Defective recruitment of motor proteins to autophagic compartments contributes to autophagic failure in aging.
Inability to preserve proteostasis with age contributes to the gradual loss of function that characterizes old organisms. Defective autophagy, a component of the proteostasis network for delivery and degradation of intracellular materials in lysosomes, has been described in multiple old organisms, while a robust autophagy response has been linked to longevity. The molecular mechanisms responsible for defective autophagic function with age remain, for the most part, poorly characterized. In this work, we have identified differences between young and old cells in the intracellular trafficking of the vesicular compartments that participate in autophagy. Failure to reposition autophagosomes and lysosomes toward the perinuclear region with age reduces the efficiency of their fusion and the subsequent degradation of the sequestered cargo. Hepatocytes from old mice display lower association of two microtubule-based minus-end-directed motor proteins, the well-characterized dynein, and the less-studied KIFC3, with autophagosomes and lysosomes, respectively. Using genetic approaches to mimic the lower levels of KIFC3 observed in old cells, we confirmed that reduced content of this motor protein in fibroblasts leads to failed lysosomal repositioning and diminished autophagic flux. Our study connects defects in intracellular trafficking with insufficient autophagy in old organisms and identifies motor proteins as a novel target for future interventions aiming at correcting autophagic activity with anti-aging purposes.


Chemotherapy induces macrophage chemoattractant protein-1 production in ovarian cancer.
Tumor infiltrating macrophages play an important role in tumor progression. Macrophage chemoattractant protein-1 (MCP-1) is one of the major chemokines responsible for inducing macrophage migration. Our objective was to investigate chemotherapy-induced modulation of MCP-1 in ovarian cancer by investigating macrophage infiltration, tumor vascularity, and MCP-1 expression after chemotherapy exposure. MA-148 ovarian cancer cells were treated with paclitaxel (43 pg/mL) and carboplatin (5 microg/mL) alone or in combination. Reverse transcription-polymerase chain reaction determined MCP-1 transcript levels and enzyme-linked immunosorbent assay evaluated MCP-1 protein production at multiple time points. The effect of kinase inhibitors on MCP-1 expression was investigated. In vivo MCP-1 production was examined in tumor-bearing mice and immunohistochemistry with fluorescein isothiocyanate conjugated anti-mouse F4/80 antibody, phycoerythrin-anti-CD31, and terminal deoxynucleotide transferase dUTP nick-end labeling assay were performed. Macrophage chemoattractant protein-1 transcript levels were up-regulated in MA-148 after treatment with paclitaxel and carboplatin individually and in combination. The greatest elevation was seen with combination therapy: 2.5-fold increase in the MCP-1 protein levels from baseline (P = 0.011) with the mitogen-activated protein kinase and janus kinases/signal transducers and activators of transcription pathways appearing to be involved in the regulation of MCP-1 production. In vivo mouse studies confirmed increased MCP-1 production after chemotherapy; however, there was no significant difference in macrophage, apoptosis, or vessel density. Macrophage chemoattractant protein-1 is up-regulated in ovarian cancer after chemotherapy in vitro and in vivo. Whether MCP-1 production is increased because of a stress-induced response or a scavenger response promoting macrophage infiltration remains unknown. Chemotherapy induction of MCP-1 in ovarian cancer suggests this chemokine plays an important role in the immune response occurring after chemotherapy exposure.


Techniques and results of portal vein/superior mesenteric vein reconstruction using femoral and saphenous vein during pancreaticoduodenectomy.
Patients with pancreatic tumors may have portal vein (PV) and/or superior mesenteric vein (SMV) invasion. In such cases, lower extremity veins can provide an autogenous conduit for PV/SMV reconstruction. Little data exist, however, describing the technique of PV/SMV reconstruction, patency of such reconstructions, and the morbidity of using lower extremity veins for PV/SMV reconstruction during pancreaticoduodenectomy. Thirty-four patients underwent PV/SMV reconstruction during pancreaticoduodenectomy using lower extremity vein. The saphenous vein was preferred for patching and femoral vein for replacement. We analyzed preoperative imaging, reconstruction patency, vein harvest morbidity, and late mortality. The mean age was 62.6 years. All 34 patients had preoperative computed tomography (CT) imaging and/or endoscopic ultrasound (EUS) scan. Fourteen of the 34 patients had evidence of PV/SMV invasion on CT or EUS scans, 14 did not, and six studies were indeterminate. Twenty-five patients had follow-up imaging, and 22 (88%) had patent reconstructions. Fifteen patients had PV/SMV replacement using femoral vein. Seven of these 15 had minor postoperative lower extremity edema that resolved over time, five had wound complications from the femoral vein harvest site, three of which required minor operative procedures for treatment. Fifteen patients had PV/SMV patching with the great saphenous vein, none had postoperative wound problems, and one had minimal postoperative lower extremity edema. Four patients had PV/SMV patching using femoral vein, none had postoperative wound problems, and one had minimal postoperative lower extremity edema. Compared with patients undergoing pancreaticoduodenectomy without PV/SMV reconstruction, by Kaplan-Meier analysis, there was no difference in late mortality. Preoperative imaging may fail to detect PV/SMV involvement in patients undergoing pancreaticoduodenectomy. The PV/SMV reconstruction with leg vein provides good patency with minimal postoperative lower extremity complications and no increase in late mortality. The lower extremities should be routinely included in the operative field of patients undergoing pancreaticoduodenectomy.


miR-183-5p regulates uterine receptivity and enhances embryo implantation.
Receptive endometrium is a prerequisite for successful embryo implantation, and it follows that poor endometrial receptivity is a leading cause of implantation failure. miRNAs play important roles as epigenetic regulators of endometrial receptivity and embryo implantation through post-transcriptional modifications. However, the mechanisms of action of many miRNAs are poorly understood. In this study, we investigated the role of the miR-183 family, comprising three miRNAs (miR-183-5p, miR-182-5p, and miR-96-5p) in endometrial receptivity and embryo implantation. The miR-183 family shows estrogen-dependent upregulation in endometrial Ishikawa (IK) cells. The miR-183 family also has a positive role in migration and proliferation of IK cells. Furthermore, JAr spheroid attachment experiments show that attachment rates were significantly decreased after treatment of IK cells with inhibitors for miR-183-5p and miR-182-5p and increased after treatment with miR-183-5p-mimic and miR-96-5p-mimic, respectively. The downstream analysis shows that catenin alpha 2 (CTNNA2) is a potential target gene for miR-183-5p, and this was confirmed in luciferase reporter assays. An in vivo mouse pregnancy model shows that inhibition of miR-183-5p significantly decreases embryo implantation rates and increases CTNNA2 expression. Downregulation of CTNNA2 in endometrial cells by miR-183-5p may be significant in mediating estrogenic effects on endometrial receptivity. In conclusion, miR-183-5p and the CTNNA2 gene may be potential biomarkers for endometrial receptivity and may be useful diagnostic and therapeutic targets for successful embryo implantation.


Determinations of personal carbon monoxide exposure and blood carboxyhemoglobin levels in Korea.
Determinant factors for personal carbon monoxide (CO) exposures were sought in Korea, where CO poisoning has been a major public health problem due to coal briquette (Yeontan) combustion for space heating and cooking. Personal 24-hr CO exposures of 15 housewives were measured by CO passive samplers on 2 days of the week (Wednesday and Sunday). Blood samples were taken to measure carboxyhemoglobin (COHb) just after the exposure sampling. Average CO exposure and COHb level were 5.6 ppm and 2.4%, respectively. Personal CO exposures as well as COHb levels were significantly increased by the use of Yeontan, especially on a weekday. Carboxyhemoglobin levels were closely related to the time between blood collection and replacement of Yeontan: the closer the blood collection was to replace Yeontan, the higher the COHb levels were. Assuming a background COHb of 1.34%, COHb increased on average by 1.8% with a 24-hr personal CO exposure of 10 ppm. The relationship between CO exposure and COHb level was provided by simultaneous direct measurements in real environment, although a measurement of COHb at the end of exposure could not represent previous 24-hr exposure thoroughly.


Mechanisms of acquired thymic tolerance in experimental autoimmune encephalomyelitis: thymic dendritic-enriched cells induce specific peripheral T cell unresponsiveness in vivo.
Experimental autoimmune encephalomyelitis (EAE), an experimental model for the study of multiple sclerosis, is an autoimmune disease of the central nervous system that can be induced in a number of species by immunization with myelin basic protein (MBP). MBP-reactive CD4+ T cells, predominantly expressing the V beta 8.2 T cell receptor (TCR), migrate from the peripheral lymphoid organs and initiate the inflammatory response in the brain. We have previously shown that a single intrathymic injection of MBP or its major encephalitogenic peptide (p71-90), but not a nonencephalitogenic peptide (p21-40), induces antigen-specific systemic tolerance and inhibits the induction of EAE in Lewis rats. In this study, we investigated the mechanisms of induction and maintenance of acquired thymic tolerance in this model. First, we investigated which thymic cell is responsible for "induction" of systemic tolerance. Thymic dendritic-enriched cells, isolated by plastic adherence, when incubated in vitro with p71-90 and injected intravenously into Lewis rats, were capable of preventing the development of EAE, but his protection was lost in thymectomized recipients. In addition, intravenous injection of thymic dendritic cells isolated from animals that had been previously injected intrathymically with p71-90 but not p21-40 also prevented the development of EAE. Second, to determine the "effector" mechanisms involved in acquired thymic tolerance, we compared TCR expression in the brains of animals with actively induced EAE with TCR expression in animals that received intrathymic injection of p71-90 or p21-40. Using a semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) technique, we found increased expression of CD4 and V beta 8.2 message in brains of immunized animals compared with those of naive animals. In animals intrathymically injected with p71-90 but not p21-40, CD4 and V beta 8.2 transcript levels were significantly reduced compared with immunized controls. Immunohistologic studies of brain tissue and spleens with specific V beta 8.2 and control V beta 10 monoclonal antibodies confirmed these observations in vivo. These findings, taken together with recent data demonstrating that activated T cells circulate through the thymus, suggest that interaction of thymic dendritic cells with specific TCR of activated peripheral T cells can lead to inactivation of these antigen-specific cells and confirm the role of V beta 8.2-expressing T cells in EAE.


Continuous wave mirrorless lasing in cholesteric liquid crystals with a pitch gradient across the cell gap.
Despite numerous efforts, continuous wave (CW) lasing in dye doped, one-dimensional (1D) photonic bandgap cholesteric liquid crystal (CLC) structures has not been previously reported, to our knowledge. Here we report on the observation of lasing in such structures under both coherent (laser) and incoherent (LED) CW light excitation. To achieve this effect, we used a 1D-photonic bandgap structure made of a polymer stabilized CLC with a pitch gradient across the cell thickness. A spectral reflectivity profile of such a CLC structure reveals local minima in the area within a photonic stopband and close to it. The realization of lasing pumped by low power CW light sources opens the possibility of all-organic, compact, tunable CW lasers for display and medical applications.


A Novel Perfusion System for Damage Control of Hyperkalemia in Swine.
The standard of care for refractory hyperkalemia is renal replacement therapy (RRT). However, traditional RRT requires specialized equipment, trained personnel, and large amounts of dialysate. It is therefore poorly suited for austere environments. We hypothesized that a simplified hemoperfusion system could control serum potassium concentration in a swine model of acute hyperkalemia. Ten pigs were anesthetized and instrumented. A dialysis catheter was inserted. After bilateral nephrectomy, animals received intravenous potassium chloride and were randomized to the control or treatment group. In both groups, blood was pumped through an extracorporeal circuit (EC) with an in-line hemodialyzer. In the treatment arm, ultrafiltrate from the hemodialyzer was diverted through cartridges containing novel potassium binding beads and returned to the EC. Blood samples were obtained every 30 min for 6 h. Serum potassium concentration was significantly lower in the treatment than in the control group over time (P = 0.02). There was no difference in serum total calcium concentration for group or time (P = 0.13 and 0.44, respectively) or platelet count between groups or over time (P = 0.28 and 1, respectively). No significant EC thrombosis occurred. Two of five animals in the control group and none in the treatment group developed arrhythmias. All animals survived until end of experiment. A simplified hemoperfusion system removed potassium in a porcine model. In austere settings, this system could be used to temporize patients with hyperkalemia until evacuation to a facility with traditional RRT.


Transitioning from Forensic Genetics to Forensic Genomics.
Due to its support of law enforcement, forensics is a conservative field; nevertheless, driven by scientific and technological progress, forensic genetics is slowly transitioning into forensic genomics. With this Special Issue of Genes we acknowledge and appreciate this rather recent development by not only introducing the field of forensics to the wider community of geneticists, but we do so by emphasizing on different topics of forensic relevance where genomic, transcriptomic, and epigenomic principles, methods, and datasets of humans and beyond are beginning to be used to answer forensic questions.


Dopamine reveals neural circuit mechanisms of fly memory.
A goal of memory research is to understand how changing the weight of specific synapses in neural circuits in the brain leads to an appropriate learned behavioral response. Finding the relevant synapses should allow investigators to probe the underlying physiological and molecular operations that encode memories and permit their retrieval. In this review I discuss recent work in Drosophila that implicates specific subsets of dopaminergic (DA) neurons in aversive reinforcement and appetitive motivation. The zonal architecture of these DA neurons is likely to reveal the functional organization of aversive and appetitive memory in the mushroom bodies. Combinations of fly DA neurons might code negative and positive value, consistent with a motivational systems role as proposed in mammals.


Phase behaviour of the symmetric binary mixture from thermodynamic perturbation theory.
We study the phase behaviour of symmetric binary mixtures of hard core Yukawa (HCY) particles via thermodynamic perturbation theory (TPT). We show that all the topologies of phase diagram reported for the symmetric binary mixtures are correctly reproduced within the TPT approach. In a second step we use the capability of TPT to be straightforwardly extended to mixtures that are nonsymmetric in size. Starting from mixtures that belong to the different topologies of symmetric binary mixtures we investigate the effect on the phase behaviour when an asymmetry in the diameters of the two components is introduced. Interestingly, when the energy of interaction between unlike particles is weaker than the interaction between like particles, the propensity for the solution to demix is found to increase strongly with size asymmetry.


Role of fragile histidine triad protein expression in pathogenesis of malignant pleural mesothelioma.
To investigate the relationship of fragile histidine triad (FHIT) and Ki-67 expression with clinicopathological variables of patients with malignant pleural mesothelioma (MPM). Formalin-fixed, paraffin-embedded tissue sections of 30 asbestos induced MPM (epithelial and biphasic) patients were examined for FHIT and Ki-67 expression using immunohistochemical techniques and results were compared with clinicopathological variables. Immunohistochemical study results were as follows: 12 (40%) cases showed low FHIT expression and 18 (60%) showed high expression. There was no significant relationship between FHIT and age, gender or histological subtypes (p > 0.05). Ki-67 expression was 'low' in 13 (43.3%) cases and 'high' in 17 (56.7%) cases. No correlation could be demonstrated between Ki-67 expression and age, gender or histological subtypes (p > 0.05). No significant association was observed between FHIT and Ki-67 expression in MPM. The results support the role of FHIT as a tumour suppressor gene in asbestos induced MPM. There is no significant correlation between FHIT and cell proliferation marker expressions in malignant pleural mesothelioma. Therefore, it can be concluded that loss of FHIT does not interfere with tumour proliferation. This can be accepted as evidence for an early role of FHIT loss in carcinogenesis; however, it needs to be strengthened by further studies.


Intrathecal enzyme replacement therapy improves motor function and survival in a preclinical mouse model of infantile neuronal ceroid lipofuscinosis.
The neuronal ceroid lipofuscinoses (NCLs) are a group of related hereditary lysosomal storage disorders characterized by progressive loss of neurons in the central nervous system resulting in dementia, loss of motor skills, seizures and blindness. A characteristic intralysosomal accumulation of autofluorescent storage material occurs in the brain and other tissues. Three major forms and nearly a dozen minor forms of NCL are recognized. Infantile-onset NCL (CLN1 disease) is caused by severe deficiency in a soluble lysosomal enzyme, palmitoyl-protein thioesterase-1 (PPT1) and no therapy beyond supportive care is available. Homozygous Ppt1 knockout mice reproduce the known features of the disease, developing signs of motor dysfunction at 5 months of age and death around 8 months. Direct delivery of lysosomal enzymes to the cerebrospinal fluid is an approach that has gained traction in small and large animal models of several other neuropathic lysosomal storage diseases, and has advanced to clinical trials. In the current study, Ppt1 knockout mice were treated with purified recombinant human PPT1 enzyme delivered to the lumbar intrathecal space on each of three consecutive days at 6 weeks of age. Untreated PPT1 knockout mice and wild-type mice served as additional controls. Four enzyme concentration levels (0, 2.6, 5.3 and 10.6 mg/ml of specific activity 20 U/mg) were administered in a volume of 80 μl infused over 8 min. Each group consisted of 16-20 mice. The treatment was well tolerated. Disease-specific survival was 233, 267, 272, and 284days for each of the four treatment groups, respectively, and the effect of treatment was highly significant (p<0.0001). The timing of motor deterioration was also delayed. Neuropathology was improved as evidenced by decreased autofluorescent storage material in the spinal cord and a decrease in CD68 staining in the cortex and spinal cord. The improvements in motor function and survival are similar to results reported for preclinical studies involving other lysosomal storage disorders, such as CLN2/TPP1 deficiency, for which intraventricular ERT is being offered in clinical trials. If ERT delivery to the CSF proves to be efficacious in these disorders, PPT1 deficiency may also be amenable to this approach.


Novel HPAIV H5N8 Reassortant (Clade 22.214.171.124b) Detected in Germany.
A novel H5N8 highly pathogenic avian influenza virus (HPAIV) was detected in a greater white-fronted goose in January 2020 in Brandenburg, Germany, and, in February 2020, in domestic chickens belonging to a smallholding in Baden-Wuerttemberg, Germany. Full-genome sequencing was conducted on the MinION platform, enabling further phylogenetic analyses. The virus of clade 22.214.171.124b holds six segments from a Eurasian/Asian/African HPAIV H5N8 reassortant and two segments from low pathogenic avian influenza H3N8 subtype viruses recently detected in wild birds in Central Russia. These new entries continue to show the reassortment potential of the clade 188.8.131.52 H5Nx viruses, underlining the necessity for full-genome sequencing and continuous surveillance.


High-sensitivity CRP discriminates HNF1A-MODY from other subtypes of diabetes.
Maturity-onset diabetes of the young (MODY) as a result of mutations in hepatocyte nuclear factor 1-α (HNF1A) is often misdiagnosed as type 1 diabetes or type 2 diabetes. Recent work has shown that high-sensitivity C-reactive protein (hs-CRP) levels are lower in HNF1A-MODY than type 1 diabetes, type 2 diabetes, or glucokinase (GCK)-MODY. We aim to replicate these findings in larger numbers and other MODY subtypes. hs-CRP levels were assessed in 750 patients (220 HNF1A, 245 GCK, 54 HNF4-α [HNF4A], 21 HNF1-β (HNF1B), 53 type 1 diabetes, and 157 type 2 diabetes). hs-CRP was lower in HNF1A-MODY (median [IQR] 0.3 [0.1-0.6] mg/L) than type 2 diabetes (1.40 [0.60-3.45] mg/L; P < 0.001) and type 1 diabetes (1.10 [0.50-1.85] mg/L; P < 0.001), HNF4A-MODY (1.45 [0.46-2.88] mg/L; P < 0.001), GCK-MODY (0.60 [0.30-1.80] mg/L; P < 0.001), and HNF1B-MODY (0.60 [0.10-2.8] mg/L; P = 0.07). hs-CRP discriminated HNF1A-MODY from type 2 diabetes with hs-CRP <0.75 mg/L showing 79% sensitivity and 70% specificity (receiver operating characteristic area under the curve = 0.84). hs-CRP levels are lower in HNF1A-MODY than other forms of diabetes and may be used as a biomarker to select patients for diagnostic HNF1A genetic testing.


Pedicled anterolateral thigh flap transfer for the reconstruction of a large gluteal defect assisted by preoperative computed tomographic angiography and intraoperative indocyanine green angiography: A case report.
The efficacy of the use of the pedicled ALT flap for buttock defects has not been established in the literature. Here, we present a case of a successful reconstruction of a large gluteal defect after sarcoma resection using a pedicled ALT flap assisted by preoperative computed tomographic angiography (CTA) and intraoperative indocyanine green (ICG) angiography. A 94-year-old man underwent resection of a large soft tissue tumor in the gluteal region resulting in a 19 × 15 cm defect. Based on three-dimensional reconstructed images of the preoperative CTA, the length of the pedicle of an ALT flap was estimated. Perfusion of the whole flap was confirmed using intraoperative ICG angiography. The flap was inserted through a subcutaneous tunnel and covered the defect completely. The postoperative course was uneventful. The patient resumed ambulation 1 week after the operation. The follow-up at 2 months showed complete flap survival, and the patient was able to walk with a cane. The pedicled ALT flap transfer may be a reliable alternative for gluteal reconstruction when guided by pre- and intraoperative imaging modalities.


[Impact of education program and clinical posting in psychiatry on medical students' stigmatizing attitudes towards psychiatry and psychiatric disorders].
The aim of the study was to explore whether a medical student education program and clinical posting in psychiatry had an impact on medical students' stigmatizing attitudes towards psychiatry and psychiatric disorders. Medical students from the University of Bordeaux were recruited during their 4-year course at the beginning of the academic education program in psychiatry. Medical students who were concomitantly in a clinical posting in wards of psychiatry or neurology were invited to participate in the study. The medical student version of the scale Mental Illness: Clinicians' Attitudes (MICA) was used to measure their attitudes towards psychiatry and persons with psychiatric disorder. This 16-item scale is designed to measure attitudes of health care professionals towards people with mental illness, a higher score indicating more stigmatizing attitudes. Items exploring history of psychiatric disorders in close persons were added at the end of the MICA scale. The questionnaire was completed twice by each student, at the beginning and the end of the 11-week clinical posting. All questionnaires were strictly anonymized. Multivariate linear regression analyses were used to identify the variables independently associated with MICA total score. At the beginning of the education program and clinical posting, 174 students completed the MICA scale: the mean MICA total score was equal to 46.4 (SD 6.9) in students in clinical posting in psychiatry (n=72) and 45.1 (SD 7.01) in those in neurology (n=102). At the end of the academic and clinical training, 138 students again completed the questionnaire, with mean MICA total scores equal to 41.4 (SD 8.1) in students in clinical posting in psychiatry (n=51) and 43.5 (SD 7.3) in those in neurology (n=87). Multivariate analyses showed that lower total MICA scores were independently associated with the time of assessment (lower scores at the end of education program and clinical posting) (b=-2.8; P=0.001), female gender (b=-1.8; P=0.03) and history of a psychiatric disorder in a close person (b=-1.92; P=0.02). Type of clinical posting (psychiatry vs. neurology) was not independently associated with MICA total scores (b=-0.02; P=0.98). A significant interaction was found between the variables "time of assessment" and "type of clinical posting" (P=0.05): stratified analyses showed that MICA total scores decreased significantly only when the clinical posting was in psychiatry (b=-4.66; P=0.001), with no significant change in medical students in neurology wards (b=-1.45; P=0.16). Stigmatizing attitudes of medical students towards psychiatry and psychiatric disorders are reduced by an education program in psychiatry, with a positive impact more marked when the education program is concomitant to a clinical posting in psychiatry. As future health professionals in charge of persons with psychiatric disorders, medical students are key targets of actions aimed at reducing stigma towards mental health disorders. It is hence of great importance to promote clinical training in psychiatric wards during medical studies for all future practitioners, irrespective of their future specialty.


SARS-CoV-2 lineage B.1.1.7 is associated with greater disease severity among hospitalised women but not men: multicentre cohort study.
SARS-CoV-2 lineage B.1.1.7 has been associated with an increased rate of transmission and disease severity among subjects testing positive in the community. Its impact on hospitalised patients is less well documented. We collected viral sequences and clinical data of patients admitted with SARS-CoV-2 and hospital-onset COVID-19 infections (HOCIs), sampled 16 November 2020 to 10 January 2021, from eight hospitals participating in the COG-UK-HOCI study. Associations between the variant and the outcomes of all-cause mortality and intensive therapy unit (ITU) admission were evaluated using mixed effects Cox models adjusted by age, sex, comorbidities, care home residence, pregnancy and ethnicity. Sequences were obtained from 2341 inpatients (HOCI cases=786) and analysis of clinical outcomes was carried out in 2147 inpatients with all data available. The HR for mortality of B.1.1.7 compared with other lineages was 1.01 (95% CI 0.79 to 1.28, p=0.94) and for ITU admission was 1.01 (95% CI 0.75 to 1.37, p=0.96). Analysis of sex-specific effects of B.1.1.7 identified increased risk of mortality (HR 1.30, 95% CI 0.95 to 1.78, p=0.096) and ITU admission (HR 1.82, 95% CI 1.15 to 2.90, p=0.011) in females infected with the variant but not males (mortality HR 0.82, 95% CI 0.61 to 1.10, p=0.177; ITU HR 0.74, 95% CI 0.52 to 1.04, p=0.086). In common with smaller studies of patients hospitalised with SARS-CoV-2, we did not find an overall increase in mortality or ITU admission associated with B.1.1.7 compared with other lineages. However, women with B.1.1.7 may be at an increased risk of admission to intensive care and at modestly increased risk of mortality.


Inferring physiological energetics of loggerhead turtle (Caretta caretta) from existing data using a general metabolic theory.
Loggerhead turtle is an endangered sea turtle species with a migratory lifestyle and worldwide distribution, experiencing markedly different habitats throughout its lifetime. Environmental conditions, especially food availability and temperature, constrain the acquisition and the use of available energy, thus affecting physiological processes such as growth, maturation, and reproduction. These physiological processes at the population level determine survival, fecundity, and ultimately the population growth rate-a key indicator of the success of conservation efforts. As a first step towards the comprehensive understanding of how environment shapes the physiology and the life cycle of a loggerhead turtle, we constructed a full life cycle model based on the principles of energy acquisition and utilization embedded in the Dynamic Energy Budget (DEB) theory. We adapted the standard DEB model using data from published and unpublished sources to obtain parameter estimates and model predictions that could be compared with data. The outcome was a successful mathematical description of ontogeny and life history traits of the loggerhead turtle. Some deviations between the model and the data existed (such as an earlier age at sexual maturity and faster growth of the post-hatchlings), yet probable causes for these deviations were found informative and discussed in great detail. Physiological traits such as the capacity to withstand starvation, trade-offs between reproduction and growth, and changes in the energy budget throughout the ontogeny were inferred from the model. The results offer new insights into physiology and ecology of loggerhead turtle with the potential to lead to novel approaches in conservation of this endangered species.


Trigeminal neurinomas.
Trigeminal neurinomas may produce atypical facial pain, loss of facial sensation, facial palsy, hearing loss, nystagmus, or vertigo, and thus may be confused with nasopharyngeal carcinomas or acoustic neurinomas. The diagnosis can be made by the clinical presentation together with radiographic features showing widening of the foramen ovale and/or smooth destruction of the anteromedial portion of the peterous apex. Tomography, pneumoencephalography, angiography, and other diagnostic procedures are sometimes helpful. Treatment is removal by transtemporal craniotomy and offers an excellent prognosis.


Abnormal transcription factor induction through the surface immunoglobulin M receptor of B-1 lymphocytes.
Populations of murine peritoneal B-1 and splenic B-2 cells, highly purified by negative selection techniques, were used to demonstrate that B-1 cells completely fail to enter cell cycle in response to surface immunoglobulin M (sIgM) crosslinking without any decrease in cell number or viability. This failure of B-1 cell responsiveness appears to represent a specific defect in sIgM-derived signaling inasmuch as stimulation to enter S phase occurs normally in response to activated and fixed T cells, and to lipopolysaccharide (LPS). The level at which sIgM signaling fails was determined by evaluating the nuclear expression of the transcription factor complex, NF-kappa B, whose sIgM-mediated induction in B-2 cells is dependent on protein kinase C (PKC) activation but is independent of protein synthesis. There was no induction of nuclear NF-kappa B in B-1 cells stimulated by sIgM crosslinking, although NF-kappa B was stimulated by phorbol myristate acetate and by LPS. In contrast, NF-kappa B was induced in B-2 cells by all three stimuli. Thus, in B-1 cells, the sIgM-mediated induction of a transcription factor that is substantially stimulated by anti-IgM in B-2 cells is blocked. However, all sIgM-derived signaling in B-1 cells was not impaired inasmuch as anti-IgM increased I-A antigen expression. These results strongly suggest that sIgM receptor-mediated signaling in B-1 cells is interrupted early in the signal transduction pathway, at a point proximal to the activation of PKC. These results further demonstrate that transcription factor induction can be used to analyze the level at which receptor-mediated signaling is blocked.


Approaches to improve cellular retention of radiohalogen labels delivered by internalising tumour-targeting proteins and peptides.
Specific targeting of radionuclides is a promising approach to improve diagnosis and treatment of tumors. Targeting vectors may be monoclonal antibodies directed toward tumour-specific antigens or regulatory peptides binding to receptors overexpressed on or by malignant cells. Depending on the aim of the procedure and the biokinetics of the targeting vectors, radionuclides with different nuclear properties (decay scheme, half-life, etc.) must be applied. Halogen radioisotopes are attractive since they exhibit a variety of nuclear properties suitable for various applications. At the same time, their chemistry shows great similarities, which enables the use of similar labelling procedures for different nuclides. A problem in using radiohalogens for labelling of tumour-targeting proteins and peptides is that the commonly used radiohalogenation methods provide labels, which, after internalisation and lysosomal digestion, rapidly "leak" from malignant cells as radiohalogenated degradation products. The main reason for such leakage is free diffusion of the radiometabolites through lysosomal and cellular membranes. This review describes current approaches in molecular design to improve cellular retention of radiohalogen labels. These approaches include the use of prosthetic groups for the attachment of radiohalogens to targeting vectors of bulky hydrophilic non-charged molecules, molecules positively charged at lysosomal pH and negatively charged molecules. The emphasis in this paper is on labelling chemistry and the results of the biological testing of labelled compounds.


Relationship of Cigarette Smoking and Time of Quitting with Incident Dementia and Cognitive Decline.
Understanding how dementia risk is impacted by timing of smoking cessation has public health implications for prevention efforts. We investigated the relationship of cigarette smoking and cessation with dementia risk and cognitive decline in the Atherosclerosis Risk in Communities (ARIC) study. Ongoing prospective cohort study. Begun in 1987-1989, ARIC was conducted in four US communities. A total of 13 002 men and women (25% African American) aged 52 to 75 years. All-cause dementia was defined using standardized algorithms incorporating longitudinal cognitive data, proxy report, and hospital and death certificate dementia codes. Cognitive decline was measured using a composite cognitive score created from three tests measured at two time points (1996-1998 and 2011-2013). Smoking and cessation status were defined by self-report using data from 1987-1989 (visit 1) and 1996-1998 (visit 4). Incident dementia risk and differences in cognitive change by smoking status were estimated with Cox proportional hazards and linear regression models, respectively. To address smoking-related attrition, cognitive scores were imputed for living participants with incomplete cognitive testing. The proportion of never, former, and current smokers was 44%, 41%, and 14%; 79% of former smokers quit 9 years or more before baseline. A total of 1347 participants developed dementia. After adjustment, compared with never smoking, the hazard ratio for all-cause dementia for current smoking was 1.33 (95% confidence interval [CI] = 1.12-1.59) and for recent quitting (<9 y before baseline) was 1.24 (95% CI = 1.01-1.52). Quitting 9 years or more before baseline was not associated with dementia. We found no differences in rates of cognitive decline by smoking status. Although quitting at any time suggested benefit, dementia risk depended on time since smoking cessation. Our study highlights the importance of early midlife cessation to decrease dementia risk. J Am Geriatr Soc 68:337-345, 2020.


Food and drink purchasing habits out of school at lunchtime: a national survey of secondary school pupils in Scotland.
Food and drink purchasing habits of pupils out of school at lunchtime may be contributing to poor dietary intakes and overweight and obesity. The aim of this study was to identify the places from which purchases were made, types of food and drinks purchased and, the reasons for purchasing food or drinks out of school. A survey of the food and drinks purchasing habits of secondary school pupils (11-16 yrs) out of school at lunchtime was conducted in Scotland in 2010. A face-to-face interview and a self-completion questionnaire was designed to identify the food outlets used at lunchtime, types of food and drinks purchased and pupils' reasons for purchasing food or drinks out of school. Height and weight were measured and BMI centiles used to classify pupils as normal weight, overweight or obese. Results were compared by age group, sex, BMI group and level of socio-economic deprivation. Of the 612 pupils who completed the survey, 97 % reported having access to places selling food or drinks out of school at lunchtime, and of these 63 % made purchases. A higher proportion of pupils from more deprived areas reported purchasing food or drinks out of school, but the proportion making purchases did not differ significantly by sex or BMI group. Supermarkets were the outlets from which pupils reported most often making purchases, with fewer purchasing food or drinks from fast food takeaways, and this did not differ significantly by socio-economic deprivation. Reasons for making purchases included availability of preferred food and drinks, some of which are restricted for sale in schools, and social reasons, such as wanting to be with friends. Sandwiches and non-diet soft drinks were items most commonly purchased, followed by confectionery and diet soft drinks. However, less than 10 % of all the secondary school pupils reported purchasing these foods every day. Supermarkets, not just fast food outlets, should be considered when developing strategies to improve the dietary habits of pupils at lunchtime. The importance of food preferences and social reasons for purchasing food and drinks need to be acknowledged and integrated in future interventions.


A meta-analysis of group cognitive behavioral therapy for insomnia.
Insomnia is the most common sleep disorder among the general population. Although cognitive behavioral therapy for insomnia (CBT-I) is the psychological treatment of choice, the availability of individual therapy is often not sufficient to meet the demand for treatment. Group treatment can increase the efficiency of delivery, but its efficacy has not been well-established. Randomized controlled trials (RCTs) comparing group CBT-I to a control group in patients with insomnia were identified. A review of 670 unique citations resulted in eight studies that met criteria for analysis. Outcome variables included both qualitative (e.g., sleep quality) and quantitative (e.g., sleep diary) outcomes, as well as depression and pain severity, at both pre- to post-treatment and follow-up (3-12 mo post-treatment). Overall, we found medium to large effect sizes for sleep onset latency, sleep efficiency, and wake after sleep onset and small effect sizes for pain outcomes. Effect sizes remained significant at follow-up, suggesting that treatment gains persist over time. Other variables, including total sleep time, sleep quality, and depression, showed significant improvements, but these findings were limited to the within treatment group analyses. It is clear that group CBT-I is an efficacious treatment. Implications for stepped care models for insomnia are discussed.


Posture affects how robots and infants map words to objects.
For infants, the first problem in learning a word is to map the word to its referent; a second problem is to remember that mapping when the word and/or referent are again encountered. Recent infant studies suggest that spatial location plays a key role in how infants solve both problems. Here we provide a new theoretical model and new empirical evidence on how the body - and its momentary posture - may be central to these processes. The present study uses a name-object mapping task in which names are either encountered in the absence of their target (experiments 1-3, 6 & 7), or when their target is present but in a location previously associated with a foil (experiments 4, 5, 8 & 9). A humanoid robot model (experiments 1-5) is used to instantiate and test the hypothesis that body-centric spatial location, and thus the bodies' momentary posture, is used to centrally bind the multimodal features of heard names and visual objects. The robot model is shown to replicate existing infant data and then to generate novel predictions, which are tested in new infant studies (experiments 6-9). Despite spatial location being task-irrelevant in this second set of experiments, infants use body-centric spatial contingency over temporal contingency to map the name to object. Both infants and the robot remember the name-object mapping even in new spatial locations. However, the robot model shows how this memory can emerge -not from separating bodily information from the word-object mapping as proposed in previous models of the role of space in word-object mapping - but through the body's momentary disposition in space.


The Small Molecule R-(-)-β-O-Methylsynephrine Binds to Nucleoporin 153 kDa and Inhibits Angiogenesis.
R-(-)-β-O-methylsynephrine (OMe-Syn) is a naturally occurring small molecule that was identified in a previous screen as an inhibitor of angiogenesis. In this study, we conducted two animal model experiments to investigate the in vivo antiangiogenic activity of OMe-Syn. OMe-Syn significantly inhibited angiogenesis in a transgenic zebrafish model as well as in a mouse retinopathy model. To elucidate the underlying mechanisms responsible for the antiangiogenic activity of OMe-Syn, we used phage display cloning to isolate potential OMe-Syn binding proteins from human cDNA libraries and identified nucleoporin 153 kDa (NUP153) as a primary binding partner of OMe-Syn. OMe-Syn competitively inhibited mRNA binding to the RNA-binding domain of NUP153. Furthermore, depletion of NUP153 in human cells or zebrafish embryos led to an inhibition of angiogenesis, in a manner similar to that seen in response to OMe-Syn treatment. These data suggest that OMe-Syn is a promising candidate for the development of a novel antiangiogenic agent and that inhibition of NUP153 is possibly responsible for the antiangiogenic activity of OMe-Syn.


Identification of Schlafen-11 as a Target of CD47 Signaling That Regulates Sensitivity to Ionizing Radiation and Topoisomerase Inhibitors.
Knockdown or gene disruption of the ubiquitously expressed cell surface receptor CD47 protects non-malignant cells from genotoxic stress caused by ionizing radiation or cytotoxic chemotherapy but sensitizes tumors in an immune competent host to genotoxic stress. The selective radioprotection of non-malignant cells is mediated in part by enhanced autophagy and protection of anabolic metabolism pathways, but differential H2AX activation kinetics suggested that the DNA damage response is also CD47-dependent. A high throughput screen of drug sensitivities indicated that CD47 expression selectively sensitizes Jurkat T cells to inhibitors of topoisomerases, which are known targets of Schlafen-11 (SLFN11). CD47 mRNA expression positively correlated with schlafen-11 mRNA expression in a subset of human cancers but not the corresponding non-malignant tissues. CD47 mRNA expression was also negatively correlated with SLFN11 promoter methylation in some cancers. CD47 knockdown, gene disruption, or treatment with a CD47 function-blocking antibody decreased SLFN11 expression in Jurkat cells. The CD47 signaling ligand thrombospondin-1 also suppressed schlafen-11 expression in wild type but not CD47-deficient T cells. Re-expressing SLFN11 restored radiosensitivity to a CD47-deficient Jurkat cells. Disruption of CD47 in PC3 prostate cancer cells similarly decreased schlafen-11 expression and was associated with a CD47-dependent decrease in acetylation and increased methylation of histone H3 in the SLFN11 promoter region. The ability of histone deacetylase or topoisomerase inhibitors to induce SLFN11 expression in PC3 cells was lost when CD47 was targeted in these cells. Disrupting CD47 in PC3 cells increased resistance to etoposide but, in contrast to Jurkat cells, not to ionizing radiation. These data identify CD47 as a context-dependent regulator of SLFN11 expression and suggest an approach to improve radiotherapy and chemotherapy responses by combining with CD47-targeted therapeutics.


I787 provides signals for c-Kit receptor internalization and functionality that control mast cell survival and development.
Mast cell (MC) differentiation, survival, and activation are controlled by the membrane tyrosine kinase c-Kit upon interaction with stem cell factor (SCF). Here we describe a single point mutation induced by N-ethyl-N-nitrosurea (ENU) mutagenesis in C57BL/6J mice-an A to T transversion at position 2388 (exon 17) of the c-Kit gene, resulting in the isoleucine 787 substitution by phenylalanine (787F), and analyze the consequences of this mutation for ligand binding, signaling, and MC development. The Kit(787F/787F) mice carrying the single amino acid exchange of c-Kit lacks both mucosal and connective tissue-type MCs. In bone marrow-derived mast cells (BMMCs), the 787F mutation does not affect SCF binding and c-Kit receptor shedding, but strongly impairs SCF-induced cytokine production, degranulation enhancement, and apoptosis rescue. Interestingly, c-Kit downstream signaling in 787F BMMCs is normally initiated (Erk1/2 and p38 activation as well as c-Kit autophosphorylation) but fails to be sustained thereafter. In addition, 787F c-Kit does not efficiently mediate Cbl activation, leading to the absence of subsequent receptor ubiquitination and impaired c-Kit internalization. Thus, I787 provides nonredundant signals for c-Kit internalization and functionality.


Maurotoxin: a potent inhibitor of intermediate conductance Ca2+-activated potassium channels.
Maurotoxin, a 34-amino acid toxin from Scorpio maurus scorpion venom, was examined for its ability to inhibit cloned human SK (SK1, SK2, and SK3), IK1, and Slo1 calcium-activated potassium (K(Ca)) channels. Maurotoxin was found to produce a potent inhibition of Ca(2+)-activated (86)Rb efflux (IC(50), 1.4 nM) and inwardly rectifying potassium currents (IC(50), 1 nM) in CHO cells stably expressing IK1. In contrast, maurotoxin produced no inhibition of SK1, SK2, and SK3 small-conductance or Slo1 large-conductance K(Ca) channels at up to 1 microM in physiologically relevant ionic strength buffers. Maurotoxin did inhibit (86)Rb efflux (IC(50), 45 nM) through, and (125)I-apamin binding (K(i), 10 nM) to SK channels in low ionic strength buffers (i.e., 18 mM sodium, 250 mM sucrose), which is consistent with previous reports of inhibition of apamin binding to brain synaptosomes. Under similar low ionic strength conditions, the potency for maurotoxin inhibition of IK1 increased by approximately 100-fold (IC(50), 14 pM). In agreement with its ability to inhibit recombinant IK1 potassium channels, maurotoxin was found to potently inhibit the Gardos channel in human red blood cells and to inhibit the K(Ca) in activated human T lymphocytes without affecting the voltage-gated potassium current encoded by Kv1.3. Maurotoxin also did not inhibit Kv1.1 potassium channels but potently blocked Kv1.2 (IC(50), 0.1 nM). Mutation analysis indicates that similar amino acid residues contribute to the blocking activity of both IK1 and Kv1.2. The results from this study show that maurotoxin is a potent inhibitor of the IK1 subclass of K(Ca) potassium channels and may serve as a useful tool for further defining the physiological role of this channel subtype.


Observing carbon cycle-climate feedbacks from space.
The impact of human emissions of carbon dioxide and methane on climate is an accepted central concern for current society. It is increasingly evident that atmospheric concentrations of carbon dioxide and methane are not simply a function of emissions but that there are myriad feedbacks forced by changes in climate that affect atmospheric concentrations. If these feedbacks change with changing climate, which is likely, then the effect of the human enterprise on climate will change. Quantifying, understanding, and articulating the feedbacks within the carbon-climate system at the process level are crucial if we are to employ Earth system models to inform effective mitigation regimes that would lead to a stable climate. Recent advances using space-based, more highly resolved measurements of carbon exchange and its component processes-photosynthesis, respiration, and biomass burning-suggest that remote sensing can add key spatial and process resolution to the existing in situ systems needed to provide enhanced understanding and advancements in Earth system models. Information about emissions and feedbacks from a long-term carbon-climate observing system is essential to better stewardship of the planet.


A self-marker-like protein governs hemocyte allorecognition in Halocynthia roretzi.
Self-incompatibility, fusion/non-fusion reactions, and contact reactions (CRs) have all been identified as allorecognition phenomena in ascidians. CR is a reaction characteristic of the hemocytes of Halocynthia roretzi, whereby they release phenol oxidase (PO) upon contact with non-self hemocytes. Thus, these cells may represent a primitive form of the vertebrate immune system. In the present study, we focused on the CR of H. roretzi hemocytes and sought to identify self-marker proteins that distinguish between self and non-self cells. We initially generated a CR-inducing monoclonal antibody against the complete hemocyte membrane-protein complement (mAb11B16B10). This antibody was identified based on the differential induction of PO activity in individual organisms. The level of PO activity induced by this antibody in individual ascidians was consistent with the observed CR-induced PO activity. mAb11B16B10 recognized a series of 12 spots corresponding to a 100-kDa protein, with differing isoelectric points (pIs). A comparison of the 2D electrophoresis gels of samples from CR-reactive/non-reactive individuals revealed that some spots in this series in hemocytes were common to the CR-non-inducible individuals, but not to CR-inducible individuals. We cloned the corresponding gene and named it Halocynthia roretzi self-marker-like protein-1 (HrSMLP1). This gene is similar to the glycoprotein DD3-3 found in Dictyostelium, and is conserved in invertebrates. We generated a CR-inducing monoclonal antibody (mAb11B16B10) that recognized a series of novel membrane proteins (HrSMLP1) in the hemocytes of H. roretzi. The combination of expressed spots of HrSMLP1 distinguishes non-self cells from self cells with respect to CR inducibility. Given that the HrSMLP1 gene is a single gene, it may represent a novel type of self-marker protein with a role in CR.


Efficacy and safety of entecavir in patients with chronic hepatitis B and advanced hepatic fibrosis or cirrhosis.
The efficacy and safety of entecavir in patients with chronic hepatitis B and advanced liver fibrosis/cirrhosis was assessed from three large, randomized, multicenter, phase III studies. These studies enrolled patients (> or = 16 yr) with chronic hepatitis B, elevated alanine aminotransferase (ALT) levels, and compensated liver disease. Two trials enrolled nucleos(t)ide-naive patients randomized to at least 48 wk of treatment with entecavir 0.5 mg/day or lamivudine 100 mg/day. The third trial randomized lamivudine-refractory patients to 48 wk of entecavir 1 mg/day or lamivudine 100 mg/day. In this post hoc descriptive analysis, the efficacy and safety in patients with advanced liver fibrosis/cirrhosis (Ishak fibrosis stages 4-6) were examined for consistency with those seen in the overall study populations. Of the 1,633 treated patients, 245 had advanced liver fibrosis/cirrhosis (120 entecavir and 125 lamivudine). Among entecavir-treated patients with advanced liver fibrosis, improvement in Ishak fibrosis was observed in 57% of nucleos(t)ide-naive hepatitis B e antigen (HBeAg)-positive patients, 59% of nucleos(t)ide-naive HBeAg-negative patients, and 43% of lamivudine-refractory HBeAg-positive patients versus 49%, 53%, and 33% of lamivudine-treated patients with advanced liver fibrosis. The overall trends in other histologic, virologic, biochemical, and serologic outcomes in entecavir- versus lamivudine-treated patients with advanced liver fibrosis/cirrhosis were consistent with those observed in the overall study populations in each trial. The treatment was well tolerated. These data confirm that the performance of entecavir relative to that of lamivudine in patients with advanced liver fibrosis/cirrhosis was consistent with the relationship observed in the overall treated population.


Randomized Controlled Trial of Interscalene Block Compared with Injectable Liposomal Bupivacaine in Shoulder Arthroplasty.
Shortcomings of interscalene brachial plexus blockade include technical failure and rebound pain. Bupivacaine liposome injectable suspension, a sustained release preparation, is used for surgical-site administration. The purpose of this study was to evaluate these 2 postoperative pain management strategies in patients undergoing shoulder arthroplasty. In a non-blinded, randomized controlled trial of participants undergoing primary shoulder arthroplasty, patients were randomized to interscalene brachial plexus blockade or intraoperative soft-tissue infiltration of bupivacaine liposome injectable suspension. The primary outcome variable was morphine equivalent units consumed over the first 24 hours postoperatively. Secondary outcomes included morphine equivalent units consumed intraoperatively and a visual analog scale (VAS) for pain at 0, 8, 16, and 24 hours. Seventy-eight patients were randomized to interscalene brachial plexus blockade treatment (the blockade group) and 78 patients were randomized to bupivacaine liposome injectable suspension treatment (the suspension group). The mean total postoperative narcotic consumption (and standard deviation) over 24 hours after the surgical procedure was 14.8 ± 11.3 morphine equivalent units in the blockade group compared with 14.4 ± 16.8 morphine equivalent units in the suspension group (p = 0.849). Intraoperative narcotics were significantly lower (p < 0.001) in the blockade group (8.9 ± 4.1 morphine equivalent units) compared with the suspension group (16.2 ± 7.0 morphine equivalent units). The mean VAS pain score was significantly lower in the blockade group than in the suspension group at 0 hours postoperatively (0.8 ± 2.2 compared with 3.3 ± 2.7 points; p < 0.001) and at 8 hours postoperatively (1.4 ± 2.4 compared with 3.2 ± 2.2 points; p < 0.001), but it was not significantly different at 16 hours postoperatively (4.3 ± 2.8 compared with 3.8 ± 2.4 points; p = 0.348). The VAS pain scores were significantly higher (p = 0.021) in the blockade group (4.9 ± 2.7 points) compared with the suspension group (3.9 ± 2.3 points) at 24 hours postoperatively. Patients treated with bupivacaine liposome injectable suspension required an equivalent amount of postoperative narcotics and greater intraoperative narcotics compared with patients treated with interscalene brachial plexus blockade. Although interscalene brachial plexus blockade provided improved pain scores for the first 8 hours after the surgical procedure, pain scores were worse at 24 hours. The optimal postoperative pain regimen for shoulder arthroplasty and the cost-effectiveness of analgesic techniques require further investigation. Therapeutic Level II. See Instructions for Authors for a complete description of levels of evidence.


Computation beyond the turing limit.
Extensive efforts have been made to prove the Church-Turing thesis, which suggests that all realizable dynamical and physical systems cannot be more powerful than classical models of computation. A simply described but highly chaotic dynamical system called the analog shift map is presented here, which has computational power beyond the Turing limit (super-Turing); it computes exactly like neural networks and analog machines. This dynamical system is conjectured to describe natural physical phenomena.


Transcriptomic profiling of Methylococcus capsulatus (Bath) during growth with two different methane monooxygenases.
Methylococcus capsulatus (Bath) is a methanotroph that possesses both a membrane-embedded (pMMO) and a soluble methane monooxygenase (sMMO). The expression of these two MMO's is tightly controlled by the availability of copper in the growth medium, but the underlying mechanisms and the number of genes involved in this switch in methane oxidation is not yet fully elucidated. Microarray analyses were used to assess the transcriptome in cells producing either pMMO or sMMO. A total of 137 genes were differentially expressed, with 87 genes showing a significant up-regulation during sMMO production. The majority of the differentially expressed genes could be assigned to functional roles in the energy metabolism and transport. Furthermore, three copper responding gene clusters were discovered, including an extended cluster that also harbors the genes for sMMO. Our data also indicates that major changes takes place in the respiratory chain between pMMO- and sMMO-producing cells, and that quinone are predominantly used as the electron donors for methane oxidation by pMMO. Intriguingly, a large proportion of the differentially expressed genes between pMMO- and sMMO-producing cells encode c-type cytochromes. By combining microarray- and mass spectrometry data, a total of 35 c-type cytochromes are apparently expressed in M. capsulatus when grown in nitrate mineral salt medium with methane as sole energy and carbon source, and the expression of 21 of these respond to the availability of copper. Interestingly, several of these c-type cytochromes are recovered from the cell surface, suggesting that extracellular electron transfers may occur in M. capsulatus.


Psychometric properties of the Community Integration Questionnaire: a systematic review of five populations.
This systematic review documents the content and the quality of the psychometric evidence concerning the utilization of the Community Integration Questionnaire for individuals living with a disability other than a traumatic brain injury. Medline, Embase, CINAHL, OTseeker and PsycINFO (searched from inception to June 2019). Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines were used for conducting and reporting this review. Studies that investigated at least one psychometric property of the Community Integration Questionnaire for individuals living with a disability other than traumatic brain injury were included. Data extraction and critical methodological appraisal of the articles (MacDermid checklist, COnsensus-based Standards for the selection of health Measurement INstruments checklist) were independently performed and validated by the first two authors. Ten studies representing 3000 individuals met the inclusion criteria. Five populations were documented. There are positive and trustable data regarding internal consistency for adults living with multiple sclerosis, spinal cord injury or burns and in mixed samples (α = 0.71-0.84). Construct validity is fairly documented for adults living with multiple sclerosis or aphasia and in mixed samples. Test-retest reliably is acceptable for adults living with multiple sclerosis (intraclass correlation coefficient = 0.91-0.97) as well as responsiveness (area under the receiver operating characteristic curve = 0.81). Other psychometric properties could not be demonstrated sufficiently solid. Many psychometric properties of the Community Integration Questionnaire are still poorly evaluated for adults living with a disability other than a traumatic brain injury. However, promising data have been documented in each population included in this review.


Refractive index measurement by interference microscopy: corrections for dispersion.
The refractive index of a particle can be determined with microscope equipment for measuring interference in transmitted light. If the path difference is several wavelengths, white illumination will used. If the particle has strong dispersion, measurements made with white light give an apparent refractive index much higher than the true one. The apparent index is related to the group velocity of light waves in the substance in the same way that the true index is related to phase velocity. Flaky crystals of hematite showed apparent index 4.52, in agreement with theory, compared with the true ordinary index around 3.09.


Predicting beef carcass meat, fat and bone proportions from carcass conformation and fat scores or hindquarter dissection.
Equations for predicting the meat, fat and bone proportions in beef carcasses using the European Union carcass classification scores for conformation and fatness, and hindquarter composition were developed and their accuracy was tested using data from 662 cattle. The animals included bulls, steers and heifers, and comprised of Holstein-Friesian, early- and late-maturing breeds × Holstein-Friesian, early-maturing × early-maturing, late-maturing × early-maturing and genotypes with 0.75 or greater late-maturing ancestry. Bulls, heifers and steers were slaughtered at 15, 20 and 24 months of age, respectively. The diet offered before slaughter includes grass silage only, grass or maize silage plus supplementary concentrates, or concentrates offered ad libitum plus 1 kg of roughage dry matter per head daily. Following the slaughter, carcasses were classified mechanically for conformation and fatness (scale 1 to 15), and the right side of each carcass was dissected into meat, fat and bone. Carcass conformation score ranged from 4.7 to 14.4, 5.4 to 10.9 and 2.0 to 12.0 for bulls, heifers and steers, respectively; the corresponding ranges for fat score were 2.7 to 11.5, 3.2 to 11.3 and 2.8 to 13.3. Prediction equations for carcass meat, fat and bone proportions were developed using multiple regression, with carcass conformation and fat score both included as continuous independent variables. In a separate series of analyses, the independent variable in the model was the proportion of the trait under investigation (meat, fat or bone) in the hindquarter. In both analyses, interactions between the independent variables and gender were tested. The predictive ability of the developed equations was assed using cross-validation on all 662 animals. Carcass classification scores accounted for 0.73, 0.67 and 0.71 of the total variation in carcass meat, fat and bone proportions, respectively, across all 662 animals. The corresponding values using hindquarter meat, fat and bone in the model were 0.93, 0.87 and 0.89, respectively. The bias of the prediction equations when applied across all animals was not different from zero, but bias did exist among some of the genotypes of animals present. In conclusion, carcass classification scores and hindquarter composition are accurate and efficient predictors of carcass meat, fat and bone proportions.


Bioethics for clinicians: 2. Disclosure.
In the context of patient consent, "disclosure" refers to the provision of relevant information by the clinician and its comprehension by the patient. Both elements are necessary for valid consent. Disclosure should inform the patient adequately about the treatment and its expected effects, relevant alternative options and their benefits and risks, and the consequences of declining or delaying treatment. The clinician's goal is to disclose information that a reasonable person in the patient's position would need in order to make an informed decision. Therefore, clinicians may need to consider how the proposed treatment (and other options) might affect the patient's employment, finances, family life and other personal concerns. Clinicians may also need to be sensitive to cultural and religious beliefs that can affect disclosure.


The role of Epstein-Barr virus in Hodgkin's disease from different geographical areas.
Recent studies have suggested that Epstein-Barr virus (EBV) may play a role in the aetiology of Hodgkin's disease. To determine the role of EBV in childhood Hodgkin's disease in different geographical areas, immunohistochemical staining and in situ hybridisation were used to analyse latent membrane protein 1 (LMP 1) and small nuclear non-transcribed RNAs (EBER-1) respectively. Testing for EBV within the Reed-Sternberg and Hodgkin's cells was carried out in childhood Hodgkin's disease from 10 different countries. The proportion of LMP 1 positive cases varied significantly, being 50% of cases from the United Kingdom (38/75), South Africa (9/18), Egypt (7/14), and Jordan (8/16), 60% from the United Arab Emirates (6/10), 70% from Australia (11/16), 81% from Costa Rica (34/42), 88% from Iran (7/8), 90% from Greece (20/22), and 100% of the 56 cases from Kenya. A sensitive polymerase chain reaction based EBV strain typing technique was established using archival tissues. EBV strain type 1 was shown to be predominant in childhood Hodgkin's disease from the United Kingdom, South Africa, Australia, and Greece. Type 2 was predominant in Egypt. EBV strain types 1 and 2 were both detected in some cases of childhood Hodgkin's disease in the United Kingdom, Costa Rica, and Kenya. The high incidence of EBV and the presence especially in developing countries of dual infection with both strain types 1 and 2 may reflect socioeconomic conditions leading to malnutrition induced immunological impairment. The possibility of HIV infection also needs to be explored.


C-reactive Protein as a Potential Biomarker of Residual Obstructive Sleep Apnea Following Adenotonsillectomy in Children.
Adenotonsillectomy (AT) is first-line treatment for pediatric obstructive sleep apnea (OSA), with most children having improvements in polysomnography (PSG). However, many children have residual OSA following AT as determined through PSG. Identification of a biomarker of residual disease would be clinically meaningful to detect children at risk. We hypothesize serum high-sensitivity C-reactive protein (hsCRP), an inflammatory biomarker, is predictive of residual OSA following AT. PSG was performed both preoperatively and postoperatively on children undergoing AT for the diagnosis of OSA. HsCRP serum concentrations were determined in all children pre-AT, and in most children post-AT. Resolution of OSA after AT was defined by a post-AT apnea-hypopnea index (AHI) < 1.5/h total sleep time (TST). Residual OSA was defined as a post-AT AHI > 5/h TST, which is considered clinically significant. AT significantly improved the AHI from 15.9 ± 16.4 to 4.1 ± 5.3/h TST in 182 children (P < 0.001). Of 182 children, residual OSA (post-AT AHI > 5) was seen in 46 children (25%). Among children who had hsCRP levels measured pre- and post-AT (n = 155), mean hsCRP levels pre-AT were 0.98 ± 1.91 mg/L and were significantly reduced post-AT (0.63 ± 2.24 mg/dL; P = 0.011). Stratification into post-AT AHI groups corresponding to < 1.5/h TST, 1.5/h TST < AHI < 5/h TST, and AHI > 5/h TST revealed post-AT hsCRP levels of 0.09 ± 0.12, 0.57 ± 2.28, and 1.49 ± 3.34 mg/L with statistical significance emerging comparing residual AHI > 5/h TST compared to post-AT AHI < 1.5/h TST (P = 0.006). Hierarchical multivariate modeling confirmed that pre-AT AHI and post-AT hsCRP levels were most significantly associated with residual OSA. Even though AT improves OSA in most children, residual OSA is frequent. Assessment of post-AT hsCRP levels emerges as a potentially useful biomarker predicting residual OSA.


Early Transcriptomic Response to LDL and oxLDL in Human Vascular Smooth Muscle Cells.
Although nowadays it is well known that the human transcriptome can importantly vary according to external or environmental condition, the reflection of this concept when studying oxidative stress and its direct relationship with gene expression profiling during the process of atherogenesis has not been thoroughly achieved. The ability to analyze genome-wide gene expression through transcriptomics has shown that the genome responds dynamically to diverse stimuli. Here, we describe the transcriptome of human vascular smooth muscle cells (hVSMC) stimulated by native and oxidized low-density lipoprotein (nLDL and oxLDL respectively), with the aim of assessing the early molecular changes that induce a response in this cell type resulting in a transcriptomic transformation. This expression has been demonstrated in atherosclerotic plaques in vivo and in vitro, particularly in the light of the oxidative modification hypothesis of atherosclerosis. Total RNA was isolated with TRIzol reagent (Life Technologies) and quality estimated using an Agilent 2100 bioanalyzer. The transcriptome of hVSMC under different experimental conditions (1,5 and 24 hours for nLDL and oxLDL) was obtained using the GeneChip Human Gene 1.0 ST (Affymetrix) designed to measure gene expression of 28,869 well-annotated genes. A fixed fold-change cut-off corresponding to ± 2 was used to identify genes exhibiting the most significant variation and statistical significance (P< 0.05), and 8 genes validated by qPCR using Taqman probes. 10 molecular processes were significantly affected in hVSMC: Apoptosis and cell cycle, extracellular matrix remodeling, DNA repair, cholesterol efflux, cGMP biosynthesis, endocytic mechanisms, calcium homeostasis, redox balance, membrane trafficking and finally, the immune response to inflammation. The evidence we present supporting the hypothesis for the involvement of oxidative modification of several processes and metabolic pathways in atherosclerosis is strengthen by the fact that gene expression patterns obtained when hVSMC are incubated for a long period of time in the presence of nLDL, correspond very much the same as when cells are incubated for a short period of time in the presence of chemically modified oxLDL. Our results indicate that under physiological conditions and directly related to specific environmental conditions, LDL particles most probably suffer chemical modifications that initially serve as an alert signal to overcome a harmful stimulus that with time might get transformed to a pathological pattern and therefore consolidate a pathological condition.


Platelet proteome and function in X-linked thrombocytopenia with thalassemia and in silico comparisons with gray platelet syndrome.
In X-linked thrombocytopenia with thalassemia (XLTT; OMIM 314050), caused by the mutation p.R216Q in exon 4 of the GATA1 gene, male hemizygous patients display macrothrombocytopenia, bleeding diathesis and a β-thalassemia trait. Herein, we describe findings in two unrelated Swedish XLTT families with a bleeding tendency exceeding what is expected from the thrombocytopenia. Blood tests revealed low P-PAI-1 and P-factor 5, and elevated S-thrombopoietin levels. Transmission electron microscopy showed diminished numbers of platelet α- and dense granules. The proteomes of isolated blood platelets from 5 male XLTT patients, compared to 5 gender- and age matched controls, were explored. Quantitative mass spectrometry showed alterations of 83 proteins (fold change ≥±1.2, q< .05). Of 46 downregulated proteins, 39 were previously reported to be associated with platelet granules. Reduced protein levels of PTGS1 and SLC35D3 were validated in megakaryocytes of XLTT bone marrow biopsies by immunohistochemistry. Platelet function testing by flow cytometry revealed low dense- and α-granule release and fibrinogen binding in response to ligation of receptors for ADP, the thrombin receptor PAR4 and the collagen receptor GPVI. Significant reductions of a number of α-granule proteins overlapped with a previous platelet proteomics investigation in the inherited macrothrombocytopenia gray platelet syndrome (GPS). In contrast, Ca2+ transporter proteins that facilitate dense granule release were downregulated in XLTT but upregulated in GPS. Ingenuity Pathway Analysis showed altered Coagulation System and Protein Ubiquitination pathways in the XLTT platelets. Collectively, the results revealed protein and functional alterations affecting platelet α- and dense granules in XLTT, probably contributing to bleeding.


Deadly progress: changes in Australian Aboriginal and Torres Strait Islander adult daily smoking, 2004-2015.
Tobacco smoking is the leading contributor to the burden of disease among Aboriginal and Torres Strait Islander Australians. Reducing tobacco use in this population is a public health priority. Precise monitoring of smoking prevalence trends is central to implementation and evaluation of effective tobacco control. The way in which trends are reported influences understanding of the extent of progress, with potential implications for policy. Our objective was to quantify absolute changes in Aboriginal and Torres Strait Islander adult (≥18 years old) daily tobacco smoking prevalence from 2004 to 2015, including comparisons with the total Australian population, and by age, sex and remoteness. We analysed multiple nationally representative surveys of the Aboriginal and Torres Strait Islander, and total Australian, population conducted from 2004 to 2015. Aligned with strength-based approaches, we applied a progress frame, focusing on absolute differences in smoking prevalence within the Aboriginal and Torres Strait Islander population. The prevalence of current daily smoking among Aboriginal and Torres Strait Islander adults nationally was 50.0% (95% confidence interval [CI] 47.9, 52.2) in 2004-05 and 41.4% (95% CI 39.1, 43.6) in 2014-15, representing an absolute prevalence decrease of 8.6 percentage points (95% CI 5.5, 11.8) over the past decade. This is comparable with the 6.8 percentage point (95% CI 5.6, 7.9) decrease in smoking prevalence in the total Australian population over the same period, from 21.3% in 2004-05 (95% CI 20.5, 22.0) to 14.5% in 2014-15 (95% CI 13.6, 15.4). Particular success in reducing Aboriginal and Torres Strait Islander daily smoking was observed among younger age groups, with a decrease of 13.2 percentage points for 18-24-year-olds (95% CI 5.9, 20.4), 9.0 percentage points for 25-34-year-olds (95% CI 2.7, 15.3) and 8.7 percentage points for 35-44-year-olds (95% CI 2.6, 14.8). Smoking prevalence in those living in urban/regional areas decreased by 10.2 percentage points (95% CI 6.2, 14.1). Substantial progress has been made in reducing smoking, with an estimated 35 000 fewer Aboriginal and Torres Strait Islander adults smoking every day in 2014-15 compared with if daily smoking remained at 2004-05 prevalence. This will lead to thousands of lives saved. The observed success in the younger age groups is encouraging. Continued resourcing and comprehensive tobacco control efforts are required to ensure positive trends continue.


Emerging battery-ingestion hazard: clinical implications.
Recent cases suggest that severe and fatal button battery ingestions are increasing and current treatment may be inadequate. The objective of this study was to identify battery ingestion outcome predictors and trends, define the urgency of intervention, and refine treatment guidelines. Data were analyzed from 3 sources: (1) National Poison Data System (56535 cases, 1985-2009); (2) National Battery Ingestion Hotline (8648 cases, July 1990-September 2008); and (3) medical literature and National Battery Ingestion Hotline cases (13 deaths and 73 major outcomes) involving esophageal or airway button battery lodgment. All 3 data sets signal worsening outcomes, with a 6.7-fold increase in the percentage of button battery ingestions with major or fatal outcomes from 1985 to 2009 (National Poison Data System). Ingestions of 20- to 25-mm-diameter cells increased from 1% to 18% of ingested button batteries (1990-2008), paralleling the rise in lithium-cell ingestions (1.3% to 24%). Outcomes were significantly worse for large-diameter lithium cells (> or = 20 mm) and children who were younger than 4 years. The 20-mm lithium cell was implicated in most severe outcomes. Severe burns with sequelae occurred in just 2 to 2.5 hours. Most fatal (92%) or major outcome (56%) ingestions were not witnessed. At least 27% of major outcome and 54% of fatal cases were misdiagnosed, usually because of nonspecific presentations. Injuries extended after removal, with unanticipated and delayed esophageal perforations, tracheoesophageal fistulas, fistulization into major vessels, and massive hemorrhage. Revised treatment guidelines promote expedited removal from the esophagus, increase vigilance for delayed complications, and identify patients who require urgent radiographs.


Dynamic frontotemporal systems process space and time in working memory.
How do we rapidly process incoming streams of information in working memory, a cognitive mechanism central to human behavior? Dominant views of working memory focus on the prefrontal cortex (PFC), but human hippocampal recordings provide a neurophysiological signature distinct from the PFC. Are these regions independent, or do they interact in the service of working memory? We addressed this core issue in behavior by recording directly from frontotemporal sites in humans performing a visuospatial working memory task that operationalizes the types of identity and spatiotemporal information we encounter every day. Theta band oscillations drove bidirectional interactions between the PFC and medial temporal lobe (MTL; including the hippocampus). MTL theta oscillations directed the PFC preferentially during the processing of spatiotemporal information, while PFC theta oscillations directed the MTL for all types of information being processed in working memory. These findings reveal an MTL theta mechanism for processing space and time and a domain-general PFC theta mechanism, providing evidence that rapid, dynamic MTL-PFC interactions underlie working memory for everyday experiences.


Estimated Effects of Different Alcohol Taxation and Price Policies on Health Inequalities: A Mathematical Modelling Study.
While evidence that alcohol pricing policies reduce alcohol-related health harm is robust, and alcohol taxation increases are a WHO "best buy" intervention, there is a lack of research comparing the scale and distribution across society of health impacts arising from alternative tax and price policy options. The aim of this study is to test whether four common alcohol taxation and pricing strategies differ in their impact on health inequalities. An econometric epidemiological model was built with England 2014/2015 as the setting. Four pricing strategies implemented on top of the current tax were equalised to give the same 4.3% population-wide reduction in total alcohol-related mortality: current tax increase, a 13.4% all-product duty increase under the current UK system; a value-based tax, a 4.0% ad valorem tax based on product price; a strength-based tax, a volumetric tax of £0.22 per UK alcohol unit (= 8 g of ethanol); and minimum unit pricing, a minimum price threshold of £0.50 per unit, below which alcohol cannot be sold. Model inputs were calculated by combining data from representative household surveys on alcohol purchasing and consumption, administrative and healthcare data on 43 alcohol-attributable diseases, and published price elasticities and relative risk functions. Outcomes were annual per capita consumption, consumer spending, and alcohol-related deaths. Uncertainty was assessed via partial probabilistic sensitivity analysis (PSA) and scenario analysis. The pricing strategies differ as to how effects are distributed across the population, and, from a public health perspective, heavy drinkers in routine/manual occupations are a key group as they are at greatest risk of health harm from their drinking. Strength-based taxation and minimum unit pricing would have greater effects on mortality among drinkers in routine/manual occupations (particularly for heavy drinkers, where the estimated policy effects on mortality rates are as follows: current tax increase, -3.2%; value-based tax, -2.9%; strength-based tax, -6.1%; minimum unit pricing, -7.8%) and lesser impacts among drinkers in professional/managerial occupations (for heavy drinkers: current tax increase, -1.3%; value-based tax, -1.4%; strength-based tax, +0.2%; minimum unit pricing, +0.8%). Results from the PSA give slightly greater mean effects for both the routine/manual (current tax increase, -3.6% [95% uncertainty interval (UI) -6.1%, -0.6%]; value-based tax, -3.3% [UI -5.1%, -1.7%]; strength-based tax, -7.5% [UI -13.7%, -3.9%]; minimum unit pricing, -10.3% [UI -10.3%, -7.0%]) and professional/managerial occupation groups (current tax increase, -1.8% [UI -4.7%, +1.6%]; value-based tax, -1.9% [UI -3.6%, +0.4%]; strength-based tax, -0.8% [UI -6.9%, +4.0%]; minimum unit pricing, -0.7% [UI -5.6%, +3.6%]). Impacts of price changes on moderate drinkers were small regardless of income or socioeconomic group. Analysis of uncertainty shows that the relative effectiveness of the four policies is fairly stable, although uncertainty in the absolute scale of effects exists. Volumetric taxation and minimum unit pricing consistently outperform increasing the current tax or adding an ad valorem tax in terms of reducing mortality among the heaviest drinkers and reducing alcohol-related health inequalities (e.g., in the routine/manual occupation group, volumetric taxation reduces deaths more than increasing the current tax in 26 out of 30 probabilistic runs, minimum unit pricing reduces deaths more than volumetric tax in 21 out of 30 runs, and minimum unit pricing reduces deaths more than increasing the current tax in 30 out of 30 runs). Study limitations include reducing model complexity by not considering a largely ineffective ban on below-tax alcohol sales, special duty rates covering only small shares of the market, and the impact of tax fraud or retailer non-compliance with minimum unit prices. Our model estimates that, compared to tax increases under the current system or introducing taxation based on product value, alcohol-content-based taxation or minimum unit pricing would lead to larger reductions in health inequalities across income groups. We also estimate that alcohol-content-based taxation and minimum unit pricing would have the largest impact on harmful drinking, with minimal effects on those drinking in moderation.


LINE-1 activity in facultative heterochromatin formation during X chromosome inactivation.
During X chromosome inactivation (XCI), Xist RNA coats and silences one of the two X chromosomes in female cells. Little is known about how XCI spreads across the chromosome, although LINE-1 elements have been proposed to play a role. Here we show that LINEs participate in creating a silent nuclear compartment into which genes become recruited. A subset of young LINE-1 elements, however, is expressed during XCI, rather than being silenced. We demonstrate that such LINE expression requires the specific heterochromatic state induced by Xist. These LINEs often lie within escape-prone regions of the X chromosome, but close to genes that are subject to XCI, and are associated with putative endo-siRNAs. LINEs may thus facilitate XCI at different levels, with silent LINEs participating in assembly of a heterochromatic nuclear compartment induced by Xist, and active LINEs participating in local propagation of XCI into regions that would otherwise be prone to escape.


Association of a Novel Intronic Variant in RPGR With Hypomorphic Phenotype of X-Linked Retinitis Pigmentosa.
Pathogenic variants in retinitis pigmentosa GTPase regulator (RPGR) gene typically lead to a severe form of X-linked retinitis pigmentosa, which is associated with early severe vision loss. To investigate an X-linked retinal degeneration family with atypical preservation of visual acuity in the presence of a novel deep intronic splice site RPGR c.779-5T>G variant. In this case series, 3 members of an X-linked retinal degeneration family were studied by in-depth phenotyping and genetic screening at a single center. Data were collected and analyzed from November 2018 to March 2020. Data were collected on full ophthalmic history, examination, and retinal imaging. A full retinitis pigmentosa gene panel was analyzed by next-generation sequencing. The pathogenicity of the RPGR c.779-5T>G variant was assessed by in silico splice prediction tools and by purpose-designed in vitro splicing assay. An 84-year-old man was referred with clinical diagnosis of choroideremia and possible inclusion into a gene therapy trial. He presented with late-stage retinal degeneration and unusually preserved visual acuity (78 and 68 ETRDS letters) that clinically resembled choroideremia. His 23-year-old grandson was still in early stages of degeneration but showed a very different clinical picture, typical of retinitis pigmentosa. Next-generation sequencing identified a sole RPGR c.779-5T>G variant of undetermined pathogenicity in both cases. The daughter of the proband showed an RPGR carrier phenotype and was confirmed to carry the same variant. The molecular analysis confirmed that the RPGR c.779-5T>G variation reduced the efficiency of intron splicing compared with wild type, leading to a population of mutant and normal transcripts. The predicted consequences of the pathogenic variant are potential use of an alternative splice acceptor site or complete skipping of exon 8, resulting in truncated forms of the RPGR protein with different levels of glutamylation. These results support the importance of careful interpretation of inconsistent clinical phenotypes between family members. Using a molecular splicing assay, a new pathogenic variant in a noncoding region of RPGR was associated with a proportion of normal and hypomorphic RPGR, where cones are likely to survive longer than expected, potentially accounting for the preserved visual acuity observed in this family.


What constitutes a serotonergic neurone in the retina?
When exposed to exogenous serotonin, specific neurones in retinas from different species (guinea pig, rabbit, frog, pigeon and goldfish) take the amine up; these same neurones do not reveal endogenous serotonin following immunocytochemistry for the localization of the amine. The use of autoradiography and immunocytochemistry to localize the uptake of radioactive or unlabelled amine gives identical results, with the exception of the guinea pig retina, where a subset of horizontal-like cells specifically takes up a metabolite of [(3)H]serotonin (presumably produced in the extracellular space of the retina). This uptake is blocked specifically by N-acetyl-serotonin. After an intraocular injection of forskolin into the rabbit eye the serotonin content of the tissue is elevated and this is reflected in the demonstration of serotonin immunoreactivity associated with a subpopulation of amacrine cells. These cells correspond in size and position to the "indoleamine-accumulating cells" of the retina. Animals (goldfish, frog, pigeon) dark-adapted over a certain period manifest an elevated level of serotonin associated with their retinas compared with that in light-adapted animals. However, immunofluorescence histochemistry used to localize serotonin showed that the same neurones in the different retinas contain the amine, although the "fluorescence" was elevated in the dark-adapted retinas. In the light of the present data it is clear that what constitutes a "serotonergic" neurone cannot be restricted to the immunohistochemical demonstration of the amine. The evidence is persuasive for "serotonergic neurones" having the property to take up serotonin specifically by a high affinity transport process. What warrants serious consideration is whether this property is a characteristic of "serotonergic neurones" rather than the endogenous localization of serotonin without pharmacological pretreatment.


Policy options for endgame planning in tobacco control: a simulation modelling study.
To investigate the potential impacts of several tobacco control interventions on adult daily smoking prevalence in the Australian state of Queensland, using a system dynamics model codeveloped with local and national stakeholders. Eight intervention scenarios were simulated and compared with a reference scenario (business as usual), in which all tobacco control measures currently in place are maintained unchanged until the end of the simulation period (31 December 2037). Under the business as usual scenario, adult daily smoking prevalence is projected to decline from 11.8% in 2017 to 5.58% in 2037. A sustained 50% increase in antismoking advertising exposure from 2018 reduces projected prevalence in 2037 by 0.80 percentage points. Similar reductions are projected with the introduction of tobacco wholesaler and retailer licensing schemes that either permit or prohibit tobacco sales by alcohol-licensed venues (0.65 and 1.73 percentage points, respectively). Increasing the minimum age of legal supply of tobacco products substantially reduces adolescent initiation, but has minimal impact on smoking prevalence in the adult population over the simulation period. Sustained reductions in antismoking advertising exposure of 50% and 100% from 2018 increase projected adult daily smoking prevalence in 2037 by 0.88 and 1.98 percentage points, respectively. These results suggest that any prudent approach to endgame planning should seek to build on rather than replace existing tobacco control measures that have proved effective to date. Additional interventions that can promote cessation are expected to be more successful in reducing smoking prevalence than interventions focussing exclusively on preventing initiation.


Cashew Nut Positioning during Stone Tool Use by Wild Bearded Capuchin Monkeys (Sapajus libidinosus).
Wild capuchin monkeys (Sapajus libidinosus) at Serra da Capivara National Park, Brazil, regularly use stone tools to break open cashew nuts (Anacardium spp.). Here we examine 2 approaches used by the capuchins to position the kidney-shaped cashew nuts on an anvil before striking with a stone tool. Lateral positioning involves placing the nut on its flatter, more stable side, therefore requiring less attention from the monkey during placement. However, the less stable and never previously described arched position, in which the nut is balanced with its curved side uppermost, requires less force to crack the outer shell. We observed cashew nut cracking in a field experimental setting. Only 6 of 20 adults, of both sexes, were observed to deliberately place cashew nuts in an arched position, which may indicate that the technique requires time and experience to learn. We also found that use of the arched position with dry nuts, but not fresh, required, in 63% of the time, an initial processing to remove one of the cashew nut lobes, creating a more stable base for the arch. This relatively rare behaviour appears to have a complex ontogeny, but further studies are required to establish the extent to which social learning is involved.


Effects of Acibenzolar-S-Methyl Application to Rotylenchulus reniformis and Meloidogyne javanica.
Effects of acibenzolar-s-methyl, an inducer of systemic acquired resistance in plants, on Rotylenchulus reniformis and Meloidogyne javanica in vitro and in vivo were determined. A single foliar application of acibenzolar at 50 mg/liter (5 ml of solution per plant) to 7-day-old cowpea or soybean seedlings decreased R. reniformis and M. javanica egg production by 50% 30 days after inoculation. The mechanism of acibenzolar on plant-parasitic nematodes was then investigated. Acibenzolar at 50 to 200 mg/liter did not affect movement of R. reniformis and M. javanica or penetration of second-stage juveniles (J2) of M. javanica on cowpea. However, M. javanica development was slowed and fecundity was reduced in plants treated with acibenzolar. On average, 50% of J2 that penetrated acibenzolar-treated cowpeas developed into mature females with eggs, whereas the other 50% exhibited arrested development. The number of eggs per egg mass was 450 in water-treated cowpeas, whereas the number declined to 250 in acibenzolar-treated plants. Acibenzolar may be responsible for stimulating the plants to express some resistance to the nematodes.


The established and future biomarkers of malignant pleural mesothelioma.
Malignant pleural mesothelioma (MPM) is an asbestos-related cancer with a median survival of 12months. The MPM incidence is 1-6/100,000 and is increasing as a result of historic asbestos exposure in industrialized countries and continued use of asbestos in developing countries. Lack of accurate biomarkers makes diagnosis, prognostication and treatment prediction of MPM challenging. The aim of this review is to identify the front line of MPM biomarkers with current or potential clinical impact. Literature search using the PubMed and PLoS One databases, the related-articles function of PubMed and the reference lists of associated publications until April 26th 2015 revealed a plethora of candidate biomarkers. The current gold standard of MPM diagnosis is a combination of two positive and two negative immunohistochemical markers in the epithelioid and biphasic type, but sarcomatous type do not have specific markers, making diagnosis more difficult. Mesothelin in serum and pleural fluid may serve as adjuvant diagnostic with high specificity but low sensitivity. Circulating proteomic and microRNA signatures, fibulin-3, tumor cell gene-ratio test, transcriptomic, lncRNA, glycopeptides, pleural fluid FISH assay, hyaluronate/N-ERC mesothelin and deformability cytometry may be important future markers. Putative predictive markers for pemetrexed-platinum are tumor TS and TYMS, for vinorelbine the ERCC1, beta-tubuline class III and BRCA1. Mutations of the BAP1 gene are potential markers of MPM susceptibility. In conclusion, the current status of MPM biomarkers is not satisfactory but encouraging as more sensitive and specific non-invasive markers are emerging. However, prospective validation is needed before clinical application.


Effect of a free healthcare policy on health services utilisation for non-malarial febrile illness by children under five years in Burkina Faso: an interrupted time series analysis.
To assess the effect of a free healthcare policy for children under five years old implemented in Burkina Faso since April 2016, on the use of health care of non-malarial febrile illnesses (NMFI). To assess the immediate and long-term effect of the free healthcare policy in place, we conducted an interrupted time series analysis of routinely collected data on febrile illnesses from three urban primary health centres of Ouagadougou between 1 January 2015 and 31 December 2016. Of the 39 046 febrile cases reported in the study period, 17 017 NMFI were included in the study. Compared to the period before the intervention, we observed an immediate, non-statistically significant increase of 7% in the number of NMFI (IRR = 1.07; 95% CI = 0.75, 1.51). Compared to the trend that would have been expected in absence of the intervention, the results showed a small but sustained increase of 6% in the trend of monthly number of NMFI during the intervention period (IRR = 1.06; 95%CI = 1.01, 1.12). Our study highlighted an increase in the uptake of healthcare services, specifically for NMFI by children under five years of age, after the implementation of a free care policy. This analysis contributes to informing decision makers on the need to strengthen the capacities of healthcare centres and to anticipate the challenges of the sustainability of this policy.


Identification of peripheral blood mononuclear cells targeted by Ovine herpesvirus-2 in sheep.
Sheep-associated malignant catarrhal fever (MCF), caused by Ovine herpesvirus 2 (OvHV-2), is usually a fatal disease of various ruminants and swine. In contrast, natural OvHV-2 infection in sheep, which are the main OvHV-2 reservoir, proceeds without any clinical symptoms. Since the range of targeted cells may be important for pathogenesis, we wanted to analyze the natural range of peripheral mononuclear blood cells (PBMC) targeted by OvHV-2. To this end, OvHV-2-free sheep were exposed to natural infection and blood samples were taken at intervals. Four different PBMC subpopulations were purified by fluorescence activated cell sorting (FACS) before being subjected to analysis for OvHV-2-DNA. After an incubation period of between 11 and 12 weeks, all exposed sheep became positive for OvHV-2. In most sheep, a first peak of OvHV-2-DNA was identified in the CD2 and CD4 double positive subpopulation. However, with time, the highest load of OvHV-2-DNA shifted to the CD2-positive and CD4-negative T-cells. Furthermore, low amounts of OvHV-2-DNA were occasionally detected also in the fractions that represented either CD14-positive monocytes or triple negative cells (CD2(-)/CD4(-)/CD14(-)). We conclude from these experiments that OvHV-2 has a similar host cellular range in sheep and cattle, respectively. Our results may be relevant in the context of comparative analysis of OvHV-2 pathogenesis in animal species that are susceptible to MCF.