [PHE]) Porton Down, was tasked by the Department of Health and Social Care with setting up a national surveillance laboratory facility to study SARS-CoV-2 antibody responses and population-level sero-surveillance in response to the growing SARS-CoV-2 outbreak. In the following 12 months, the laboratory tested more than 160,000 samples, facilitating a wide range of research and informing UKHSA, DHSC, and UK government policy. Here we describe the implementation and use of the Euroimmun anti-SARS-CoV-2 IgG assay and provide an extended evaluation of its performance. We present a markedly improved overall sensitivity of 91.39% (≥14 days 92.74%, ≥21 days 93.59%) compared to our small-scale early study, and a specificity of 98.56%. In addition, we detail extended characteristics of the Euroimmun assay: intra- and interassay precision, correlation to neutralization, and assay linearity. IMPORTANCE Serology assays have been useful in determining those with previous SARS-CoV-2 infection in a wide range of research and serosurveillance projects. However, assays vary in their sensitivity at detecting SARS-CoV-2 antibodies. Here, we detail an extended evaluation and characterization of the Euroimmun anti-SARS-CoV-2 IgG assay, one that has been widely used within the United Kingdom on over 160,000 samples to date.


Stroke prevention in atrial fibrillation: a systematic review.
Atrial fibrillation (AF) is associated with an increase in mortality and morbidity, with a substantial increase in stroke and systemic thromboembolism. Strokes related to AF are associated with higher mortality, greater disability, longer hospital stays, and lower chance of being discharged home than strokes unrelated to AF. To provide an overview of current concepts and recent developments in stroke prevention in AF, with suggestions for practical management. A comprehensive structured literature search was performed using MEDLINE for studies published through March 11, 2015, that reported on AF and stroke, bleeding risk factors, and stroke prevention. The risk of stroke in AF is reduced by anticoagulant therapy. Thromboprophylaxis can be obtained with vitamin K antagonists (VKA, eg, warfarin) or a non-VKA oral anticoagulant (NOAC). Major guidelines emphasize the important role of oral anticoagulation (OAC) for effective stroke prevention in AF. Initially, clinicians should identify low-risk AF patients who do not require antithrombotic therapy (ie, CHA2DS2-VASc score, 0 for men; 1 for women). Subsequently, patients with at least 1 stroke risk factor (except when the only risk is being a woman) should be offered OAC. A patient's individual risk of bleeding from antithrombotic therapy should be assessed, and modifiable risk factors for bleeding should be addressed (blood pressure control, discontinuing unnecessary medications such as aspirin or nonsteroidal anti-inflammatory drugs). The international normalized ratio should be tightly controlled for patients receiving VKAs. Stroke prevention is central to the management of AF, irrespective of a rate or rhythm control strategy. Following the initial focus on identifying low-risk patients, all others with 1 or more stroke risk factors should be offered OAC.


Multi-level inhibition of coronavirus replication by chemical ER stress.
Coronaviruses (CoVs) are important human pathogens for which no specific treatment is available. Here, we provide evidence that pharmacological reprogramming of ER stress pathways can be exploited to suppress CoV replication. The ER stress inducer thapsigargin efficiently inhibits coronavirus (HCoV-229E, MERS-CoV, SARS-CoV-2) replication in different cell types including primary differentiated human bronchial epithelial cells, (partially) reverses the virus-induced translational shut-down, improves viability of infected cells and counteracts the CoV-mediated downregulation of IRE1α and the ER chaperone BiP. Proteome-wide analyses revealed specific pathways, protein networks and components that likely mediate the thapsigargin-induced antiviral state, including essential (HERPUD1) or novel (UBA6 and ZNF622) factors of ER quality control, and ER-associated protein degradation complexes. Additionally, thapsigargin blocks the CoV-induced selective autophagic flux involving p62/SQSTM1. The data show that thapsigargin hits several central mechanisms required for CoV replication, suggesting that this compound (or derivatives thereof) may be developed into broad-spectrum anti-CoV drugs.


Targeting radioresistant breast cancer cells by single agent CHK1 inhibitor via enhancing replication stress.
Radiotherapy (RT) remains a standard therapeutic modality for breast cancer patients. However, intrinsic or acquired resistance limits the efficacy of RT. Here, we demonstrate that CHK1 inhibitor AZD7762 alone significantly inhibited the growth of radioresistant breast cancer cells (RBCC). Given the critical role of ATR/CHK1 signaling in suppressing oncogene-induced replication stress (RS), we hypothesize that CHK1 inhibition leads to the specific killing for RBCC due to its abrogation in the suppression of RS induced by oncogenes. In agreement, the expression of oncogenes c-Myc/CDC25A/c-Src/H-ras/E2F1 and DNA damage response (DDR) proteins ATR/CHK1/BRCA1/CtIP were elevated in RBCC. AZD7762 exposure led to significantly higher levels of RS in RBCC, compared to the parental cells. The mechanisms by which CHK1 inhibition led to specific increase of RS in RBCC were related to the interruptions in the replication fork dynamics and the homologous recombination (HR). In summary, RBCC activate oncogenic pathways and thus depend upon mechanisms controlled by CHK1 signaling to maintain RS under control for survival. Our study provided the first example where upregulating RS by CHK1 inhibitor contributes to the specific killing of RBCC, and highlight the importance of the CHK1 as a potential target for treatment of radioresistant cancer cells.


Estimating the proportion of HIV transmissions from main sex partners among men who have sex with men in five US cities.
HIV incidence in the United States among men who have sex with men (MSM) has been increasing since 2000, and MSM remain the most heavily impacted risk group in the US HIV epidemic. We modeled HIV transmissions, using data from MSM in five US cities from the National HIV Behavioral Surveillance System, the HIVNET Vaccine Preparedness Study, and other published data. Annual HIV transmissions were estimated by partner type (main or casual) and by sex type (receptive anal intercourse, insertive anal intercourse, or oral sex). Sixty-eight percent [95% confidence interval (CI) 58-78) of HIV transmissions were from main sex partners because of a higher number of sex acts with main partners, more frequent receptive roles in anal sex with main partners, and lower condom use during anal sex with main partners. By sex type, 69% (95% CI 59-79) of infections were from receptive anal intercourse, 28% (95% CI 19-38) were from insertive anal intercourse, and 2% (95% CI 0-5) were from oral sex. The model-based estimated HIV incidence rate was 2.2% (95% CI 1.7-2.7) per year. Sensitivity analyses demonstrated estimates of transmission from main sex partners as low as 52% (95% CI 41-62) and as high as 74% (95% CI 68-80). According to our model, most HIV transmissions among MSM in five US cities are from main sex partners. HIV prevention efforts should take into account the risks of HIV transmissions in male partnerships, and couples-based HIV prevention interventions for MSM should be given high priority in the US HIV prevention research portfolio.


Impact of oral vitamin C on histamine levels and seasickness.
Seasickness is a risk aboard a ship. Histamine is postulated as a causative agent, inversely related to the intake of vitamin C. Persons with mastocytosis experienced improvement of nausea after the intake of vitamin C. To determine whether vitamin C suppresses nausea in 70 volunteers who spent 20 minutes in a life raft, exposed to one-meter-high waves in an indoor pool. Double-blind placebo-controlled crossover study. Two grams of vitamin C or placebo was taken one hour before exposure. Blood samples were taken one hour before and after exposure to determine histamine, diamine oxidase, tryptase, and vitamin C levels. Symptom scores were noted on a visual analog scale. On the second day the test persons were asked which day they had felt better. Seven persons without symptoms were excluded from the analysis. Test persons had less severe symptoms after the intake of vitamin C (p < 0.01). Scores on the visual analog scale were in favor of vitamin C, but the difference was not significant. Twenty-three of 63 persons wished to leave the raft earlier: 17 after the intake of placebo and 6 after the intake of vitamin C (p < 0.03). Women (p < 0.02) and men below 27 years of age (p < 0.02) had less pronounced symptoms after the intake of vitamin C. Histamine (p < 0.01) and DAO levels were increased after the intake of vitamin C (p < 0.001) and after placebo (n.s.). The fact that the second test day was rated less stressful by most volunteers is indicative of habituation. Some of the data show that vitamin C is effective in suppressing symptoms of seasickness, particularly in women and men younger than 27 years of age, and is devoid of side effects. Histamine levels were initially increased after the test persons had been exposed to waves.


Geothermal System at 21{degrees}N, East Pacific Rise: Physical Limits on Geothermal Fluid and Role of Adiabatic Expansion.
Pressure-volume-temperature relations for water at the depth of the magma chamber at 21 degrees N on the East Pacific Rise suggest that the maximum subsurface temperature of the geothermal fluid is about 420 degrees C. Both the chemistry of the discharging fluid and thermal balance considerations indicate that the effective water/rock ratios in the geothermal system are between 7 and 16. Such low ratios preclude effective metal transport at temperatures below 350 degrees C, but metal solubilization at 400 degrees C and above is effective even at such low ratios. It is proposed that the 420 degrees C fluid ascends essentially adiabatically and in the process expands, cools, and precipitates metal sulfides within the upper few hundred meters of the sea floor and on the sea floor itself.


The Role of Melatonin in Electroacupuncture Alleviating Lung Injury Induced by Limb Ischemia-Reperfusion in Rabbits.
BACKGROUND Our previous studies have shown that electroacupuncture (EA) can alleviate lung injury induced by limb ischemia-reperfusion, but the specific mechanism is still unclear. MATERIAL AND METHODS The animals were randomly divided into sham operation group (Sham), model group (IR), electroacupuncture group (EA), sham electroacupuncture group (SEA), and EA+luzindole group (EA+luzindole). The limb ischemia-reperfusion model was established according to previously described, the rabbits in the EA and EA+luzindole groups were given EA at ST36 and BL13 for 7 days before the model preparation and during the model implementation, however, sham EA was mainly used to stimulate the rabbits in the SEA group with shallow needling at the points 0.5 cm near ST36 and BL13. Then, 30 mg/kg of luzindole was intraperitoneally injected 30 minutes before the model preparation in the EA+luzindole group. RESULTS The wet weight/dry weight (W/D) ratio, lung injury score, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1ß, IL-6, and malondialdehyde (MDA) contents in the EA group at 4 hours after reperfusion were significantly lower than those in the IR, SEA, and EA+luzindole groups. The levels of serum melatonin at T0 in the EA and EA+luzindole groups were significantly higher than those in the Sham group. The levels of serum melatonin at T1 and T2 in the IR group were significantly lower than those in the Sham group. There was no significant difference in the expression levels of melatonin receptor 1 (MR-1) and MR-2 in lung tissues among the 5 groups. CONCLUSIONS EA could alleviate the lung injury induced by limb ischemia-reperfusion by promoting the secretion of melatonin, while having no effect on the expression of melatonin receptor in lung tissues.


The indirect antioxidant sulforaphane protects against thiopurine-mediated photooxidative stress.
Long-term treatment with thiopurines, such as the widely used anticancer, immunosuppressive and anti-inflammatory agent azathioprine, combined with exposure to ultraviolet (UV) radiation is associated with increased oxidative stress, hyperphotosensitivity and high risk for development of aggressive squamous cell carcinomas of the skin. Sulforaphane, an isothiocyanate derived from broccoli, is a potent inducer of endogenous cellular defenses regulated by transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), including cytoprotective enzymes and glutathione, which in turn act as efficient indirect and direct antioxidants that have long-lasting effects. Treatment with 6-thioguanine, a surrogate for azathioprine, leads to profound sensitization to oxidative stress and glutathione depletion upon exposure to UVA radiation, the damaging effects of which are primarily mediated by generation of reactive oxygen species. The degree of sensitization is greater for irradiation exposures spanning the absorption spectrum of 6-thioguanine, and is dependent on the length of treatment and the level of guanine substitution with 6-thioguanine, suggesting that the 6-thioguanine that is incorporated in genomic DNA is largely responsible for this sensitization. Sulforaphane provides protection against UVA, but not UVB, radiation without affecting the levels of 6-thioguanine incorporation into DNA. The protective effect is lost under conditions of Nrf2 deficiency, implying that it is due to induction of Nrf2-dependent cytoprotective proteins, and that this strategy could provide protection against any potentially photosensitizing drugs that generate electrophilic or reactive oxygen species. Thus, our findings support the development of Nrf2 activators as protectors against drug-mediated photooxidative stress and encourage future clinical trials in populations at high risk for cutaneous photodamage and photocarcinogenesis.


Mechanisms of immune tolerance to allergens.
In allergic diseases, immune responses are induced by normally well-tolerated allergens, which result in chronic inflammation characterized by antibody secretion and T cell activation. For almost 100 years, allergen-specific immunotherapy (allergen-SIT) has been the potentially curative and antigen-specific method for the treatment of allergic diseases. Allergen-SIT alters the course of allergic diseases and can reduce allergic symptoms and medication use. The key mechanism behind allergen-SIT is the induction of peripheral T cell tolerance by altering the balance between Th cells and regulatory T cells. Both naturally occurring thymus-derived FOXP3(+)CD4(+)CD25(+) regulatory T cells and inducible type 1 regulatory T cells suppress the development of allergic diseases via several mechanisms including suppression of dendritic cells, Th cells, mast cells, eosinophils and basophils; suppression of inflammatory cell migration to tissues; and decrease of the ratio between allergen-specific IgE and IgG4 antibodies. These effects are mainly mediated by the suppressive cytokines IL-10 and TGF-β. Knowledge of this molecular basis is crucial to understanding the regulation of the immune response and their possible therapeutic applications for allergic diseases.


Growth of Sinclair swine melanoma as a function of age, histopathological staging, and gonadal status.
We have analyzed the association between host hormonal status and growth rate of congenital exophytic melanomas of Sinclair swine. The growth of multiple exophytic lesions during the first year of life of intact males, orchiectomized males, intact females, and ovariectomized females was quantitated using a proliferative index which assigned a numerical value to fixed increments of gains or losses in tumor volume. The proliferative index from 6 wk (gonadectomy at 6 wk) to 52 wk of age of each treatment group was statistically increased from 0 (P less than 0.01) except that of gonadectomized females. The proliferative indices from lesions in gonadectomized females were significantly lower than those from intact males, intact females, and gonadectomized males. A total of 93 exophytic tumors from 63 swine were biopsied and histopathologically staged according to the degree of progression or regression and analyzed as a function of animal age. There were no Stage I and only two Stage II lesions at the time biopsies were taken. Twenty-six of 32 (81.2%) Stage III tumors were found in swine of both sexes less than 26 wk of age of which 71.8% were found less than 10 wk of age, while 20 of 29 (68.9%) were Stage IV, and only 3 of 32 (9%) were Stage V lesions present in this age group (P less than 0.001, chi 2). Only 20.7% of Stage IV tumors were present prior to 6 wk of age. Preliminary results suggest that castration of either sex also altered tumor histopathology. Our data suggest that a reduction in gonadal steroid secretion was associated with a decrease in exophytic tumor growth rate and regression in animals of both sexes during the first year of life in Sinclair swine. The effect in female swine was due to significant reduction in the proliferative index over the first 6 mo of age.


Inequalities in maternal health: national cohort study of ethnic variation in severe maternal morbidities.
To describe on a national basis ethnic differences in severe maternal morbidity in the United Kingdom. National cohort study using the UK Obstetric Surveillance System (UKOSS). All hospitals with consultant led maternity units in the UK. 686 women with severe maternal morbidity between February 2005 and February 2006. Rates, risk ratios, and odds ratios of severe maternal morbidity in different ethnic groups. 686 cases of severe maternal morbidity were reported in an estimated 775 186 maternities, representing an estimated incidence of 89 (95% confidence interval 82 to 95) cases per 100 000 maternities. 74% of women were white, and 26% were non-white. The estimated risk of severe maternal morbidity in white women was 80 cases per 100 000 maternities, and that in non-white women was 126 cases per 100,000 (risk difference 46 (27 to 66) cases per 100 000; risk ratio 1.58, 95% confidence interval 1.33 to 1.87). Black African women (risk difference 108 (18 to 197) cases per 100,000 maternities; risk ratio 2.35, 1.45 to 3.81) and black Caribbean women (risk difference 116 (59 to 172) cases per 100 000 maternities; risk ratio 2.45, 1.81 to 3.31) had the highest risk compared with white women. The risk in non-white women remained high after adjustment for differences in age, socioeconomic and smoking status, body mass index, and parity (odds ratio 1.50, 1.15 to 1.96). Severe maternal morbidity is significantly more common among non-white women than among white women in the UK, particularly in black African and Caribbean ethnic groups. This pattern is very similar to reported ethnic differences in maternal death rates. These differences may be due to the presence of pre-existing maternal medical factors or to factors related to care during pregnancy, labour, and birth; they are unlikely to be due to differences in age, socioeconomic or smoking status, body mass index, or parity. This highlights to clinicians and policy makers the importance of tailored maternity services and improved access to care for women from ethnic minorities. National information on the ethnicity of women giving birth in the UK is needed to enable ongoing accurate study of these inequalities.


Epilepsy and driving.
The status of the epileptic applicant for a licence to drive a private motor vehicle has changed from total prohibition through a stage of partial approval subject to medical assessment to the present conditional right to a driving licence. We report a study of a series of patients which shows that many and probably most epileptics have obtained driving licences by concealment of their condition, yet the new regulations continue to expect true declarations. It is suggested that, in common with applicants for a public service vehicle or a heavy goods vehicle driving licence, the applicant for a licence to drive a private motor vehicle should provide a medical report of fitness. The epileptic should be included in a general category of medically restricted drivers subject to individual medical assessment.


Landlord attitudes and behaviors regarding smoke-free policies: implications for voluntary policy change.
The study purpose was to describe multiunit landlord attitudes and behaviors toward smoke-free policies. This was a descriptive, cross-sectional survey of multiunit landlords in Douglas County (N=392). A 25-item survey was developed and pilot tested. It was administered by telephone (n=143) and mail (n=249) to multiunit landlords. Combined response rate was 30.1% (81/143 telephone, 37/249 mail) representing 24,080 units on 974 properties with 34,399 tenants. Most respondents (73.7%) allowed smoking. Reasons for not implementing smoke-free policies were potential enforcement problems (57.0%), tenant objections (43.0%), loss of market share (39.5%). Respondents without smoke-free policies expected vacancy (53.6%) and turnover (50.0%) rates to increase, which was significantly different (p <.0001) than respondents with smoke-free policies where only 10.7% reported increased vacancy and only 3.7% reported increased turnover. Expected adverse impacts of smoke-free policies do not reflect real experiences of smoke-free policy implementation. Public health advocates can use these study findings to develop community-based education and social marketing messages directed at voluntary smoke-free policy changes. Respondents without smoke-free policies expressed interest at the end of the survey in learning how to implement smoke-free policies indicating a readiness for change.


Targeted Learning of the Mean Outcome under an Optimal Dynamic Treatment Rule.
We consider estimation of and inference for the mean outcome under the optimal dynamic two time-point treatment rule defined as the rule that maximizes the mean outcome under the dynamic treatment, where the candidate rules are restricted to depend only on a user-supplied subset of the baseline and intermediate covariates. This estimation problem is addressed in a statistical model for the data distribution that is nonparametric beyond possible knowledge about the treatment and censoring mechanism. This contrasts from the current literature that relies on parametric assumptions. We establish that the mean of the counterfactual outcome under the optimal dynamic treatment is a pathwise differentiable parameter under conditions, and develop a targeted minimum loss-based estimator (TMLE) of this target parameter. We establish asymptotic linearity and statistical inference for this estimator under specified conditions. In a sequentially randomized trial the statistical inference relies upon a second-order difference between the estimator of the optimal dynamic treatment and the optimal dynamic treatment to be asymptotically negligible, which may be a problematic condition when the rule is based on multivariate time-dependent covariates. To avoid this condition, we also develop TMLEs and statistical inference for data adaptive target parameters that are defined in terms of the mean outcome under the estimate of the optimal dynamic treatment. In particular, we develop a novel cross-validated TMLE approach that provides asymptotic inference under minimal conditions, avoiding the need for any empirical process conditions. We offer simulation results to support our theoretical findings.


Origins of HIV and the evolution of resistance to AIDS.
The cross-species transmission of lentiviruses from African primates to humans has selected viral adaptations which have subsequently facilitated human-to-human transmission. HIV adapts not only by positive selection through mutation but also by recombination of segments of its genome in individuals who become multiply infected. Naturally infected nonhuman primates are relatively resistant to AIDS-like disease despite high plasma viral loads and sustained viral evolution. Further understanding of host resistance factors and the mechanisms of disease in natural primate hosts may provide insight into unexplored therapeutic avenues for the prevention of AIDS.


Identification of the fliI and fliJ components of the Caulobacter flagellar type III protein secretion system.
Caulobacter crescentus is motile by virtue of a polar flagellum assembled during the predivisional stage of the cell cycle. Three mutant strains in which flagellar assembly was blocked at an early stage were isolated. The mutations in these strains mapped to an operon of two genes, fliI and fliJ, both of which are necessary for motility. fliI encodes a 50-kDa polypeptide whose sequence is closely related to that of the Salmonella typhimurium FliI protein, an ATPase thought to energize the export of flagellar subunits across the cytoplasmic membrane through a type III protein secretion system. fliJ encodes a 16-kDa hydrophilic protein of unknown function. Epistasis experiments demonstrated that the fliIJ operon is located in class II of the C. crescentus flagellar regulatory hierarchy, suggesting that the gene products act at an early stage in flagellar assembly. The expression of fliIJ is induced midway through the cell cycle, coincident with other class II operons, but the FliI protein remains present throughout the cell cycle. Subcellular fractionation showed that FliI is present both in the cytoplasm and in association with the membrane. Mutational analysis of FliI showed that two highly conserved amino acid residues in a bipartite ATP binding motif are necessary for flagellar assembly.


A poly(ethylene glycol)-based surfactant for formulation of drug-loaded mucus penetrating particles.
Mucosal surfaces are protected by a highly viscoelastic and adhesive mucus layer that traps most foreign particles, including conventional drug and gene carriers. Trapped particles are eliminated on the order of seconds to hours by mucus clearance mechanisms, precluding sustained and targeted drug and nucleic acid delivery to mucosal tissues. We have previously shown that polymeric coatings that minimize adhesive interactions with mucus constituents lead to particles that rapidly penetrate human mucus secretions. Nevertheless, a particular challenge in formulating drug-loaded mucus penetrating particles (MPP) is that many commonly used surfactants are either mucoadhesive, or do not facilitate efficient drug encapsulation. We tested a novel surfactant molecule for particle formulation composed of Vitamin E conjugated to 5 kDa poly(ethylene glycol) (VP5k). We show that VP5k-coated poly(lactide-co-glycolide) (PLGA) nanoparticles rapidly penetrate human cervicovaginal mucus, whereas PLGA nanoparticles coated with polyvinyl alcohol or Vitamin E conjugated to 1 kDa PEG were trapped. Importantly, VP5k facilitated high loading of paclitaxel, a frontline chemo drug, into PLGA MPP, with controlled release for at least 4 days and negligible burst release. Our results offer a promising new method for engineering biodegradable, drug-loaded MPP for sustained and targeted delivery of therapeutics at mucosal surfaces.


Dexmedetomidine Maintains Its Direct Cardioprotective Effect Against Ischemia/Reperfusion Injury in Hypertensive Hypertrophied Myocardium.
Dexmedetomidine (DEX) has a direct cardioprotective effect against ischemia/reperfusion injury through endothelial nitric oxide synthase (eNOS) phosphorylation via α2-adrenoreceptor (α2-AR). By using spontaneously hypertensive rat (SHR) and Wistar-Kyoto (WKY) rat models, the cardioprotective effect of DEX in hypertrophied myocardium and the differential characteristics of cardiac α2-AR and the I1 imidazoline receptor (I1R) were examined. Langendorff-perfused rat hearts underwent 40 minutes of global ischemia followed by 120 minutes of reperfusion in the presence or absence of DEX before ischemia. Infarct size was measured, and eNOS phosphorylation was assessed by Western blotting. The presence and expression of the receptors were assessed by immunohistochemistry, real-time reverse transcriptase polymerase chain reaction, and Western blotting. In WKY, DEX significantly decreased infarct size and increased phosphorylated-eNOS/eNOS. These effects were counteracted by yohimbine (α2-AR antagonist) and efaroxan (α2-AR and I1R antagonist). In SHR, DEX significantly decreased infarct size, and the effect was counteracted by efaroxan but not yohimbine. DEX did not alter phosphorylated-eNOS/eNOS in SHR. α2-AR and I1R were observed in WKY and SHR hearts. Although alpha2A-AR and alpha2B-AR messenger RNA and protein levels were upregulated in SHR, I1R expression was comparable between the 2 species. In the hypertrophied heart, DEX maintains its direct cardioprotective effect against ischemia/reperfusion injury via I1R in an eNOS-nondependent manner despite upregulation of α2-AR.


Working hours and the onset of depressive disorder: a systematic review and meta-analysis.
: This systematic review and meta-analysis aimed to examine whether working beyond the standard working hours was associated with a greater risk of depressive disorder among workers included in published prospective studies. This manuscript was prepared according to the PRISMA guideline checklist. A database search was conducted using MEDLINE (PubMed), PsycINFO, and PsycARTICLES using a relevant set of keywords. The eligibility criteria were as follows: (1) participants were adult workers; (2) exposure was defined as overtime work; (3) outcome were depressive disorders clinically diagnosed or assessed by a structured interview and (4) the study design was prospective or cohort. 7 studies were identified in the systematic review and meta-analysis. Overtime work was associated with a small, non-significant, elevated risk of depressive disorder (pooled relative risk=1.075; 95% CI 0.834 to 1.387; p=0.575) in a random effects model. The association tended to be greater for women. The risk of working 50 or more hours per week was slightly but not significantly increased (pooled relative risk=1.241; 95% CI 0.880 to 1.750; p=0.218). The effect of overtime work on depressive disorder remains inconclusive and may be small if not negligible. Sex differences and the effect of longer working hours on depressive disorder should be addressed in the future. Prospero CRD42015020003; Results.


Characterizing Bonding Patterns in Diradicals and Triradicals by Density-Based Wave Function Analysis: A Uniform Approach.
Density-based wave function analysis enables unambiguous comparisons of the electronic structure computed by different methods and removes ambiguity of orbital choices. We use this tool to investigate the performance of different spin-flip methods for several prototypical diradicals and triradicals. In contrast to previous calibration studies that focused on energy gaps between high- and low spin-states, we focus on the properties of the underlying wave functions, such as the number of effectively unpaired electrons. Comparison of different density functional and wave function theory results provides insight into the performance of the different methods when applied to strongly correlated systems such as polyradicals. We show that canonical molecular orbitals for species like large copper-containing diradicals fail to correctly represent the underlying electronic structure due to highly non-Koopmans character, while density-based analysis of the same wave function delivers a clear picture of the bonding pattern.


Role of Dietary Habits in the Prevention of Diverticular Disease Complications: A Systematic Review.
Recent evidence showed that dietary habits play a role as risk factors for the development of diverticular complications. This systematic review aims to assess the effect of dietary habits in the prevention of diverticula complications (i.e., acute diverticulitis and diverticula bleeding) in patients with diverticula disease. PubMed and Scopus databases were searched up to 19 January 2021, 330 records were identified, and 8 articles met the eligibility criteria and were subjected to data extraction. The quality of the studies was evaluated by the Newcastle-Ottawa quality assessment form. No study meets the criteria for being a high-quality study. A high intake of fiber was associated to a decreased risk of diverticulitis or hospitalization due to diverticular disease, with a protective effect for fruits and cereal fiber, but not for vegetable fiber; whereas, a high red meat consumption and a generally Western dietary pattern were associated with an increased risk of diverticulitis. Alcohol use seemed to be associated to diverticular bleeding, but not to recurrent diverticulitis or diverticular complications. Further high-quality studies are needed to better define these associations. It is mandatory to ascertain the role of dietary habits for the development of recurrent acute diverticulitis and diverticular bleeding.


Spatial summation of pain in humans investigated using transcutaneous electrical stimulation.
Spatial summation of pain is well accepted but surprisingly understudied. Area-based summation refers to the increase in pain evoked by increasing the area of stimulation. Distance-based summation refers to the increase in pain evoked by increasing the distance between multiple stimuli. Although transcutaneous electrical stimulation has several advantages over other experimental pain paradigms, whether or not this modality evokes spatial summation remains unknown. We aimed to answer this question in order to lay the foundation for critical studies of spatial summation. Twenty-five healthy participants received stimuli on their forearm, and the primary outcome, pain intensity, was compared across 5 spatial configurations-1 with a single stimulus and 4 paired configurations at 0-, 5-, 10-, and 20-cm separations. Importantly, the potential confounder of a proximal-distal gradient in nociceptive sensitivity was removed in this study. Pain intensity was higher in response to the paired stimuli than in response to the single stimulus (P < .001), and the paired stimuli separated by 5, 10 and 20 cm, evoked greater pain than stimuli at a separation of 0 cm (P < .001), thus confirming both area- and distance-based summation, respectively. We conclude that transcutaneous electrical stimulation is appropriate for future investigations of spatial summation. Distance-based summation is likely implicated in some clinical pain. However, current understanding for spatial summation is limited. This study demonstrates that transcutaneous electrical stimulation is safe, feasible, and valid for future investigations of spatial summation and will allow critical questions to be answered.


Quantification of the electrophilic reactivities of aldehydes, imines, and enones.
The rates of the epoxidation reactions of aldehydes, of the aziridination reactions of aldimines, and of the cyclopropanation reactions of α,β-unsaturated ketones with aryl-stabilized dimethylsulfonium ylides have been determined photometrically in dimethyl sulfoxide (DMSO). All of these sulfur ylide-mediated cyclization reactions as well as the addition reactions of stabilized carbanions to N-tosyl-activated aldimines have been shown to follow a second-order rate law, where the rate constants reflect the (initial) CC bond formation between nucleophile and electrophile. The derived second-order rate constants (log k(2)) have been combined with the known nucleophilicity parameters (N, s(N)) of the aryl-stabilized sulfur ylides 4a,b and of the acceptor-substituted carbanions 4c-h to calculate the electrophilicity parameters E of aromatic and aliphatic aldehydes (1a-i), N-acceptor-substituted aromatic aldimines (2a-e), and α,β-unsaturated ketones (3a-f) according to the linear free-energy relationship log k(2) = s(N)(N + E) as defined in J. Am. Chem. Soc.2001, 123, 9500-9512. The data reported in this work provide the first quantitative comparison of the electrophilic reactivities of aldehydes, imines, and simple Michael acceptors in DMSO with carbocations and cationic metal-π complexes within our comprehensive electrophilicity scale.


Homophilic PECAM-1(CD31) interactions prevent endothelial cell apoptosis but do not support cell spreading or migration.
PECAM-1 (CD31) is a highly abundant cell surface glycoprotein expressed on haemopoietic and endothelial cells. As well as mediating homophilic (PECAM-1/PECAM-1) adhesion, PECAM-1 can also bind the integrin alphavbeta3. Both PECAM-1 and alphavbeta3 have been shown to have roles in regulating angiogenesis, endothelial tube formation and in the case of alphavbeta3, endothelial cell apoptosis. In this study we show that despite being expressed at equivalent levels, endothelial alphavbeta3 is not a ligand for PECAM-1. Rather, PECAM-1 supports homophilic binding on HUVEC with similar characteristics to those we have previously reported for leukocytes and becomes tyrosine phosphorylated after homophilic PECAM-1 and integrin/fibronectin engagement. Immunoprecipitation studies show that in addition to SHP-2, tyrosine phosphorylated PECAM-1 can interact with at least four other phosphoproteins in pervanadate stimulated HUVEC. While PECAM-1/PECAM-1 interactions support robust endothelial cell adhesion, they do not support cell spreading or migration. In addition PECAM-1 homophilic adhesion rescues HUVEC from serum deprivation-induced apoptosis. Taken together our results indicate that PECAM-1 homophilic interactions play an important role in interendothelial cell adhesion, survival and signalling.


Appropriate nonmycorrhizal controls in arbuscular mycorrhiza research: a microbiome perspective.
Establishment of nonmycorrhizal controls is a "classic and recurrent theme" in mycorrhizal research. For decades, authors reported mycorrhizal plant growth/nutrition as compared to various nonmycorrhizal controls. In such studies, uncertainties remain about which nonmycorrhizal controls are most appropriate and, in particular, what effects the control inoculations have on substrate and root microbiomes. Here, different types of control and mycorrhizal inoculations were compared with respect to plant growth and nutrition, as well as the structure of root and substrate microbiomes, assessed by next-generation sequencing. We compared uninoculated ("absolute") control to inoculation with blank pot culture lacking arbuscular mycorrhizal fungi, filtrate of that blank inoculum, and filtrate of complex pot-produced mycorrhizal inoculum. Those treatments were compared to a standard mycorrhizal treatment, where the previously sterilized substrate was inoculated with complex pot-produced inoculum containing Rhizophagus irregularis SYM5. Besides this, monoxenically produced inoculum of the same fungus was applied either alone or in combination with blank inoculum. The results indicate that the presence of mycorrhizal fungus always resulted in stimulation of Andropogon gerardii plant biomass as well as in elevated phosphorus content of the plants. The microbial (bacterial and fungal) communities developing in the differently inoculated treatments, however, differed substantially from each other and no control could be obtained comparable with the treatment inoculated with complex mycorrhizal inoculum. Soil microorganisms with significant biological competences that could potentially contribute to the effects of the various inoculants on the plants were detected in roots and in plant cultivation substrate in some of the treatments.


PML nuclear bodies are general targets for inflammation and cell proliferation.
Acute promyelocytic leukemia is associated with a t(15;17) translocation that generates a fusion product between PML and the retinoic acid receptor alpha. Recently, PML was shown to concentrate within subnuclear domains, referred to as nuclear bodies, that are disorganized in acute promyelocytic leukemia cells. This observation provided the first evidence that alteration of a nuclear structure may play a role in human pathogenesis. In an attempt to clarify the role of PML and, more generally, of the associated nuclear bodies, we used immunohistochemistry to explore the expression of PML in normal, inflammatory, and neoplastic human tissues. With the exception of endothelial cells and macrophages that contain a high amount of PML protein, a weak speckled labeling pattern was observed in the nucleus of all cell types analyzed. By contrast to normal tissues, the level of PML expression was considerably enhanced in inflammatory tissues, predominantly around the mononuclear cell infiltrate, as well as during either normal or pathological proliferative states, in particular in tumoral pathology. Surprisingly, in most hepatocellular carcinoma, a cytoplasmic delocalization of PML was observed. Finally, the number of PML nuclear bodies increased up to twice their normal value as quiescent cultured cells were stimulated to grow upon serum addition. Altogether these results strongly suggest that the PML-associated nuclear bodies are implicated both in the inflammatory process and in cell growth control.