Implementation of Perioperative Music Using the Consolidated Framework for Implementation Research.
Complementary integrative health therapies have a perioperative role in the reduction of pain, analgesic use, and anxiety, and increasing patient satisfaction. However, long implementation lags have been quantified. The Consolidated Framework for Implementation Research (CFIR) can help mitigate this translational problem. We reviewed evidence for several nonpharmacological treatments (CFIR domain: characteristics of interventions) and studied external context and organizational readiness for change by surveying providers at 11 Veterans Affairs (VA) hospitals (domains: outer and inner settings). We asked patients about their willingness to receive music and studied the association between this and known risk factors for opioid use (domain: characteristics of individuals). We implemented a protocol for the perioperative use of digital music players loaded with veteran-preferred playlists and evaluated its penetration in a subgroup of patients undergoing joint replacements over a 6-month period (domain: process of implementation). We then extracted data on postoperative recovery time and other outcomes, comparing them with historic and contemporary cohorts. Evidence varied from strong and direct for perioperative music and acupuncture, to modest or weak and indirect for mindfulness, yoga, and tai chi, respectively. Readiness for change surveys completed by 97 perioperative providers showed overall positive scores (mean >0 on a scale from -2 to +2, equivalent to >2.5 on the 5-point Likert scale). Readiness was higher at Durham (+0.47) versus most other VA hospitals (range +0.05 to +0.63). Of 3307 veterans asked about willingness to receive music, approximately 68% (n = 2252) answered "yes." In multivariable analyses, a positive response (acceptability) was independently predicted by younger age and higher mean preoperative pain scores (>4 out of 10 over 90 days before admission), factors associated with opioid overuse. Penetration was modest in the targeted subset (39 received music out of a possible 81 recipients), potentially reduced by device nonavailability due to diffusion into nontargeted populations. Postoperative recovery time was not changed, suggesting smooth integration into workflow. CFIR-guided implementation of perioperative music was feasible at a tertiary VA hospital, with moderate penetration in a high-risk subset of patients. Use of digital music players with preferred playlists was supported by strong evidence, tension for change, modest readiness among providers, good acceptability among patients (especially those at risk for opioid overuse), and a protocolized approach. Further study is needed to identify similar frameworks for effective knowledge-translation activities.


Leishmania promastigotes are recognized by the macrophage receptor for advanced glycosylation endproducts.
In this paper we demonstrate the involvement of the macrophage receptor for advanced glycosylation endproducts (AGE) in the phagocytosis of Leishmania major promastigotes. Blocking of this receptor with the ligand, AGE-BSA, leads to a 50% decrease in phagocytosis relative to controls, and a comparable decrease in the respiratory burst. The inhibition of phagocytosis by AGE-BSA was specific to leishmania. The binding of zymosan or C3bi-RBC and the phagocytosis of IgG-RBC or latex beads was not affected by the presence of AGE-BSA. Blocking of both the AGE receptor and CR3 decreases leishmania binding by nearly 90%, and reduces the respiratory burst by 80%, indicating that the two receptors account for the bulk of L. tropica promastigote recognition and uptake by the macrophage.


Recommendations for high-priority research on cancer-related fatigue in children and adults.
Over the past decades, some scientific progress has been made in understanding and treating cancer-related fatigue (CRF). However, three major problems have limited further progress: lack of agreement about measurement, inadequate understanding of the underlying biology, and problems in the conduct of clinical trials for CRF. This commentary reports the recommendations of a National Cancer Institute Clinical Trials Planning Meeting and an ongoing National Cancer Institute working group to address these problems so that high-priority research and clinical trials can be conducted to advance the science of CRF and its treatment. Recommendations to address measurement issues included revising the current case definition to reflect more rigorous criteria, adopting the Patient Reported Outcomes Measurement Information System fatigue scales as standard measures of CRF, and linking legacy measures to the scales. With regard to the biology of CRF, the group identified the need for longitudinal research to examine biobehavioral mechanisms underlying CRF and testing mechanistic hypotheses within the context of intervention research. To address clinical trial issues, recommendations included using only placebo-controlled trial designs. setting eligibility to minimize sample heterogeneity or enable subgroup analysis, establishing a CRF severity threshold for participation in clinical trials, conducting dissemination trials of efficacious interventions (such as exercise), and combining nonpharmacologic and pharmacologic interventions to exploit the potential synergy between these approaches. Accomplishing these goals has the potential to advance the science of CRF and improve the clinical management of this troubling symptom.


Remobilization of Sleeping Beauty transposons in the germline of Xenopus tropicalis.
The Sleeping Beauty (SB) transposon system has been used for germline transgenesis of the diploid frog, Xenopus tropicalis. Injecting one-cell embryos with plasmid DNA harboring an SB transposon substrate together with mRNA encoding the SB transposase enzyme resulted in non-canonical integration of small-order concatemers of the transposon. Here, we demonstrate that SB transposons stably integrated into the frog genome are effective substrates for remobilization. Transgenic frogs that express the SB10 transposase were bred with SB transposon-harboring animals to yield double-transgenic 'hopper' frogs. Remobilization events were observed in the progeny of the hopper frogs and were verified by Southern blot analysis and cloning of the novel integrations sites. Unlike the co-injection method used to generate founder lines, transgenic remobilization resulted in canonical transposition of the SB transposons. The remobilized SB transposons frequently integrated near the site of the donor locus; approximately 80% re-integrated with 3 Mb of the donor locus, a phenomenon known as 'local hopping'. In this study, we demonstrate that SB transposons integrated into the X. tropicalis genome are effective substrates for excision and re-integration, and that the remobilized transposons are transmitted through the germline. This is an important step in the development of large-scale transposon-mediated gene- and enhancer-trap strategies in this highly tractable developmental model system.


Randomized, controlled clinical trial of safety and plasma concentrations of diclofenac in healthy neonatal foals after repeated topical application of 1% diclofenac sodium cream.
OBJECTIVE To determine the plasma pharmacokinetics and safety of 1% diclofenac sodium cream applied topically to neonatal foals every 12 hours for 7 days. ANIMALS Twelve 2- to 14-day old healthy Arabian and Arabian-pony cross neonatal foals. PROCEDURES A 1.27-cm strip of cream containing 7.3 mg of diclofenac sodium (n = 6 foals) or an equivalent amount of placebo cream (6 foals) was applied topically to a 5-cm square of shaved skin over the anterolateral aspect of the left tarsometatarsal region every 12 hours for 7 days. Physical examination, CBC, serum biochemistry, urinalysis, gastric endoscopy, and ultrasonographic examination of the kidneys and right dorsal colon were performed before and after cream application. Venous blood samples were collected at predefined intervals following application of the diclofenac cream, and plasma diclofenac concentrations were determined by liquid chromatography-mass spectrometry. RESULTS No foal developed any adverse effects attributed to diclofenac application, and no significant differences in values of evaluated variables were identified between treatment groups. Plasma diclofenac concentrations peaked rapidly following application of the diclofenac cream, reaching a maximum of < 1 ng/mL within 2 hours, and declined rapidly after application ceased. CONCLUSIONS AND CLINICAL RELEVANCE Topical application of the 1% diclofenac sodium cream to foals as described appeared safe, and low plasma concentrations of diclofenac suggested minimal systemic absorption. Practitioners may consider use of this medication to treat focal areas of pain and inflammation in neonatal foals.


Quality of life, healthcare use and costs in 'at-risk' children after early antibiotic treatment versus placebo for influenza-like illness: within-trial descriptive economic analyses of the ARCHIE randomised controlled trial.
To characterise the quality of life, healthcare use and costs associated with early antibiotic treatment of influenza-like illness (ILI) in 'at-risk' children. Economic analysis of a two-arm double-blind parallel group pragmatic randomised controlled trial. Children were recruited from community-based healthcare settings, including general practices, walk-in centres and hospital ambulatory care. Children with risk factors for influenza-related complications, including respiratory, cardiac and neurological conditions, who presented within the first 5 days of an ILI. Co-amoxiclav 400/57 suspension or placebo. This economic analysis focused on quality of life measured by the EQ-5D-Y, symptoms assessed by the Canadian Acute Respiratory Infection and Flu Scale (CARIFS), healthcare use and costs including medication, hospital visits and admissions, general practitioner and nurse contacts. Outcomes were assessed for up to 28 days post randomisation. Information on resource use, EQ-5D-Y (day 28) and CARIFS (day 7) was available for 265 (98%), 72 (27%) and 123 (45%) out of 271 participants, respectively. Average costs in the co-amoxiclav group were £25 lower (95% CI -£113 to £65), but this difference was not statistically significant (p=0.566). The difference in EQ-5D-Y scores between groups was also not statistically significant (-0.014 (95% CI -0.124 to 0.096), p=0.798). However, day 7 CARIFS scores were 3.5 points lower in the co-amoxiclav arm (95% CI -6.9 to -0.1, p=0.044). Our findings did not show evidence that early co-amoxiclav treatment improves quality of life or reduces healthcare use and costs in 'at-risk' children with ILI, but may reduce symptom severity though confirmation from further research would be important. Reliable data collection from children's parents/carers was challenging, and resulted in high levels of missing data, which is common in pragmatic trials involving children with acute respiratory tract infections. ISRCTN70714783; EudraCT 2013-002822-21.


Cholecystectomy and changing mortality from gallbladder cancer.
Over the past decade, mortality from gallbladder cancer has declined sharply in England and Wales, Scotland, the United States, and Canada, but has risen by 33% in Sweden. In all these countries, gallbladder cancer mortality rates are inversely related to cholecystectomy rates in the previous year. It is suggested that changes in cholecystectomy rates produce changes in the prevalence of cholelithiasis, and consequently in gallbladder cancer incidence and mortality. It is estimated that 1 fewer death from gallbladder cancer occurs for about every 100 cholecystectomies done during the preceding year. The possibility of preventing death from gallbladder cancer in the patient with asymptomatic cholelithiasis must be balanced, however, against the risks of surgery.


Investigation of multilevel amplitude modulation for a dual-wavelength free-space optical communications system using realistic channel estimation and minimum mean-squared-error linear equalization.
Fog is a highly dispersive medium at optical wavelengths, and the received pulse waveform may suffer significant distortion. Thus it is desirable to have the impulse response of the propagation channel to recover data transmitted through fog. The fog particle density and the particle size distribution both strongly influence the channel impulse response, yet it is difficult to estimate these parameters. We present a method using a dual-wavelength free-space optical system for estimating the average particle diameter and the particle number density and for approximating the particle distribution function. These parameters serve as inputs to estimate the atmospheric channel impulse response using simulation based on the modified vector radiative transfer theory. The estimated channel response is used to design a minimum mean-square-error equalization filter to improve the bit error rate by correcting distortion in the received signal waveform due to intersymbol interference and additive white Gaussian noise.


A Critical Examination of the Use of Trained Health Coaches to Decrease the Metabolic Syndrome for Participants of a Community-Based Diabetes Prevention and Management Program.
The epidemic of obesity and diabetes in the United States poses major challenge to the prevention and management of chronic diseases. Furthermore, when this is viewed in other components of the metabolic syndrome (i.e., the burden of high cholesterol and hypertension), the prevalence of the metabolic syndrome continues to rise in the USA continued challenge is how to deal with this epidemic from a medical and public health standpoint. Community Based Participatory Research (CBPR) is a unique approach and offers a novel perspective for answering this challenge. A critical set of goals for CBPR is to address health disparities and social inequalities while getting community members engaged in all aspects of the research process. Utilizing the West Virginia Diabetes Prevention and Management Program and trained Health Coaches as a model, we discuss topics of consideration related to CBPR, involving trained health coaches, optimizing early adoption of healthy lifestyle behaviors, and enhancing participation. Through careful project planning and design, questions regarding disparities increasing susceptibility and preventive efforts within the community can be addressed successfully. These topics are part of a broader integration of theories such as participatory research, community engagement, and outcomes measurement. The understanding of the pathophysiology and epidemiology of the metabolic syndrome can help frame an appropriate strategy for establishing long-term community-wide changes that promote health. In order to continue to improve investigations for preventing the metabolic syndrome, it will be necessary to have aggressive efforts at the individual and population level for developing culturally sensitive programs that start early and are sustainable in practical environments such as the workplace. In this comprehensive review, we will discuss practical considerations related to project design, implementation, and how to measure effectiveness in regards to reducing the burden of the metabolic syndrome.


1.5D Egocentric Dynamic Network Visualization.
Dynamic network visualization has been a challenging research topic due to the visual and computational complexity introduced by the extra time dimension. Existing solutions are usually good for overview and presentation tasks, but not for the interactive analysis of a large dynamic network. We introduce in this paper a new approach which considers only the dynamic network central to a focus node, also known as the egocentric dynamic network. Our major contribution is a novel 1.5D visualization design which greatly reduces the visual complexity of the dynamic network without sacrificing the topological and temporal context central to the focus node. In our design, the egocentric dynamic network is presented in a single static view, supporting rich analysis through user interactions on both time and network. We propose a general framework for the 1.5D visualization approach, including the data processing pipeline, the visualization algorithm design, and customized interaction methods. Finally, we demonstrate the effectiveness of our approach on egocentric dynamic network analysis tasks, through case studies and a controlled user experiment comparing with three baseline dynamic network visualization methods.


Why do couples drop-out from IVF treatment? A prospective cohort study.
Cumulative IVF pregnancy rates are compromised by the large number of couples who drop-out of treatment before achieving pregnancy. The aim of this study was to identify the role of the treatment strategy applied, and potential other factors that influence the decision of couples to discontinue treatment. The incidence of drop-out from IVF treatment and factors related to drop-out were studied in a cohort of IVF patients aged <38 years embarking on IVF treatment either with a mild or a standard treatment strategy for a planned maximum number of treatment cycles. Of the 384 couples studied, 17% dropped out of IVF treatment. The physical or psychological burden of treatment was the most frequent cause of drop-out (28%). The application of a mild treatment strategy (mild ovarian stimulation along with the transfer of a single embryo) significantly reduced the chance of drop-out (hazard ratio (HR) 0.55; 95% confidence interval (CI), 0.31-0.96). When a mild IVF strategy was employed, the association between the baseline anxiety score and drop-out was reduced by >50%. The presence of severe male subfertility (HR 4.80; 95% CI, 1.63-14.13) and the failure to achieve embryo transfer (odds ratio 0.41; 95% CI, 0.24-0.72) were also related to drop-out. Reducing drop-out rate is crucial to further improve the efficacy and cost-effectiveness of IVF treatment. An important factor determining the risk of drop-out is the burden of the treatment strategy. The application of a mild treatment strategy and managing patient's expectations might reduce drop-out rates.


Control of MHC restriction by TCR Valpha CDR1 and CDR2.
Individual T cell receptor (TCR) Valpha elements are expressed preferentially in CD4 or CD8 peripheral T cell subsets. The closely related Valpha3.1 and Valpha3.2 elements show reciprocal selection into CD4 and CD8 subsets, respectively. Transgenic mice expressing site-directed mutants of a Valpha3.1 gene were used to show that individual residues in either the complementarity-determining region 1 (CDR1) or CDR2 were sufficient to change selection from the CD4 subset to the CD8 subset. Thus, the germline-encoded Valpha elements are a major influence on major histocompatibility class complex (MHC) restriction, most likely by a preferential interaction with one or the other class of MHC molecule.


Obesity related risk of sudden cardiac death in the atherosclerosis risk in communities study.
To examine the association of body mass index (BMI), waist circumference (WC) and waist hip ratio (WHR) with sudden cardiac death (SCD) in community dwelling individuals. Data from a multicentre, prospective, cohort study of 14 941 men and women (African American, and white), aged 45-64 years, participating in the Atherosclerosis Risk in Communities study was analysed. Obesity measures were assessed at baseline (1987-1989). SCD was adjudicated by a committee. At enrolment mean±SD age of the participants was 54±6 years (55% female; 26% African American). During 12.6±2.5 years of follow-up, 253 SCD occurred (incidence rate 1.34/100 person-years). The association between obesity and SCD differed by smoking status (interaction p≤0.01). In models adjusting for age, sex, race, study centre and education level, SCD risk was positively associated (p<0.001) with BMI, WC and WHR in non-smokers, but not in smokers. WHR was more strongly associated with SCD in non-smokers than was BMI or WC (HR per SD increment (95% CI) 2.00 (1.65 to 2.42); 1.34 (1.15 to 1.56) and 1.49 (1.28 to 1.74), respectively). After adjustment for potential mediators (hypertension, diabetes, lipid profile, prevalent coronary heart disease, heart failure, and LV hypertrophy), non-smokers in the highest WHR category (>0.95 in women; >1.01 in men) had double the risk of SCD (HR 2.03, 95% CI 1.19 to 3.46; incidence rate 1.43/1000 person-years) versus those with normal WHR. General obesity is associated with increased risk of SCD in middle-aged, non-smoking individuals, mediated by traditional cardiovascular risk factors. Central obesity, however, is independently associated with SCD by pathways that remain to be elucidated.


What Is the Meaning of Increased Myocardial Injury Enzymes during Hemodialysis? A Tissue Doppler Imaging Study.
Cardiovascular death is decreasing in the general population; however, it appears in still higher rates and even increases gradually in hemodialysis (HD) patients. This situation has led to a debate about cardiovascular adverse effects of HD which lead to significant changes in cardiac and hemodynamic events. It is known that troponins are often elevated in HD patients, and high levels of troponin are associated with increased mortality. Therefore, it is difficult to interpret the value of elevations in chronic kidney disease patients. Echocardiographic and biochemical parameters of 41 patients treated with HD were evaluated before and after a HD session. HD led to an increased heart rate, and tissue Doppler imaging parameters such as early diastolic mitral peak velocity (E)/early diastolic myocardial peak velocity (é) and septal é decreased significantly after HD. HD caused an increase in troponin I, myoglobin and cardiac creatine kinase (CK MB) levels (p = 0.019, p < 0.001 and p = 0.018, respectively). A decrease in the left ventricular peak systolic myocardial (LV S') velocity (p = 0.011) was detected in patients with increased levels of cardiac damage markers (group 2) compared to those without increased levels of cardiac damage markers (group 1) in HD. A decrease in LV S' velocity was found to be an independent predictor of an increase of myocardial injury enzymes in HD (odds ratio = 1.099; p = 0.039). We concluded that HD may lead to significant acute stress upon the myocardium.


Secreted proteins of Neisseria meningitidis protect mice against infection.
We addressed the hypothesis that meningococcal secreted proteins (MSPs) can elicit protective immunity against meningococcal disease. Endotoxin-depleted MSP preparations were used to immunise a group of 15 six-week-old BALB/c mice (25microg MSPs/dose mixed with Freund's complete adjuvant) on days 0, 14 and 21. Mice were challenged 2 weeks later with 10(7) colony forming units of live Neisseria meningitidis strain MC58 (serogroup B, ET-5). Negative and positive control groups of 15 mice each were injected with adjuvant only, or a live attenuated strain of MC58, respectively. Seven out of 15 mice (47%) from the negative control group died after 72h of challenge, whereas none of test or positive control group died. Protection afforded by the anti-MSP immune response can be at least partly attributed to complement-mediated bacterial lysis, detectable in vitro using the serum of immunised mice. Murine anti-MC58 MSP sera were bactericidal against homologous and five unrelated ET-5 serogroup B strains. However, failed to kill strains from other hypervirulent clonal lineages belonging to the same or different serogroups, despite the presence of cross-reactive antibodies detectable by immunoblotting. Similar sera raised against MSPs from an isolate belonging to the ET-37 electropherotype lineage were bactericidal against all tested isolates of this lineage and, in addition, against some but not all isolates belonging to the ET-5 lineage. FACS analysis of intact bacteria treated with anti-MSPs confirmed surface-binding of antibodies.


The effects of corneal endothelium on graft survival in a murine model of lamellar keratoplasty.
Here we investigate the role of donor endothelium on allograft rejection in a lamellar keratoplasty (LK) model using grafts with or without donor endothelium. Corneal buttons of donor C57BL/6 mice (2.0 mm) were transplanted to lamellar recipient beds (1.5 mm) in BALB/c mice. Two variations of the LK procedure were performed: (1) standard LK (SLK) (n = 13) without donor endothelium and (2) modified LK (MLK) (n = 14) with retained donor endothelium. The graft status was assessed by slit lamp biomicroscopy and scored for stromal opacity, corneal edema, neovascularization, and anterior chamber reaction up to 46 days post-transplantation. Corneas were also observed histologically. The presence of a grafted corneal endothelium promoted graft rejection; 92.9% (13/14) of grafts were rejected in MLK after an average of 8.3 days, while 69.2% (9/13) of grafts were rejected in SLK on average 10.8 days after transplantation. The former's stromal opacity was significantly greater at all time points after day 14 except for day 21 (p = 0.77) and day 32 (p = 0.25). Corneal edema was significantly greater in the former at all time points after day 10 except for day 21 (p = 0.16). Neovascularization was significantly greater in the former at all time points after day 10 except for day 25 (p = 0.22). Variations of this model of LK may be useful for studies of immunological mechanisms in corneal transplantation. The donor corneal endothelium may serve as a target of the immune response which promotes inflammation, neovascularization, and graft rejection.


Control of risk factors for cardiovascular disease among adults with previously diagnosed type 2 diabetes mellitus: a descriptive study from a middle eastern arab population.
Despite the high burden of type 2 diabetes mellitus (T2DM) in Oman, there are scarce data from a nationally representative sample on the level of glycaemia and other cardiovascular (CVD) risk factor control. To estimate the proportion of patients with T2DM at goal for glycaemia and CVD risk factors using the National Diabetes Guidelines (NDG) and the American Diabetes Association (ADA) clinical care guidelines; and to assess the quality of selected services provided to patients with T2DM. A sample of 2,551 patients (47% men) aged ≥20 years with T2DM treated at primary health care centers was selected. Patient characteristics, medical history and treatment were collected from case notes, Diabetes Registers and computer frameworks including the use of the last 3 laboratory investigations results and blood pressure (BP) readings recorded in 2007. The overall mean age of the cohort was 54±13 years with an average median duration of diabetes of 4 (range 2 to 6) years. Over 80% of patients were overweight or obese (body mass index (BMI) of ≥25 Kg/m(2)). Sixty-nine percent were on oral anti-diabetic medication, 52% on anti-hypertensives and 40% on lipid lowering drugs. Thirty percent of patients were at goal for glycosylated haemoglobin level (<7%), 26% for BP (systolic/diastolic <130/80 mmHg), 55% for total cholesterol (<5.2 mmol/l), 4.5% for low-density lipoprotein cholesterol (<1.8 mmol/l), 52% for high-density lipoprotein cholesterol (>1 mmol/l for men, >1.3 mmol/l for women), and 61% for triglycerides (<1.7 mmol/l). Over 37% had micro-albuminuria and 5% had diabetic nephropathy. Control of hyperglycaemia and other CVD risk factor appears to be suboptimal in Omani patients with T2DM and need to be addressed in the triad of patient, physician and health system.


The effects of ring annuloplasty on mitral leaflet geometry during acute left ventricular ischemia.
The perturbed mitral leaflet geometry that leads to acute ischemic mitral regurgitation during acute left ventricular ischemia has not been quantified, nor is it known whether annuloplasty rings affect these detrimental changes in leaflet geometry. Radiopaque markers were implanted on both mitral leaflets and around the anulus in 3 groups of sheep: one group without rings served as the control group (n = 7); the others underwent Duran (n = 6; Medtronic Heart Valve Division, Minneapolis, Minn) or Carpentier-Edwards Physio (n = 5; Baxter Cardiovascular Division, Santa Ana, Calif) ring annuloplasty. After recovery, 3-dimensional marker coordinates were obtained by means of biplane videofluoroscopy before and during acute posterolateral left ventricular ischemia. Leaflet geometry was defined by measuring distances between annular and leaflet markers and perpendicular distances to the leaflet markers from a best-fit annular plane. In all control animals, left ventricular ischemia was associated with acute ischemic mitral regurgitation and apical displacement (away from the annular plane) of the posterior leaflet edge and base markers by 0.6 +/- 0.4 mm (P =.01) and 0.7 +/- 0.2 mm (P <.001), respectively. The distance between the posterior leaflet markers and the mid-posterior anulus did not change significantly during ischemia. The anterior leaflet edge marker extended 1.0 +/- 0. 5 mm (P =.01) away from the mid-anterior anulus during ischemia, but compared with its nonischemic position, the anterior leaflet was not displaced apically away from the annular plane. In all animals in the Duran and Physio groups, leaflet geometry was unchanged during ischemia, and acute ischemic mitral regurgitation was not detected. Acute ischemic mitral regurgitation was associated with restricted motion of the posterior leaflet and extension of the anterior leaflet. Annuloplasty rings prevented these geometric perturbations of the mitral leaflets during acute left ventricular ischemia and preserved valvular competence.


Stage-specific signaling through TGFβ family members and WNT regulates patterning and pancreatic specification of human pluripotent stem cells.
The generation of insulin-producing β-cells from human pluripotent stem cells is dependent on efficient endoderm induction and appropriate patterning and specification of this germ layer to a pancreatic fate. In this study, we elucidated the temporal requirements for TGFβ family members and canonical WNT signaling at these developmental stages and show that the duration of nodal/activin A signaling plays a pivotal role in establishing an appropriate definitive endoderm population for specification to the pancreatic lineage. WNT signaling was found to induce a posterior endoderm fate and at optimal concentrations enhanced the development of pancreatic lineage cells. Inhibition of the BMP signaling pathway at specific stages was essential for the generation of insulin-expressing cells and the extent of BMP inhibition required varied widely among the cell lines tested. Optimal stage-specific manipulation of these pathways resulted in a striking 250-fold increase in the levels of insulin expression and yielded populations containing up to 25% C-peptide+ cells.


Effects of phenolic monoamines on release of luteinizing hormone stimulated by gonadotropin-releasing hormone and on plasma adrenocorticotropic hormone, norepinephrine, and cortisol concentrations in wethers.
Eighteen Suffolk and Suffolk x Hampshire wethers (56.3 +/- 1.3 kg) were used to determine the effects of naturally occurring amines, N-methyl-beta-phenethylamine (NMP) and tyramine (T), on plasma cortisol, norepinephrine (NE), ACTH, and GnRH-stimulated LH concentrations. In each experiment, wethers were assigned randomly to treatments with six replicates/treatment. In Exp. 1, treatments consisted of saline, 2 mg of NMP/kg BW (NMP2), or 4 mg of NMP/kg BW (NMP4). In Exp. 2, treatments consisted of saline, 4 mg of NMP/kg BW (N), or 2 mg of NMP + 4 mg of T/kg BW (NT). All treatments were given via indwelling jugular catheters in single doses of 2 mL. In Exp. 1, no effects of time after treatment on response were detected. The NMP2 wethers had lower cortisol and LH peak amplitudes (PKAMP) than other wethers (P < .03 and .05, respectively). Time to peak (PKT) concentration of NE was significantly shorter in NMP4-treated wethers (P < .004). In Exp. 2, significant interactions between time and treatment were detected for cortisol, NE, and LH. Cortisol and NE concentrations were increased (P < .0001 and P < .03, respectively) in N- and NT-treated wethers compared with controls, whereas LH was reduced (P < .003). The N and NT treatments increased cortisol PKT, peak height (PKHT), and PKAMP above that of controls (P < .02, < .006, and < .02, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)


Investigating the genetic architecture of dementia with Lewy bodies: a two-stage genome-wide association study.
Dementia with Lewy bodies is the second most common form of dementia in elderly people but has been overshadowed in the research field, partly because of similarities between dementia with Lewy bodies, Parkinson's disease, and Alzheimer's disease. So far, to our knowledge, no large-scale genetic study of dementia with Lewy bodies has been done. To better understand the genetic basis of dementia with Lewy bodies, we have done a genome-wide association study with the aim of identifying genetic risk factors for this disorder. In this two-stage genome-wide association study, we collected samples from white participants of European ancestry who had been diagnosed with dementia with Lewy bodies according to established clinical or pathological criteria. In the discovery stage (with the case cohort recruited from 22 centres in ten countries and the controls derived from two publicly available database of Genotypes and Phenotypes studies [phs000404.v1.p1 and phs000982.v1.p1] in the USA), we performed genotyping and exploited the recently established Haplotype Reference Consortium panel as the basis for imputation. Pathological samples were ascertained following autopsy in each individual brain bank, whereas clinical samples were collected after participant examination. There was no specific timeframe for collection of samples. We did association analyses in all participants with dementia with Lewy bodies, and also only in participants with pathological diagnosis. In the replication stage, we performed genotyping of significant and suggestive results from the discovery stage. Lastly, we did a meta-analysis of both stages under a fixed-effects model and used logistic regression to test for association in each stage. This study included 1743 patients with dementia with Lewy bodies (1324 with pathological diagnosis) and 4454 controls (1216 patients with dementia with Lewy bodies vs 3791 controls in the discovery stage; 527 vs 663 in the replication stage). Results confirm previously reported associations: APOE (rs429358; odds ratio [OR] 2·40, 95% CI 2·14-2·70; p=1·05 × 10-48), SNCA (rs7681440; OR 0·73, 0·66-0·81; p=6·39 × 10-10), an GBA (rs35749011; OR 2·55, 1·88-3·46; p=1·78 × 10-9). They also provide some evidence for a novel candidate locus, namely CNTN1 (rs7314908; OR 1·51, 1·27-1·79; p=2·32 × 10-6); further replication will be important. Additionally, we estimate the heritable component of dementia with Lewy bodies to be about 36%. Despite the small sample size for a genome-wide association study, and acknowledging the potential biases from ascertaining samples from multiple locations, we present the most comprehensive and well powered genetic study in dementia with Lewy bodies so far. These data show that common genetic variability has a role in the disease. The Alzheimer's Society and the Lewy Body Society.


Optical constants of absorbing materials: a new approach.
A detailed study of a new method of determining the optical properties of absorbing materials is presented. It makes use of normal incidence reflectances from the specimen itself (R(s)) and from the specimen coated with a transparent film of two different thicknesses but of the same refractive index (R(1s) and R(2s)) in the form of R(1s)/R(s) and R(2s)/R(s)- It is seen that a simple goniometer can be easily adopted for measuring the reflectance ratios over a wide spectral range. The versatility of the method has been proved by the fact that it has been successfully adopted for specimens with surface structures varying from atomically smooth to rough surfaces.


Identification of EEG events in the MR scanner: the problem of pulse artifact and a method for its subtraction.
Triggering functional MRI (fMRI) image acquisition immediately after an EEG event can provide information on the location of the event generator. However, EEG artifact associated with pulsatile blood flow in a subject inside the scanner may obscure EEG events. This pulse artifact (PA) has been widely recognized as a significant problem, although its characteristics are unpredictable. We have investigated the amplitude, distribution on the scalp, and frequency of occurrence of this artifact. This showed large interindividual variations in amplitude, although PA is normally largest in the frontal region. In five of six subjects, PA was greater than 50 microV in at least one of the temporal, parasagittal, and central channels analyzed. Therefore, we developed and validated a method for removing PA. This subtracts an averaged PA waveform calculated for each electrode during the previous 10 s. Particular attention has been given to reliable ECG peak detection and ensuring that the average PA waveform is free of other EEG artifacts. Comparison of frequency spectra for EEG recorded outside and inside the scanner, with and without PA subtraction, showed a clear reduction in artifact after PA subtraction for all four frequency ranges analyzed. As further validation, lateralized epileptiform spikes were added to recordings from inside and outside the scanner: PA subtraction significantly increased the proportion of these spikes that were correctly identified and decreased the number of false spike detections. We conclude that in some subjects, EEG/fMRI studies will be feasible only using PA subtraction.


Insights into the Evolution of Host Association through the Isolation and Characterization of a Novel Human Periodontal Pathobiont, Desulfobulbus oralis.
The human oral microbiota encompasses representatives of many bacterial lineages that have not yet been cultured. Here we describe the isolation and characterization of previously uncultured Desulfobulbus oralis, the first human-associated representative of its genus. As mammalian-associated microbes rarely have free-living close relatives, D. oralis provides opportunities to study how bacteria adapt and evolve within a host. This sulfate-reducing deltaproteobacterium has adapted to the human oral subgingival niche by curtailing its physiological repertoire, losing some biosynthetic abilities and metabolic independence, and by dramatically reducing environmental sensing and signaling capabilities. The genes that enable free-living Desulfobulbus to synthesize the potent neurotoxin methylmercury were also lost by D. oralis, a notably positive outcome of host association. However, horizontal gene acquisitions from other members of the microbiota provided novel mechanisms of interaction with the human host, including toxins like leukotoxin and hemolysins. Proteomic and transcriptomic analysis revealed that most of those factors are actively expressed, including in the subgingival environment, and some are secreted. Similar to other known oral pathobionts, D. oralis can trigger a proinflammatory response in oral epithelial cells, suggesting a direct role in the development of periodontal disease.IMPORTANCE Animal-associated microbiota likely assembled as a result of numerous independent colonization events by free-living microbes followed by coevolution with their host and other microbes. Through specific adaptation to various body sites and physiological niches, microbes have a wide range of contributions, from beneficial to disease causing. Desulfobulbus oralis provides insights into genomic and physiological transformations associated with transition from an open environment to a host-dependent lifestyle and the emergence of pathogenicity. Through a multifaceted mechanism triggering a proinflammatory response, D. oralis is a novel periodontal pathobiont. Even though culture-independent approaches can provide insights into the potential role of the human microbiome "dark matter," cultivation and experimental characterization remain important to studying the roles of individual organisms in health and disease.


Drug induction of hepatic glutathione S-transferases in male and female rats*.
The induction of the glutathione S-transferases by phenobarbital and polycyclic hydrocarbons was studied in male and female rats. Administration of phenobarbital resulted in 60-80% increase in S-aryl and S-aralkyl enzyme specific activities, whereas the S-epoxide and S-alkyl activities were increased by 30-40%. In following the sequence of induction, the former two activities were noted to reach peak activities before an increase in the latter two activities was observed. Both 3-methylcholanthrene and 3,4-benzopyrene were shown toi nduce these four enzymic activities, although without the discrimination between pairs of activities noted with phenobarbital. No change in Km accompanied the increase in Vmax. after induction by drugs, and no change occurred in Ki for sulphobromophthalein inhibition. Significantly lower enzyme specific activities were found for three of the activities studied in female rats but no difference was observed in the S-alkyltransferase activity. However, the proportional increase in the enzymic activities in response to phenobarbital was the same in males and females. These studies demonstrate the drug induction of a group of cytosolic drug-metabolizing enzymes as well as the identification of sex differences in these activities.


Pupil Dilation Progression Modulates Aberrant Social Cognition in Autism Spectrum Disorder.
Progression of pupil dilation (PD) in response to visual stimuli may indicate distinct internal processes. No study has been performed on PD progression during a social cognition task. Here, we describe PD progression during the Movie for the Assessment of Social Cognition (MASC) test in n = 23 adolescents with Autism Spectrum Disorder (ASD) and n = 24 age, IQ and sex-matched neurotypical controls (NTC). The MASC consists of 43 video sequences depicting human social interactions, each followed by a multiple-choice question concerning characters' mental states. PD progression data were extracted by eye tracking and controlled for fixation behavior. Segmenting PD progression during video sequences by principal component analysis, three sequential PD components were unveiled. In ASD compared with NTC, a distinct PD progression was observed with increased constriction amplitude, increased dilation latency, and increased dilation amplitude that correlated with PD progression components. These components predicted social cognition performance. The first and second PD components correlated positively with MASC behavioral performance in ASD but negatively in NTC. These PD components may be interpreted as indicators of sensory-perceptual processing and attention function. In ASD, aberrant sensory-perceptual processing and attention function could contribute to attenuated social cognition performance. This needs to be tested by additional studies combining the respective cognitive tests and the outlined PD progression analysis. Phasic activity of the locus coeruleus-norepinephrine system is discussed as putatively shared underlying mechanism. Autism Res2019. © 2019 The Authors. Autism Research published by International Society for Autism Research published by Wiley Periodicals, Inc. LAY SUMMARY: In adolescents with autism, we found an altered pupil dilation during watching scenes of human interactions. Early pupil dilation correlated positively with the number of correct answers to questions about the shown human interactions. Our findings suggest that aberrant sensory processing and attention function may contribute to altered social cognition in autism.


A fluorescent dipyrrinone oxime for the detection of pesticides and other organophosphates.
An N,N-carbonyl-bridged dipyrrinone oxime has been synthesized and studied as a potential sensor for organophosphates. The molecular sensor underwent a drastic colorimetric response upon formation of the adduct. The pesticide dimethoate was found to produce the biggest spectral response, with a limit of detection equal to 4.0 ppm using UV-visible spectroscopy. Minimal fluorescence "turn on" via a PET mechanism was seen, and molecular modeling studies were used to explain the lower than expected PET response. The X-ray crystal structure of the fluorescent dipyrrinone oxime was also obtained.


The decline of the impetigo epidemic caused by the epidemic European fusidic acid-resistant impetigo clone: an 11.5-year population-based incidence study from a community in Western Norway.
From around the year 2000, Northern Europe experienced a rise in impetigo caused by Staphylococcus aureus resistant to fusidic acid. A single clone of S. aureus was found to be the bacterial pathogen involved in the impetigo outbreak in Norway, Sweden, the UK and Ireland, termed 'the epidemic European fusidic acid-resistant impetigo clone' (EEFIC). We have followed the incidence of impetigo during the years 2001-2012 based on all patients in general practice in the island community of Austevoll, Western Norway. We previously reported a marked decline of impetigo incidence in Austevoll, from 0.0260 cases per person-year in 2002 to 0.0038 in 2009. This article explores indications of an end to the impetigo epidemic caused by the EEFIC clone. All four general practitioners (GPs) in the community (mean population = 4400) were asked to diagnose impetigo in a uniform way and to take bacterial specimens from all impetigo cases. Phenotypic characteristics of specimen bacteria were determined for the whole period and molecular analyses were performed on isolates in the period 2008-2012. We observed a further decline in incidence of impetigo in Austevoll in the study period. The proportion of fusidic acid-resistant S. aureus isolates decreased during the period 2002-2012, with a mean of 80% in the epidemic years of 2002-2004, 55% in 2005-2009, and 6% in 2010-2012. In total, 44 S. aureus isolates from impetigo were subject to molecular analyses in the period 2008-2012, and 11 were found to be related to the EEFIC. All EEFIC isolates were found in 2008-2009, with no new isolates in 2010-2012. There is an apparent end to the impetigo epidemic related to the EEFIC in this population in Western Norway.


Radiation recall myelitis following paclitaxel chemotherapy: The first reported case.
Stereotactic body radiotherapy (SBRT) of the spine has become an increasingly utilized modality in the United States, most commonly for metastatic disease (McClelland et al., 2017). Spinal SBRT in patients with spinal instrumentation has been sparsely examined. We report a patient who developed myelitis following spinal SBRT to a region with existing hardware. A 55-year-old woman with Stage IV breast cancer developed a T4 vertebral body metastasis and underwent tumor debulking with posteriorly instrumented T3-T5 fusion. Postoperatively she proceeded with SBRT to the T3-T5 vertebral bodies, receiving 30 Gy in 6 Gy/fraction. Seven months later, she required paclitaxel chemotherapy (80 mg/m2 per cycle) for new liver metastases. Eight months following spine SBRT, four weeks after having started chemotherapy she developed intractable back pain and right lower extremity numbness which improved upon receiving steroids for weekly chemotherapy; the numbness subsequently spread to her left leg. Thoracic spine MRI revealed a 1.7 cm ovoid focus of T4-T5 spinal cord enhancement with extensive surrounding cord edema extending superiorly to C6-C7, consistent with radiation myelitis. Hyperbaric oxygen moderately improved her symptoms; fortunately, she never developed motor symptomatology or bowel/bladder dysfunction. Thorough re-evaluation of the original thoracic spine SBRT plan revealed no deviations from the standard of care, nor did re-planning with alternate treatment planning software demonstrate any significant difference in maximum cord dosage than the original plan. The timing of symptomatology related to chemotherapy administration is consistent with radiation recall myelitis, which has yet to be reported following SBRT. Given the potentially disastrous consequences of myelitis, patients with metastatic disease previously treated with spine SBRT may be susceptible to developing myelitis if treated with paclitaxel chemotherapy.


Physiological and medical monitoring for en route care of combat casualties.
Most prehospital medical interventions during civilian and military trauma casualty transport fail to utilize advanced decision-support systems for treatment and delivery of medical interventions, particularly intravenous fluids and oxygen. Current treatment protocols are usually based on standard vital signs (eg, blood pressure, arterial oxygen saturation) which have proven to be of limited value in detecting the need to implement an intervention before cardiovascular collapse. A primary objective of the US Army combat casualty care research program is to reduce mortality and morbidity during casualty transport from the battlefield through advanced development of a semiautomated decision-support capability for closed-loop resuscitation and oxygen delivery. To accomplish this goal, the Trauma Informatics Research Team at the US Army Institute of Surgical Research has developed two models for evidence-based decision support 1) a trauma patient database for capture and analysis of prehospital vital signs for identification of early, novel physiologic measurements that could improve the control of closed-loop systems in trauma patients; and, 2) a human experimental model of central hypovolemia using lower body negative pressure to improve the understanding and identification of physiologic signals for advancing closed-loop capabilities with simulated hemodynamic responses to hemorrhage. In the trauma patient database and lower body negative pressure studies, traditional vital sign measurements such as systolic blood pressure and oxygen saturation fail to predict mortality or indicate the need for life saving interventions or reductions in central blood volume until after the onset of cardiovascular collapse. We have evidence from preliminary analyses, however, that indicators of reduced central blood volume in the presence of stable vital signs include 1) reductions in pulse pressure; 2) changes in indices of autonomic balance derived from calculation of heart period variability (ie, linear and non-linear analyses of R-R intervals); and 3) reductions in tissue oxygenation. We propose that derived indices based on currently available technology for continuous monitoring of specific hemodynamic, autonomic, and/or metabolic responses could provide earlier recognition of hemorrhage than current standard vital signs and allow intervention before the onset of circulatory shock. Because of this, such indices could provide improved feedback for closed-loop control of patient resuscitation and oxygen delivery. These technological advances could prove instrumental in advancing decision-support capabilities for prehospital trauma care during transport to higher levels of care in both the military and civilian environments.


Knowing loved ones' end-of-life health care wishes: attachment security predicts caregivers' accuracy.
At times, caregivers make life-and-death decisions for loved ones. Yet very little is known about the factors that make caregivers more or less accurate as surrogate decision makers for their loved ones. Previous research suggests that in low stress situations, individuals with high attachment-related anxiety are attentive to their relationship partners' wishes and concerns, but get overwhelmed by stressful situations. Individuals with high attachment-related avoidance are likely to avoid intimacy and stressful situations altogether. We hypothesized that both of these insecure attachment patterns limit surrogates' ability to process distressing information and should therefore be associated with lower accuracy in the stressful task of predicting their loved ones' end-of-life health care wishes. Older patients visiting a medical clinic stated their preferences toward end-of-life health care in different health contexts, and surrogate decision makers independently predicted those preferences. For comparison purposes, surrogates also predicted patients' perceptions of everyday living conditions so that surrogates' accuracy of their loved ones' perceptions in nonstressful situations could be assessed. Surrogates high on either type of insecure attachment dimension were less accurate in predicting their loved ones' end-of-life health care wishes. It is interesting to note that even though surrogates' attachment-related anxiety was associated with lower accuracy of end-of-life health care wishes of their loved ones, it was associated with higher accuracy in the nonstressful task of predicting their loved ones' everyday living conditions. Attachment orientation plays an important role in accuracy about loved ones' end-of-life health care wishes. Interventions may target emotion regulation strategies associated with insecure attachment orientations.


A statistical approach to understanding reproductive isolation in two sympatric species of tree crickets.
In acoustically communicating animals, reproductive isolation between sympatric species is usually maintained through species-specific calls. This requires that the receiver be tuned to the conspecific signal. Mapping the response space of the receiver onto the signal space of the conspecific investigates this tuning. A combinatorial approach to investigating the response space is more informative as the influence on the receiver of the interactions between the features is also elucidated. However, most studies have examined individual preference functions rather than the multivariate response space. We studied the maintenance of reproductive isolation between two sympatric tree cricket species (Oecanthus henryi and Oecanthus indicus) through the temporal features of the calls. Individual response functions were determined experimentally for O. henryi, the results from which were combined in a statistical framework to generate a multivariate quantitative receiver response space. The predicted response was higher for the signals of the conspecific than for signals of the sympatric heterospecific, indicating maintenance of reproductive isolation through songs. The model allows prediction of response to untested combinations of temporal features as well as delineation of the evolutionary constraints on the signal space. The model can also be used to predict the response of O. henryi to other heterospecific signals, making it a useful tool for the study of the evolution and maintenance of reproductive isolation via long-range acoustic signals.


Enzyme plus light therapy to repair DNA damage in ultraviolet-B-irradiated human skin.
Ultraviolet-B (UVB) (290-320 nm) radiation-induced cyclobutane pyrimidine dimers within the DNA of epidermal cells are detrimental to human health by causing mutations and immunosuppressive effects that presumably contribute to photocarcinogenesis. Conventional photoprotection by sunscreens is exclusively prophylactic in nature and of no value once DNA damage has occurred. In this paper, we have therefore assessed whether it is possible to repair UVB radiation-induced DNA damage through topical application of the DNA-repair enzyme photolyase, derived from Anacystis nidulans, that specifically converts cyclobutane dimers into their original DNA structure after exposure to photoreactivating light. When a dose of UVB radiation sufficient to induce erythema was administered to the skin of healthy subjects, significant numbers of dimers were formed within epidermal cells. Topical application of photolyase-containing liposomes to UVB-irradiated skin and subsequent exposure to photoreactivating light decreased the number of UVB radiation-induced dimers by 40-45%. No reduction was observed if the liposomes were not filled with photolyase or if photoreactivating exposure preceded the application of filled liposomes. The UVB dose administered resulted in suppression of intercellular adhesion molecule-1 (ICAM-1), a molecule required for immunity and inflammatory events in the epidermis. In addition, in subjects hypersensitive to nickel sulfate, elicitation of the hypersensitivity reaction in irradiated skin areas was prevented. Photolyase-induced dimer repair completely prevented these UVB radiation-induced immunosuppressive effects as well as erythema and sunburn-cell formation. These studies demonstrate that topical application of photolyase is effective in dimer reversal and thereby leads to immunoprotection.


Somatostatin release, electrical activity, membrane currents and exocytosis in human pancreatic delta cells.
The aim of this study was to characterise electrical activity, ion channels, exocytosis and somatostatin release in human delta cells/pancreatic islets. Glucose-stimulated somatostatin release was measured from intact human islets. Membrane potential, currents and changes in membrane capacitance (reflecting exocytosis) were recorded from individual human delta cells identified by immunocytochemistry. Somatostatin secretion from human islets was stimulated by glucose and tolbutamide and inhibited by diazoxide. Human delta cells generated bursting or sporadic electrical activity, which was enhanced by tolbutamide but unaffected by glucose. Delta cells contained a tolbutamide-insensitive, Ba(2+)-sensitive inwardly rectifying K(+) current and two types of voltage-gated K(+) currents, sensitive to tetraethylammonium/stromatoxin (delayed rectifying, Kv2.1/2.2) and 4-aminopyridine (A current). Voltage-gated tetrodotoxin (TTX)-sensitive Na(+) currents contributed to the action potential upstroke but TTX had no effect on somatostatin release. Delta cells are equipped with Ca(2+) channels blocked by isradipine (L), omega-agatoxin (P/Q) and NNC 55-0396 (T). Blockade of any of these channels interferes with delta cell electrical activity and abolishes glucose-stimulated somatostatin release. Capacitance measurements revealed a slow component of depolarisation-evoked exocytosis sensitive to omega-agatoxin. Action potential firing in delta cells is modulated by ATP-sensitive K(+)-channel activity. The membrane potential is stabilised by Ba(2+)-sensitive inwardly rectifying K(+) channels. Voltage-gated L- and T-type Ca(2+) channels are required for electrical activity, whereas Na(+) currents and P/Q-type Ca(2+) channels contribute to (but are not necessary for) the upstroke of the action potential. Action potential repolarisation is mediated by A-type and Kv2.1/2.2 K(+) channels. Exocytosis is tightly linked to Ca(2+)-influx via P/Q-type Ca(2+) channels. Glucose stimulation of somatostatin secretion involves both K(ATP) channel-dependent and -independent processes.


Binding properties of YjeQ (RsgA), RbfA, RimM and Era to assembly intermediates of the 30S subunit.
Our understanding regarding the function of YjeQ (also called RsgA), RbfA, RimM and Era in ribosome biogenesis has been derived in part from the study of immature 30S particles that accumulate in null strains lacking one of these factors. However, their mechanistic details are still unknown. Here, we demonstrate that these immature particles are not dead-end products of assembly, but progress into mature 30S subunits. Mass spectrometry analysis revealed that in vivo the occupancy level of these factors in these immature 30S particles is below 10% and that the concentration of factors does not increase when immature particles accumulate in cells. We measured by microscale thermophoresis that YjeQ and Era binds to the mature 30S subunit with high affinity. However, the binding affinity of these factors to the immature particles and of RimM and RbfA to mature or immature particles was weak, suggesting that binding is not occurring at physiological concentrations. These results suggest that in the absence of these factors, the immature particles evolve into a thermodynamically stable intermediate that exhibits low affinity for the assembly factors. These results imply that the true substrates of YjeQ, RbfA, RimM and Era are immature particles that precede the ribosomal particles accumulating in the knockouts strains.


In vivo calcium deposition on polyvinyl alcohol matrix used in hollow fiber cell macroencapsulation devices.
The encapsulation of genetically modified cells represents a promising approach for the delivery of therapeutic proteins. The functionality of the device is dependent on the characteristics of the biomaterials, the procedures used in its confection and the adaptability of the encapsulated cells in the host. We report conditions leading to the development of calcifications on the polyvinyl alcohol (PVA) matrix introduced in hollow fiber devices for the encapsulation of primary human fibroblasts implanted in mice. The manufacturing procedures, batches of PVA matrix and cell lineages were assessed for their respective role in the development of the phenomenon. The results showed that the calcification is totally prevented by substituting phosphate-buffer saline with ultra-pure sterile water in the rinsing procedure of the matrix. Moreover, a positive correlation was found, when comparing two fibroblast cell lineages, between the level of lactate dehydrogenase (LDH) activity measured in the cells and the degree of calcium deposition. Higher LDH activity may decrease calcium depositions because it generates in the device a more acidic microenvironment inhibiting calcium precipitation. The present study defines optimized conditions for the encapsulation of primary human fibroblasts in order to avoid potentially detrimental calcifications and to allow long-term survival of encapsulated cells.


Folinic acid treatment for schizophrenia associated with folate receptor autoantibodies.
Auto-antibodies against folate receptor alpha (FRα) at the choroid plexus that block N(5)-methyltetrahydrofolate (MTHF) transfer to the brain were identified in catatonic schizophrenia. Acoustic hallucinations disappeared following folinic acid treatment. Folate transport to the CNS prevents homocysteine accumulation and delivers one-carbon units for methyl-transfer reactions and synthesis of purines. The guanosine derivative tetrahydrobiopterin acts as common co-factor for the enzymes producing dopamine, serotonin and nitric oxide. Our study selected patients with schizophrenia unresponsive to conventional treatment. Serum from these patients with normal plasma homocysteine, folate and vitamin B12 was tested for FR autoantibodies of the blocking type on serial samples each week. Spinal fluid was analyzed for MTHF and the metabolites of pterins, dopamine and serotonin. The clinical response to folinic acid treatment was evaluated. Fifteen of 18 patients (83.3%) had positive serum FR auto-antibodies compared to only 1 in 30 controls (3.3%) (χ(2)=21.6; p<0.0001). FRα antibody titers in patients fluctuated over time varying between negative and high titers, modulating folate flux to the CNS, which explained low CSF folate values in 6 and normal values in 7 patients. The mean±SD for CSF MTHF was diminished compared to previously established controls (t-test: 3.90; p=0.0002). A positive linear correlation existed between CSF MTHF and biopterin levels. CSF dopamine and serotonin metabolites were low or in the lower normal range. Administration of folinic acid (0.3-1mg/kg/day) to 7 participating patients during at least six months resulted in clinical improvement. Assessment of FR auto-antibodies in serum is recommended for schizophrenic patients. Clinical negative or positive symptoms are speculated to be influenced by the level and evolution of FRα antibody titers which determine folate flux to the brain with up- or down-regulation of brain folate intermediates linked to metabolic processes affecting homocysteine levels, synthesis of tetrahydrobiopterin and neurotransmitters. Folinic acid intervention appears to stabilize the disease process.


Quantitation of anti-Pythium insidiosum antibodies before and after administration of an immunotherapeutic product to healthy dogs.
OBJECTIVE To evaluate the effect of an immunotherapeutic product on concentrations of anti-Pythium insidiosum antibodies in dogs. ANIMALS 7 healthy hound-crossbreds. PROCEDURES Antibody concentrations were evaluated before (day 0) and after administration of the immunotherapeutic product. The immunotherapeutic product was administered on days 0, 7, and 21. Serum was obtained on days 0, 7, 14, 21, 28, 35, 42, 49, and 56. Anti-P insidiosum antibody concentrations were measured and reported as the percentage positivity relative to results for a strongly positive control serum. RESULTS Mean ± SD percentage positivity before administration of the immunotherapeutic product was 7.45 ± 3.02%. There was no significant change in anti-P insidiosum antibody concentrations after administration of the product, with percentage positivity values in all dogs remaining within the range expected for healthy dogs (3% to 15%). CONCLUSIONS AND CLINICAL RELEVANCE Administration of the immunotherapeutic product to healthy dogs in accordance with the manufacturer's suggested protocol did not induce a significant change in anti-P insidiosum antibody concentrations. These results suggested that administration of the immunotherapeutic product may not interfere with postadministration serologic monitoring. However, further investigations will be required to determine whether there is a similar effect in naturally infected dogs.


The major human pregnane X receptor (PXR) splice variant, PXR.2, exhibits significantly diminished ligand-activated transcriptional regulation.
The pregnane X receptor (PXR; PXR.1) can be activated by structurally diverse lipophilic ligands. PXR.2, an alternatively spliced form of PXR, lacks 111 nucleotides encoding 37 amino acids in the ligand binding domain. PXR.2 bound a classic CYP3A4 PXR response element (PXRE) in electrophoretic mobility shift assays, but transfected PXR.2 failed to transactivate a CYP3A4-promoter-luciferase reporter plasmid in HepG2 cells treated with various PXR ligands. Cotransfection experiments showed that PXR.2 behaved as a dominant negative, interfering with PXR.1/rifampin activation of CYP3A4-PXRE-LUC. In HepG2 and LS180 cells stably transduced with PXR.1, PXR target genes (CYP3A4, MDR1, CYP2B6, and UGT1A1) were higher than mock-transduced cells in the absence of ligand and were further induced in the presence of rifampin. In contrast, PXR.2 stably introduced into the same host cells failed to induce target genes over levels in mock-transfected cells after drug treatment. Our homology modeling suggests that ligands bind PXR.1 more favorably, probably because of the presence of a key disordered loop region, which is missing in PXR.2. Yeast two-hybrid assays revealed that, even in the presence of ligand, the corepressors remain tightly bound to PXR.2, and coactivators are unable to bind at helix 12. In summary, PXR.2 can bind to PXREs but fails to transactivate target genes because ligands do not bind the ligand binding domain of PXR.2 productively, corepressors remain tightly bound, and coactivators are not recruited to PXR.2.


A comparison of anti-type II collagen antibody titres and isotype profiles in pregnant and virgin DBA/1 mice with collagen-induced arthritis.
In order to discover whether changes in the nature of the humoral response to type II collagen occur during pregnancy we compared, by enzyme-linked immunosorbent assay (ELISA), the anti-type II collagen antibody titres and isotype profiles of pregnant and virgin DBA/1 mice immunized with type II collagen. Significant reductions in the mean clinical scores and anti-collagen antibody titres were observed during late pregnancy. Titres of all of the anti-collagen isotypes tended to be lower in pregnant than non-pregnant mice, although only in the case of IgG3 was there a significant difference between the two groups. We conclude that while pregnancy may cause changes to the isotype distribution of anti-collagen antibodies it is not possible on the basis of this study to relate such changes to the remission of arthritis observed during pregnancy.


Antigenic typing of Brazilian rabies virus samples isolated from animals and humans, 1989-2000.
Animal and human rabies samples isolated between 1989 and 2000 were typified by means of a monoclonal antibody panel against the viral nucleoprotein. The panel had been previously established to study the molecular epidemiology of rabies virus in the Americas. Samples were isolated in the Diagnostic Laboratory of the Pasteur Institute and in other rabies diagnostic centers in Brazil. In addition to the fixed virus samples CVS-31/96-IP, preserved in mouse brain, and PV-BHK/97, preserved in cell culture, a total of 330 rabies virus samples were isolated from dogs, cats, cattle, horses, bats, sheep, goat, swine, foxes, marmosets, coati and humans. Six antigenic variants that were compatible with the pre-established monoclonal antibodies panel were defined: numbers 2 (dog), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (vampire bat from Venezuela), 6 (Lasiurus cinereus) and Lab (reacted to all used antibodies). Six unknown profiles, not compatible with the panel, were also found. Samples isolated from insectivore bats showed the greatest variability and the most commonly isolated variant was variant-3 (Desmodus rotundus). These findings may be related to the existence of multiple independent transmission cycles, involving different bat species.


Towards linkage analysis with markers in linkage disequilibrium by graphical modelling.
We review recent developments of MCMC integration methods for computations on graphical models for two applications in statistical genetics: modelling allelic association and pedigree based linkage analysis. We discuss and illustrate estimation of graphical models from haploid and diploid genotypes, and the importance of MCMC updating schemes beyond what is strictly necessary for irreducibility. We then outline an approach combining these methods to compute linkage statistics when alleles at the marker loci are in linkage disequilibrium. Other extensions suitable for analysis of SNP genotype data in pedigrees are also discussed and programs that implement these methods, and which are available from the author's web site, are described. We conclude with a discussion of how this still experimental approach might be further developed.


Treatment of granuloma annulare with the 585 nm pulsed dye laser.
Granuloma annulare is a chronic asymptomatic dermatosis that is typically manifested by annular papules arising on the dorsa of the hands, feet, elbows, and knees. Multiple treatment modalities have been used with variable efficacy, but no known cure exists. Obtaining long-term resolution of involved areas is often a therapeutic challenge. We report the results of treatment of a patient with localized erythematous granuloma annulare with a 585 nm flashlamp-pumped pulsed dye laser. A single granuloma annulare plaque present on the left wrist for 3 years was treated on three occasions with a 585 nm flashlamp-pumped pulsed dye laser initially and repeated at months 5 and 13. After initial pulsed dye laser treatment, significant flattening and reduction of erythema were evident within the treated area. After a second treatment at month 5 and a third treatment at month 13, further improvement was evident, and long-term remission was achieved. Localized granuloma annulare may be difficult to manage, presenting a therapeutic challenge. Thorough understanding of available treatment options, their relative efficacies, and risks of complication can facilitate successful management of this chronic disease. Pulsed dye laser therapy may be an effective and relatively safe treatment option for erythematous granuloma annulare, although prospective studies in additional patients would be desirable. Improvement of the condition in a single patient does not imply that others might improve, and variability in response is more likely.


Near-infrared spectroscopy and spectral mapping of Jupiter and the Galilean satellites: results from Galileo's initial orbit.
The Near Infrared Mapping Spectrometer performed spectral studies of Jupiter and the Galilean satellites during the June 1996 perijove pass of the Galileo spacecraft. Spectra for a 5-micrometer hot spot on Jupiter are consistent with the absence of a significant water cloud above 8 bars and with a depletion of water compared to that predicted for solar composition, corroborating results from the Galileo probe. Great Red Spot (GRS) spectral images show that parts of this feature extend upward to 240 millibars, although considerable altitude-dependent structure is found within it. A ring of dense clouds surrounds the GRS and is lower than it by 3 to 7 kilometers. Spectra of Callisto and Ganymede reveal a feature at 4. 25 micrometers, attributed to the presence of hydrated minerals or possibly carbon dioxide on their surfaces. Spectra of Europa's high latitudes imply that fine-grained water frost overlies larger grains. Several active volcanic regions were found on Io, with temperatures of 420 to 620 kelvin and projected areas of 5 to 70 square kilometers.


Gastrointestinal stability of urolithins: an in vitro approach.
Urolithins are bioactive ellagitannin-derived metabolites showing a wide phenotypic variation in their production by the gut microbiota. This work represents a first in vitro step toward the development of new strategies focused on the oral supplementation of urolithins with the aim of overcoming their selective production and making their putative health benefits available for the whole population. In order to study their gastrointestinal stability, urolithin A, urolithin B, and urolithin B-glucuronide, as well as ellagic acid, were subjected to a simulated gastrointestinal digestion model consisting of oral, gastric, and pancreatic steps followed by a 24-h fecal fermentation. The effect of the entero-hepatic recirculation on urolithin B-glucuronide, a phase II metabolite, was also investigated. Urolithin B was the molecule able to resist to a greater extent the conditions of the gastrointestinal tract, while urolithin A and ellagic acid were drastically unstable during the colonic step. Conjugation with glucuronic acid, ideally occurring in the liver, conferred to urolithin B an increased stability, which may be interesting in the framework of entero-hepatic recirculation. This set of experiments lets hypothesize that orally supplemented urolithins may come into contact with the colonic epithelium and become accessible for uptake or exert local anti-inflammatory activity, overcoming the limitations of enterotypes unable to convert ellagitannins into these putatively beneficial metabolites.


Angiotensin II analogues and aldosterone secretion in sodium-deficient sheep.
1. The angiotensin analogues Sar1-Ala8-angiotensin II (AII), Sar1-Ile8-AII, Sar1-Leu8-AII, Sar1-Thr8-AII, [Des1-Asp]-Ile8-AII and [Des1-Asp]-Sar2-Ile8-AII and converting enzyme inhibitor (SQ 80221) infused by intra-adrenal arterial infusion had no effect on aldosterone secretion in sodium-deficient sheep at doses in excess of those shown to block exogenous angiotensin II or III infusion. 2. It is suggested that the intrinsic agonist activity of the analogues may fulfil the requirements for a permissive role for angiotensin in the aldosterone response to sodium deficiency.


A multivariate geostatistical framework for combining multiple indices of abundance for disease vectors and reservoirs: a case study of rattiness in a low-income urban Brazilian community.
A key requirement in studies of endemic vector-borne or zoonotic disease is an estimate of the spatial variation in vector or reservoir host abundance. For many vector species, multiple indices of abundance are available, but current approaches to choosing between or combining these indices do not fully exploit the potential inferential benefits that might accrue from modelling their joint spatial distribution. Here, we develop a class of multivariate generalized linear geostatistical models for multiple indices of abundance. We illustrate this novel methodology with a case study on Norway rats in a low-income urban Brazilian community, where rat abundance is a likely risk factor for human leptospirosis. We combine three indices of rat abundance to draw predictive inferences on a spatially continuous latent process, rattiness, that acts as a proxy for abundance. We show how to explore the association between rattiness and spatially varying environmental factors, evaluate the relative importance of each of the three contributing indices and assess the presence of residual, unexplained spatial variation, and identify rattiness hotspots. The proposed methodology is applicable more generally as a tool for understanding the role of vector or reservoir host abundance in predicting spatial variation in the risk of human disease.


Prediction of the severity of obstructive sleep apnea by anthropometric features via support vector machine.
To develop an applicable prediction for obstructive sleep apnea (OSA) is still a challenge in clinical practice. We apply a modern machine learning method, the support vector machine to establish a predicting model for the severity of OSA. The support vector machine was applied to build up a prediction model based on three anthropometric features (neck circumference, waist circumference, and body mass index) and age on the first database. The established model was then valided independently on the second database. The anthropometric features and age were combined to generate powerful predictors for OSA. Following the common practice, we predict if a subject has the apnea-hypopnea index greater then 15 or not as well as 30 or not. Dividing by genders and age, for the AHI threhosld 15 (respectively 30), the cross validation and testing accuracy for the prediction were 85.3% and 76.7% (respectively 83.7% and 75.5%) in young female, while the negative likelihood ratio for the AHI threhosld 15 (respectively 30) for the cross validation and testing were 0.2 and 0.32 (respectively 0.06 and 0.1) in young female. The more accurate results with lower negative likelihood ratio in the younger patients, especially the female subgroup, reflect the potential of the proposed model for the screening purpose and the importance of approaching by different genders and the effects of aging.


Feline immunodeficiency virus reverse transcriptase: expression, functional characterization, and reconstitution of the 66- and 51-kilodalton subunits.
The two subunits of the feline immunodeficiency virus (FIV) reverse transcriptase (RT) were cloned and functionally expressed in Escherichia coli. The recombinant proteins are enzymatically active as homodimers (p66 and p51) as well as a heterodimer p66/p51. The biochemical properties of the FIV RT are very similar to those of the counterpart of the human immunodeficiency virus type 1 in being an RNA-dependent and DNA-dependent DNA polymerase. When a double-stranded DNA containing a small gap of 26 nucleotides was tested, we found a new activity of the FIV RT p66/p51 heterodimer--the cat viral enzyme could perform strand displacement DNA synthesis of approximately 300 bases. The FIV RT homodimer p66 alone could carry out limited strand displacement DNA synthesis, but this activity was stimulated by the p51 subunit at a molar ratio of one molecule of p66 to five molecules of p51. On the other hand, the homodimeric p51 itself was unable to fill a small gap of 26 nucleotides in a double-stranded DNA substrate and was not active by itself in strand displacement DNA synthesis. These data are in agreement with an earlier finding of strand displacement DNA synthesis by human immunodeficiency virus type 1 RT (M. Hottiger, V.N. Podust, R.L. Thimmig, C.S. McHenry, and U. Hübscher. J. Biol. Chem. 269:986-991, 1994). Our data therefore suggest a general and important function of lentiviral p51 subunits in strand displacement DNA synthesis which appears to be required in later stages of the lentiviral replication cycle, when DNA-dependent DNA synthesis occurs on double-stranded DNA.


Compensation of PVT Variations in ToF Imagers with In-Pixel TDC.
The design of a direct time-of-flight complementary metal-oxide-semiconductor (CMOS) image sensor (dToF-CIS) based on a single-photon avalanche-diode (SPAD) array with an in-pixel time-to-digital converter (TDC) must contemplate system-level aspects that affect its overall performance. This paper provides a detailed analysis of the impact of process parameters, voltage supply, and temperature (PVT) variations on the time bin of the TDC array. Moreover, the design and characterization of a global compensation loop is presented. It is based on a phase locked loop (PLL) that is integrated on-chip. The main building block of the PLL is a voltage-controlled ring-oscillator (VCRO) that is identical to the ones employed for the in-pixel TDCs. The reference voltage that drives the master VCRO is distributed to the voltage control inputs of the slave VCROs such that their multiphase outputs become invariant to PVT changes. These outputs act as time interpolators for the TDCs. Therefore the compensation scheme prevents the time bin of the TDCs from drifting over time due to the aforementioned factors. Moreover, the same scheme is used to program different time resolutions of the direct time-of-flight (ToF) imager aimed at 3D ranging or depth map imaging. Experimental results that validate the analysis are provided as well. The compensation loop proves to be remarkably effective. The spreading of the TDCs time bin is lowered from: (i) 20% down to 2.4% while the temperature ranges from 0 °C to 100 °C; (ii) 27% down to 0.27%, when the voltage supply changes within ±10% of the nominal value; (iii) 5.2 ps to 2 ps standard deviation over 30 sample chips, due to process parameters' variation.


Variables affecting immediate pain tolerance in X-ray hysterosalpingography.
To determine the variables, which affect pain tolerance in x-ray hysterosalpingography (HSG). Cross-sectional comparative study. Radiology departments of Civil and Bismillah Taqee Hospitals and Delhi Mercantile Medical Centre, Karachi from January 1999 to June 2003. The study included 250 patients who underwent x-ray HSG using small sized Leech Wilkinson cannula by the same radiologist. Informed verbal consent was obtained from all the patients. Pain was graded into tolerable and intolerable using a four-point visual analog scale (VAS) as well as the Pakistani coin scale after pre-medication with diclofenac (generic name). Standard procedures using day-light fluoroscopy was performed. Variables considered were age, indication of the procedure, type of infertility, whether primary or secondary, emotional state, motivation for investigation, presence or absence of cystocele, duration of procedure, type of contrast used, whether ionic or non-ionic, tubal patency and lastly the amount of contrast used in the procedure. Those patients having infection, cervical erosion, use of larger size of cannula and technical complications including excessive bleeding requiring the use of sanitary napkins and vascular intravasation were excluded. The obtained data was analyzed on SPSS version 10.0 using cross tabulation and Pearson's correlation coefficient. Significance was taken at p<0.05. The mean age of the patients was 30 years. Amenorrhea, secondary infertility, apprehension, prolonged procedure time (>15 minutes) and the use of ionic contrast media were the variables significantly associated with intolerable pain during the procedure. Primary infertility, calm emotional state, self-motivation, absence of cystocele and short procedure duration were associated with good pain tolerance. The amount of contrast used and tubal patency status had no significant association. In the setting of base line pre-medication with diclofenac and minimal cervical trauma, amenorrhea, primary infertility, emotional state, self-motivation, procedure duration, and the presence or absence of cystocele significantly affect pain tolerance during the procedure. Amount of contrast used and tubal patency does not affect pain tolerance. Effect of the type of contrast medium requires further confirmation on a larger number of patients.


Nucleotide binding kinetics and conformational change analysis of tissue transglutaminase with switchSENSE.
Function, activity, and interactions of proteins crucially depend on their three-dimensional structure and are often regulated by effector binding and environmental changes. Tissue transglutaminase (Transglutaminase 2, TG2) is a multifunctional protein, allosterically regulated by nucleotides and Ca2+ ions, which trigger opposing conformational changes. Here we introduce switchSENSE as a versatile tool for TG2 characterization and provide novel insights into protein conformation as well as analyte binding kinetics. For the first time, we succeeded in measuring the kinetic rate constants and affinities (kon, koff, KD) for guanosine nucleotides (GMP, GDP, GTP, GTPγS). Further, the conformational changes induced by GDP, Ca2+ and the covalent inhibitor Z-DON were observed by changes in TG2's hydrodynamic diameter. We confirmed the well-known compaction by guanosine nucleotides and extension by Ca2+, and provide evidence for TG2 conformations so far not described by structural analysis. Moreover, we analyze the influence of the peptidic Z-DON inhibitor and the R580A mutation on the conformational responsiveness of TG2 to its natural effectors. In summary, this work shows how the combination of structural and kinetic information obtained by switchSENSE opens new perspectives for the characterization of conformationally active proteins and their interactions with ligands, e.g. potential drug candidates.


Structure-activity relationships of C-17-substituted estratriene-3-O-sulfamates as anticancer agents.
The synthesis and antiproliferative activities of analogues of 2-substituted estradiol-3,17-O,O-bis-sulfamates (E2bisMATEs) are discussed. Modifications of the C-17 substituent confirm that an H-bond acceptor is essential for high activity; its optimal linkage to C-17 and the local environment in which it resides are defined. In the non-sulfamoylated series 17β-acyl substitution delivers 48b, the most potent compound identified to date. In the sulfamate series a number of permutations of linker and H-bond acceptor deliver excellent activity, with 55, 61, 65, 49a, and 49b proving especially promising. The in vivo potential of these compounds was explored in the NCI hollow fiber assay and also in a mouse Matrigel model of antiangiogenesis in which 49 and 55 show significant inhibitory activity.


Non-rigid registration of breast images using feature points.
We present a feature point detection algorithm which we use for non-rigid registration, illustrated for breast images (mammography, MRI). By associating the continuous intrinsic dimensionality of image structure with the output of a scale saliency algorithm, breast boundary points can be separated from internal feature points. Correspondences established for the breast boundary and internal feature points respectively are used to drive two recent non-rigid registration techniques: polyaffine transformation and coherent point drift registration. Experimental results are presented for digital breast tomosynthesis and 3D breast MRI, and in all case achieve good spatial alignments.


Robotic resection of huge presacral tumors: case series and comparison with an open resection.
Clinical case series and analysis. The purpose of the present study is to evaluate the advantages and disadvantages of robotic presacral tumor resection compared with conventional open approach. Conventional open approach for huge presacral tumors in the retroperitoneal space often demands excessive hospitalization and poor cosmesis. Furthermore, narrow surgical field sometimes interrupt delicate procedures. Nine patients with huge (diameter >10 cm) presacral tumors underwent surgery. Five patients among them had robotic procedure and the others had open transperitoneal tumor resection. Operation time, blood loss, hospitalization, and complications were analyzed. Robotic presacral tumor resection showed shorter operation time, less bleeding, and shorter hospitalization. Moreover, there was no complication related to abdominal adhesion. Although robotic resection for presacral tumor still has limitations technically and economically, robotic resection for huge presacral tumors demonstrated advantages over open resection specifically for benign neurogenic tumors.


Linagliptin: a novel dipeptidyl peptidase 4 inhibitor with a unique place in therapy.
The dipeptidyl peptidase 4 (DPP-4) inhibitors comprise a promising new class of agent for the management of type 2 diabetes. They possess a range of physiological effects associated with improved glycemic control including stimulation of glucose-dependent insulin secretion and suppression of glucagon secretion, and lower blood glucose levels through different, but potentially complementary, mechanisms to standard oral therapies. Linagliptin is the latest DPP-4 inhibitor to complete pivotal phase 3 trials. The data show that linagliptin provides significant, clinically meaningful and sustained improvements in glycemic control, with an incidence of adverse events similar to placebo and an excellent tolerability profile. In addition, linagliptin has been shown to be weight neutral and, importantly, there was no increased risk of hypoglycemia attributed to linagliptin use in monotherapy or combination therapy with metformin or pioglitazone. A unique characteristic of linagliptin that differentiates it from other members of the class is its primarily nonrenal route of excretion. The linagliptin phase 3 program included several hundred patients with type 2 diabetes and different stages of renal disease and the data suggest that the drug would not need dose adjustment, regardless of the degree of renal impairment. There is a particular need for safe and effective therapeutic agents that can be used when renal function declines. Linagliptin has recently been approved by the US Food and Drug Administration and may find a place in therapy as a treatment option for the significant number of patients in whom metformin and the other DPP-4 inhibitors are either contraindicated or require dose adjustment because of moderate to severe renal impairment.


Biomineralization control related to population density under ocean acidification.
Anthropogenic CO2 is a major driver of current environmental change in most ecosystems1, and the related ocean acidification (OA) is threatening marine biota2. With increasing pCO2, calcification rates of several species decrease3, although cases of up-regulation are observed4. Here, we show that biological control over mineralization relates to species abundance along a natural pH gradient. As pCO2 increased, the mineralogy of a scleractinian coral (Balanophyllia europaea) and a mollusc (Vermetus triqueter) did not change. In contrast, two calcifying algae (Padina pavonica and Acetabularia acetabulum) reduced and changed mineralization with increasing pCO2, from aragonite to the less soluble calcium sulphates and whewellite, respectively. As pCO2 increased, the coral and mollusc abundance was severely reduced, with both species disappearing at pH < 7.8. Conversely, the two calcifying and a non-calcifying algae (Lobophora variegata) showed less severe or no reductions with increasing pCO2, and were all found at the lowest pH site. The mineralization response to decreasing pH suggests a link with the degree of control over the biomineralization process by the organism, as only species with lower control managed to thrive in the lowest pH.


Epalrestat, an aldose reductase inhibitor, improves an impaired generation of oxygen-derived free radicals by neutrophils from poorly controlled NIDDM patients.
To study the in vivo effect of epalrestat (Epa), an aldose reductase inhibitor, on the generation of oxygen-derived free radicals by neutrophils from poorly controlled NIDDM patients (HbA1c > 10%). A total of 31 diabetic patients were randomly divided into two groups: an Epa(+) group of 16 patients treated with 150 mg/day epalrestat and an Epa(-) group of 15 patients treated without epalrestat. A control group of 20 age- and sex-matched normal healthy subjects also participated. HbA1c, postprandial plasma glucose (PPG), and neutrophil bactericidal function were measured before and at the end of the drug treatment period (4 weeks). Neutrophil bactericidal function was measured as chemilu-minescence amplified by a Cypridina luciferin analog (CLA), which is dependent on O2- generation, and by luminol (L), which is highly dependent on OCl- generation, in response to formyl-methonyl-leucyl-phenylalanine (fMLP). At the start of the experiment, both CLA-dependent chemiluminescence (CLA-DCL) and L-dependent chemiluminescence (L-DCL) were clearly decreased in diabetic subjects (64 and 54%, respectively; P < 0.05) compared with control subjects (2,182 +/- 144 and 3,221 +/- 173 kc.min-1.10(-6) cells, respectively). At the end of the experiment, CLA-DCL and L-DCL in the Epa(+) group were significantly improved by 44 and 46%, respectively; however, these values were still lower than the corresponding results in the control group. HbA1c and PPG in both the Epa(+) and Epa (-) groups were significantly higher than in the control group, and treatment had no effect on either HbA1c or PPG. These data suggest that epalrestat may be a useful drug to prevent infection by improving the impaired O2- and OCl- generation by neutrophils from poorly controlled NIDDM patients.


Sub-chronic effects of AgNPs and AuNPs on Gammarus fossarum (Crustacea Amphipoda): From molecular to behavioural responses.
The aim of the present study was the assessment of the sub-chronic effects of silver (AgNPs) and gold nanoparticles (AuNPs) of 40 nm primary size either stabilised with citrate (CIT) or coated with polyethylene glycol (PEG) on the freshwater invertebrate Gammarus fossarum. Silver nitrate (AgNO3) was used as a positive control in order to study the contribution of silver ions potentially released from AgNPs on the observed effects. A multibiomarker approach was used to assess the long-term effects of AgNPs and AuNPs 40 nm on molecular, cellular, physiological and behavioural responses of G. fossarum. Specimen of G. fossarum were exposed for 15 days to 0.5 and 5 µgL-1 of CIT and PEG AgNPs and AuNPs 40 nm in the presence of food. A significant uptake of both Ag and Au was observed in exposed animals but was under the toxic threshold leading to mortality of G. fossarum. Silver nanoparticles (CIT-AgNPs and PEG-AgNPs 40 nm) led to an up-regulation of Na+K+ATPase gene expression. An up-regulation of Catalse and Chitinase gene expressions due to exposure to PEG-AgNPs 40 nm was also observed. Gold nanoparticles (CIT and PEG-AuNPs 40 nm) led to an increase of CuZnSOD gene expression. Furthermore, both AgNPs and AuNPs led to a more developed digestive lysosomal system indicating a general stress response in G. fossarum. Both AgNPs and AuNPs 40 nm significantly affected locomotor activity of G. fossarum while no effects were observed on haemolymphatic ions and ventilation.


Career destinations, views and future plans of the UK medical qualifiers of 1988.
To report the career destinations, views and future plans of a cohort of senior doctors who qualified in the 1980s. Postal questionnaire survey of all doctors who qualified from all UK medical schools in 1988. The response rate was 69%. We estimated that 81% of the total cohort was working in the NHS, 16 years after qualification; and that at least 94% of graduates who, when students, were from UK homes, were working in medicine. Of NHS doctors, 30% worked part-time. NHS doctors rated their job satisfaction highly (median score 19.9, scale 5-25) but were less satisfied with the amount of leisure time available to them (median score 5.4, scale 1-10). NHS doctors were very positive about their careers, but were less positive about working hours and some other aspects of the NHS. Women were more positive than men about working conditions; general practitioners were more positive than hospital doctors. Twenty-five percent reported unmet needs for further training or career-related advice, particularly about career development. Twenty-nine percent intended to reduce their hours in future, while 6%, mainly part-time women, planned to increase their hours. Overall, 10% of NHS doctors planned to do more service work in future and 24% planned to do less; among part-time women, 18% planned to do more and only 14% less. These NHS doctors, now in their 40s, had a high level of satisfaction with their jobs and their careers but were less satisfied with some other aspects of their working environment. A substantial percentage had expectations about future career development and change.


A Nomogram to Predict Prognosis in Malignant Pleural Mesothelioma.
Malignant pleural mesothelioma (MPM) is a rare malignancy with heterogeneous outcomes. This study was aimed to develop a nomogram to precisely and visually predict survival of MPM patients. Data from Surveillance, Epidemiology, and End Results database (1973-2014) on MPM were screened and retrieved. The prognostic effects of variables, including age, sex, race, year of diagnosis, laterality, histology, tumor stage, surgery, chemotherapy, and radiotherapy were analyzed using Kaplan-Meier method and Cox proportional hazard model. A nomogram was formulated to predict overall survival of MPM patients. A total of 1092 cases who met inclusion criteria were included in this study. The overall 1-, 2-, and 3-year survival rate in the entire cohort was 45.1, 23.0, and 12.1%, with median survival of 11 months. Cox regression analysis showed that age (P < 0.001), race (P = 0.003), histology type (P < 0.001), T stage (P < 0.001), M stage (P < 0.001), TNM stage (P < 0.001), cancer-directed surgery (P < 0.001), and chemotherapy (P < 0.001) were all independent prognostic factors of MPM patients. A nomogram was established based on the results of multivariate analysis. The internal bootstrap resampling approach suggested the nomogram had sufficient discriminatory power with the C-index of 0.705 (95% CI 0.681-0.729). The calibration plots also demonstrated good consistence between the prediction and the observation. We developed a nomogram to accurately predict clinical outcomes of MPM patients based on individual characteristics. Risk stratification by the survival nomogram could optimize individual therapies and follow-up.