Non-Technical Skills for Surgeons (NOTSS): Critical appraisal of its measurement properties.
To critically appraise the development and measurement properties, including sensibility, reliability, and validity of the Non-Technical Skills of Surgeons (NOTSS) system. Articles that described development process of the NOTSS system were identified. Relevant primary studies that presented evidence of reliability and validity were identified through a comprehensive literature review. NOTSS was developed through robust item generation and reduction strategies. It was shown to have good content validity, acceptability, and feasibility. Inter-rater reliability increased with greater expertise and number of assessors. Studies demonstrated evidence of cross-sectional construct validity, in that the tool was able to differentiate known groups of varied non-technical skill levels. Evidence of longitudinal construct validity also existed to demonstrate that NOTSS detected changes in non-technical skills before and after targeted training. In populations and settings presented in our critical appraisal, NOTSS provided reliable and valid measurements of intraoperative non-technical skills of surgeons.


Protective socks for people with diabetes: a systematic review and narrative analysis.
Padded socks to protect the at-risk diabetic foot have been available for a number of years. However, the evidence base to support their use is not well known. We aimed to undertake a systematic review of padded socks for people with diabetes. Additionally, a narrative analysis of knitted stitch structures, yarn and fibres used together with the proposed benefits fibre properties may add to the sock. Assessment of the methodological quality was undertaken using a quality tool to assess non-randomised trials. From the 81 articles identified only seven met the inclusion criteria. The evidence to support to use of padded socks is limited. There is a suggestion these simple-to-use interventions could be of value, particularly in terms of plantar pressure reduction. However, the range of methods used and limited methodological quality limits direct comparison between studies. The socks were generally of a sophisticated design with complex use of knit patterns and yarn content. This systematic review provides limited support for the use of padded socks in the diabetic population to protect vulnerable feet. More high quality studies are needed; including qualitative components of sock wear and sock design, prospective randomized controlled trials and analysis of the cost-effectiveness of protective socks as a non-surgical intervention.


Search for tb resonances in proton-proton collisions at √s=7 TeV with the ATLAS detector.
This Letter presents a search for tb resonances in 1.04 fb(-1) of LHC proton-proton collision data collected by the ATLAS detector at a center-of-mass energy of 7 TeV. Events with a lepton, missing transverse momentum, and two jets are selected and the invariant mass of the corresponding final state is reconstructed. The search exploits the shape of the tb invariant mass distribution compared to the expected standard model backgrounds. The model of a right-handed W(R)' with standard model-like couplings is chosen as the benchmark model for this search. No statistically significant excess of events is observed in the data, and upper limits on the cross section times the branching ratio of W(R)' resonances at 95% C.L. lie in the range of 6.1-1.0 pb for W(R)' masses ranging from 0.5 to 2.0 TeV. These limits are translated into a lower bound on the allowed right-handed W(R)' mass, giving m(W(R)'))>1.13 TeV at 95% C.L.


Long-term exposure to ozone and sulfur dioxide increases the incidence of type 2 diabetes mellitus among aged 30 to 50 adult population.
Worldwide, the information regarding the associations between long-term exposure to ozone (O3) and sulfur dioxide (SO2) and the development of type 2 diabetes remains scarce, especially in Asia. This study aimed to investigate the long-term effects of exposure to ambient O3 and SO2 on the incidence of type 2 diabetes with consideration of other air pollutants in Taiwanese adults aged 30 to 50 years. A total of 6,426,802 non-diabetic participants aged between 30 and 50 years old were obtained from the National Health Insurance Research Database between 2005 and 2016. Incident type 2 diabetes was the main diagnosis at medical visits. Air quality data were provided by the Taiwan Environmental Protection Administration. The air pollutant concentrations for each participant were estimated using the ordinary kriging method to interpolate daily concentrations of O3, SO2, carbon monoxide (CO), nitrogen dioxide (NO2), suspended fine particles (with an aerodynamic diameter less than 2.5 μm; PM2.5), and suspended particles (with an aerodynamic diameter less than 10 μm; PM10) in residential districts across Taiwan. Six-year average concentrations of pollutants were calculated from January 1, 2005 to December 31, 2010, and data were categorized into quartiles. We performed Cox regression models to analyze the long-term effects of exposure to O3 and SO2 on the incidence of type 2 diabetes. The hazard ratio (HR) for the incidence of diabetes per each interquartile range (IQR) increase in ozone exposure (3.30 ppb) was 1.058 (95% confidence interval (CI): 1.053, 1.064) and 1.011 (95% CI: 1.007, 1.015) for SO2 exposure (1.77 ppb) after adjusting for age, sex, socioeconomic status, urbanization level, temperature, humidity, and chronic comorbidities (Model 3). Furthermore, for every 3.30 ppb increase of O3, the HR for incident type 2 diabetes was 1.093 (95% CI: 1.087, 1.100) after controlling factors shown in Model 3 plus SO2 and PM2.5. On the other hand, for every 1.77 ppb increase of SO2, the HR for incident type 2 diabetes was 1.073 (95% CI: 1.068, 1.079) after controlling factors shown in Model 3 plus NO2 and PM2.5. Long-term exposure to ambient O3 and SO2 was associated with a higher risk of developing type 2 diabetes for Taiwanese population. Exposure to O3 and SO2 may play a role in the adult early-onset type 2 diabetes.


Numerical analysis of a high-resolution fast tunable filter based on an intracavity Bragg grating.
A fast tunable filtering technique is proposed associating a diffraction grating with an intracavity Bragg grating. The bandwidth and the tuning range of this filter can be easily adapted by changing the diffraction grating's orientation, or its period, and its response is uniform over the whole tuning range. A numerical simulation of the filter response to a Gaussian beam has been developed, and it fits the experimental results allowing a calculation of the performances that could be obtained with more specific elements. For example, using a commercial acousto-optic deflector would allow a separation of 500 frequencies. It would then be possible to have a tuning range of 100 nm with a bandwidth of 0.2 nm for optical telecommunications.


Environmental determinants of polychlorinated biphenyl concentrations in residential carpet dust.
Polychlorinated biphenyls (PCBs), banned in the United Sates in the late 1970s, are still found in indoor and outdoor environments. Little is known about the determinants of PCB levels in homes. We measured concentrations of five PCB congeners (105, 138, 153, 170, and 180) in carpet dust collected between 1998 and 2000 from 1187 homes in four sites: Detroit, Iowa, Los Angeles, and Seattle. Home characteristics, occupational history, and demographic information were obtained by interview. We used a geographic information system to geocode addresses and determine distances to the nearest major road, freight route, and railroad; percentage of developed land; number of industrial facilities within 2 km of residences; and population density. Ordinal logistic regression was used to estimate the associations between the covariates of interest and the odds of PCB detection in each site separately. Total PCB levels [all congeners < maximum practical quantitation limit (MPQL) vs at least one congener ≥ MPQL to < median concentration vs at least one congener > median concentration] were positively associated with either percentage of developed land [odds ratio (OR) range 1.01-1.04 for each percentage increase] or population density (OR 1.08 for every 1000/mi(2)) in each site. The number of industrial facilities within 2 km of a home was associated with PCB concentrations; however, facility type and direction of the association varied by site. Our findings suggest that outdoor sources of PCBs may be significant determinants of indoor concentrations.


Halothiobacillus neapolitanus carboxysomes sequester heterologous and chimeric RubisCO species.
The carboxysome is a bacterial microcompartment that consists of a polyhedral protein shell filled with ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO), the enzyme that catalyzes the first step of CO2 fixation via the Calvin-Benson-Bassham cycle. To analyze the role of RubisCO in carboxysome biogenesis in vivo we have created a series of Halothiobacillus neapolitanus RubisCO mutants. We identified the large subunit of the enzyme as an important determinant for its sequestration into alpha-carboxysomes and found that the carboxysomes of H. neapolitanus readily incorporate chimeric and heterologous RubisCO species. Intriguingly, a mutant lacking carboxysomal RubisCO assembles empty carboxysome shells of apparently normal shape and composition. These results indicate that carboxysome shell architecture is not determined by the enzyme they normally sequester. Our study provides, for the first time, clear evidence that carboxysome contents can be manipulated and suggests future nanotechnological applications that are based upon engineered protein microcompartments.


First Report of Bacterial Blight of Crucifers Caused by Pseudomonas cannabina pv. alisalensis in Minnesota on Arugula (Eruca vesicaria subsp. sativa).
In 2011, bacterial blight of arugula (Eruca vesicaria subsp. sativa; cv. Roquette) was observed in organically grown plants under overhead irrigation in a field near Delano, MN. Approximately 80 to 100% of each planting was affected, with greater rates of infection occurring after periods of high humidity. Small, water-soaked, angular spots apparent on both sides of the leaves comprised the initial symptoms, which sometimes expanded and coalesced. Lesions maintained a dark water-soaked appearance or dried and turned a brown/tan color. Additionally, some lesions were outlined by a purple margin. Blue-green fluorescent pseudomonads were isolated consistently on King's Medium B agar (KMB) from symptomatic leaf tissue surface-disinfested with sodium hypochlorite (0.525%). The isolates nucleated ice and produced levan. Isolates were oxidase and arginine dihydrolase negative. They did not rot potato slices but did induce a hypersensitive reaction in tobacco (Nicotiana tabacum cv. Samsun). These data indicated that the bacteria belonged to Lelliott's LOPAT group 1 (2). DNA fragment banding patterns generated by amplifying DNA of the arugula isolates using repetitive extragenic palindromic sequence-polymerase chain reaction (rep-PCR) and the BOX A1R primer were identical and nearly identical to the banding patterns of the Pseudomonas cannabina pv. alisalensis (formerly P. syringae pv. alisalensis) (1) strain (CFBP1637) and the pathotype strain (CFBP 6866PT), respectively. Pathogenicity was confirmed on the arugula cv. My Way in two independent experiments, each with three replicate plants per treatment. Four isolates were grown on KMB for 48 h at 27°C, suspended in 0.01M potassium phosphate buffer (pH 7.0), and adjusted to 0.6 optical density at 600 nm (approximately 1 × 108 CFU/ml). Five- to six-week old plants were spray-inoculated until run-off, incubated in a humidity chamber for 48 h, and then placed in a greenhouse at 20 to 25°C for symptom development. For negative and positive control treatments, a similar number of plants each were sprayed with sterile buffer or P. cannabina pv. alisalensis strains CFBP1637 and CFBP 6866PT, respectively. Water-soaked and brown/tan lesions similar to the original symptoms appeared on plants inoculated with the arugula isolates and P. cannabina pv. alisalensis strains 7 to 14 days postinoculation. No symptoms developed on plants treated with sterile buffer. The bacterial strains re-isolated from surface-disinfested symptomatic tissue were identical by rep-PCR to the isolates used to inoculate the plants, thus, confirming Koch's postulates. Identical replicated experiments conducted on broccoli raab indicated that the arugula isolates were also pathogens of broccoli raab (Brassica rapa subsp. rapa, the original host from which P. cannabina pv. alisalensis was isolated). To our knowledge, this is the first report of bacterial blight of crucifers caused by P. cannabina pv. alisalensis in Minnesota. Arugula germplasm is being evaluated for resistance to this pathogen as an acceptable management method for organic cropping systems. References: (1) C. T. Bull et al. Syst. Appl. Microbiol. 33:105, 2010. (2) R. A. Lelliott. J. Appl. Bacteriol. 29:470, 1966.


Acute stressors stimulate ion secretion and increase epithelial permeability in rat intestine.
Wistar-Kyoto rats were subjected to 4 h restraint stress (RS) or cold restraint stress (CRS), and jejunal tissues were examined in Ussing chambers for alterations in transport functions compared with tissues from unstressed control rats. The baseline short-circuit current (Isc) was significantly elevated in tissues from RS (approximately 50%) and CRS (100%). Substitution of Cl- eliminated the abnormality, suggesting that stress stimulates Cl- secretion. Electrical transmural stimulation of enteric nerves caused a transient increase in Isc in all tissues. The magnitude of this response was significantly less in tissues from CRS than from control rats; however, the ability of the epithelium to secrete in response to exogenous stimulation with bethanechol or vasoactive intestinal polypeptide was unimpaired, implicating a neural change. Tissue conductance was higher in jejunum from RS and CRS rats than from controls. Increased intestinal epithelial permeability in stressed rats was confirmed by significantly greater fluxes of the inert radiolabeled probes, [3H]mannitol and 51Cr-labeled EDTA. No structural changes were observed. We conclude that acute stressors have profound effects on intestinal epithelial physiology, stimulating ion secretion and reducing barrier function.


Serum ferritin as risk factor for sinusoidal obstruction syndrome of the liver in patients undergoing hematopoietic stem cell transplantation.
Hepatic sinusoidal obstruction syndrome (SOS) is a serious complication in hematopoietic stem cell transplant (HSCT) recipients. To determine the impact of pretransplantation hyperferritinemia on the risk of SOS after HSC transplantation, we retrospectively studied 427 HSCT recipients (179 autologous and 248 allogeneic). Serum ferritin levels were measured before transplantation. Patients with and without a diagnosis of SOS were compared regarding demographics; underlying disease; transplant characteristics; receipt of imatinib, busulfan, total body irradiation, gemtuzumab, vancomycin, acyclovir, or methotrexate; and baseline serum ferritin. Univariate and multivariate (stepwise logistic regression) analyses were performed. SOS was diagnosed in 88 patients (21%) at a median of 10 days (range, 2-29 days) after transplantation. By multivariate analysis, allogeneic HSC transplantation (odds ratio [OR] = 8.25; 95% confidence interval [95% CI], 3.31-20.57), receipt of imatinib (OR = 2.60; 95% CI, 1.16-5.84), receipt of busulfan (OR = 2.18; 95% CI, 1.25-3.80), and ferritin serum level higher than 1000 ng/dL (OR = 1.78; 95% CI, 1.02-3.08) were risk factors for SOS. A ferritin serum level higher than 1000 ng/dL in the pretransplantation period is an independent risk factor for SOS. The results suggest the need for prospective studies addressing the use of iron chelation in the pretransplantation period.


Dose effects in behavioural treatment of post-stroke aphasia: a systematic review and meta-analysis.
Aphasia is a debilitating chronic acquired language disorder that impacts heavily on a person's life. Behavioural treatments aim to remediate language processing skills or to enhance communication between the person with aphasia and others, and a number of different treatments are efficacious. However, it is unclear how much of a particular treatment a person needs in order to optimise recovery of language and communication skills following stroke. Systematic search for and meta-analysis of experimental studies that directly compared different amounts of the same behavioural aphasia treatment, following PRISMA guidelines. Treatment dose research in aphasia is an emerging area. Just six studies comparing different doses of the same intervention met all criteria for inclusion. Evidence from these studies was synthesised and meta-analysed, where possible. Meta-analyses were inconclusive due to limited data; however, there are indications that suggest increased dose may confer greater improvement on language and communication measures, but with diminishing returns over time. Aphasia severity and chronicity may affect dose-response relationships. There is currently insufficient evidence to determine the effect of dose on treatment response. A dedicated and coordinated research agenda is required to systematically explore dose-response relationships in post-stroke aphasia interventions.A video abstract is available in the Supplementary Material.Implications for rehabilitationThe investigation of the effect of dose on treatment outcomes in post-stroke aphasia is an emerging research area with few studies reporting comparison of different amounts of the same intervention.In the acute phase of recovery following stroke, higher doses of treatment provided over short periods may not be preferable, tolerable, or superior to lower doses of the same treatment.In the chronic phase, providing additional blocks of treatment may confer additional benefit for some people with aphasia but with diminishing returns.People with chronic aphasia can achieve and maintain significant gains in picture naming after a relatively brief period of high-dose treatment.


Leveraging the Domain of Work to Improve Migrant Health.
Work is a principal driver of current international migration, a primary social determinant of health, and a fundamental point of articulation between migrants and their host society. Efforts by international organizations to promote migrant health have traditionally focused on infectious diseases and access to healthcare, while international labor organizations have largely focused on issues of occupational health. The underutilization of the domain of work in addressing the health of migrants is truly a missed opportunity for influencing worker well-being and reducing societal economic burden. Understanding of the relationships among migration, work, and health would facilitate further integration of migrant health concerns into the policy agenda of governments and international agencies that work at the nexus of labor, health and development. The domain of work offers an opportunity to capitalize on the existing health and development infrastructure and leverage technical resources, programs and research to promote migrant health. It also provides the opportunity to advance migrant health through new and innovative approaches and partnerships.


A unique pattern of expression of the four muscle regulatory factor proteins distinguishes somitic from embryonic, fetal and newborn mouse myogenic cells.
A unique pattern of expression of the four muscle regulatory factor (MRF) proteins was found to distinguish early somitic from embryonic, fetal and newborn limb myogenic cells in vitro. Expression of the myosin heavy chain (MHC), MyoD, myogenin, Myf-5, and MRF4 proteins was examined by immunocytochemistry in cultures of four distinct types of mouse myogenic cells: somitic (E8.5), embryonic (E11.5), fetal (E16.5) and newborn limb. In embryonic, fetal and newborn cultures, the MRF proteins were expressed in generally similar patterns: MyoD was the first MRF expressed; MyoD and myogenin were expressed by more cells than Myf-5 or MRF4; and each of the four MRFs was found both in cells that expressed MHC and in cells that did not express MHC. In cultures of somitic cells, in contrast, Myf-5 was expressed first and by more cells than MyoD or myogenin; MRF4 was not detected; and the MRFs were never found to be coexpressed with MHC in the same cell. Thus, some somitic cells had the unexpected ability to maintain MHC expression in the absence of detectable MRF protein expression. The different myogenic programs of embryonic, fetal and newborn myogenic cells are not, therefore, a simple result of qualitatively different MRF expression patterns, whereas myogenesis by somitic cells does include a unique pattern of MRF expression.


Physical properties of phase-change emulsions.
Phase-change emulsions (PCE) are important in a variety of applications, from ultrasound imaging to the explosive material used in the mining industry, but until now there has been no adequate theory to describe their activation properties. The PCE consists of a low-boiling-point liquid, known as the volatile phase, dispersed in an aqueous phase. The volatile phase boils as a result of an increase in the temperature of the emulsion. The volume of the emulsion will increase during this phase transition, with the transition temperature and final volume of the emulsion highly dependent on the initial radius of the liquid droplets. Here a description of the change in boiling point and freezing point of the volatile phase, as well as the volume change of a droplet in the emulsion as a function of the initial droplet radius, is presented. The influence of volatile phase solubility, liquid-liquid interfacial tension, and final temperature are explored, accounting for the influence of confinement on the properties of the volatile phase. Beyond this, a means by which the diffusivity of the gas in the continuous liquid phase can be measured is derived.


Factors associated with control of type I diabetes in Malaysian adolescents and young adults.
To examine the association between ethnicity, depression, quality of life, and diabetic control in Malaysian adolescents and young adults with type I diabetes mellitus. Fifty-two outpatients with type I diabetes (mean age 15.5 years) who attended a Diabetes Clinic were included. The level of HbA1c was the measure of diabetes control used (better control defined as HbA1c < 10%). Other variables were measured through questionnaires (e.g., depressive symptoms, quality of life), computerized diagnostic interviews (major depression), and medical records (e.g., demographic, family circumstances, compliance with treatment). Ethnic Chinese youth showed better diabetic control than Malays and Indians (mean HbA1c 9.1%, 10.3%, and 11.0% respectively). Young people with better diabetic control (HbA1c < 10%) were more likely to have better quality of life and less likely to live in problematic families. When the cut-off for diabetic control was stricter (HbA1c < or = 8%), the young person's compliance was the main predictor of poor control. Family problems were also associated with poor control but to a lesser extent. The initial association between poorer diabetes control and depression became non-significant when quality of life was taken into account. There are ethnic differences in juvenile diabetic control in this Malaysian sample which need to be understood further. Previous findings of an association between quality of life and glycemic control were verified but different definitions of good control showed different associations with individual and environmental variables. Clinicians' awareness and early intervention for psychosocial problems (for example, inadequate family support) could improve diabetes control.


Neoadjuvant treatment: a novel standard?
The aim of this study was to summarize developments in the adjuvant/neoadjuvant chemotherapy of high-risk adult-type soft tissue sarcomas (STS). The role of adjuvant/neaodjuvant chemotherapy in these patients is controversial, with a meta-analysis suggesting a 10% survival benefit. Recently, a randomized controlled trial in high-risk STS of extremities and trunk wall showed a 20% improvement in progression-free and overall survival after three preoperative cycles of epirubicin along with ifosfamide compared with a histology-tailored chemotherapy. This study has major strengths, including the selected high-risk population and the full-dose chemotherapy regimen. However, this was an interim analysis with a short follow-up in a trial originally planned to test the superiority of a histology-driven chemotherapy. As to high-risk patient selection, the new AJCC TNM staging system adds primary tumour site as a stratifying factor, while available prognostic nomograms account for additional criteria. A recent trial strengthens perioperative chemotherapy as an option for high-risk STS patients within a shared decision-making process. If the final analysis of this trial confirms the currently observed progression-free and overall survival benefits, perioperative chemotherapy may become a standard. Also, new staging tools may refine our ability to select patients with a risk high enough as to deserve chemotherapy.


Docking Screens for Dual Inhibitors of Disparate Drug Targets for Parkinson's Disease.
Modulation of multiple biological targets with a single drug can lead to synergistic therapeutic effects and has been demonstrated to be essential for efficient treatment of CNS disorders. However, rational design of compounds that interact with several targets is very challenging. Here, we demonstrate that structure-based virtual screening can guide the discovery of multi-target ligands of unrelated proteins relevant for Parkinson's disease. A library with 5.4 million molecules was docked to crystal structures of the A2A adenosine receptor (A2AAR) and monoamine oxidase B (MAO-B). Twenty-four compounds that were among the highest ranked for both binding sites were evaluated experimentally, resulting in the discovery of four dual-target ligands. The most potent compound was an A2AAR antagonist with nanomolar affinity ( Ki = 19 nM) and inhibited MAO-B with an IC50 of 100 nM. Optimization guided by the predicted binding modes led to the identification of a second potent dual-target scaffold. The two discovered scaffolds were shown to counteract 6-hydroxydopamine-induced neurotoxicity in dopaminergic neuronal-like SH-SY5Y cells. Structure-based screening can hence be used to identify ligands with specific polypharmacological profiles, providing new avenues for drug development against complex diseases.


Atypical Gait Cycles in Parkinson's Disease.
It is important to find objective biomarkers for evaluating gait in Parkinson's Disease (PD), especially related to the foot and lower leg segments. Foot-switch signals, analyzed through Statistical Gait Analysis (SGA), allow the foot-floor contact sequence to be characterized during a walking session lasting five-minutes, which includes turnings. Gait parameters were compared between 20 PD patients and 20 age-matched controls. PDs showed similar straight-line speed, cadence, and double-support compared to controls, as well as typical gait-phase durations, except for a small decrease in the flat-foot contact duration (-4% of the gait cycle, p = 0.04). However, they showed a significant increase in atypical gait cycles (+42%, p = 0.006), during both walking straight and turning. A forefoot strike, instead of a "normal" heel strike, characterized the large majority of PD's atypical cycles, whose total percentage was 25.4% on the most-affected and 15.5% on the least-affected side. Moreover, we found a strong correlation between the atypical cycles and the motor clinical score UPDRS-III (r = 0.91, p = 0.002), in the subset of PD patients showing an abnormal number of atypical cycles, while we found a moderate correlation (r = 0.60, p = 0.005), considering the whole PD population. Atypical cycles have proved to be a valid biomarker to quantify subtle gait dysfunctions in PD patients.


Interrelations between cognitive dysfunction and motor symptoms of Parkinson's disease: behavioral and neural studies.
Parkinson's disease (PD) is characterized by a range of motor symptoms. Besides the cardinal symptoms (tremor, bradykinesia/akinesia, and rigidity), PD patients also show other motor deficits, including gait disturbance, speech deficits, and impaired handwriting. However, along with these key motor symptoms, PD patients also experience cognitive deficits in attention, executive function, working memory, and learning. Recent evidence suggests that these motor and cognitive deficits of PD are not completely dissociable, as aspects of cognitive dysfunction can impact motor performance in PD. In this article, we provide a review of behavioral and neural studies on the associations between motor symptoms and cognitive deficits in PD, specifically akinesia/bradykinesia, tremor, gait, handwriting, precision grip, and speech production. This review paves the way for providing a framework for understanding how treatment of cognitive dysfunction, for example cognitive rehabilitation programs, may in turn influence the motor symptoms of PD.


The Fugu tyrp1 promoter directs specific GFP expression in zebrafish: tools to study the RPE and the neural crest-derived melanophores.
In vertebrates, pigment cells account for a small percentage of the total cell population and they intermingle with other cell types. This makes it difficult to isolate them for analyzes of their functions in the context of development. To alleviate such difficulty, we generated two stable transgenic zebrafish lines (pt101 and pt102) that express green fluorescent protein (GFP) in melanophores under the control of the 1 kb Fugu tyrp1 promoter. In pt101, GFP is expressed in both retinal pigment epithelium (RPE) cells and the neural crest-derived melanophores (NCDM), whereas in pt102, GFP is predominately expressed in the NCDM. Our results indicate that the Fugu tyrp1 promoter can direct transgene expression in a cell-type-specific manner in zebrafish. In addition, our findings provide evidence supporting differential regulations of melanin-synthesizing genes in RPE cells and the NCDM in zebrafish. Utilizing the varying GFP expression levels in these fish, we have isolated melanophores via flow cytometry and revealed the capability of sorting the NCDM from RPE cells as well. Thus, these transgenic lines are useful tools to study melanophores in zebrafish.


Novel Anti-CRR9/CLPTM1L Antibodies with Antitumorigenic Activity Inhibit Cell Surface Accumulation, PI3K Interaction, and Survival Signaling.
We and others have recently shown cisplatin resistance-related protein 9 (CRR9)/Cleft Lip and Palate Transmembrane 1-Like (CLPTM1L) to affect survival and proliferation in lung and pancreatic tumor cells. Our research has indicated that CLPTM1L affects multiple survival signaling pathways in tumor cells under oncogenic, genotoxic, and microenvironmental stress. We have confirmed the association of CLPTM1L with pancreatic cancer by demonstrating overexpression of CLPTM1L in pancreatic tumors and poor survival in patients with high tumor expression of CLPTM1L. Predicting a transmembrane structure, we determined that CLPTM1L could be targeted at the plasma membrane. Herein, we describe the development of mAbs targeting CLPTM1L. Lead antibodies inhibited surface accumulation of CLPTM1L, Akt phosphorylation, anchorage-independent growth, and chemotherapeutic resistance in lung and pancreatic tumor cells. Gemcitabine promoted a physical interaction between CLPTM1L and p110α in pancreatic tumor cells, which was inhibited by anti-CLPTM1L. In vivo treatment with anti-CLPTM1L robustly inhibited the growth of both lung and pancreatic adenocarcinoma xenografts. The efficacy of anti-CLPTM1L correlated with specific epitopes representing important targets in human cancers, particularly those driven by KRas, for which effective targeted therapies have been elusive. This study is the first to report cell-surface exposure of the tumor survival protein CLPTM1L and inhibition of the function of surface CLPTM1L with novel, systematically developed inhibitory mAbs establishing proof of concept of clinically practical agents inhibiting this compelling new tumor survival target in cancer. Mol Cancer Ther; 15(5); 985-97. ©2016 AACR.


Abnormal keratinocyte differentiation in the nasal planum of Labrador Retrievers with hereditary nasal parakeratosis (HNPK).
Hereditary nasal parakeratosis (HNPK) is an inherited disorder described in Labrador Retrievers and Greyhounds. It has been associated with breed-specific variants in the SUV39H2 gene encoding a histone 3 methyltransferase involved in epigenetic silencing. Formalin-fixed biopsies of the nasal planum of Labrador Retrievers were screened by immunofluorescence microscopy for the presence and distribution of epidermal proliferation and differentiation markers. Gene expression of these markers was further analysed using RNA sequencing (RNA-seq) and ultrastructural epidermal differences were investigated by electron microscopy. Differentiation of the nasal planum in the basal and suprabasal epidermal layers of HNPK-affected dogs (n = 6) was similar compared to control dogs (n = 6). In the upper epidermal layers, clear modifications were noticed. Loricrin protein was absent in HNPK-affected nasal planum sections in contrast to sections of the same location of control dogs. However, loricrin was present in the epidermis of paw pads and abdominal skin from HNPK dogs and healthy control dogs. The patterns of keratins K1, K10 and K14, were not markedly altered in the nasal planum of HNPK-affected dogs while the expression of the terminal differentiation marker involucrin appeared less regular. Based on RNA-seq, LOR and IVL expression levels were significantly decreased, while KRT1, KRT10 and KRT14 levels were up-regulated (log2fold-changes of 2.67, 3.19 and 1.71, respectively) in HNPK-affected nasal planum (n = 3) compared to control dogs (n = 3). Electron microscopical analysis revealed structural alterations in keratinocytes and stratum corneum, and disrupted keratinocyte adhesions and distended intercellular spaces in lesional samples (n = 3) compared to a sample of a healthy control dog (n = 1). Our findings demonstrate aberrant keratinocyte terminal differentiation of the nasal planum of HNPK-affected Labrador Retrievers and provide insights into biological consequences of this inactive SUV39H2 gene variant.


Identifying Smoking Environments From Images of Daily Life With Deep Learning.
Environments associated with smoking increase a smoker's craving to smoke and may provoke lapses during a quit attempt. Identifying smoking risk environments from images of a smoker's daily life provides a basis for environment-based interventions. To apply a deep learning approach to the clinically relevant identification of smoking environments among settings that smokers encounter in daily life. In this cross-sectional study, 4902 images of smoking (n = 2457) and nonsmoking (n = 2445) locations were photographed by 169 smokers from Durham, North Carolina, and Pittsburgh, Pennsylvania, areas from 2010 to 2016. These images were used to develop a probabilistic classifier to predict the location type (smoking or nonsmoking location), thus relating objects and settings in daily environments to established smoking patterns. The classifier combines a deep convolutional neural network with an interpretable logistic regression model and was trained and evaluated via nested cross-validation with participant-wise partitions (ie, out-of-sample prediction). To contextualize model performance, images taken by 25 randomly selected participants were also classified by smoking cessation experts. As secondary validation, craving levels reported by participants when viewing unfamiliar environments were compared with the model's predictions. Data analysis was performed from September 2017 to May 2018. Classifier performance (accuracy and area under the receiver operating characteristic curve [AUC]), comparison with 4 smoking cessation experts, contribution of objects and settings to smoking environment status (standardized model coefficients), and correlation with participant-reported craving. Of 169 participants, 106 (62.7%) were from Durham (53 [50.0%] female; mean [SD] age, 41.4 [12.0] years) and 63 (37.3%) were from Pittsburgh (31 [51.7%] female; mean [SD] age, 35.2 [13.8] years). A total of 4902 images were available for analysis, including 3386 from Durham (mean [SD], 31.9 [1.3] images per participant) and 1516 from Pittsburgh (mean [SD], 24.1 [0.5] images per participant). Images were evenly split between the 2 classes, with 2457 smoking images (50.1%) and 2445 nonsmoking images (49.9%). The final model discriminated smoking vs nonsmoking environments with a mean (SD) AUC of 0.840 (0.024) (accuracy [SD], 76.5% [1.6%]). A model trained only with images from Durham participants effectively classified images from Pittsburgh participants (AUC, 0.757; accuracy, 69.2%), and a model trained only with images from Pittsburgh participants effectively classified images from Durham participants (AUC, 0.821; accuracy, 75.0%), suggesting good generalizability between geographic areas. Only 1 expert's performance was a statistically significant improvement compared with the classifier (α = .05). Median self-reported craving was significantly correlated with model-predicted smoking environment status (ρ = 0.894; P = .003). In this study, features of daily environments predicted smoking vs nonsmoking status consistently across participants. The findings suggest that a deep learning approach can identify environments associated with smoking, can predict the probability that any image of daily life represents a smoking environment, and can potentially trigger environment-based interventions. This work demonstrates a framework for predicting how daily environments may influence target behaviors or symptoms that may have broad applications in mental and physical health.


Structural constraints identified with covariation analysis in ribosomal RNA.
Covariation analysis is used to identify those positions with similar patterns of sequence variation in an alignment of RNA sequences. These constraints on the evolution of two positions are usually associated with a base pair in a helix. While mutual information (MI) has been used to accurately predict an RNA secondary structure and a few of its tertiary interactions, early studies revealed that phylogenetic event counting methods are more sensitive and provide extra confidence in the prediction of base pairs. We developed a novel and powerful phylogenetic events counting method (PEC) for quantifying positional covariation with the Gutell lab's new RNA Comparative Analysis Database (rCAD). The PEC and MI-based methods each identify unique base pairs, and jointly identify many other base pairs. In total, both methods in combination with an N-best and helix-extension strategy identify the maximal number of base pairs. While covariation methods have effectively and accurately predicted RNAs secondary structure, only a few tertiary structure base pairs have been identified. Analysis presented herein and at the Gutell lab's Comparative RNA Web (CRW) Site reveal that the majority of these latter base pairs do not covary with one another. However, covariation analysis does reveal a weaker although significant covariation between sets of nucleotides that are in proximity in the three-dimensional RNA structure. This reveals that covariation analysis identifies other types of structural constraints beyond the two nucleotides that form a base pair.


The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3.
We investigated whether the affinity of tissue inhibitor of metalloproteinases (TIMP)-3 for adamalysins with thrombospondin motifs (ADAMTS)-4 and ADAMTS-5 is affected by the non-catalytic ancillary domains of the enzymes. For this purpose, we first established a novel method of purifying recombinant FLAG-tagged TIMP-3 and its inhibitory N-terminal domain (N-TIMP-3) by treating transfected HEK293 cells with sodium chlorate to prevent heparan sulfate proteoglycan-mediated TIMP-3 internalization. TIMP-3 and N-TIMP-3 affinity for selected matrix metalloproteinases and forms of ADAMTS-4 and -5 lacking sequential C-terminal domains was determined. TIMP-3 and N-TIMP-3 displayed similar affinity for various matrix metalloproteinases as has been previously reported for E. coli-expressed N-TIMP-3. ADAMTS-4 and -5 were inhibited more strongly by N-TIMP-3 than by full-length TIMP-3. The C-terminal domains of the enzymes enhanced interaction with N-TIMP-3 and to a lesser extent with the full-length inhibitor. For example, N-TIMP-3 had 7.5-fold better K(i) value for full-length ADAMTS-5 than for the catalytic and disintegrin domain alone. We propose that the C-terminal domains of the enzymes affect the structure around the active site, favouring interaction with TIMP-3.


Occupational airborne exposure of the general population of a Norwegian county.
Occupational airborne exposure was examined for a stratified sample (N = 1275) of the general population aged 18-73 years in Hordaland County, Norway. The subjects identified all jobs of more than six months since leaving school and stated whether they had been occupationally exposed to specific agents and work processes potentially harmful to the lungs. The prevalence in the population ever having been exposed was 18% for asbestos, 9% for quartz, 5% for aluminum dust, 6% for wood dust, 12% for metal gases, 12% for welding, 9% for soldering, and 1% for hairdressing. According to occupational title (last job), 3% of the population had held a job with a high degree of airborne exposure, 26% a job with moderate exposure, and 70% a job with no airborne exposure. During their worklife both the men and the women tended to leave polluted jobs more often than unpolluted jobs. Occupational exposure to airborne pollutants potentially harmful to the lungs is widespread in this Norwegian general population.


Mycobacterium tuberculosis mutation rate estimates from different lineages predict substantial differences in the emergence of drug-resistant tuberculosis.
A key question in tuberculosis control is why some strains of M. tuberculosis are preferentially associated with resistance to multiple drugs. We demonstrate that M. tuberculosis strains from lineage 2 (East Asian lineage and Beijing sublineage) acquire drug resistances in vitro more rapidly than M. tuberculosis strains from lineage 4 (Euro-American lineage) and that this higher rate can be attributed to a higher mutation rate. Moreover, the in vitro mutation rate correlates well with the bacterial mutation rate in humans as determined by whole-genome sequencing of clinical isolates. Finally, using a stochastic mathematical model, we demonstrate that the observed differences in mutation rate predict a substantially higher probability that patients infected with a drug-susceptible lineage 2 strain will harbor multidrug-resistant bacteria at the time of diagnosis. These data suggest that interventions to prevent the emergence of drug-resistant tuberculosis should target bacterial as well as treatment-related risk factors.


Stepwise optimization of the procedure for assessment of circulating progenitor cells in patients with myocardial infarction.
The number and functional activity of circulating progenitor cells (CPCs) is altered in diabetic patients. Furthermore, reduced CPC count has been shown to independently predict cardiovascular events. Validation of CPCs as a biomarker for cardiovascular risk stratification requires rigorous methodology. Before a standard operation protocol (SOP) can be designed for such a trial, a variety of technical issues have to be addressed fundamentally, which include the appropriate type of red blood cell lysis buffer, FMO or isotype controls to identify rare cell populations from background noise, optimal antibody dilutions and conditions of sample storage. We herein propose improvements in critical steps of CPC isolation, antigenic characterization and determination of functional competence for final application in a prospective investigation of CPCs as a biomarker of outcome following acute myocardial infarction. In this validation study, we refined the standard operating procedure (SOP) for flow cytometry characterisation and functional analysis of CPCs from the first 18 patients of the Progenitor Cell Response after Myocardial Infarction Study (ProMIS). ProMIS aims to verify the prognostic value of CPCs in patients with either ST elevation or non-ST elevation myocardial infarction with or without diabetes mellitus, using cardiac magnetic resonance imaging (MRI) for assessment of ventricular function as a primary endpoint. Results indicate crucial steps for SOP implementation, namely timely cell isolation after sampling, use of appropriate lysis buffer to separate blood cell types and minimize the acquisition events during flow cytometry, adoption of proper fluorophore combination and antibody titration for multiple antigenic detection and introduction of counting beads for precise quantification of functional CPC activity in migration assay. With systematic specification of factors influencing the enumeration of CPC by flow cytometry, the abundance and migration capacity of CPCs can be correctly assessed. Adoption of validated SOP is essential for refined comparison of patients with different comorbidities in the analysis of risk stratification.


Does cold loop hysteroscopic myomectomy reduce intrauterine adhesions? A retrospective study.
To estimate the prevalence and the characteristics of intrauterine adhesions after cold loop resectoscopic myomectomy. Retrospective study. Endoscopic gynecologic center. 688 women with one or more G1-G2 myomas. Cold loop resectoscopic myomectomy and diagnostic hysteroscopy in all patients 2 months after surgery. Integrity of the uterine cavity and prevalence of intrauterine synechiae. A total of 806 myomas were removed, ranging from a minimum of one to a maximum of five fibroids removed for each surgical procedure. Complications were reported in eight cases (1.16%). No hemorrhage, intravasation clinical syndrome, or perforation with the thermal loop were registered. Synechiae were found in 29 patients (4.23%): in 2 patients a new surgical hysteroscopic treatment was required to remove fibrous synechiae, and in 27 patients light adhesions were removed with the tip of the instrument in outpatient hysteroscopy. Neither intrauterine device nor anti-adherence mixtures were used at the end of surgery. The cold loop hysteroscopic myomectomy is a safe and effective procedure that seems to be associated with a lower rate of intrauterine adhesions in comparison with the reported literature. The issue appears to be of notable importance for fertility patients.


Genetic characterization and sequence heterogeneity of a canadian isolate of Swine hepatitis E virus.
Swine hepatitis E virus (HEV) is a newly identified potentially zoonotic agent that is possibly transmitted to humans from pigs. Swine HEV is prevalent in pig populations and does not cause abnormal clinical symptoms in infected pigs, further implicating a likelihood of a risk of transmission to humans by normal contact. To date in North America, only one strain of swine HEV (strain US swine) has been fully sequenced. In the present study, we identified a swine HEV isolate from pigs in Canada, designated the Arkell strain, and determined the full length of the genomic sequence. The genome of Canadian strain Arkell consisted of 7,242 nucleotides, excluding the poly(A) tail of at least 15 A residues. The genome contained three open reading frames (ORFs), ORF1, ORF2, and ORF3, which had coding capacities for proteins of 1,708, 660, and 122 amino acids, respectively. Comparative analysis of the full-length genomic sequence indicated that the sequence of strain Arkell was distinct from those of all other known HEV isolates by 13 to 27% and shared the highest degrees of identity with human HEV isolates US-1 and US-2, HEV isolate US swine, and the human and swine HEV isolates recently isolated in Japan. On the basis of sequence similarities and phylogenetic analyses, HEV strain Arkell was grouped into genotype 3. The sequence of the Arkell swine HEV isolate differed from those of HEV isolate US swine and HEV isolate Japan swine by 13 and 14%, respectively. To date, two isolates of swine HEV (isolates Arkell and SK3 [D. Yoo et al., Clin. Diagn. Lab. Immunol. 8:1213-1219, 2001]) have been identified in Canadian pigs, and their sequences also differ from each other by 11.8%. Our studies indicate that, as with human HEV strains, swine HEV isolates exhibit extensive genetic heterogeneity.


Clinical and Laboratory Profiles of Idiopathic Small Fiber Neuropathy in Children: Case Series.
The role of autoimmune mechanisms in idiopathic small fiber neuropathy (SFN) is not completely understood. Serum IgM binding to trisulfated disaccharide IdoA2S-GlcNS-6S (TS-HDS) and IgG to fibroblast growth factor receptor 3 were associated with sensory motor polyneuropathies and sensory neuronopathy among others. In this retrospective case review, we describe the clinical and laboratory findings of idiopathic SFN in a small cohort of pediatric patients. Eight children were diagnosed with SFN clinically and confirmed by reduced epidermal nerve fiber density. No involvement of large fibers was confirmed by clinical examination and electrophysiological tests. Possible triggering factors were infectious mononucleosis in 4 patients and human papilloma virus vaccination in 1 patient. Tilt table test was positive in 1 patient, and clinical autonomic dysfunctions were noted in 6 patients. Five patients had positive IgM against TS-HDS, 3 of whom had lower extremity predominant paresthesia. In conclusion, a high proportion of patients with idiopathic SFN in our cohort had a positive IgM TS-HDS antibody.


The chemoattractant chemerin suppresses melanoma by recruiting natural killer cell antitumor defenses.
Infiltration of specialized immune cells regulates the growth and survival of neoplasia. Here, in a survey of public whole genome expression datasets we found that the gene for chemerin, a widely expressed endogenous chemoattractant protein, is down-regulated in melanoma as well as other human tumors. Moreover, high chemerin messenger RNA expression in tumors correlated with improved outcome in human melanoma. In experiments using the B16 transplantable mouse melanoma, tumor-expressed chemerin inhibited in vivo tumor growth without altering in vitro proliferation. Growth inhibition was associated with an altered profile of tumor-infiltrating cells with an increase in natural killer (NK) cells and a relative reduction in myeloid-derived suppressor cells and putative immune inhibitory plasmacytoid dendritic cells. Tumor inhibition required host expression of CMKLR1 (chemokine-like receptor 1), the chemoattractant receptor for chemerin, and was abrogated by NK cell depletion. Intratumoral injection of chemerin also inhibited tumor growth, suggesting the potential for therapeutic application. These results show that chemerin, whether expressed by tumor cells or within the tumor environment, can recruit host immune defenses that inhibit tumorigenesis and suggest that down-regulation of chemerin may be an important mechanism of tumor immune evasion.


Induction of CD4(+) and CD8(+) anti-tumor effector T cell responses by bacteria mediated tumor therapy.
Facultative anaerobic bacteria like E. coli can colonize solid tumors often resulting in tumor growth retardation or even clearance. Little mechanistic knowledge is available for this phenomenon which is however crucial for optimization and further implementation in the clinic. Here, we show that intravenous injections with E. coli TOP10 can induce clearance of CT26 tumors in BALB/c mice. Importantly, re-challenging mice which had cleared tumors showed that clearance was due to a specific immune reaction. Accordingly, lymphopenic mice never showed tumor clearance after infection. Depletion experiments revealed that during induction phase, CD8(+) T cells are the sole effectors responsible for tumor clearance while in the memory phase CD8(+) and CD4(+) T cells were involved. This was confirmed by adoptive transfer. CD4(+) and CD8(+) T cells could reject newly set tumors while CD8(+) T cells could even reject established tumors. Detailed analysis of adoptively transferred CD4(+) T cells during tumor challenge revealed expression of granzyme B, FasL, TNF-α and IFN-γ in such T cells that might be involved in the anti-tumor activity. Our findings should pave the way for further optimization steps of this promising therapy.


Genetic variation in response to 6-mercaptopurine for childhood acute lymphoblastic leukaemia.
6-mercaptopurine (6-MP) can be inactivated by S-methylation, which is catalysed by thiopurine methyltransferase (TPMT). An alternative metabolic route leads to the formation of cytotoxic 6-thioguanine nucleotides (6-TGN). To investigate whether these two pathways compete with each other to affect the therapeutic response to 6-MP, 6-TGN concentrations and TPMT enzymatic activity were measured in erythrocytes (RBC) from 95 children on long-term 6-MP therapy for lymphoblastic leukaemia (ALL). RBC TPMT activities were also measured in 130 control children and 104 long-term survivors of ALL no longer on treatment. The 95 children on 6-MP showed wide interindividual differences in RBC 6-TGN concentrations at the full protocol dose of 75 mg/m2, and RBC 6-TGN concentrations correlated negatively with RBC TPMT activity. Children with 6-TGN concentrations below the group median had higher TPMT activities and a higher subsequent relapse rate. 50 of the 104 long-term survivors had been treated with "gentle" low-dose protocols, and this subgroup contained an excess of children with lower TPMT activities compared with normal controls. These results indicate that genetically determined TPMT activity may be a substantial regulator of the cytotoxic effect of 6-MP, an effect which in turn could be important in influencing the outcome of therapy for childhood ALL.


Differential expression of cell cycle regulators in CDK5-dependent medullary thyroid carcinoma tumorigenesis.
Medullary thyroid carcinoma (MTC) is a neuroendocrine cancer of thyroid C-cells, for which few treatment options are available. We have recently reported a role for cyclin-dependent kinase 5 (CDK5) in MTC pathogenesis. We have generated a mouse model, in which MTC proliferation is induced upon conditional overexpression of the CDK5 activator, p25, in C-cells, and arrested by interrupting p25 overexpression. Here, we identify genes and proteins that are differentially expressed in proliferating versus arrested benign mouse MTC. We find that downstream target genes of the tumor suppressor, retinoblastoma protein, including genes encoding cell cycle regulators such as CDKs, cyclins and CDK inhibitors, are significantly upregulated in malignant mouse tumors in a CDK5-dependent manner. Reducing CDK5 activity in human MTC cells down-regulated these cell cycle regulators suggesting that CDK5 activity is critical for cell cycle progression and MTC proliferation. Finally, the same set of cell cycle proteins was consistently overexpressed in human sporadic MTC but not in hereditary MTC. Together these findings suggest that aberrant CDK5 activity precedes cell cycle initiation and thus may function as a tumor-promoting factor facilitating cell cycle protein expression in MTC. Targeting aberrant CDK5 or its downstream effectors may be a strategy to halt MTC tumorigenesis.


Pulsating Radio Sources near the Crab Nebula.
Two new pulsating radio sources, designated NP 0527 and NP 0532, were found near the Crab Nebula and could be coincident with it. Both sources are sporadic, and no periodicities are evident. The pulse dispersions indicate that 1.58 +/- 0.03 and 1.74 +/- 0.02 x 10(20) electrons per square centimeter lie in the direction of NP 0527 and NP 0532, respectively.


Accurate calculation of optically induced birefringences in chiral systems using efficient polarized basis sets.
Using state-of-the-art ab initio methodology, we evaluate universal molecular parameters entering the expressions for various optically induced birefringences in chiral fluids. For this, we use the single and double excitation coupled cluster (CCSD) theory together with Dunning's augmented correlation consistent polarized basis sets of increasing size. As this is the first time these parameters are evaluated for chiral molecules using the CCSD approach, we choose possibly small test systems: a model asymmetric methane molecule, and (R)-fluoro-oxirane. With this choice, the convergence of the molecular parameters with the increase of the basis set size is investigated in detail. The results are compared to those obtained with the LPol-n (n = ds, dl, fs) and the ORP basis sets, and to the corresponding Density Functional Theory (DFT) counterparts. We can conclude that the DFT medium constant values are considerably far from coupled cluster, and therefore, aware of the known excellent performance of the CCSD method in the evaluation of various dynamic responses, we do not recommend the former methodology for accurate evaluation of the present properties. Regarding basis set convergence, the performace of the LPol-ds basis set is more efficient than that of the d-aug-cc-pVDZ set, and therefore, the former basis set can be a good choice when dealing with the study of larger systems.


Pandemic vulnerability, policy feedback and support for immigration: Evidence from Asia.
Past studies have shown that disease threat increases people's hostility towards immigrants. However, in our survey (N = 9571) conducted in five advanced Asian economies during the outbreak of COVID-19, we found that COVID-19 vulnerability was positively associated with support for immigration. Drawing on insight from policy feedback theories, we propose that the positive association is caused by the presence of widespread border crossing restrictions, which have changed the meaning and cost implications of COVID-19. As the outbreak expands, the pandemic has become not just a threat to people's health but also a barrier to globalization. Consequently, people who are worried about the disease may see globalization processes, including migration, as signs of pandemic relief. We find supportive evidence in our analysis. First, the positive association between COVID-19 vulnerability and support for immigration is more salient among respondents who considered restrictions on international travel to be stringent. Second, the positive association between COVID-19 vulnerability and immigration attitude was mediated by perceived economic threat from the pandemic and contribution by immigrants towards the containment of the pandemic. These findings suggest that disease control measures adopted at the global level may alter certain widely accepted effects of disease threat on immigration attitudes.


Comparative genomics reveals new single-nucleotide polymorphisms that can assist in identification of adherent-invasive Escherichia coli.
Adherent-invasive Escherichia coli (AIEC) have been involved in Crohn's disease (CD). Currently, AIEC are identified by time-consuming techniques based on in vitro infection of cell lines to determine their ability to adhere to and invade intestinal epithelial cells as well as to survive and replicate within macrophages. Our aim was to find signature sequences that can be used to identify the AIEC pathotype. Comparative genomics was performed between three E. coli strain pairs, each pair comprised one AIEC and one non-AIEC with identical pulsotype, sequence type and virulence gene carriage. Genetic differences were further analysed in 22 AIEC and 28 non-AIEC isolated from CD patients and controls. The strain pairs showed similar genome structures, and no gene was specific to AIEC. Three single nucleotide polymorphisms displayed different nucleotide distributions between AIEC and non-AIEC, and four correlated with increased adhesion and/or invasion indices. Here, we present a classification algorithm based on the identification of three allelic variants that can predict the AIEC phenotype with 84% accuracy. Our study corroborates the absence of an AIEC-specific genetic marker distributed across all AIEC strains. Nonetheless, point mutations putatively involved in the AIEC phenotype can be used for the molecular identification of the AIEC pathotype.


Polyclonal catalytic antibodies.
Polyclonal catalytic antibodies offer advantages in the evaluation of immunogens and in the ease of production of large quantities of antibodies. They comprise the entire immune response of an animal to an immunogen where monoclonals represent a subset. Polyclonal antibodies are consequently particularly suitable for evaluating catalytic antibody responses generated by different haptens or a group of structurally related haptens. The authors reported the first polyclonal catalytic antibodies in 1990. An unexpected finding is that polyclonal catalytic antibodies show single-site kinetic behaviour, i.e. whatever structural heterogeneity exists, the kinetic behaviour is homogeneous. Many groups worldwide have since published work in this area. Three groups are prominent. The authors' group, a group based in Austin, Texas, and led by Iverson, and a Shanghai group. The authors' group works with sheep antibodies and has published mechanistic studies and, more recently, specificity studies that revealed the catalysis of a beta-lactam. Most of this work over a 10-year period was performed by using a single bleed from a single sheep, which gives an indication of the ease of production and utility of such catalytic antibodies. Iverson's group works with rabbit antibodies and has published much work on the evaluation of catalytic antibodies generated by structurally related haptens. The Shanghai group has concentrated on the studies of polyclonal catalytic antibodies for electrocyclic reactions where any danger of contaminating enzymes is reduced. The use of immunisation to generate therapeutically useful catalytic antibodies (necessarily polyclonal) in a host animal is an attractive target. Several groups are working towards this. The authors themselves have published in this area and promising recent studies come from a group working in France on immunisation to protect against nerve agents and a Texas group (based in Houston) that has demonstrated active immunisation to generate antibodies that catalyse the hydrolysis of a carbamate insecticide.


Assessment of the health impact of occupational risk in Africa: current situation and methodological issues.
This paper presents information from the current monitoring systems in Africa, mainly Southern Africa, for occupational illness and injury and discusses the quality of the reported data in estimating the health impact of occupational risk. The paper presents and discusses the current profile of reported injury and fatalities for those countries for which data are available, in particular for the countries of the Southern African Development Community. These data indicate that the reported annual injury rates for wage workers in the Southern African Development Community region range widely from 0.35 to 49.42 injuries per 1,000 workers, and reported occupational fatality in the region ranges from 0.85 to 21.6 fatalities per 100,000 workers. Despite wide variability in reported rates (probably caused by variability in coverage and accuracy of reporting systems), transport, agriculture, mining and, to a lesser extent, construction consistently make up about three-quarters of all fatalities, with vehicle- or transport-related causes accounting for high proportions of fatal accidents. The paper identifies and discusses major sources and direction of bias and error in the reported data and suggests approaches for a better assessment of the health impact of occupational illness, injury, and mortality in African countries.


Human neural progenitors express functional lysophospholipid receptors that regulate cell growth and morphology.
Lysophospholipids regulate the morphology and growth of neurons, neural cell lines, and neural progenitors. A stable human neural progenitor cell line is not currently available in which to study the role of lysophospholipids in human neural development. We recently established a stable, adherent human embryonic stem cell-derived neuroepithelial (hES-NEP) cell line which recapitulates morphological and phenotypic features of neural progenitor cells isolated from fetal tissue. The goal of this study was to determine if hES-NEP cells express functional lysophospholipid receptors, and if activation of these receptors mediates cellular responses critical for neural development. Our results demonstrate that Lysophosphatidic Acid (LPA) and Sphingosine-1-phosphate (S1P) receptors are functionally expressed in hES-NEP cells and are coupled to multiple cellular signaling pathways. We have shown that transcript levels for S1P1 receptor increased significantly in the transition from embryonic stem cell to hES-NEP. hES-NEP cells express LPA and S1P receptors coupled to G i/o G-proteins that inhibit adenylyl cyclase and to G q-like phospholipase C activity. LPA and S1P also induce p44/42 ERK MAP kinase phosphorylation in these cells and stimulate cell proliferation via G i/o coupled receptors in an Epidermal Growth Factor Receptor (EGFR)- and ERK-dependent pathway. In contrast, LPA and S1P stimulate transient cell rounding and aggregation that is independent of EGFR and ERK, but dependent on the Rho effector p160 ROCK. Thus, lysophospholipids regulate neural progenitor growth and morphology through distinct mechanisms. These findings establish human ES cell-derived NEP cells as a model system for studying the role of lysophospholipids in neural progenitors.


Radiologists' preferences for digital mammographic display. The International Digital Mammography Development Group.
To determine the preferences of radiologists among eight different image processing algorithms applied to digital mammograms obtained for screening and diagnostic imaging tasks. Twenty-eight images representing histologically proved masses or calcifications were obtained by using three clinically available digital mammographic units. Images were processed and printed on film by using manual intensity windowing, histogram-based intensity windowing, mixture model intensity windowing, peripheral equalization, multiscale image contrast amplification (MUSICA), contrast-limited adaptive histogram equalization, Trex processing, and unsharp masking. Twelve radiologists compared the processed digital images with screen-film mammograms obtained in the same patient for breast cancer screening and breast lesion diagnosis. For the screening task, screen-film mammograms were preferred to all digital presentations, but the acceptability of images processed with Trex and MUSICA algorithms were not significantly different. All printed digital images were preferred to screen-film radiographs in the diagnosis of masses; mammograms processed with unsharp masking were significantly preferred. For the diagnosis of calcifications, no processed digital mammogram was preferred to screen-film mammograms. When digital mammograms were preferred to screen-film mammograms, radiologists selected different digital processing algorithms for each of three mammographic reading tasks and for different lesion types. Soft-copy display will eventually allow radiologists to select among these options more easily.


Cranberry for Urinary Tract Infection: From Bench to Bedside.
Urinary tract infections are common infectious diseases which can occur in any part of the urinary tract such as bladder, kidney, ureters, and urethra. They are commonly caused by bacteria that enter through the urethra. Urinary tract infections commonly develop in the bladder and spread to renal tissues. Up to now, there are different antimicrobial agents which have beneficial role on urinary tract infections. However, most of them cause different adverse effects and therefore, much attention has been paid to the search for effective therapeutic agents with negligible adverse effects. Cranberry is known as one of the most important edible plants, which possesses potent antimicrobial effects against the bacteria responsible for urinary tract infections. Growing evidence has shown that cranberry suppresses urinary tract infections and eradicates the bacteria. Therefore, the aim of this study is to critically review the available literature regarding the antimicrobial activities of cranberry against urinary tract infection microorganisms. In addition, we discuss etiology, epidemiology, risk factors, and current drugs of urinary tract infections to provide a more complete picture of this disease.


Matrix-isolation and comparative far-IR investigation of free linear [Cl3]- and a series of alkali trichlorides.
Here we discuss the reaction products of laser ablated alkali chlorides and elemental chlorine. Salt ablation using this technique combined with matrix-isolation spectroscopy allows for the formation and characterization of novel anionic species. The laser ablation of solid MCl with M = Cs, Rb, and K in the presence of Cl2 produced free [Cl3]- ions which were isolated in solid noble-gas matrices. For M = Cs, Rb, K, and Na, the ion pairs M+[Cl3]- are the main reaction products. Trends in the formation and bonding of these trichloride anions will be discussed. In contrast to the trifluoride analogues, the isolated ion pairs M+[Cl3]- feature a systematic distortion due to metal coordination.


Difficulty assisting with health care tasks among caregivers of multimorbid older adults.
Family caregivers provide assistance with health care tasks for many older adults with chronic illnesses. The difficulty they experience in providing this assistance, and related implications for their well-being, have not been well described. The objectives of this study are: (1) to describe caregiver's health care task difficulty (HCTD), (2) determine the characteristics associated with HCTD, and (3) explore the association between HCTD and caregiver well-being. This is a cross-sectional study. Baseline sample of caregivers to older (aged 65+ years) multimorbid adults enrolled in an ongoing cluster-randomized controlled trial (N = 308). The HCTD scale (0-16) is comprised of questions measuring self-reported difficulty in assisting older adults with eight health care tasks, including taking medication, visiting health care providers, and managing medical bills. Caregivers were categorized using this scale into no, low, medium, and high HCTD groups. We used ordinal logistic regression and multivariate linear regression analyses to examine the relationships between HCTD, caregiver self-efficacy, caregiver strain (Caregiver Strain Index), and depression (Center for Epidemiological Studies Depression Scale), controlling for patient and caregiver socio-demographic and health factors. Caregiver age and number of health care tasks performed were positively associated with increased HCTD. The quality of the caregiver's relationship with the patient, and self-efficacy were inversely associated with increased HCTD. A one-point increase in self-efficacy was associated with a significant lower odds of reporting high HCTD (OR, 0.64; 95% CI, 0.54, 0.77).Adjusted linear regression models indicated that high HCTD was independently associated with significantly greater caregiver strain (B, 2.7; 95% CI, 1.12, 4.29) and depression (B, 3.01; 95% CI, 1.06, 4.96). This study demonstrates that greater HCTD is associated with increased strain and depression among caregivers of multimorbid older adults. That caregiver self-efficacy was strongly associated with HCTD suggests health-system-based educational and empowering interventions might improve caregiver well-being.


Identification of abundant mRNAs from the third stage larvae of the parasitic nematode, ostertagia ostertagi.
Reverse transcriptase-PCR (RT-PCR) was carried out on total RNA prepared from the third-stage larvae (L3) of Ostertagia ostertagi in order to clone and characterize the major transcripts expressed in this larval stage, as an initial investigation of arrested larval development in the parasite. Distinct bands were visible on an agarose gel and four of these were cloned and sequenced. Three of the bands represented multiple transcripts, while the fourth band encoded the enzyme GTP cyclohydrolase I (GTP-CH), which catalyses the first and rate-limiting step in pteridine biosynthesis. Northern blot analysis and RT-PCR demonstrated that GTP-CH is highly up-regulated in the L3 stage and undetectable in either the L2 or adult stages. Using immunogold electron microscopy, GTP-CH was shown to be predominantly localized to the cell body of the body wall muscles and the cells of the intestine in the L3.


Toward a two-tier clinical warning system for hospitalized patients.
Clinical study has found early detection and intervention to be essential for preventing clinical deterioration in patients at general hospital units. In this paper, we envision a two-tiered early warning system designed to identify the signs of clinical deterioration and provide early warning of serious clinical events. The first tier of the system automatically identifies patients at risk of clinical deterioration from existing electronic medical record databases. The second tier performs real-time clinical event detection based on real-time vital sign data collected from on-body wireless sensors attached to those high-risk patients. We employ machine-learning techniques to analyze data from both tiers, assigning scores to patients in real time. The assigned scores can then be used to trigger early-intervention alerts. Preliminary study of an early warning system component and a wireless clinical monitoring system component demonstrate the feasibility of this two-tiered approach.


Construction of Novel Gene Signature-Based Predictive Model for the Diagnosis of Acute Myocardial Infarction by Combining Random Forest With Artificial Neural Network.
Acute myocardial infarction (AMI) is one of the most common causes of mortality around the world. Early diagnosis of AMI contributes to improving prognosis. In our study, we aimed to construct a novel predictive model for the diagnosis of AMI using an artificial neural network (ANN), and we verified its diagnostic value via constructing the receiver operating characteristic (ROC). We downloaded three publicly available datasets (training sets GSE48060, GSE60993, and GSE66360) from Gene Expression Omnibus (GEO) database, and differentially expressed genes (DEGs) were identified between 87 AMI and 78 control samples. We applied the random forest (RF) and ANN algorithms to further identify novel gene signatures and construct a model to predict the possibility of AMI. Besides, the diagnostic value of our model was further validated in the validation sets GSE61144 (7 AMI patients and 10 controls), GSE34198 (49 AMI patients and 48 controls), and GSE97320 (3 AMI patients and 3 controls). A total of 71 DEGs were identified, of which 68 were upregulated and 3 were downregulated. Firstly, 11 key genes in 71 DEGs were screened with RF classifier for the classification of AMI and control samples. Then, we calculated the weight of each key gene using ANN. Furthermore, the diagnostic model was constructed and named neuralAMI, with significant predictive power (area under the curve [AUC] = 0.980). Finally, our model was validated with the independent datasets GSE61144 (AUC = 0.900), GSE34198 (AUC = 0.882), and GSE97320 (AUC = 1.00). Machine learning was used to develop a reliable predictive model for the diagnosis of AMI. The results of our study provide potential gene biomarkers for early disease screening.


Portal vein afferents are critical for the sympathoadrenal response to hypoglycemia.
We sought to elucidate the role of the portal vein afferents in the sympathetic response to hypoglycemia. Laparotomy was performed on 27 male Wistar rats. Portal veins were painted with either 90% phenol (denervation group [PDN]) or 0.9% saline solution (sham-operated group [SHAM]). Rats were chronically cannulated in the carotid artery (sampling), jugular vein (infusion), and portal vein (infusion). After a recovery period of 5 days, animals were exposed to a hyperinsulinemic-hypoglycemic clamp, with glucose infused either portally (POR) or peripherally (PER). In all animals, systemic hypoglycemia (2.48+/-0.09 mmol/l) was induced via jugular vein insulin infusion (50 mU x kg(-1) x min(-1)). Arterial plasma catecholamines were assessed at basal (-30 and 0 min) and during sustained hypoglycemia (60, 75, 90, and 105 min). By design, portal vein glucose concentrations were significantly elevated during POR versus PER (4.4+/-0.14 vs. 2.5+/-0.07 mmol/l; P<0.01, respectively) for both PDN and SHAM. There were no significant differences in arterial glucose or insulin concentration between the four experimental conditions at any point in time. When portal glycemia and systemic glycemia fell concomitantly (SHAM-PER), epinephrine increased 12-fold above basal (3.75+/-0.34 and 44.56+/-6.1 nmol/l; P<0.001). However, maintenance of portal normoglycemia (SHAM-POR) caused a 50% suppression of the epinephrine response, despite cerebral hypoglycemia (22.2+/-3.1 nmol/l, P<0.001). Portal denervation resulted in a significant blunting of the sympathoadrenal response to whole-body hypoglycemia (PDN-PER 27.6+/-3.8 nmol/l vs. SHAM-PER; P<0.002). In contrast to the sham experiments, there was no further suppression in arterial epinephrine concentrations observed during PDN-POR versus PDN-PER (P = 0.8). These findings indicate that portal vein afferent innervation is critical for hypoglycemic detection and normal sympathoadrenal counterregulation.


Characteristics of individuals with integrated pensions.
Employer pensions that integrate benefits with Social Security have been the focus of relatively little research. Since changes in Social Security benefit levels and other program characteristics can affect the benefit levels and other features of integrated pension plans, it is important to know who is covered by these plans. This article examines the characteristics of workers covered by integrated pension plans, compared to those with nonintegrated plans and those with no pension coverage. Integrated pension plans are those that explicitly adjust their benefit structure to help compensate for the employer's contributions to the Social Security program. There are two basic integration methods used by defined benefit (DB) plans. The offset method causes a reduction in employer pension benefits by up to half of the Social Security retirement benefit; the excess rate method is characterized by an accrual rate that is lower for earnings below the Social Security taxable maximum than above it. Defined contribution (DC) pension plans can be integrated along the lines of the excess rate method. To date, research on integrated pensions has focused on plan characteristics, as reported to the Bureau of Labor Statistics (BLS) through its Employee Benefits Survey (EBS). This research has examined the prevalence of integration among full-time, private sector workers by industry, firm size, and broad occupational categories. However, because the EBS provides virtually no data on worker characteristics, analyses of the effects of pension integration on retirement benefits have used hypothetical workers, varying according to assumed levels of earnings and job tenure. This kind of analysis is not particularly helpful in examining the potential effects of changes in the Social Security program on workers' pension benefits. However, data on pension integration at the individual level are available, most recently from the Health and Retirement Study (HRS), a nationally representative survey of individuals aged 51-61 in 1992. This dataset provides the basis for the analysis presented here. The following are some of the major findings from this analysis. The incidence of pension integration in the HRS sample is 32 percent of all workers with a pension (14 percent of all workers). The HRS can also identify integrated DC plans, a statistic that is not available from BLS data. The rate of integration for workers with only DC plans is 8 percent. After controlling for other variables, several socio-demographic characteristics are significantly related to the incidence of integration. The probability of having an integrated pension is 4.6 percentage points less for men compared to women. Non-Hispanic blacks are 6.4 percentage points less likely than non-Hispanic whites to have integrated pensions. Union members are 14 percentage points less likely to have integrated pensions, while workers with less than a graduate level education are at least 15 percentage points more likely to have a pension that is integrated. Some earnings and pension characteristics are also significantly correlated with pension integration. Earnings are positively related, with the probability of having an integrated pension increasing by 2 percentage points for an increase of $1,000 in annual pay. An even larger effect comes from earning at or above the Social Security taxable maximum. Workers at or above this income level are 10 percentage points more likely to have an integrated plan, but for those with more than one plan the probability of pension integration goes up by 13 percentage points.


FXYD5 (dysadherin) regulates the paracellular permeability in cultured kidney collecting duct cells.
FXYD5 (dysadherin or RIC) is a member of the FXYD family of single-span transmembrane proteins associated with the Na(+)-K(+)-ATPase. Several studies have demonstrated enhanced expression of FXYD5 during metastasis and effects on cell adhesion and motility. The current study examines effects of FXYD5 on the paracellular permeability in the mouse kidney collecting duct cell line M1. Expressing FXYD5 in these cells leads to a large decrease in amiloride-insensitive transepithelial electrical resistance as well as increased permeability to 4-kDa dextran. Impairment of cell-cell contact was also demonstrated by staining cells for the tight and adherence junction markers zonula occludens-1 and β-catenin, respectively. This is further supported by large expansions of the interstitial spaces, visualized in electron microscope images. Expressing FXYD5 in M1 cells resulted in a decrease in N-glycosylation of β1 Na(+)-K(+)-ATPase, while silencing it in H1299 cells had an opposite effect. This may provide a mechanism for the above effects, since normal glycosylation of β1 plays an important role in cell-cell contact formation (Vagin O, Tokhtaeva E, Sachs G. J Biol Chem 281: 39573-39587, 2006).


Analysis of metagene portraits reveals distinct transitions during kidney organogenesis.
Organogenesis is a multistage process, but it has been difficult, by conventional analysis, to separate stages and identify points of transition in developmentally complex organs or define genetic pathways that regulate pattern formation. We performed a detailed time-series examination of global gene expression during kidney development and then represented the resulting data as self-organizing maps (SOMs), which reduced more than 30,000 genes to 650 metagenes. Further clustering of these maps identified potential stages of development and suggested points of stability and transition during kidney organogenesis that are not obvious from either standard morphological analyses or conventional microarray clustering algorithms. We also performed entropy calculations of SOMs generated for each day of development and found correlations with morphometric parameters and expression of candidate genes that may help in orchestrating the transitions between stages of kidney development, as well as macro- and micropatterning of the organ.


Host range and cell cycle activation properties of polyomavirus large T-antigen mutants defective in pRB binding.
We have examined the growth properties of polyomavirus large T-antigen mutants that are unable to bind pRB, the product of the retinoblastoma tumor suppressor gene. These mutants grow poorly on primary mouse cells yet grow well on NIH 3T3 and other established mouse cell lines. Preinfection of primary baby mouse kidney (BMK) epithelial cells with wild-type simian virus 40 renders these cells permissive to growth of pRB-binding polyomavirus mutants. Conversely, NIH 3T3 cells transfected by and expressing wild-type human pRB become nonpermissive. Primary fibroblasts from mouse embryos that carry a homozygous knockout of the RB gene are permissive, while those from normal littermates are nonpermissive. The host range of polyomavirus pRB-binding mutants is thus determined by expression or lack of expression of functional pRB by the host. These results demonstrate the importance of pRB binding by large T antigen for productive viral infection in primary cells. Failure of pRB-binding mutants to grow well in BMK cells correlates with their failure to induce progression from G0 or G1 through the S phase of the cell cycle. Time course studies show delayed synthesis and lower levels of accumulation of large T antigen, viral DNA, and VP1 in mutant compared with wild-type virus-infected BMK cells. These results support a model in which productive infection by polyomavirus in normal mouse cells is tightly coupled to the induction and progression of the cell cycle.


Study on the influence of slope shape with numerical calculation models on slope safety during slope excavation.
Under the influence of natural weathering and excavation in human engineering, slopes in nature have various slope shapes. In human engineering activities, the stability of different types of slopes formed by manual excavation must be affected by multiple factors such as geologic setting, lithology and environment. Therefore, to understand the impact of slope shape, geologic setting, and other conditions on slope stability of artificial slopes, calculation models for straight slope, concave slope, and convex slope are constructed based on the three slope shape characteristics. By changing the angles of upward and downward slope angles and analysing the parameters of slope shape, joint spacing, and joint angle, discrete element software is used to calculate the slope safety factor. The calculation results show that the slope shape, joint spacing, and joint inclination affect the safety of slopes. In straight slopes with large joint spacing, the smaller the slope angle, the greater the safety factor. However, in the interval of small joint spacing, the safety coefficient of slopes with slight joint inclination has the opposite variation characteristics. When a<90°, the straight slope has a dominant joint inclination angle that minimises the slope safety factor. In concave slopes, the more concave the slope shape is, the smaller the safety factor is; For concave slopes with small joint spacing, the slope with slight joint inclination has a more significant safety factor; Under the condition of large joint spacing, there is a dominant joint inclination in the joint inclination range of 30° to 70° for concave slopes, which minimises the safety factor of the slope. In convex slopes, the smaller the joint inclination angle of the slope, the smaller the safety factor of the slope, and the smaller the upslope angle of the slope, the greater the safety factor of the slope.


ER stress inhibits neuronal death by promoting autophagy.
Endoplasmic reticulum (ER) stress has been implicated in neurodegenerative diseases but its relationship and role in disease progression remain unclear. Using genetic and pharmacological approaches, we showed that mild ER stress ("preconditioning") is neuroprotective in Drosophila and mouse models of Parkinson disease. In addition, we found that the combination of mild ER stress and apoptotic signals triggers an autophagic response both in vivo and in vitro. We showed that when autophagy is impaired, ER-mediated protection is lost. We further demonstrated that autophagy inhibits caspase activation and apoptosis. Based on our findings, we conclude that autophagy is required for the neuroprotection mediated by mild ER stress, and therefore ER preconditioning has potential therapeutic value for the treatment of neurodegenerative diseases.


Drug repositioning of mevalonate pathway inhibitors as antitumor agents for ovarian cancer.
Drug repositioning is an alternative strategy redirecting existing drugs for new disease. We have previously reported an antitumor effect of statins, antidyslipidemic drugs, on ovarian cancer in vitro and in vivo. In this study, we investigated the antitumor effects of other mevalonate pathway inhibitors and the mechanism of the antitumor effect from a metabolic perspective. The effects of inhibitors of the mevalonate pathway on tumor cell growth were evaluated in vitro. Bisphosphonates that inhibit this pathway are commonly used as antiosteoporotic drugs, and antitumor effects of the bisphosphonate were examined in vitro and in vivo. Metabolites in SKOV3 ovarian cancer cells were analyzed before and after lovastatin treatment, using capillary electrophoresis-mass spectrometry. All mevalonate pathway inhibitors showed concentration-dependent inhibitory effects on tumor cell growth. Particularly marked effects were obtained with inhibitors of farnesyltransferase and geranylgeranyltransferase. The bisphosphonate was also shown to have an antitumor effect in vivo. The expression of autophagy marker LC3A/3B was increased in cells after treatment. In metabolomics analysis, lovastatin treatment increased the metabolites involved in the tricarboxylic acid cycle while reducing the metabolites associated with glycolysis. Also it decreased glutathione and resulted to work with chemotherapeutic agents synergistically. Inhibition at any point in the mevalonate pathway, and especially of farnesyl pyrophosphate and geranylgeranyl pyrophosphate, suppresses growth of ovarian cancer cells. Inhibition of this pathway may induce autophagy, cause a shift to activation of the tricarboxylic acid cycle and enhance susceptibility to chemotherapy. Drug repositioning targeting mevalonate pathway for ovarian cancer deserves consideration for clinical application.


Brain-stem structures and catecholamines in the control of arterial blood pressure in the rat.
1. Noradrenaline, adrenaline and alpha-methylnoradrenaline administration into the nucleus tractus solitarii (NTS) of anaesthetized rats decreased blood pressure and heart rate in a dose-dependent fashion. 2. Bilateral injections were effective in lower doses than unilateral administration. alpha-Methylnoradrenaline given bilaterally produced hypotension in a dose of 0-08 nmol whereas after unilateral injection a dose of 0-32 nmol was needed to obtain the same degree of hypotension. 3. Electrical stimulation of the NTS caused hypotension and bradycardia. Conversely, bilateral electrolytic lesions or deafferentation of the NTS led to acute hypertension. Chronically such lesions caused neurogenic hypertension. 4. In spontaneously hypertensive rats increased concentrations of noradrenaline, adrenaline and dopamine were measured in the part of the NTS located just caudal to the obex (A2 region).


Denture use and Risk for Cardiometabolic Disease: Observational and Mendelian Randomization analyses.
Denture use may potentially increase the risk of cardiometabolic diseases (CMDs), but the casual relevance and strength of the associations are currently unknown. 495 938 participants from the UK Biobank were included in the observational analyses. Linkage disequilibrium score (LDSC) regression and Mendelian randomization analyses were employed to estimate genetic correlation and the associations between the genetic liability for denture use with coronary artery disease, myocardial infarction, heart failure (HF), any stroke (AS), ischemic stroke, hemorrhagic stroke, type 2 diabetes (T2D), and related clinical risk factors. In observational analysis, denture use was associated with 14-25% higher risks of various CMDs. LDSC analysis found that denture use showed a positive genetic correlation with CMDs (rg 0.21-0.38). Genetic liability for denture use was associated with an elevated risk of HF [odds ratio:1.49 (1.20-1.83)] and T2D [1.11 (1.01-1.24)]. By integrating genetic summary data of denture use with the sum of Decayed, Missing, and Filled tooth Surfaces (DMFS), a clinical measure of dental caries obtained from an independent source, genetically-determined denture use/DMFS was also associated with an elevated risk of AS [1.21 (1.04-1.40)]. Furthermore, genetically-predicted denture use/DMFS was significantly associated with established cardiometabolic risk factors, including high-density lipoprotein cholesterol, triglycerides, waist circumference, waist-to-hip ratio, and height. Our study supported potential causal associations between the genetic liability for denture use and risks for HF, AS, and T2D, and related clinical risk factors. These findings may inform prevention and intervention strategies targeting dental diseases and cardiometabolic diseases.


WS-SNPs&GO: a web server for predicting the deleterious effect of human protein variants using functional annotation.
SNPs&GO is a method for the prediction of deleterious Single Amino acid Polymorphisms (SAPs) using protein functional annotation. In this work, we present the web server implementation of SNPs&GO (WS-SNPs&GO). The server is based on Support Vector Machines (SVM) and for a given protein, its input comprises: the sequence and/or its three-dimensional structure (when available), a set of target variations and its functional Gene Ontology (GO) terms. The output of the server provides, for each protein variation, the probabilities to be associated to human diseases. The server consists of two main components, including updated versions of the sequence-based SNPs&GO (recently scored as one of the best algorithms for predicting deleterious SAPs) and of the structure-based SNPs&GO(3d) programs. Sequence and structure based algorithms are extensively tested on a large set of annotated variations extracted from the SwissVar database. Selecting a balanced dataset with more than 38,000 SAPs, the sequence-based approach achieves 81% overall accuracy, 0.61 correlation coefficient and an Area Under the Curve (AUC) of the Receiver Operating Characteristic (ROC) curve of 0.88. For the subset of ~6,600 variations mapped on protein structures available at the Protein Data Bank (PDB), the structure-based method scores with 84% overall accuracy, 0.68 correlation coefficient, and 0.91 AUC. When tested on a new blind set of variations, the results of the server are 79% and 83% overall accuracy for the sequence-based and structure-based inputs, respectively. WS-SNPs&GO is a valuable tool that includes in a unique framework information derived from protein sequence, structure, evolutionary profile, and protein function. WS-SNPs&GO is freely available at http://snps.biofold.org/snps-and-go.


Swimming performance assessment in fishes.
Swimming performance tests of fish have been integral to studies of muscle energetics, swimming mechanics, gas exchange, cardiac physiology, disease, pollution, hypoxia and temperature. This paper describes a flexible protocol to assess fish swimming performance using equipment in which water velocity can be controlled. The protocol involves one to several stepped increases in flow speed that are intended to cause fish to fatigue. Step speeds and their duration can be set to capture swimming abilities of different physiological and ecological relevance. Most frequently step size is set to determine critical swimming velocity (U(crit;)), which is intended to capture maximum sustained swimming ability. Traditionally this test has consisted of approximately ten steps each of 20 min duration. However, steps of shorter duration (e.g. 1 min) are increasingly being utilized to capture acceleration ability or burst swimming performance. Regardless of step size, swimming tests can be repeated over time to gauge individual variation and recovery ability. Endpoints related to swimming such as measures of metabolic rate, fin use, ventilation rate, and of behavior, such as the distance between schooling fish, are often included before, during and after swimming tests. Given the diversity of fish species, the number of unexplored research questions, and the importance of many species to global ecology and economic health, studies of fish swimming performance will remain popular and invaluable for the foreseeable future.


Determinants of gastroesophageal reflux disease, including hookah smoking and opium use- a cross-sectional analysis of 50,000 individuals.
Gastroesophageal reflux disease (GERD) is a common cause of discomfort and morbidity worldwide. However, information on determinants of GERD from large-scale studies in low- to medium-income countries is limited. We investigated the factors associated with different measures of GERD symptoms, including frequency, patient-perceived severity, and onset time. We performed a cross-sectional analysis of the baseline data from a population-based cohort study of ∼ 50,000 individuals in in Golestan Province, Iran. GERD symptoms in this study included regurgitation and/or heartburn. Approximately 20% of participants reported at least weekly symptoms. Daily symptoms were less commonly reported by men, those of Turkmen ethnicity, and nass chewers. On the other hand, age, body mass index, alcohol drinking, cigarette smoking, opium use, lower socioeconomic status, and lower physical activity were associated with daily symptoms. Most of these factors showed similar associations with severe symptoms. Women with higher BMI and waist to hip ratio were more likely to report frequent and severe GERD symptoms. Hookah smoking (OR 1.34, 95% CI 1.02-1.75) and opium use (OR 1.70, 95% CI 1.55-1.87) were associated with severe symptoms, whereas nass chewing had an inverse association (OR 0.87, 95% CI 0.76-0.99). After exclusion of cigarette smokers, hookah smoking was still positively associated and nass chewing was inversely associated with GERD symptoms (all frequencies combined). GERD is common in this population. The associations of hookah and opium use and inverse association of nass use with GERD symptoms are reported for the first time. Further studies are required to investigate the nature of these associations. Other determinants of GERD were mostly comparable to those reported elsewhere.