Assessing the functional relationship between dung beetle traits and dung removal, burial, and seedling emergence.
The relationship between biodiversity and ecosystem functioning is often assessed through trait diversity. However, the relationship between traits and functions is typically assumed but seldom tested. We analyze the relationship between dung beetle traits and three ecological functions: dung removal, dung burial, and seedling emergence. We set up a laboratory experiment using nine Scarabaeidae species (three endocoprids, four paracoprids, and two telecoprids). We placed a sexual pair of beetles in each experimental unit, together with a mixture of dung and seeds, and measured the amount of dung removed and buried, burial depth, and the number of emerged seedlings. Sixteen morphological traits related to dung removal and burial were measured in each individual. Results indicate that these traits were related to dung beetle performance in dung removal and burial. Most traits were positively related to dung removal, indicating the existence of a general trait syndrome associated with dung manipulation and digging capability. Dung exploitation strategies did not provide further explanatory power. Seedling emergence showed a negative but weak relationship with dung burial amount and depth and species identity. This implies that specific differences in dung-soil interface activity may be important in secondary seed dispersal by dung beetles.


Mitochondrial ATAD3A combines with GRP78 to regulate the WASF3 metastasis-promoting protein.
AAA domain containing 3A (ATAD3A) is an integral mitochondrial membrane protein with unknown function, although we now show that high-level expression is associated with poor survival in breast cancer patients. Using a mass spectrometry approach we have demonstrated that ATAD3A interacts with the WASF3 metastasis-promoting protein. Knockdown of ATAD3A leads to decreased WASF3 protein levels in breast and colon cancer cells. Silencing ATAD3A also results in loss of both cell anchorage-independent growth and invasion and suppression of tumor growth and metastasis in vivo using immuno-compromised mice. HSP70 is responsible for stabilizing WASF3 in the cytoplasm, but inactivation of HSP70 does not lead to the loss of WASF3 stability at the mitochondrial membrane, where presumably it is protected through its interaction with ATAD3A. In response to endoplasmic reticulum (ER) stress, increases in the GRP78 protein level leads to increased WASF3 protein levels. We also show that ATAD3A was present in a WASF3-GRP78 complex, and suppression of GRP78 led to destabilization of WASF3 at the mitochondrial membrane, which was ATAD3A dependent. Furthermore, ATAD3A-mediated suppression of CDH1/E-cadherin occurs through its regulation of GRP78-mediated WASF3 stability. Proteolysis experiments using isolated mitochondria demonstrates the presence of the N-terminal end of WASF3 within the mitochondria, which is the interaction site with the N-terminal end of ATAD3A. It appears, therefore, that stabilization of WASF3 function occurs through its interaction with ATAD3A and GRP78, which may provide a bridge between the ER and mitochondria, allowing communication between the two organelles. These findings also suggest that pharmacologic inhibition of ATAD3A could be an effective therapeutic strategy to treat human cancer.


Training Pre-Service Early Childhood Educators in Physical Activity (TEACH): Protocol for a Quasi-Experimental Study.
Early childhood educators (ECEs) influence young children's early uptake of positive health behaviours in childcare settings and serve as important daytime role models. As such, it is imperative that post-secondary early childhood education programs provide students with the foundational knowledge and professional training required to confidently facilitate quality active play opportunities for young children. The primary objective of the Training pre-service EArly CHildhood educators in physical activity (TEACH) study is to develop and implement an e-Learning course in physical activity and sedentary behaviour to facilitate improvements in: pre-service ECEs' self-efficacy and knowledge to lead physical activity and outdoor play opportunities and minimize sedentary behaviours in childcare. This study will also explore pre-service ECEs' behavioural intention and perceived control to promote physical activity and outdoor play, and minimize sedentary behaviour in childcare, and the implementation of the e-Learning course. A mixed-methods quasi-experimental design with three data collection time points (baseline, post-course completion, 3-month follow-up) will be employed to test the e-Learning course in early childhood education programs (n = 18; 9 experimental, 9 comparison) across Canada. Pre-service ECEs enrolled in colleges/universities assigned to the experimental group will be required to complete a 4-module e-Learning course, while programs in the comparison group will maintain their typical curriculum. Pre-service ECEs' self-efficacy, knowledge, as well as behavioural intention and perceived behavioural control will be assessed via online surveys and module completion rates will be documented using website metrics. Group differences across timepoints will be assessed using linear mixed effects modelling and common themes will be identified through thematic analysis. The TEACH study represents a novel, evidence-informed approach to address the existing gap in physical activity and sedentary behaviour-related education in Canadian post-secondary early childhood education programs. Moreover, e-Learning platforms, can be employed as an innovative, standardized, and scalable way to provide ECEs with consistent training across jurisdictions.


Beta-blocker therapy for tremor in Parkinson's disease.
The tremor of Parkinson's disease can cause considerable disability for the individual concerned. Traditional antiparkinsonian therapies such as levodopa have only a minor effect on tremor. Beta-blockers are used to attenuate other forms of tremor such as Essential Tremor or the tremor associated with anxiety. It is thought that beta-blockers may be of use in controlling the tremor of Parkinson's disease. To compare the efficacy and safety of adjuvant beta-blocker therapy against placebo for the treatment of tremor in patients with Parkinson's disease. Electronic searches of MEDLINE, EMBASE, SCISEARCH, BIOSIS, GEROLIT, OLDMEDLINE, LILACS, MedCarib, PASCAL, JICST-EPLUS, RUSSMED, DISSERTATION ABSTRACTS, SIGLE, ISI-ISTP, Aslib Index to Theses, The Cochrane Controlled Trials Register, Clinicaltrials.gov, metaRegister of Controlled Trials, NIDRR, NRR and CENTRAL were conducted. Grey literature was hand searched and the reference lists of identified studies and reviews examined. The manufacturers of beta-blockers were contacted. Randomised controlled trials of adjuvant beta-blocker therapy versus placebo in patients with a clinical diagnosis of idiopathic Parkinson's disease. Data was abstracted independently by two of the authors onto standardised forms and disagreements were resolved by discussion. Four randomised controlled trials were found comparing beta-blocker therapy with placebo in patients with idiopathic Parkinson's disease. These were double-blind cross-over studies involving a total of 72 patients. Three studies did not present data from the first arm, instead presenting results as combined data from both treatment arms and both placebo arms. The risk of a carry-over effect into the second arm meant that these results were not analysed. The fourth study presented data from each arm. This was in the form of a mean total score for tremor for each group. Details of the baseline scores, the numbers of patients in each group and standard deviations were not provided, meaning that the magnitude and significance of any changes due to therapy could not be calculated. One study reported a substantial fall in heart rate in 14 of the 22 patients, with one patient withdrawing after his heart rate dropped to 56 beats per minute (baseline heart rate was not reported). In view of this lack of evidence, it is impossible to determine whether beta-blocker therapy is effective and safe for the treatment of tremor in Parkinson's disease. The high frequency of bradycardia in one trial raises some concerns about the prescription of beta-blockers to normotensive elderly patients but the study was too small for the true degree of risk to be calculated.


[Efficacy of behavioural lifestyle interventions for weight management of psychiatric patients].
Significant weight gain is common in patients with psychiatric disorders taking antipsychotics, mood stabilisers and/or antidepressants. Psychiatric patients are at a higher risk of suffering from obesity and from the metabolic syndrome than the general population. One promising alternative to limiting weight gain involves modifying lifestyle behaviors. We reviewed the literature on behavioral weight management programs for this clientele. Of the 17 identified studies, 12 reported a significant impact of the program on weight gain compared to usual care. The importance of offering weight management programs to psychiatric patients in outpatient settings is stressed.


Whipple's disease: misdiagnosed as sarcoidosis with further tricuspid valve endocarditis and pulmonary embolism - a case report.
GH Whipple described a 36-year-old physician in 1907 with gradual loss of weight and strength, stools consisting chiefly of neutral fat and fatty acids, indefinite abdominal signs and a peculiar multiple arthritis. The patient died of this progressive illness. Whipple called it intestinal lipodystrophy since he observed accumulation of large masses of neutral fats and fatty acids in the lymph spaces. It was renamed Whipple's disease in 1949. An infectious aetiology was suspected as early as Whipple's initial report. However, successful treatment with antibiotics was not reported until 1952, which resulted in dramatic clinical responses. The cause is now known to be Tropheryma whipplei. Light and electron microscopy of infected tissue identified a gram-positive, non-acid-fast, periodic acid-Schiff (PAS) positive bacillus with a characteristic trilamellar plasma membrane resembling that of gram-negative bacteria. Whipple's disease is extremely rare. It is a systemic infectious disorder affecting mostly middle-aged white men. The clinical presentation is often non-specific, which may make its diagnosis difficult. The four cardinal clinical manifestations are arthralgias, weight loss, diarrhoea and abdominal pain. The frequently vague articular symptoms can precede the diagnosis of Whipple's disease by an average of 6-8 years. Lymph nodes and other tissues may present diagnostic problems, since the changes in routinely stained sections may mimic those of sarcoidosis. The detection of PAS-positive histiocytes in the small intestine remains the mainstay of the diagnosis, although Whipple's disease without gastrointestinal involvement is described. We illustrate a case in which, retrospectively, the clinical presentation would have been typical for Whipple's disease. However, the clinical presentation and the histological examinations of lymph nodes, liver biopsies and ascites initially were misinterpreted as sarcoidosis with consecutive immunosuppressive therapy and progressive worsening of the patient's health presenting at least as sepsis with endocarditis.


Characterization of Polyphenolic Content in the Aquatic Plants Ruppia cirrhosa and Ruppia maritima -A Source of Nutritional Natural Products.
Herein, the polyphenolic content in extracts of Ruppia cirrhosa (Petagna) Grande and Ruppia maritima L.was fully characterized for the first time. High amounts of the main compound chicoric acid (CA) (≤30.2 ± 4.3 mg/g) were found in both Ruppia species. In addition, eight flavonoids, namely the 3-O-glucopyranosides and 3-O-galactopyranosides, as well as malonylated 3-O-glycosides of quercetin and isorhamnetin, were isolated and identified. The antioxidant activity of Ruppia cirrhosa extracts and isolated compounds was investigated spectrophotometrically by a 1,1-diphenyl-2-picrylhydrazyl (DPPH·) radical scavenging assay. IC50 values were 31.8-175.7 μg/mL for Ruppia cirrhosa extracts and 12.1-88.4 μg/mL for isolated flavonoids. Both individual and total phenolic and flavonoid content were quantified in crude extracts using analytical HPLC. The relative high amount of total flavonoids ranged from 5.9 to 14.7 mg/g in both species, with concentrations of individual flavonoids ranging from 0.4 to 2.9 mg/g dry weight. The content of chicoric acid was twofold more in Ruppia maritima than in Ruppia cirrhosa. Seasonal variation of the quantitative content in Ruppia cirrhosa was examined. Total flavonoid content ranged from 8.4 mg/g in October to 14.7 mg/g in August, whereas the highest concentration of chicoric acid was observed in March (29.2 mg/g).


Neglected diseases of neglected populations: thinking to reshape the determinants of health in Latin America and the Caribbean.
People living in poverty throughout the developing world are heavily burdened with neglected communicable diseases and often marginalized by the health sector. These diseases are currently referred to as Neglected Diseases of Neglected Populations. The neglected diseases create social and financial burdens to the individual, the family, the community, and the nation. Numerous studies of successful individual interventions to manage communicable disease determinants in various types of communities have been published, but few have applied multiple interventions in an integrated, coordinated manner. We have identified a series of successful interventions and developed three hypothetical scenarios where such interventions could be applied in an integrated, multi-disease, inter-programmatic, and/or inter-sectoral approach for prevention and control of neglected diseases in three different populations: a slum, an indigenous community, and a city with a mix of populations. The objective of this paper is to identify new opportunities to address neglected diseases, improve community health and promote sustainable development in neglected populations by highlighting examples of key risk and protective factors for neglected diseases which can be managed and implemented through multi-disease-based, integrated, inter-programmatic, and/or inter-sectoral approaches. Based on a literature review, analysis and development of scenarios we visualize how multiple interventions could manage multiple disease problems and propose these as possible strategies to be tested. We seek to stimulate intra- and inter-sectoral dialogue which will help in the construction of new strategies for neglected diseases (particularly for the parasitic diseases) which could benefit the poor and marginalized based on the principle of sustainability and understanding of key determinants of health, and lead to the establishment of pilot projects and activities which can contribute to the achievement of the Millennium Development Goals.


Feasibility of neuromuscular training in patients with severe hip or knee OA: the individualized goal-based NEMEX-TJR training program.
Although improvements are achieved by general exercise, training to improve sensorimotor control may be needed for people with osteoarthritis (OA). The aim was to apply the principles of neuromuscular training, which have been successfully used in younger and middle-aged patients with knee injuries, to older patients with severe hip or knee OA. We hypothesized that the training program was feasible, determined as: 1) at most acceptable self-reported pain following training; 2) decreased or unchanged pain during the training period; 3) few joint specific adverse events related to training, and 4) achieved progression of training level during the training period. Seventy-six patients, between 60 and 77 years, with severe hip (n = 38, 55% women) or knee OA (n = 38, 61% women) underwent an individualized, goal-based neuromuscular training program (NEMEX-TJR) in groups for a median of 11 weeks (quartiles 7 to 15) prior to total joint replacement (TJR). Pain was self-reported immediately after each training session on a 0 to 10 cm, no pain to pain as bad as it could be, scale, where 0-2 indicates safe, > 2 to 5 acceptable and > 5 high risk pain. Joint specific adverse events were: not attending or ceasing training because of increased pain/problems in the index joint related to training, and self-reported pain > 5 after training. The level of difficulty of training was registered. Patients with severe OA of the hip or knee reported safe pain (median 2 cm) after training. Self-reported pain was lower at training sessions 10 and 20 (p = 0.04) and unchanged at training sessions 5 and 15 (p = 0.170, p = 0.161) compared with training session 1. There were no joint specific adverse events in terms of not attending or ceasing training. Few patients (n = 17, 22%) reported adverse events in terms of self-reported pain > 5 after one or more training sessions. Progression of training level was achieved over time (p < 0.001). The NEMEX-TJR training program is feasible in patients with severe hip or knee OA, in terms of safe self-reported pain following training, decreased or unchanged pain during the training period, few joint specific adverse events, and achieved progression of training level during the training period.


Anti-tumor effects of genetic vaccines against HPV major oncogenes.
Expression of HPV E5, E6 and E7 oncogenes are likely to overcome the regulation of cell proliferation and to escape immunological control, allowing uncontrolled growth and providing the potential for malignant transformation. Thus, their three oncogenic products may represent ideal target antigens for immunotherapeutic strategies. In previous attempts, we demonstrated that genetic vaccines against recombinant HPV16 E7 antigen were able to affect the tumor growth in a pre-clinical mouse model. To improve this anti-HPV strategy we developed a novel approach in which we explored the effects of E5-based genetic immunization. We designed novel HPV16 E5 genetic vaccines based on two different gene versions: whole E5 gene and E5Multi. The last one is a long multi epitope gene designed as a harmless E5 version. Both E5 genes were codon optimized for mammalian expression. In addition, we demonstrated that HPV 16 E5 oncogene is expressed in C3 mouse cell line making it an elective model for the study of E5 based vaccine. In this mouse model the immunological and biological activity of the E5 vaccines were assessed in parallel with the activity of anti-E7 and anti-E6 vaccines already reported to be effective in an immunotherapeutic setting. These E7 and E6 vaccines were made with mutated oncogenes, the E7GGG mutant that does not bind pRb and the E6F47R mutant that is less effective in inhibiting p53, respectively. Results confirmed the immunological activity of genetic formulations based on attenuated HPV16 oncogenes and showed that E5-based genetic immunization provided notable anti-tumor effects.


Transporters buffer synaptically released glutamate on a submillisecond time scale.
The role of transporters in clearing free glutamate from the synaptic cleft was studied in rat CA1 hippocampal neurons cultured on glial microislands. The time course of free glutamate in the cleft during a synaptic event was estimated by measuring the extent to which the rapidly dissociating AMPA receptor antagonist kynurenate (KYN) was replaced by glutamate during a synaptic response. Dose inhibition of the AMPA receptor EPSC by KYN was less than predicted by the equilibrium affinity of the antagonist, and the rise time of AMPA receptor miniature EPSCs (mEPSCs) was slowed by KYN. Both results indicated that KYN dissociated from AMPA receptors and was replaced by synaptically released transmitter. When transporters were blocked by D,L-threo-beta-hydroxyaspartic acid (THA) or Li+, the mEPSC rise time in the presence of KYN was slowed further, indicating that transporters affect the glutamate concentration in the first few hundred microseconds of the synaptic response. The glutamate transient necessary to cause these effects was determined by developing a detailed kinetic model of the AMPA receptor. The model replicated the effects of KYN on the amplitude and rise time of the synaptic responses when driven by glutamate transients that were similar to previous estimates (; ). The effects of THA were replicated by slowing and enlarging the slower phase of the dual component transient by about 20% or by prolonging the single component by almost 40%. Because transport is too slow to account for these effects, it is concluded that transporters buffer glutamate in the synaptic cleft.


Management of bullous pemphigoid: the European Dermatology Forum consensus in collaboration with the European Academy of Dermatology and Venereology.
Bullous pemphigoid is the most common autoimmune subepidermal blistering disease of the skin and mucous membranes. This disease typically affects the elderly and presents with itch and localized or generalized bullous lesions. In up to 20% of affected patients, bullae may be completely absent, and only excoriations, prurigo-like lesions, eczematous lesions, urticated lesions and/or infiltrated plaques are observed. The disease is significantly associated with neurological disorders. The morbidity of bullous pemphigoid and its impact on quality of life are significant. So far, a limited number of national treatment guidelines have been proposed, but no common European consensus has emerged. Our consensus for the treatment of bullous pemphigoid has been developed under the guidance of the European Dermatology Forum in collaboration with the European Academy of Dermatology and Venereology. It summarizes evidence-based and expert-based recommendations.


RhoGDI: multiple functions in the regulation of Rho family GTPase activities.
RhoGDI (Rho GDP-dissociation inhibitor) was identified as a down-regulator of Rho family GTPases typified by its ability to prevent nucleotide exchange and membrane association. Structural studies on GTPase-RhoGDI complexes, in combination with biochemical and cell biological results, have provided insight as to how RhoGDI exerts its effects on nucleotide binding, the membrane association-dissociation cycling of the GTPase and how these activities are controlled. Despite the initial negative roles attributed to RhoGDI, recent evidence has come to suggest that it may also act as a positive regulator necessary for the correct targeting and regulation of Rho activities by conferring cues for spatial restriction, guidance and availability to effectors. These potential functions are discussed in the context of RhoGDI-associated multimolecular complexes, the newly emerged shuttling capability and the importance of the particular membrane microenvironment that represents the site of action for GTPases. All these results point to a wider role for RhoGDI than initially perceived, making it a binding partner that can tightly control Rho GTPases, but which also allows them to reach their full spectrum of activities.


SIVmac251 is inefficiently transmitted to rhesus macaques by penile inoculation with a single SIVenv variant found in ramp-up phase plasma.
Abstract Despite the fact that approximately half of all HIV patients acquire infection through penile exposure, there have been no recent studies of penile SIV transmission in rhesus macaques and the nature of the virus variants transmitted, target cells, and pathways of virus dissemination to systemic lymphoid tissues are not known. Single genome amplification (SGA) and sequencing of HIV-1 RNA in plasma of acutely infected humans allows the identification and enumeration of transmitted/founder viruses responsible for productive systemic infection. Studies using the SGA strategy have shown that intrarectal and intravaginal SIV transmission to macaques recapitulates key features of human HIV transmission. To date, no studies have used the SGA assay to identify transmitted/founder virus(es) in macaques infected after penile SIV exposure. Here we report that SIV can be transmitted by penile SIV exposure. However, similar exposure to a high-dose inoculum infects only about half the animals, which is about 50% less efficient transmission than occurs after vaginal SIV challenge. In addition, only a single SIV env variant established the systemic infection in all five animals that became infected after penile exposure, a result that is consistent with low incidence and few transmitted HIV variants in heterosexually infected men. Our results suggest that the penile transmission of SIVmac251 in rhesus macaques recapitulates the key features of penile HIV-1 transmission and may provide insight into host or viral factors that permit penile transmission and dissemination. Furthermore, this SIV challenge exposure route will be useful in testing vaccines and other prophylactic approaches.


Ethanol inhibits kainate responses of glutamate receptors expressed in Xenopus oocytes: role of calcium and protein kinase C.
Recombinant alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate receptors expressed in oocytes are inhibited by ethanol and the sensitivity to ethanol depends on the kainate concentration and the subunit(s) expressed. For example, GluR3 kainate channels are more sensitive to inhibition by ethanol than GluR6 channels in the presence of maximally effective kainate concentrations. To determine if the ethanol inhibition was influenced by the cation permeability (Na+ vs Na+ and Ca2+) of the channels expressed, we compared ethanol inhibition of Ca(2+)-permeable glutamate receptors (GluRs) in oocytes perfused with normal- and high-Ca2+ buffers. The ethanol inhibition was much greater when Ca2+ was the only permeant cation. When Ba2+ was substituted for Ca2+, the ethanol inhibition was reduced, although it was still greater than with normal buffer. The enhanced ethanol inhibition of kainate-stimulated Ca2+ currents was reduced in oocytes injected with the Ca2+ chelator BAPTA, suggesting a role for intracellular Ca2+ in mediating enhanced ethanol sensitivity of kainate channels. The enhanced ethanol inhibition of Ca2+ currents was not due to a direct ethanol inhibition of Ca(2+)-stimulated Cl- currents in the oocyte because ethanol produced no effect on Ca(2+)-stimulated Cl-currents induced by injection of myo-inositol-1,4,5-trisphosphate. Because Ca2+ activates protein kinase C (PKC) and because we found that the PKC activator phorbol 12-myristate 13-acetate inhibits kainate responses (Dildy-Mayfield and Harris, 1994), we examined the role of PKC in mediating the enhanced ethanol inhibition of kainate responses produced by increased Ca2+. Inhibition of PKC by injection of the PKC inhibitor peptide or calphostin C prevented the enhanced ethanol inhibition of kainate-induced Ca2+ responses without altering ethanol inhibition in normal buffer. Thus, ethanol inhibition of kainate channels may involve two mechanisms, one that is independent of PKC and a second type that is due to activation of PKC under conditions of elevated Ca2+, resulting in enhanced inhibition of kainate responses.


Effects of mechanical strain on the function of Gap junctions in osteocytes are mediated through the prostaglandin EP2 receptor.
Osteocytes embedded in the matrix of bone are thought to be mechanosensory cells that translate mechanical strain into biochemical signals that regulate bone modeling and remodeling. We have shown previously that fluid flow shear stress dramatically induces prostaglandin release and COX-2 mRNA expression in osteocyte-like MLO-Y4 cells, and that prostaglandin E2 (PGE2) released by these cells functions in an autocrine manner to regulate gap junction function and connexin 43 (Cx43) expression. Here we show that fluid flow regulates gap junctions through the PGE2 receptor EP2 activation of cAMP-dependent protein kinase A (PKA) signaling. The expression of the EP2 receptor, but not the subtypes EP1,EP3, and EP4, increased in response to fluid flow. Application of PGE2 or conditioned medium from fluid flow-treated cells to non-stressed MLO-Y4 cells increased expression of the EP2 receptor. The EP2 receptor antagonist, AH6809, suppressed the stimulatory effects of PGE2 and fluid flow-conditioned medium on the expression of the EP2 receptor, on Cx43 protein expression, and on gap junction-mediated intercellular coupling. In contrast, the EP2 receptor agonist butaprost, not the E1/E3 receptor agonist sulprostone, stimulated the expression of Cx43 and gap junction function. Fluid flow conditioned medium and PGE2 stimulated cAMP production and PKA activity suggesting that PGE2 released by mechanically stimulated cells is responsible for the activation of cAMP and PKA. The adenylate cyclase activators, forskolin and 8-bromo-cAMP, enhanced intercellular connectivity, the number of functional gap junctions, and Cx43 protein expression, whereas the PKA inhibitor, H89, inhibited the stimulatory effect of PGE2 on gap junctions. These studies suggest that the EP2 receptor mediates the effects of autocrine PGE2 on the osteocyte gap junction in response to fluid flow-induced shear stress. These data support the hypothesis that the EP2 receptor, cAMP, and PKA are critical components of the signaling cascade between mechanical strain and gap junction-mediated communication between osteocytes.


Introduction of p130cas signaling complex formation upon integrin-mediated cell adhesion: a role for Src family kinases.
Integrin-mediated cell adhesion triggers intracellular signaling cascades, including tyrosine phosphorylation of intracellular proteins. Among these are the focal adhesion proteins p130cas (Cas) and focal adhesion kinase (FAK). Here we identify the kinase(s) mediating integrin-induced Cas phosphorylation and characterize protein-protein interactions mediated by phosphorylated Cas. We found that expression of a constitutively active FAK in fibroblasts results in a consecutive tyrosine phosphorylation of Cas. This effect required the autophosphorylation site of FAK, which is a binding site for Src family kinases. Integrin-mediated phosphorylation of Cas was not, however, compromised in fibroblasts lacking FAK. In contrast, adhesion-induced tyrosine phosphorylation of Cas was reduced in cells lacking Src, whereas enhanced phosphorylation of Cas was observed Csk- cells, in which Src kinases are activated. These results suggest that Src kinases are responsible for the integrin-mediated tyrosine phosphorylation of Cas. FAK seems not to be necessary for phosphorylation of Cas, but when autophosphorylated, FAK may recruit Src family kinases to phosphorylate Cas. Cas was found to form complexes with Src homology 2 (SH2) domain-containing signaling molecules, such as the SH2/SH3 adapter protein Crk, following integrin-induced tyrosine phosphorylation. Guanine nucleotide exchange factors C3G and Sos were found in the Cas-Crk complex upon integrin ligand binding. These observations suggest that Cas serves as a docking protein and may transduce signals to downstream signaling pathways following integrin-mediated cell adhesion.


Cell surface antigens of human melanoma identified by monoclonal antibody.
Mouse NS-1 myeloma cells were fused with spleen cells from mice that had been immunized with cells from a human melanoma, M1804. Hybrid cells were grown in selective medium and tested for production of antibody to surface antigens of M1804 cells. Three hybrids that produced antibodies that bound to the melanoma cells but not to autologous skin fibroblasts were cloned. Antibodies produced by two of the clones were cytotoxic to M1804 cells in the presence of rabbit complement. Extensive specificity tests showed that the antibodies produced by the clones bound strongly only to M1804 cells; significant, although weaker, binding occurred with 2 of 11 allogeneic melanomas. Apart from weak binding of the antibody produced by one of the clones to a breast carcinoma, binding assays of five carcinomas, one sarcoma, and fibroblasts from 17 individuals were negative, as were cytotoxic tests of 10 lymphoblastoid cell lines and peripheral blood lymphocytes from 68 normal donors and 12 chronic lymphocytic leukemia patients. This suggests that we have identified one or more determinants of a melanoma-associated antigen(s), whose expression is limited to a small proportion of melanomas.


[Surgical treatment of thoracic and thoracolumbar spinal tuberculosis].
To explore the surgical measurements and principles in the treatment of thoracic and thoracolumbar spinal tuberculosis. From 2001 to 2008, 232 cases of thoracic or thoracolumbar spinal tuberculosis were treated by operations in the study, including 148 males and 84 females with an average age of 37.8 years ranging from 20 to 76 years. Preoperative assessment displayed as follow: Cobb angles of kyphosis < 30 degrees in 65 cases, 30 degrees to 60 degrees in 147 cases, > 60 degrees in 20 cases; Frankel B grade in 13 cases, C in 12 cases, D in 41 cases, E in 166 cases. Among them, 48 cases were performed with one-stage transpedicular screw system and anterolateral debridement by single incision, 184 cases with one-stage anterior approach (debridement, fusion, and plate-screw fixation) routinely. The tissues and liquor puris debrided from focus were sent for pathological examination, Bacillus tuberculosis detection and culture, and drug sensitivity test. The patients were given anti-tuberculosis therapy according the results of drug sensitivity test for 1 to 1.5 years. The followed-up included relapse rate, fusion of the bone graft, the status of neurological restoring, kyphosis correction etc. All 232 cases recovered from perioperation and 230 cases got primary wound healing, only 2 cases performed with single incision one-stage posterior instrumentation and anterolateral debridement got complications of wound healing problems and the sinus formation,which delayed healed by changing dressings. The complications included intercostals neuralgia in 135 cases and pneumothorax or hydrothorax in 13 cases, which needed not special handling. All the patients in this series got the followed-up ranging from 1.0 to 4.5 years (means 2.6 years). No recurrence within followed-up period and bone union was found in all cases. All 66 cases with the neurological deficits recovered partially or totally. Kyphosis correction were achieved by 27.5 degrees on average postoperatively and showed a mild loss of 4.2 degrees on average during followed-up period. All cases were confirmed with Bacillus tuberculosis infection by pathology. Bacillus tuberculosis was detected and culture successfully in 107 cases (46.1%), 40 strains (37.4%) were drug resistant and in which 8 strains (7.5%) were multi-drug resistant. For the treatment of thoracic and thoracolumbar spinal tuberculosis, the best treatment include directional chemotherapy, one-stage anterior approach with thorough debridement, auto-rib or Ti-mesh fusion, and plate-screw fixation.


The Rx homeobox gene is essential for vertebrate eye development.
Development of the vertebrate eye requires a series of steps including specification of the anterior neural plate, evagination of the optic vesicles from the ventral forebrain, and the cellular differentiation of the lens and retina. Homeobox-containing genes, especially the transcription regulator Pax6, play a critical role in vertebrate and invertebrate eye formation. Mutations in Pax6 function result in eye malformations known as Aniridia in humans and Small eye syndrome in mice. The Drosophila homologue of Pax6, eyeless, is also necessary for correct invertebrate eye development, and its misexpression leads to formation of ectopic eyes in Drosophila. Here we show that a conserved vertebrate homeobox gene, Rx, is essential for normal eye development, and that its misexpression has profound effects on eye morphology. Xenopus embryos injected with synthetic Rx RNA develop ectopic retinal tissue and display hyperproliferation in the neuroretina. Mouse embryos carrying a null allele of this gene do not form optic cups and so do not develop eyes. The Rx gene family plays an important role in the establishment and/or proliferation of retinal progenitor cells.


Condenser-free contrast methods for transmitted-light microscopy.
Phase contrast microscopy allows the study of highly transparent yet detail-rich specimens by producing intensity contrast from phase objects within the sample. Presented here is a generalized phase contrast illumination schema in which condenser optics are entirely abrogated, yielding a condenser-free yet highly effective method of obtaining phase contrast in transmitted-light microscopy. A ring of light emitting diodes (LEDs) is positioned within the light-path such that observation of the objective back focal plane places the illuminating ring in appropriate conjunction with the phase ring. It is demonstrated that true Zernike phase contrast is obtained, whose geometry can be flexibly manipulated to provide an arbitrary working distance between illuminator and sample. Condenser-free phase contrast is demonstrated across a range of magnifications (4-100×), numerical apertures (0.13-1.65NA) and conventional phase positions. Also demonstrated is condenser-free darkfield microscopy as well as combinatorial contrast including Rheinberg illumination and simultaneous, colour-contrasted, brightfield, darkfield and Zernike phase contrast. By providing enhanced and arbitrary working space above the preparation, a range of concurrent imaging and electrophysiological techniques will be technically facilitated. Condenser-free phase contrast is demonstrated in conjunction with scanning ion conductance microscopy (SICM), using a notched ring to admit the scanned probe. The compact, versatile LED illumination schema will further lend itself to novel next-generation transmitted-light microscopy designs. The condenser-free illumination method, using rings of independent or radially-scanned emitters, may be exploited in future in other electromagnetic wavebands, including X-rays or the infrared.


A multiphysics model of the Pacinian corpuscle.
The Pacinian corpuscle (PC) is a dermal mechanoreceptor that responds to high-frequency (20-1000 Hz) vibrations. The PC's structure allows transmission of vibrations through its layers (lamellae) to the centrally-located nerve fiber (neurite). This work combines mechanical models of the PC with an electrochemical model of peripheral nerves to simulate the tactile response of the entire system. A three-stage model of response to a vibratory input was developed, consisting of (1) outer core mechanics, (2) inner core mechanics, and (3) neurite electrochemistry. The model correctly predicts the band-pass nature of the PC's frequency response, showing that the PC structure can amplify oscillatory strains within its target frequency band. Specifically, strain induced by a vibratory stimulus is amplified by a factor of 8-12 from the PC surface to the neurite. Our results also support the hypothesis that PC rapid adaptation is affected by the lamellar structures without requiring neuronal adaptivity. Simulated different-sized PCs showed a shift in frequency response, suggesting that clusters of different-sized PCs could enable more nuanced tactile encoding than uniform clusters. By modeling the PC's mechano-to-neural transduction, we can begin to characterize the mechanosensation of other receptors to understand how multiple receptors interact to create our sensation of touch.


Experimental infection of rainbow trout Oncorhynchus mykiss with viral haemorrhagic septicaemia virus isolates from European marine and farmed fishes.
The susceptibility of rainbow trout Oncorhynchus mykiss to infection with various isolates of viral haemorrhagic septicaemia virus (VHSV) was examined. A total of 8 experiments with rainbow trout ranging from 0.6 to 6.2 g was conducted for 139 isolates originating from wild marine fishes in European waters (115 isolates), farmed turbot from Scotland and Ireland (2 isolates), and farmed rainbow trout (22 isolates). The isolates were tested by immersion and/or intraperitoneal injection either as pooled or single isolates. The isolates from wild marine fishes did not cause mortality by immersion while some of the isolates caused mortality when injected. All VHSV isolates from farmed rainbow trout caused significant mortality by immersion. Currently, pathogenicity trials are the only way to differentiate VHSV isolates from wild marine fishes and farmed rainbow trout. The 2 farmed turbot isolates did not cause mortality by immersion, supporting the view that they originated from the marine environment.


Structural and functional characterization of an arylamine N-acetyltransferase from the pathogen Mycobacterium abscessus: differences from other mycobacterial isoforms and implications for selective inhibition.
Mycobacterium abscessus is the most pathogenic rapid-growing mycobacterium and is one of the most resistant organisms to chemotherapeutic agents. However, structural and functional studies of M. abscessus proteins that could modify/inactivate antibiotics remain nonexistent. Here, the structural and functional characterization of an arylamine N-acetyltransferase (NAT) from M. abscessus [(MYCAB)NAT1] are reported. This novel prokaryotic NAT displays significant N-acetyltransferase activity towards aromatic substrates, including antibiotics such as isoniazid and p-aminosalicylate. The enzyme is endogenously expressed and functional in both the rough and smooth M. abscessus morphotypes. The crystal structure of (MYCAB)NAT1 at 1.8 Å resolution reveals that it is more closely related to Nocardia farcinica NAT than to mycobacterial isoforms. In particular, structural and physicochemical differences from other mycobacterial NATs were found in the active site. Peculiarities of (MYCAB)NAT1 were further supported by kinetic and docking studies showing that the enzyme was poorly inhibited by the piperidinol inhibitor of mycobacterial NATs. This study describes the first structure of an antibiotic-modifying enzyme from M. abscessus and provides bases to better understand the substrate/inhibitor-binding specificities among mycobacterial NATs and to identify/optimize specific inhibitors. These data should also contribute to the understanding of the mechanisms that are responsible for the pathogenicity and extensive chemotherapeutic resistance of M. abscessus.


Simultaneous object classification and segmentation with high-order multiple shape models.
Shape models (SMs), capturing the common features of a set of training shapes, represent a new incoming object based on its projection onto the corresponding model. Given a set of learned SMs representing different objects classes, and an image with a new shape, this work introduces a joint classification-segmentation framework with a twofold goal. First, to automatically select the SM that best represents the object, and second, to accurately segment the image taking into account both the image information and the features and variations learned from the online selected model. A new energy functional is introduced that simultaneously accomplishes both goals. Model selection is performed based on a shape similarity measure, online determining which model to use at each iteration of the steepest descent minimization, allowing for model switching and adaptation to the data. High-order SMs are used in order to deal with very similar object classes and natural variability within them. Position and transformation invariance is included as part of the modeling as well. The presentation of the framework is complemented with examples for the difficult task of simultaneously classifying and segmenting closely related shapes, such as stages of human activities, in images with severe occlusions.


Dynamic simulation of three dimensional architectural and mechanical alterations in human trabecular bone during menopause.
A three dimensional (3D) computational simulation of dynamic process of trabecular bone remodeling was developed with all the parameters derived from physiological and clinical data. Contributions of the microstructural bone formation deficits: trabecular plate perforations, trabecular rod breakages, and isolated bone fragments, to the rapid bone loss and disruption of trabecular microarchitecture during menopause were studied. Eighteen human trabecular bone samples from femoral neck (FN) and spine were scanned using a micro computed tomography (microCT) system. Bone resorption and formation were simulated as a computational cycle corresponding to 40-day resorption/160-day formation. Resorption cavities were randomly created over the bone surface according to the activation frequency, which was strictly based on clinical data. Every resorption cavity was refilled during formation unless it caused trabecular plate perforation, trabecular rod breakage or isolated fragments. A 20-year-period starting 5 years before and ending 15 years after menopause was simulated for each specimen. Elastic moduli, standard and individual trabeculae segmentation (ITS)-based morphological parameters were evaluated for each simulated 3D image. For both spine and FN groups, the time courses of predicted bone loss pattern by microstructural bone formation deficits were fairly consistent with the clinical measurements. The percentage of bone loss due to trabecular plate perforation, trabecular rod breakage, and isolated bone fragments were 73.2%, 18.9% and 7.9% at the simulated 15 years after menopause. The ITS-based plate fraction (pBV/BV), mean plate surface area (pTb.S), plate number density (pTb.N), and mean rod thickness (rTb.Th) decreased while rod fraction (rBV/BV) and rod number density (rTb.N) increased after the simulated menopause. The dynamic bone remodeling simulation based on microstructural bone formation deficits predicted the time course of menopausal bone loss pattern of spine and FN. Microstructural plate perforation could be the primary cause of menopausal trabecular bone loss. The combined effect of trabeculae perforation, breakage, and isolated fragments resulted in fewer and smaller trabecular plates and more but thinner trabecular rods.


Total hip arthroplasty with an uncemented tapered femoral component.
Excellent intermediate-term results with use of tapered femoral components in primary total hip arthroplasty have been reported. The purpose of this study was to update our previous report and to evaluate the outcome of total hip arthroplasty with use of the porous Taperloc femoral component in patients who had been followed for a minimum of eighteen years postoperatively. One hundred and forty-five consecutive uncemented total hip arthroplasties in 138 patients were performed between 1983 and 1985, by a single surgeon, with use of the Taperloc femoral component. The outcome of every femoral component with regard to stem fixation, retention, or revision was determined for all 145 total hip replacements. At a mean of twenty years (range, eighteen to 22.6 years) postoperatively, fifty-eight patients (sixty-five hips) were living. In fifty living patients (fifty-seven hips), the femoral component had not undergone revision surgery. Evaluation of the living patients included clinical and radiographic analysis and recording of complications. Of the eighty patients (eighty hips) who had died, five hips had undergone revision of the femoral component. Only one stem had been revised for aseptic loosening, and no femoral component had been definitely loose by radiographic criteria. In the remaining sixty-five hips in the fifty-eight living patients, eight femoral components were revised. No femoral component underwent revision for aseptic loosening. Definite radiographic evidence of femoral component loosening occurred in one hip. In the fifty living patients (fifty-seven hips) who had not undergone femoral component revision, there was a significant improvement in the mean Harris hip score from 49.3 points preoperatively to 85.4 points at the time of latest follow-up (p = 0.001). Survivorship analysis of all 145 hips, with revision for any reason as the end point, estimated that the survival rate for the femoral component was 87% (95% confidence interval, 79% to 93%) at twenty-two years. Primary total hip arthroplasty with the Taperloc femoral component is associated with a low rate of aseptic loosening at a mean follow-up of twenty years.


Pathways towards true catalysts: computational modelling and structural transformations of Zn-polyoxotungstates.
Current catalysis undergoes a paradigm shift from molecular and heterogeneous realms towards new dynamic catalyst concepts. This calls for innovative strategies to understand the essential catalytic motifs and true catalysts emerging from oxidative transformation processes. Polyoxometalate (POM) clusters offer an inexhaustible reservoir for new noble metal-free catalysts and excellent model systems whose structure-activity relationships and mechanisms remain to be explored. Here, we first introduce a new {ZnnNa6-n(B-α-SbW9O33)2} (n = 3-6) catalyst family with remarkable tuning options of the Zn-based core structure and high activity in H2O2-assisted catalytic alcohol oxidation as a representative reaction. Next, high level solution-based computational modelling of the intermediates and transition states was carried out for [Zn6Cl6(SbW9O33)2]12- as a representative well-defined case. The results indicate a radical-based oxidation process with the involvement of tungsten and adjacent zinc metal centers. The {ZnnNa6-n(B-α-SbW9O33)2} series indeed efficiently catalyses alcohol oxidation via peroxotungstate intermediates, in agreement with strong spectroscopic support and other experimental evidence for the radical mechanism. Finally, the high performance of [Zn6Cl6(SbW9O33)2]12- was traced back to its transformation into a highly active and robust disordered Zn/W-POM catalyst. The atomic short-range structure of this resting pre-catalyst was elucidated by RMC modelling of the experimental W-L3 and Zn-K edge EXAFS spectra and supported with further analytical methods. We demonstrate that computational identification of the reactive sites combined with the analytical tracking of their dynamic transformations provides essential input to expedite cluster-based molecular catalyst design.


Attitudes toward methadone maintenance: implications for HIV prevention.
Attitudes of opioid users toward methadone maintenance were studied using semi-structured field interviews. One hundred and sixty-one heroin and speedball users in San Antonio, Texas, were interviewed between 1989 and 1992. Users were classified according to whether or not they had ever been on methadone maintenance. Opioid users who had never been on methadone maintenance were more likely to express a negative attitude toward methadone maintenance than users who had been on it (50% versus 30%). Sources of negative attitudes fell into the following categories: (1) general societal disapproval of addictive drugs, including methadone; (2) prior experience with 12 Step groups or abstinence-based treatment programs; (3) previous forced rapid detoxification from methadone in jail; and (4) observation of methadone maintained peers who continued to use drugs. Very few respondents reported adverse effects from methadone itself as a source of negative attitudes. Sources of positive attitudes included: (1) prior successful treatment with methadone; and (2) observation of methadone patients who stopped using drugs.


Changes in technique within a sprint hurdle run.
The aim of this study was to understand changes in technique within an athlete's own performance during a sprint hurdles run. Four athletes performed a training session containing four trials each over 10 hurdles. Clearances at hurdles three and nine were videotaped from a side view and manually digitised. All athletes in this study yielded a lower running speed over the hurdle at the ninth hurdle in comparison to the third hurdle in each run. All athletes also showed further signs of potential tiredness in the clearances of the ninth hurdle. Interestingly, these changes in technique varied among the athletes. This poses challenges to coaches, as they need to evaluate individually what changes in training should be introduced to keep the quality of clearances as high as possible throughout training. To match more closely the velocities to those in competitions, consideration could be given in training to shorten gradually the distances between the hurdles more during the latter part of the run, although this should be individually checked, based on the athlete. This way, athletes could learn to clear the hurdle with a higher horizontal velocity, even when fatigue is potentially influencing the performance.


Sub-cellular temporal and spatial distribution of electrotransferred LNA/DNA oligomer.
Low biological activity and inefficient targeted delivery in vivo have hindered RNA interference (RNAi)-based therapy from realising its full clinical potential. To overcome these hurdles, progresses have been made to develop new technologies optimizing oligonucleotides chemistry on one hand and achieving its effective delivery on the other hand. In this report, we achieved, by using the electropulsation technique (EP), efficient cellular delivery of chemically-modified oligonucleotide: The locked nucleic acid (LNA)/DNA oligomer. We used single cell level confocal fluorescence microscopy to follow the spatial and temporal distribution of electrotransferred cyanine 5 (Cy5)-labeled LNA/DNA oligomer. We observed that EP allowed LNA/DNA oligomer cellular uptake providing the oligomer a rapid access to the cytoplasm of HeLa cells. Within a few minutes after electrotransfer, Cy5-LNA/DNA oligomers shuttle from cytoplasm to nucleus whereas in absence of pulses application, Cy5-LNA/DNA oligomers were not detected. We then observed a redistribution of the Cy5 fluorescence that accumulated over time into cytoplasmic organelles. To go further and to identify these compartments, we used the HeLa GFP-Rab7 cell line to visualise late endosomes, and lysosomal or mitochondrial specific markers. Our results showed that the EP technique allowed direct entry into the cytoplasm of the Cy5-LNA/DNA oligomer bypassing the endocytosic pathway. However, in absence of pulses application, Cy5-LNA/DNA oligomer were able to enter cells through the endocytosic pathway. We demonstrated that EP is an efficient technique for LNA-based oligonucleotides delivery offering strong advantages by avoiding the endolysosomal compartmentalization, giving a rapid and free access to the cytoplasm and the nucleus where they can find their targets.


Serum endostatin is a genetically determined predictor of survival in pulmonary arterial hypertension.
Pulmonary arterial hypertension (PAH) is a medically incurable disease resulting in death from right ventricular (RV) failure. Both pulmonary vascular and RV remodeling are linked to dynamic changes in the microvasculature. Therefore, we hypothesized that circulating angiostatic factors could be linked to outcomes and represent novel biomarkers of disease severity in PAH. We sought to determine the relationship of a potent angiostatic factor, endostatin (ES), with disease severity and mortality in PAH. Furthermore, we assessed genetic predictors of ES expression and/or function and their association with outcomes in PAH. We measured levels of serum ES in two independent cohorts of patients with PAH. Contemporaneous clinical data included New York Heart Association functional class, 6-minute-walk distance, invasive hemodynamics, and laboratory chemistries. Serum ES correlated with poor functional status, decreased exercise tolerance, and invasive hemodynamics variables. Furthermore, serum ES was a strong predictor of mortality. A loss-of-function, missense variant in the gene encoding ES, Col18a1, was linked to lower circulating protein and was independently associated with reduced mortality. Our data link increased expression of ES to disease severity in PAH and demonstrate a significant relationship with adverse outcomes. Circulating ES levels can be genetically influenced, implicating ES as a genetically determined modifier of disease severity impacting on survival. These observations support serum ES as a potential biomarker in PAH with the capacity to predict poor outcomes. More importantly, this study implicates Col18a1/ES as a potential new therapeutic target in PAH.


Calcineurin/nuclear factor of activated T cells (NFAT) signaling in cobalt-chromium-molybdenum (CoCrMo) particles-induced tumor necrosis factor-α (TNFα) secretion in MLO-Y4 osteocytes.
Aseptic loosening is the devastating long term complication of total hip arthroplasty and orthopedic implant debris has been shown to trigger an intense inflammatory reaction leading to resorption of the bone matrix. Inflammatory cytokines, such as tumor necrosis factor-α (TNFα), have been implicated in this process and osteocytes may play a role in its production. We previously demonstrated that cobalt-chromium-molybdenum (CoCrMo) particles upregulate TNFα production by MLO-Y4 osteocytes in vitro, but the underlying mechanism has not been elucidated. Based on previous studies by others, we hypothesized that the calcineurin-nuclear factor of activated T cells (NFAT) pathway mediates CoCrMo particle-induced TNFα production in MLO-Y4 osteocytes. MLO-Y4 osteocytes exposed to CoCrMo particle treatment resulted in a rapid and significant increase in calcineurin activity. We also demonstrate that CoCrMo particle-induced upregulation of TNFα is reduced to control levels with calcineurin-NFAT inhibitors and this was also confirmed at mRNA level. Moreover, we demonstrate the localization of NFATs in MLO-Y4 osteocytes and that NFAT1 and 2 translocate to the nucleus upon CoCrMo particle treatment. Our results suggest that calcineurin-NFAT signaling is involved in TNFα production by MLO-Y4 osteocytes after CoCrMo particle treatment.


Modeling food webs: exploring unexplained structure using latent traits.
Several stochastic models have tried to capture the architecture of food webs. This approach is interesting, but it is limited by the fact that different assumptions can yield similar results. To overcome this limitation, we develop a purely statistical approach. Body size in terms of an optimal ratio between prey and predator is used as explanatory variable. In 12 observed food webs, this model predicts, on average, 20% of interactions. To analyze the unexplained part, we introduce a latent term: each species is described by two latent traits, foraging and vulnerability, that represent nonmeasured characteristics of species once the optimal body size has been accounted for. The model now correctly predicts an average of 73% of links. The key features of our approach are that latent traits quantify the structure that is left unexplained by the explanatory variable and that this quantification allows a test of whether independent biological information, such as microhabitat use, camouflage, or phylogeny, explains this structure. We illustrate this method with phylogeny and find that it is linked to one or both latent traits in nine of 12 food webs. Our approach opens the door to the formulation of more complex models that can be applied to any kind of biological network.


Short-term treatment with flumazenil restores long-term object memory in a mouse model of Down syndrome.
Down syndrome (DS) is a common genetic cause of intellectual disability yet no pro-cognitive drug therapies are approved for human use. Mechanistic studies in a mouse model of DS (Ts65Dn mice) demonstrate that impaired cognitive function is due to excessive neuronal inhibitory tone. These deficits are normalized by chronic, short-term low doses of GABAA receptor (GABAAR) antagonists in adult animals, but none of the compounds investigated are approved for human use. We explored the therapeutic potential of flumazenil (FLUM), a GABAAR antagonist working at the benzodiazepine binding site that has FDA approval. Long-term memory was assessed by the Novel Object Recognition (NOR) testing in Ts65Dn mice after acute or short-term chronic treatment with FLUM. Short-term, low, chronic dose regimens of FLUM elicit long-lasting (>1week) normalization of cognitive function in both young and aged mice. FLUM at low dosages produces long lasting cognitive improvements and has the potential of fulfilling an unmet therapeutic need in DS.


Structural basis for human coronavirus attachment to sialic acid receptors.
Coronaviruses cause respiratory tract infections in humans and outbreaks of deadly pneumonia worldwide. Infections are initiated by the transmembrane spike (S) glycoprotein, which binds to host receptors and fuses the viral and cellular membranes. To understand the molecular basis of coronavirus attachment to oligosaccharide receptors, we determined cryo-EM structures of coronavirus OC43 S glycoprotein trimer in isolation and in complex with a 9-O-acetylated sialic acid. We show that the ligand binds with fast kinetics to a surface-exposed groove and that interactions at the identified site are essential for S-mediated viral entry into host cells, but free monosaccharide does not trigger fusogenic conformational changes. The receptor-interacting site is conserved in all coronavirus S glycoproteins that engage 9-O-acetyl-sialogycans, with an architecture similar to those of the ligand-binding pockets of coronavirus hemagglutinin esterases and influenza virus C/D hemagglutinin-esterase fusion glycoproteins. Our results demonstrate these viruses evolved similar strategies to engage sialoglycans at the surface of target cells.


Toward optimal early management after whiplash injury to lessen the rate of transition to chronicity: discussion paper 5.
Expert debate and synthesis of research to inform future management approaches for acute whiplash disorders. To identify a research agenda toward improving outcomes for acute whiplash-injured individuals to lessen the incidence of transition to chronicity. International figures are concordant, estimating that 50% of individuals recover from pain and disability within 3 to 6 months of a whiplash injury. The remainder report continuing symptoms up to 1 to 2 years or longer postinjury. As no management approach to date has improved recovery rates, new clinical/research directions are required for early management of whiplash-injured patients. A group of multidisciplinary researchers critically debated evidence and current research concerning whiplash from biological, psychological, and social perspectives toward informing future research directions for management of acute whiplash. It was recognized that effective treatments for acute whiplash are constrained by a limited understanding of causes of whiplash-associated disorders. Acute whiplash presentations are heterogeneous leading to the proposal that a research priority was development of a triage system based on modifiable prognostic indicators and clinical features to better inform individualized early management decisions. Other priorities identified included researching effective early pain management for individuals presenting with moderate to high levels of pain; development of best education/information for acute whiplash; testing the efficacy of stratified and individualized rehabilitation, researching modes of delivery considering psychosocial modulators of pain and disability; and the timing, nature, and mode of delivery of cognitive-behavioral therapies. Directions were highlighted for future biomechanical research into injury prevention. The burden of whiplash injuries, the high rate of transition to chronicity, and evidence of limited effects of current management on transition rates demand new directions in evaluation and management. Several directions have been proposed for future research, which reflect the potential multifaceted dimensions of an acute whiplash disorder.


Anxiety, inhibition, efficiency, and effectiveness. An investigation using antisaccade task.
Effects of anxiety on the antisaccade task were assessed. Performance effectiveness on this task (indexed by error rate) reflects a conflict between volitional and reflexive responses resolved by inhibitory processes (Hutton, S. B., & Ettinger, U. (2006). The antisaccade task as a research tool in psychopathology: A critical review. Psychophysiology, 43, 302-313). However, latency of the first correct saccade reflects processing efficiency (relationship between performance effectiveness and use of resources). In two experiments, high-anxious participants had longer correct antisaccade latencies than low-anxious participants and this effect was greater with threatening cues than positive or neutral ones. The high- and low-anxious groups did not differ in terms of error rate in the antisaccade task. No group differences were found in terms of latency or error rate in the prosaccade task. These results indicate that anxiety affects performance efficiency but not performance effectiveness. The findings are interpreted within the context of attentional control theory (Eysenck, M. W., Derakshan, N., Santos, R., & Calvo, M. G. (2007). Anxiety and cognitive performance: Attentional control theory. Emotion, 7 (2), 336-353).


Differential Links Between Expressive Suppression and Well-Being Among Chinese and Mexican American College Students.
Previous research on culture and emotion regulation has focused primarily on comparing participants from individualistic and collectivistic backgrounds (e.g., European Americans vs. Asians/Asian Americans). However, ethnic groups that are equally individualistic or collectivistic can still vary notably in cultural norms and practices regarding emotion regulation. The present study examined the association between expressive suppression and well-being in two collectivistic ethnic groups (i.e., Chinese Americans and Mexican Americans). Results indicated that suppression of positive emotions was related to lower hedonic and eudaimonic well-being among Mexican Americans but not among Chinese Americans. Moreover, post hoc analysis revealed that Mexican Americans with a stronger collective identity reported lower eudaimonic well-being when suppressing positive emotions than Mexican Americans with a weaker collective identity. Suppression of negative emotions, by contrast, was unrelated to hedonic and eudaimonic well-being for both ethnic groups. Overall, our findings underscore the importance of taking into account the role that culture and the characteristics of emotion (e.g., valence) may play in the link between emotion regulation and well-being.


Mutations in KATNB1 cause complex cerebral malformations by disrupting asymmetrically dividing neural progenitors.
Exome sequencing analysis of over 2,000 children with complex malformations of cortical development identified five independent (four homozygous and one compound heterozygous) deleterious mutations in KATNB1, encoding the regulatory subunit of the microtubule-severing enzyme Katanin. Mitotic spindle formation is defective in patient-derived fibroblasts, a consequence of disrupted interactions of mutant KATNB1 with KATNA1, the catalytic subunit of Katanin, and other microtubule-associated proteins. Loss of KATNB1 orthologs in zebrafish (katnb1) and flies (kat80) results in microcephaly, recapitulating the human phenotype. In the developing Drosophila optic lobe, kat80 loss specifically affects the asymmetrically dividing neuroblasts, which display supernumerary centrosomes and spindle abnormalities during mitosis, leading to cell cycle progression delays and reduced cell numbers. Furthermore, kat80 depletion results in dendritic arborization defects in sensory and motor neurons, affecting neural architecture. Taken together, we provide insight into the mechanisms by which KATNB1 mutations cause human cerebral cortical malformations, demonstrating its fundamental role during brain development.


OCT Angiography-Based Detection and Quantification of the Neovascular Network in Exudative AMD.
To investigate the potential of optical coherence tomography (OCT) angiography to detect and quantify the neovascular network in exudative AMD. Treatment-naïve eyes that were diagnosed with exudative AMD were prospectively examined by OCT angiography (OCT-A). The extent of the neovascular network was measured by three independent readers. Interclass-correlation coefficient and area overlap coefficients (OC) were calculated to assess locally precise agreement between measurements. As a reference for interreader agreement, the extent of the neovascular network was further measured on fluorescein angiography (FA) images. A total of 31 eyes (27 patients, mean age 82.5 years, 15 female) were included in the study. Neovascularization subtype was classified as type I in 5, type II in 11, type III in 9, and mixed in 6 eyes, respectively. Interreader agreement for measurements of the neovascular network was 0.884 for OCT-A and 0.636 for FA. Overlap coefficient was 0.705 (interquartile [IQR]: 0.450-0.76) for OCT-A and 0.704 (IQR: 0.673-0.750) for FA, respectively. Area agreement was weaker in type III and mixed lesions. Optical coherence tomography angiography-based measurements of the new vessel complex in neovascular AMD are feasible with interreader agreement comparable with the values obtained for FA. The results underscore the potential of OCT-A as a noninvasive diagnostic tool in neovascular AMD. Yet, further studies will be required to reveal the origin of poor agreement observed in single eyes and to advance OCT-A toward dependable use (e.g., in a reading center context).


Recent perspectives on the global epidemiology of childhood eczema.
The International Study of Asthma and Allergies in Childhood (ISAAC) is the largest epidemiological study ever performed and the only truly global allergy study. This review summarises the childhood eczema-related findings from ISAAC and discusses how these fit into our current understanding of eczema aetiology, with particular emphasis on worldwide time trends in eczema prevalence, climatic and dietary risk factors, breastfeeding, the role of skin barrier impairment and allergic sensitisation.


New anticoagulants.
The limitations of heparin and warfarin have prompted the development of new anticoagulant drugs for prevention and treatment of venous and arterial thromboembolism. Novel parenteral agents include synthetic analogs of the pentasaccharide sequence of heparin that mediates its interaction with antithrombin. Fondaparinux, the first synthetic pentasaccharide, is licensed for prevention of venous thromboembolism (VTE) after major orthopedic surgery and for initial treatment of patients with VTE. Idraparinux, a long-acting pentasaccharide that is administered subcutaneously once-weekly, is being compared with warfarin for treatment of VTE and for prevention of cardioembolic events in patients with atrial fibrillation. New oral anticoagulants include direct inhibitors of thrombin, factor Xa and factor IXa. Designed to provide more streamlined anticoagulation than warfarin, these agents can be given without routine coagulation monitoring. Ximelagatran, the first oral direct thrombin inhibitor, is as effective and safe as warfarin for prevention of cardioembolic events in patients with atrial fibrillation. However, ximelagatran produces a three-fold elevation in alanine transaminase levels in 7.9% of patients treated for more than a month, the long-term significance of which is uncertain. Whether other direct thrombin inhibitors or inhibitors of factors Xa or IXa also have this problem is under investigation. After a brief review of coagulation pathways, this paper focuses on new anticoagulants in advanced stages of clinical testing.


An innately dangerous balancing act: intestinal homeostasis, inflammation, and colitis-associated cancer.
Inflammatory bowel disease (IBD) is characterized by dysregulated immune responses to the intestinal microbiota, and by chronic intestinal inflammation. Several recent studies demonstrate the importance of innate microbial recognition by immune and nonimmune cells in the gut. Paradoxically, either diminished or exacerbated innate immune signaling may trigger the breakdown of intestinal homeostasis, leading to IBD and colitis-associated cancer (CAC). This dichotomy may reflect divergent functional roles for immune sensing in intestinal epithelial cells and leukocytes, which may vary with distinct disease mechanisms.


Stimulation of adenylate cyclase by Ca2+ and calmodulin in rat islets of langerhans: explanation for the glucose-induced increase in cyclic AMP levels.
The effect of Ca2+ and calmodulin has been studied on adenylate cyclase activity in homogenates of rat islets of Langerhans. EGTA had a stimulatory effect on the enzyme in accord with the known inhibitory effect of Ca2+. In contrast, the addition of Ca2+ together with calmodulin is stimulatory and demonstrates the existence of a Ca2+-dependent adenylate cyclase in islets of Langerhans. It is suggested that the glucose-induced increase in cyclic AMP concentrations in intact islets is a secondary consequence of the glucose-induced increase in cytosol free-Ca2+ concentrations which, with calmodulin, causes an increase in the activity of adenylate cyclase.


Polymorphisms in genes in the SREBP1 signalling pathway and SCD are associated with milk fatty acid composition in Holstein cattle.
Genes in the sterol regulatory element-binding protein-1 (SREBP1) pathway play a central role in regulation of milk fat synthesis, especially the de-novo synthesis of saturated fatty acids. SCD, a SREBP-responsive gene, is the key enzyme in the synthesis of monounsaturated fatty acids in the mammary gland. In the present study, we discovered SNP in candidate genes associated with this signalling pathway and SCD to identify genetic markers that can be used for genetic and metabolically directed selection in cattle. We resequenced six candidate genes in the SREBP1 pathway (SREBP1, SCAP, INSIG1, INSIG2, MBTPS1, MBTPS2) and two genes for SCD (SCD1 and SCD5) and discovered 47 Tag SNP that were used in a marker-trait association study. Milk and blood samples were collected from Holstein cows in their 1st or 2nd parity at 100-150 days of lactation. Individual fatty acids from C4 to C20, saturated fatty acid (SFA) content, monounsaturated fatty acid content, polyunsaturated fatty acid content and desaturase indexes were measured and used to perform the asociation analysis. Polymorphisms in the SCD5 and INSIG2 genes were the most representative markers associated with SFA/unsaturated fatty acid (UFA) ratio in milk. The analysis of desaturation activity determined that markers in the SCD1 and SCD5 genes showed the most significant effects. DGAT1 K232A marker was included in the study to examine the effect of this marker on the variation of milk fatty acids in our Holstein population. The percentage of variance explained by DGAT1 in the analysis was only 6% of SFA/UFA ratio. Milk fat depression was observed in one of the dairy herds and in this particular dairy one SNP in the SREBP1 gene (rs41912290) accounted for 40% of the phenotypic variance. Our results provide detailed SNP information for key genes in the SREBP1 signalling pathway and SCD that can be used to change milk fat composition by marker-assisted breeding to meet consumer demands regarding human health, as well as furthering understanding of technological aspects of cows' milk.


Accelerated cryo-EM structure determination with parallelisation using GPUs in RELION-2.
By reaching near-atomic resolution for a wide range of specimens, single-particle cryo-EM structure determination is transforming structural biology. However, the necessary calculations come at large computational costs, which has introduced a bottleneck that is currently limiting throughput and the development of new methods. Here, we present an implementation of the RELION image processing software that uses graphics processors (GPUs) to address the most computationally intensive steps of its cryo-EM structure determination workflow. Both image classification and high-resolution refinement have been accelerated more than an order-of-magnitude, and template-based particle selection has been accelerated well over two orders-of-magnitude on desktop hardware. Memory requirements on GPUs have been reduced to fit widely available hardware, and we show that the use of single precision arithmetic does not adversely affect results. This enables high-resolution cryo-EM structure determination in a matter of days on a single workstation.


Borrelia burgdorferi persists in the brain in chronic lyme neuroborreliosis and may be associated with Alzheimer disease.
The cause, or causes, of the vast majority of Alzheimer's disease cases are unknown. A number of contributing factors have been postulated, including infection. It has long been known that the spirochete Treponema pallidum, which is the infective agent for syphilis, can in its late stages cause dementia, chronic inflammation, cortical atrophy and amyloid deposition. Spirochetes of unidentified types and strains have previously been observed in the blood, CSF and brain of 14 AD patients tested and absent in 13 controls. In three of these AD cases spirochetes were grown in a medium selective for Borrelia burgdorferi. In the present study, the phylogenetic analysis of these spirochetes was made. Positive identification of the agent as Borrelia burgdorferi sensu stricto was based on genetic and molecular analyses. Borrelia antigens and genes were co-localized with beta-amyloid deposits in these AD cases. The data indicate that Borrelia burgdorferi may persist in the brain and be associated with amyloid plaques in AD. They suggest that these spirochetes, perhaps in an analogous fashion to Treponema pallidum, may contribute to dementia, cortical atrophy and amyloid deposition. Further in vitro and in vivo studies may bring more insight into the potential role of spirochetes in AD.


Crossmodal correspondences in product packaging. Assessing color-flavor correspondences for potato chips (crisps).
We report a study designed to investigate consumers' crossmodal associations between the color of packaging and flavor varieties in crisps (potato chips). This product category was chosen because of the long-established but conflicting color-flavor conventions that exist for the salt and vinegar and cheese and onion flavor varieties in the UK. The use of both implicit and explicit measures of this crossmodal association revealed that consumers responded more slowly, and made more errors, when they had to pair the color and flavor that they implicitly thought of as being "incongruent" with the same response key. Furthermore, clustering consumers by the brand that they normally purchased revealed that the main reason why this pattern of results was observed could be their differing acquaintance with one brand versus another. In addition, when participants tried the two types of crisps from "congruently" and "incongruently" colored packets, some were unable to guess the flavor correctly in the latter case. These strong crossmodal associations did not have a significant effect on participants' hedonic appraisal of the crisps, but did arouse confusion. These results are relevant in terms of R&D, since ascertaining the appropriate color of the packaging across flavor varieties ought normally to help achieve immediate product recognition and consumer satisfaction.


Cigarette smoking cessation attempts among current US smokers who also use smokeless tobacco.
Concurrent use of cigarettes and smokeless tobacco is common, but little is known regarding the association of smokeless tobacco use with cigarette smoking cessation. Dual users may have lower cigarette consumption levels, which may also play a role in smoking cessation. The 2010-2011 Tobacco Use Supplement to the Current Population Survey included 26,760 current cigarette smokers, of which 675 concurrently used smokeless tobacco. We compared characteristics of the most recent cigarette smoking quit attempt of the past year between dual users and exclusive smokers, using multivariate regression. Dual users (45%) were more likely than exclusive smokers (37%) to have made a cigarette smoking quit attempt during the previous year (p<0.01), even after adjusting for demographic differences and cigarette dependence levels (ORadj 1.33, 95% CI 1.15-1.53). Half (48%) of dual users who made a quit attempt tried to quit "by switching to smokeless tobacco". However, once in a quit attempt, dual users relapsed more quickly than exclusive smokers (Cox regression HRadj 1.13, 95% CI 1.02-1.26). There was no difference in 30-day abstinence rates on the most recent quit attempt (ORadj 1.09, 95% CI 0.88-1.37). For both groups, the best predictor of past 30-day abstinence was cigarette consumption level. Current cigarette smokers who also use smokeless tobacco are more likely to have tried to quit, but relapse more quickly than exclusive smokers, and are not more likely to have attained 30day smoking cessation. Prospective studies at the population level are needed.


Coding movement direction by burst firing in electrosensory neurons.
Directional selectivity, in which neurons respond strongly to an object moving in a given direction ("preferred") but respond weakly or not at all to an object moving in the opposite direction ("null"), is a critical computation achieved in brain circuits. Previous measures of direction selectivity have compared the numbers of action potentials elicited by each direction of movement, but most sensory neurons display patterning, such as bursting, in their spike trains. To examine the contribution of patterned responses to direction selectivity, we recorded from midbrain neurons in weakly electric fish and found that most neurons responded with a combination of both bursts and isolated spikes to moving object stimuli. In these neurons, we separated bursts and isolated spikes using an interspike interval (ISI) threshold. The directional bias of bursts was significantly higher than that of either the full spike train or the isolated spike train. To examine the encoding and decoding of bursts, we built biologically plausible models that examine 1) the upstream mechanisms that generate these spiking patterns and 2) downstream decoders of bursts. Our model of upstream mechanisms uses an interaction between afferent input and subthreshold calcium channels to give rise to burst firing that occurs preferentially for one direction of movement. We tested this model in vivo by application of calcium antagonists, which reduced burst firing and eliminated the differences in direction selectivity between bursts, isolated spikes, and the full spike train. Our model of downstream decoders used strong synaptic facilitation to achieve qualitatively similar results to those obtained using the ISI threshold criterion. This model shows that direction selective information carried by bursts can be decoded by downstream neurons using biophysically plausible mechanisms.


'Keeping It Real': women's Enactments of Lay Health Knowledges and Expertise on Facebook.
This article presents findings from a qualitative study concerning Australian women's use of Facebook for health and medical information and support and the implications for understanding modes of lay knowledge and expertise. Thinking with feminist new materialism theory, we identify the relational connections, affective forces and agential capacities described by participants as technological affordances came together with human bodily affordances. Affective forces were a dominant feature in users' accounts. Women were able to make relational connections with peers based on how valid or relevant they found other group members' expertise and experiences, how supportive other members were, how strong they wanted their personal connection to be and how much privacy they wanted to preserve. We identified three modes of engagement: 1) expertise claims based on appropriation and distribution of biomedical knowledge and experience; 2) sharing experiential knowledge without claiming expertise and 3) evaluation and use of knowledge presented by others principally through observing. We conclude that an 'expert patient' is someone who is familiar with the rules of engagement on sites such as Facebook and is able to negotiate and understand the affects and levels of disclosure and intimacy that such engagement demands.


Changes in sebum levels and the development of acneiform rash in patients with non-small cell lung cancer after treatment with EGFR inhibitors.
It has recently been shown that patients treated with epidermal growth factor receptor (EGFR) inhibitors often develop various cutaneous adverse events. While the pathogenesis underlying these events remains unclear, the relationship between skin toxicity induced by EGFR inhibitors and the sebaceous glands that express EGFR has been previously reported. The primary aim of this study was to determine the relationship between cutaneous sebum levels and acneiform rash, a typical skin toxicity of EGFR inhibitors, by measuring the sebum levels before and after EGFR inhibitor treatment. Eight patients diagnosed with non-small cell lung cancer (NSCLC) (three men and five women with an average age of 69.3 years) who were initiated on treatment with EGFR inhibitors (either gefitinib [Iressa(®)] or erlotinib [Tarceva(®)]) were enrolled. Using a Sebumeter(®), sebum levels in the face, chest, and back of each patient were measured before and after EGFR inhibitor treatment. The development of acneiform rash in each skin region was also assessed. Changes in sebum level along with the development of an acneiform rash were observed after patients were started on EGFR inhibitor treatment. Patients who developed an EGFR inhibitor-induced acneiform rash tended to have higher pretreatment sebum levels (baseline) than did patients who did not experience an acneiform rash. At each time point measurement, sebum levels were found to be significantly higher in patients who had developed an acneiform rash at that time. Patients who developed rash during treatment showed greater differences in sebum level compared with pretreatment baseline. Patients who had increased levels of sebum or whose sebum levels showed greater change from pretreatment baseline developed an acneiform rash, suggesting that sebaceous gland activity may be involved in the mechanism underlying the development of acneiform rash, in patients treated with EGFR inhibitors.


Effects of TGFbeta and bFGF on the differentiation of human bone marrow stromal fibroblasts.
Adipocytes and osteoblasts have common origins from fibroblastic stem cells. Consequently, modulation of the processes of adipogenesis and osteogenesis has implications for the possible treatment of metabolic bone diseases, such as osteoporosis, in which medullary fat accumulates and trabecular bone volume decreases. It is likely that the balance between these two systems is affected by particular endogenous growth factors which are known to affect bone metabolism. We have therefore investigated the effects of transforming growth factor beta (TGFbeta), basic fibroblast growth factor (bFGF) and dexamethasone (Dex) on cultured human bone marrow (HBM) fibroblastic cells to observe the effects on adipogenesis and osteogenesis. In the absence of fetal calf serum (FCS), TGFbeta caused a dose-dependent increase in cell growth and alkaline phosphatase activity (AP); however, in the presence of FCS growth was inhibited at high concentrations and AP unaffected. TGFbeta increased matrix proteoglycan and collagen synthesis. bFGF inhibited AP and increased colony number and size, while Dex treatment increased AP activity and colony number, and both factors in combination resulted in an additive increase in growth. Dex-induced adipocyte formation was accelerated but not increased by bFGF. A significant inhibition of adipogenesis by TGFbeta was observed within 7 days. These results demonstrate the importance of biological factors known to be involved in bone remodelling in the regulation of osteogenesis and adipogenesis.


Podoconiosis - non-filarial geochemical elephantiasis - a neglected tropical disease?
Podoconiosis or mossy foot is a form of non-filarial lymphedema. This geochemical elephantiasis is a disabling condition caused by the passage of microparticles of silica and aluminum silicates through the skin of people walking barefoot in areas with a high content of soil of volcanic origin. Podoconiosis is widespread in tropical Africa, Central America and North India, yet it remains a neglected and under-researched condition. The disabling effects of podoconiosis cause great hardship to patients. It adversely affects the economic (reduced productivity and absenteeism), social (marriage, education, etc.) and psychological (social stigma) well-being of those affected. Podoconiosis can be prevented; the main primary preventive measure is protective footwear. Secondary measures include a strict hygiene regimen and compression therapy, which can reverse initial lesions. Tertiary approaches include surgical management, such as shaving operations to reduce hyperplastic and verrucous elephantiasis.


A review of capture and post-release mortality of elasmobranchs.
There is a need to better understand the survivorship of discarded fishes, both for commercial stocks and species of conservation concern. Within European waters, the landing obligations that are currently being phased in as part of the European Union's reformed common fisheries policy means that an increasing number of fish stocks, with certain exceptions, should not be discarded unless it can be demonstrated that there is a high probability of survival. This study reviews the various approaches that have been used to examine the discard survival of elasmobranchs, both in terms of at-vessel mortality (AVM) and post-release mortality (PRM), with relevant findings summarized for both the main types of fishing gear used and by taxonomic group. Discard survival varies with a range of biological attributes (species, size, sex and mode of gill ventilation) as well as the range of factors associated with capture (e.g. gear type, soak time, catch mass and composition, handling practices and the degree of exposure to air and any associated change in ambient temperature). In general, demersal species with buccal-pump ventilation have a higher survival than obligate ram ventilators. Several studies have indicated that females may have a higher survival than males. Certain taxa (including hammerhead sharks Sphyrna spp. and thresher sharks Alopias spp.) may be particularly prone to higher rates of mortality when caught.


Maximising return on conservation investment in the conterminous USA.
Efficient conservation planning requires knowledge about conservation targets, threats to those targets, costs of conservation and the marginal return to additional conservation efforts. Systematic conservation planning typically only takes a small piece of this complex puzzle into account. Here, we use a return-on-investment (ROI) approach to prioritise lands for conservation at the county level in the conterminous USA. Our approach accounts for species richness, county area, the proportion of species' ranges already protected, the threat of land conversion and land costs. Areas selected by a complementarity-based greedy heuristic using our full ROI approach provided greater averted species losses per dollar spent compared with areas selected by heuristics accounting for richness alone or richness and cost, and avoided acquiring lands not threatened with conversion. In contrast to traditional prioritisation approaches, our results highlight conservation bargains, opportunities to avert the threat of development and places where conservation efforts are currently lacking.


Circadian rhythm in QT interval is preserved in mice deficient of potassium channel interacting protein 2.
Potassium Channel Interacting Protein 2 (KChIP2) is suggested to be responsible for the circadian rhythm in repolarization duration, ventricular arrhythmias, and sudden cardiac death. We investigated the hypothesis that there is no circadian rhythm in QT interval in the absence of KChIP2. Implanted telemetric devices recorded electrocardiogram continuously for 5 days in conscious wild-type mice (WT, n = 9) and KChIP2-/- mice (n = 9) in light:dark periods and in complete darkness. QT intervals were determined from all RR intervals and corrected for heart rate (QT100 = QT/(RR/100)1/2). Moreover, QT intervals were determined from complexes within the RR range of mean-RR ± 1% in the individual mouse (QTmean-RR). We find that RR intervals are 125 ± 5 ms in WT and 123 ± 4 ms in KChIP2-/- (p = 0.81), and QT intervals are 52 ± 1 and 52 ± 1 ms, respectively(p = 0.89). No ventricular arrhythmias or sudden cardiac deaths were observed. We find similar diurnal (light:dark) and circadian (darkness) rhythms of RR intervals in WT and KChIP2-/- mice. Circadian rhythms in QT100 intervals are present in both groups, but at physiological small amplitudes: 1.6 ± 0.2 and 1.0 ± 0.3 ms in WT and KChIP2-/-, respectively (p = 0.15). A diurnal rhythm in QT100 intervals was only found in WT mice. QTmean-RR intervals display clear diurnal and circadian rhythms in both WT and KChIP2-/-. The amplitude of the circadian rhythm in QTmean-RR is 4.0 ± 0.3 and 3.1 ± 0.5 ms in WT and KChIP2-/-, respectively (p = 0.16). In conclusion, KChIP2 expression does not appear to underlie the circadian rhythm in repolarization duration.


Influence of Japan's 2004 postgraduate training on ophthalmologist location choice, supply and distribution.
Highly-competent patient care is paramount to medicine. Quality training and patient accessibility to physicians with a wide range of specializations is essential. Yet, poor quality of life for physicians cannot be ignored, being detrimental to patient care and leading to personnel leaving the medical profession. In 2004, the Japanese government reformed postgraduate training for medical graduates, adding a 2-year, hands-on rotation through different specialties before the specialization residency was begun. Residents could now choose practice location, but it sparked concerns that physician distribution disparities had been created. Japanese media reported that residents were choosing specialties deemed to offer a higher quality of life, like Ophthalmology or Dermatology, over underserved areas like Obstetrics or Cardiology. To explore the consequences of Japan's policy efforts, through the residency reform in 2004, to improve physician training, analyzing ophthalmologist supply and distribution in the context of providing the best possible patient care and access while maintaining physician quality of life. Using secondary data, we analyzed changes in ophthalmologist supply at the secondary tier of medical care (STM). We applied ordinary least-squares regression models to ophthalmologist density to reflect community factors such as residential quality and access to further professional development, to serve as predictors of ophthalmologist supply. Coefficient equality tests examined predictor differences before and after 2004. Similar analyses were conducted for all physicians excluding ophthalmologists (other physicians). Ophthalmologist coverage in top and bottom 10% of STMs revealed supply inequalities. Change in ophthalmologist supply was inversely associated with baseline ophthalmologist density before (P < .01) and after (P = .01) 2004. Changes in other physician supply were not associated with baseline other physician density before 2004 (P = 0.5), but positively associated after 2004 (P < .01). Inequalities between top and bottom 10% of ophthalmologist supply in STMs were large, with best-served areas maintaining roughly five times greater coverage than least-served areas. However, inequalities gradually declined between 1998 and 2012. Ophthalmologist supply increased both before and after the 2004 reform, yet contrary to media reports, proceeded at a lesser rate than supply increases for other physicians. After 2004, geographical disparities decreased for ophthalmologists, while increasing for other physicians.


Human ELG1 regulates the level of ubiquitinated proliferating cell nuclear antigen (PCNA) through Its interactions with PCNA and USP1.
The level of monoubiquitinated proliferating cell nuclear antigen (PCNA) is closely linked with DNA damage bypass to protect cells from a high level of mutagenesis. However, it remains unclear how the level of monoubiquitinated PCNA is regulated. Here, we demonstrate that human ELG1 protein, which comprises an alternative replication factor C (RFC) complex and plays an important role in preserving genomic stability, as an interacting partner for the USP1 (ubiquitin-specific protease 1)-UAF1 (USP1-associated factor 1) complex, a deubiquitinating enzyme complex for PCNA and FANCD2. ELG1 protein interacts with PCNAs that are localized at stalled replication forks. ELG1 knockdown specifically resulted in an increase in the level of PCNA monoubiquitination without affecting the level of FANCD2 ubiquitination. It is a novel function of ELG1 distinct from its role as an alternative RFC complex because knockdowns of any other RFC subunits or other alternative RFCs did not affect PCNA monoubiquitination. Lastly, we identified a highly conserved N-terminal domain in ELG1 that was responsible for the USP1-UAF1 interaction as well as the activity to down-regulate PCNA monoubiquitination. Taken together, ELG1 specifically directs USP1-UAF1 complex for PCNA deubiquitination.