Computed tomography perfusion prognostic maps do not predict reversible and irreversible neurological dysfunction following reperfusion therapies.
We aimed to evaluate the ability of commercially available computed tomography perfusion (CTP) prognostic maps software to identify reversibly and irreversibly damaged brain functions in the best case scenario: patients who achieved early and complete tissue reperfusion. Consecutive ischemic stroke patients who received reperfusion therapies, those with early (less than two-hours from treatment initiation) and complete Thrombolysis in Myocardial Infarction grade III or equivalent reperfusion were included in the analysis. Computed tomography perfusion prognostic maps were assigned as 'red,' or irreversible if cerebral blood volume declined below 2 ml/100 g and 'green,' or recoverable if the affected/unaffected mean transit time ratio was >1.45. Only patients with middle cerebral artery territory affected were included and subcomponents of the National Institutes of Health Stroke Scale scale pre- and posttreatment were analyzed based on anatomical correlation of the affected CTP areas and corresponding neurological functions. Among 109 consecutive patients who had intra-arterial reperfusion procedures, 16 (age 60 ± 17 years, 56% men, median National Institutes of Health Stroke Scale 13 . 5, interquartile range 7-18) had pretreatment CTP and had early complete reperfusion. We identified 44 affected areas on CTP (red 12 (27%), green 32 (73%)) with corresponding measurable neurological deficits including aphasia, arm, face weakness, and inattention. Red areas correctly identified 5/12 (42%) of functions that did not recover despite early reperfusion. Green areas correctly identified 18/32 (56%) of functions that recover after early reperfusion (OR 0.92, 95% CI 0.25-3.39, P = 1.0). In-patients achieving early and complete reperfusion, pretreatment CTP prognostic maps were not predictive for irreversibly or reversibly lost neurologic functions.


When Life Got in the Way: How Danish and Norwegian Immigrant Women in Sweden Reason about Cervical Screening and Why They Postpone Attendance.
Danish and Norwegian immigrant women in Sweden have an increased risk of cervical cancer compared to Swedish-born women. In addition, Danish and Norwegian immigrant women follow the national recommendations for attendance at cervical screening to much lesser extent than Swedish-born women. The aim of this study was to explore how Danish and Norwegian immigrant women in Sweden reason about attending cervical screening, focusing on women's perceptions as to why they and their compatriots do not attend. Eight focus group discussions (FGDs) were conducted with Danish and Norwegian immigrant women living in Stockholm. The women were between 26 and 66 years of age at the time of the FGDs, and were aged between <1 and 48 years old when they immigrated to Sweden. A FGD guide was used, which included questions related to cervical screening, and obstacles and motivators to attend cervical screening. The FGDs were tape recorded and transcribed, and the results analysed according to the principles of qualitative content analysis. The main theme was "Women have a comprehensive rationale for postponing cervical screening, yet do not view themselves as non-attenders". Investigation of women's rationale for non-attendance after being invited to cervical screening revealed some complex reasons related to immigration itself, including competing needs, organisational and structural factors and differences in mentality, but also reasons stemming from other factors. Postponing attendance at cervical screening was the category that linked all these factors as the reasons to why women did not attend to cervical screening according to the recommendations of the authorities. The rationale used to postpone cervical screening, in combination with the fact that women do not consider themselves to be non-attenders, indicates that they have not actively taken a stance against cervical screening, and reveals an opportunity to motivate these women to attend.


MiR-33a Controls hMSCS Osteoblast Commitment Modulating the Yap/Taz Expression Through EGFR Signaling Regulation.
Mesenchymal stromal cells (hMSCs) display a pleiotropic function in bone regeneration. The signaling involved in osteoblast commitment is still not completely understood, and that determines the failure of current therapies being used. In our recent studies, we identified two miRNAs as regulators of hMSCs osteoblast differentiation driving hypoxia signaling and cytoskeletal reorganization. Other signalings involved in this process are epithelial to mesenchymal transition (EMT) and epidermal growth factor receptor (EGFR) signalings through the regulation of Yes-associated protein (YAP)/PDZ-binding motif (TAZ) expression. In the current study, we investigated the role of miR-33a family as a (i) modulator of YAP/TAZ expression and (ii) a regulator of EGFR signaling during osteoblast commitments. Starting from the observation on hMSCs and primary osteoblast cell lines (Nh-Ost) in which EMT genes and miR-33a displayed a specific expression, we performed a gain and loss of function study with miR-33a-5p and 3p on hMSCs cells and Nh-Ost. After 24 h of transfections, we evaluated the modulation of EMT and osteoblast genes expression by qRT-PCR, Western blot, and Osteoimage assays. Through bioinformatic analysis, we identified YAP as the putative target of miR-33a-3p. Its role was investigated by gain and loss of function studies with miR-33a-3p on hMSCs; qRT-PCR and Western blot analyses were also carried out. Finally, the possible role of EGFR signaling in YAP/TAZ modulation by miR-33a-3p expression was evaluated. Human MSCs were treated with EGF-2 and EGFR inhibitor for different time points, and qRT-PCR and Western blot analyses were performed. The above-mentioned methods revealed a balance between miR-33a-5p and miR-33a-3p expression during hMSCs osteoblast differentiation. The human MSCs phenotype was maintained by miR-33a-5p, while the maintenance of the osteoblast phenotype in the Nh-Ost cell model was permitted by miR-33a-3p expression, which regulated YAP/TAZ through the modulation of EGFR signaling. The inhibition of EGFR blocked the effects of miR-33a-3p on YAP/TAZ modulation, favoring the maintenance of hMSCs in a committed phenotype. A new possible personalized therapeutic approach to bone regeneration was discussed, which might be mediated by customizing delivery of miR-33a in simultaneously targeting EGFR and YAP signaling with combined use of drugs.


The incidental finding of a persistent left superior vena cava: implications for primary care providers-case and review.
Persistent left superior vena cava (PLSVC) is the most common thoracic venous anomaly and is a persistent congenital remnant of the vena caval system from early cardiac development. Patients with congenital anomalous venous return are at increased risk of developing various cardiac arrhythmias, due to derangement of embryologic conductive tissue during the early development of the heart. Previously this discovery was commonly made during the placement of pacemakers or defibrillators for the treatment of the arrhythmias, when the operator encountered difficulty with proper lead deployment. However, in today's world of various easily obtainable imaging modalities, PLSVC is being discovered more and more by primary care providers during routine testing or screening for other ailments. Given the known association between anomalous venous return and the propensity for cardiac arrhythmias, we review the embryology of PLSVC and the mechanisms by which it leads to conduction abnormalities. We also provide the practitioner with recommendations for certain baseline cardiac observations and suggestions for proper surveillance in hopes that better understanding will reduce unnecessary and potentially harmful testing, premature subspecialty referral, and unneeded patient anxiety.


Clinical safety of brain magnetic resonance imaging with implanted deep brain stimulation hardware: large case series and review of the literature.
Over 75,000 patients have undergone deep brain stimulation (DBS) procedures worldwide. Magnetic resonance imaging (MRI) is an important clinical and research tool in analyzing electrode location, documenting postoperative complications, and investigating novel symptoms in DBS patients. Functional MRI may shed light on the mechanism of action of DBS. MRI safety in DBS patients is therefore an important consideration. We report our experience with MRI in patients with implanted DBS hardware and examine the literature for clinical reports on MRI safety with implanted DBS hardware. A total of 262 MRI examinations were performed in 223 patients with intracranial DBS hardware, including 45 in patients with an implanted pulse generator. Only 1 temporary adverse event occurred related to patient agitation and movement during immediate postoperative MR imaging. Agitation resolved after a few hours, and an MRI obtained before implanted pulse generator implantation revealed edema around both electrodes. Over 4000 MRI examinations in patients with implanted DBS hardware have been reported in the literature. Only 4 led to adverse events, including 2 hardware failures, 1 temporary and 1 permanent neurological deficit. Adverse neurological events occurred in a unique set of circumstances where appropriate safety protocols were not followed. MRI guidelines provided by DBS hardware manufacturers are inconsistent and vary among devices. The importance of MRI in modern medicine places pressure on industry to develop fully MRI-compatible DBS devices. Until then, the literature suggests that, when observing certain precautions, cranial MR images can be obtained with an extremely low risk in patients with implanted DBS hardware.


Antitumor effects of cannabidiol, a nonpsychoactive cannabinoid, on human glioma cell lines.
Recently, cannabinoids (CBs) have been shown to possess antitumor properties. Because the psychoactivity of cannabinoid compounds limits their medicinal usage, we undertook the present study to evaluate the in vitro antiproliferative ability of cannabidiol (CBD), a nonpsychoactive cannabinoid compound, on U87 and U373 human glioma cell lines. The addition of CBD to the culture medium led to a dramatic drop of mitochondrial oxidative metabolism [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide test] and viability in glioma cells, in a concentration-dependent manner that was already evident 24 h after CBD exposure, with an apparent IC(50) of 25 microM. The antiproliferative effect of CBD was partially prevented by the CB2 receptor antagonist N-[(1S)-endo-1,3,3-trimethylbicyclo[2,2,1]heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide (SR144528; SR2) and alpha-tocopherol. By contrast, the CB1 cannabinoid receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboximide hydrochloride (SR141716; SR1), capsazepine (vanilloid receptor antagonist), the inhibitors of ceramide generation, or pertussis toxin did not counteract CBD effects. We also show, for the first time, that the antiproliferative effect of CBD was correlated to induction of apoptosis, as determined by cytofluorimetric analysis and single-strand DNA staining, which was not reverted by cannabinoid antagonists. Finally, CBD, administered s.c. to nude mice at the dose of 0.5 mg/mouse, significantly inhibited the growth of subcutaneously implanted U87 human glioma cells. In conclusion, the nonpsychoactive CBD was able to produce a significant antitumor activity both in vitro and in vivo, thus suggesting a possible application of CBD as an antineoplastic agent.


Barriers and facilitators to targeted anxiety prevention programmes in families at risk: a qualitative interview study.
Anxiety disorders are the most common psychiatric disorder in children and young people. They can be prevented in those at risk, but families do not always take up opportunities to participate in prevention programmes. This qualitative study aimed to understand what families with children who were at prospective risk of anxiety disorders perceived to be the barriers to access to targeted anxiety prevention programmes, and to explore what would help facilitate access. We used Information Power to determine our sample size, and individually interviewed seven young people (14-17 years) who had anxiety disorders and their mothers, each of whom had pre-natal anxiety disorders. We transcribed all interviews and thematically analyzed them to identify perceived barriers and facilitators to targeted anxiety prevention programmes. Perceived potential barriers to access included possible negative consequences of anxiety prevention, difficulties in identifying anxiety as a problem and concerns about how professions would respond to raising concerns about anxiety. Possible facilitators included promoting awareness of anxiety prevention programmes and involvement of schools in promotion and delivery of prevention. Our findings illustrate that implementation of targeted anxiety prevention could be improved through (i) the provision of tools for parents to recognize anxiety in their children as a problem, (ii) promotion of awareness, as well as delivery, of anxiety prevention via schools and (iii) the involvement of parents and possibly adolescents in the intervention programme, but not younger children.


Comparison of allogeneic T cell-depleted peripheral blood stem cell and bone marrow transplantation: effect of stem cell source on short- and long-term outcome.
We report the results of a retrospective single-center study comparing engraftment, acute and chronic GVHD, relapse and survival in patients with malignant hematological disorders transplanted with allogeneic peripheral blood stem cells (alloPBSCT, n = 40) or bone marrow cells (alloBMT, n = 42). All transplants were T cell depleted by in vitro incubation with the Campath-1 monoclonal antibody. Primary graft failure occurred in none of the patients receiving an alloPBSCT compared with 3/42 of the recipients of an alloBMT. In addition, two patients in the alloBMT group showed no platelet engraftment. Recipients of PBSC had a more rapid recovery of neutrophils (median 14 days) compared to BM transplant recipients (median 32 days). Platelet recovery was also accelerated in PBSC recipients compared to BM recipients (11 vs 38 days). There was an increase in the incidence of grade II acute GVHD and chronic GVHD in patients after alloPBSCT (18% and 23%, respectively) compared to patients receiving alloBMT (5% and 8%, respectively). The 2-year cumulative incidence of relapse was similar in both groups (47%). At 6 months after transplantation, transplant-related mortality (TRM) was lower in PBSCT recipients than in BMT recipients. However, at a follow-up of 3 years TRM was similar in both groups. The disease-free survival rate at 3 years after transplantation did not differ between the groups (42% for PBSCT and 41% for BMT recipients). Our results indicate that T cell-depleted alloPBSCT compared to alloBMT is associated with a more rapid hematopoietic reconstitution and a decreased TRM at 6 months follow-up after transplantation. However, at a follow-up of 3 years, no sustained survival benefits were observed.


Prevalence of chronic medical conditions in Switzerland: exploring estimates validity by comparing complementary data sources.
Prevalence estimates of chronic medical conditions and their multiples (multimorbidity) in the general population are scarce and often rather speculative in Switzerland. Using complementary data sources, we assessed estimates validity of population-based prevalence rates of four common chronic medical conditions with high impact on cardiovascular health (diabetes mellitus, hypertension, dyslipidemia, obesity). We restricted our analyses to patients 15-94 years old living in the German speaking part of Switzerland. Data sources were: Swiss Health Survey (SHS, 2007, n = 13,580); Family Medicine ICPC Research using Electronic Medical Record Database (FIRE, 2010-12, n = 99,441); and hospital discharge statistics (MEDSTAT, 2009-10, n = 883,936). We defined chronic medical conditions based on use of drugs, diagnoses, and measurements. After a careful harmonization of the definitions, a high degree of concordance, especially regarding the age- and gender-specific distribution patterns, was found for diabetes mellitus (defined as drug use or diagnosis in SHS, drug use or diagnosis or blood glucose measurement in FIRE, and ICD-10 codes E10-14 as secondary diagnosis in MEDSTAT) and for hypertension (defined as drug use alone in SHS and FIRE, and ICD-10 codes I10-15 or I67.4 as secondary diagnosis in MEDSTAT). A lesser degree of concordance was found for dyslipidemia (defined as drug use alone in SHS and FIRE, and ICD-10 code E78 in MEDSTAT), and for obesity (defined as BMI ≥ 30 kg/m(2) derived from self-reported height and weight in SHS, from measured height and weight or diagnosis of obesity in FIRE, and ICD-10 code E66 as secondary diagnosis in MEDSTAT). MEDSTAT performed well for clearly defined diagnoses (diabetes, hypertension), but underrepresented systematically more symptomatic conditions (dyslipidemia, obesity). Complementary data sources can provide different prevalence estimates of chronic medical conditions in the general population. However, common age and sex patterns indicate that a careful harmonization of the definition of each chronic medical condition permits a high degree of concordance.


Activity assays and immunoassays for plasma Renin and prorenin: information provided and precautions necessary for accurate measurement.
Measurement of plasma renin is important for the clinical assessment of hypertensive patients. The most common methods for measuring plasma renin are the plasma renin activity (PRA) assay and the renin immunoassay. The clinical application of renin inhibitor therapy has thrown into focus the differences in information provided by activity assays and immunoassays for renin and prorenin measurement and has drawn attention to the need for precautions to ensure their accurate measurement. Renin activity assays and immunoassays provide related but different information. Whereas activity assays measure only active renin, immunoassays measure both active and inhibited renin. Particular care must be taken in the collection and processing of blood samples and in the performance of these assays to avoid errors in renin measurement. Both activity assays and immunoassays are susceptible to renin overestimation due to prorenin activation. In addition, activity assays performed with peptidase inhibitors may overestimate the degree of inhibition of PRA by renin inhibitor therapy. Moreover, immunoassays may overestimate the reactive increase in plasma renin concentration in response to renin inhibitor therapy, owing to the inhibitor promoting conversion of prorenin to an open conformation that is recognized by renin immunoassays. The successful application of renin assays to patient care requires that the clinician and the clinical chemist understand the information provided by these assays and of the precautions necessary to ensure their accuracy.


Leveraging The Affordable Care Act To Enroll Justice-Involved Populations In Medicaid: State And Local Efforts.
The Affordable Care Act provides an unprecedented opportunity to enroll criminal justice-involved populations in health insurance, particularly Medicaid. As a result, many state and county corrections departments have launched programs that incorporate Medicaid enrollment in discharge planning. Our study characterizes the national landscape of programs enrolling criminal justice-involved populations in Medicaid as of January 2015. We provide an overview of sixty-four programs operating in jails, prisons, or community probation and parole systems that enroll individuals during detention, incarceration, and the release process. We describe the variation among the programs in terms of settings, personnel, timing of eligibility screening, and target populations. Seventy-seven percent of the programs are located in jails, and 56 percent use personnel from public health or social service agencies. We describe four practices that have facilitated the Medicaid enrollment process: suspending instead of terminating Medicaid benefits upon incarceration, presuming that an individual is eligible for Medicaid before the process is completed, allowing enrollment during incarceration, and accepting alternative forms of identification for enrollment. The criminal justice system is a complex one that requires a variety of approaches to enroll individuals in Medicaid. Future research should examine how these approaches influence health and criminal justice outcomes.


Experimental evaluation of speckle suppression efficiency using a moving 2D Barker code DOE.
This paper reports the findings from an experimental evaluation of speckle suppression efficiency using a method based on a moving 2D Barker code diffractive optical element (DOE). The optical setup and the optical scheme parameters of the method are presented. A speckle contrast of ~4.4-5.3% and speckle suppression coefficient (coefficient of speckle contrast reduction) of k>8 was obtained in experiments. However, the experimentally obtained speckle suppression coefficient was approximately 1.5 times smaller than the theoretical prediction. It is speculated that the discrepancy between the theoretical and the experimental data is due to an inexact match between the optical setup and the optimal optical parameters of the method. Analysis of the experimental data revealed that once the optical scheme is optimized, it will be possible to obtain a speckle suppression that is closer to the theoretical prediction.


Analysis of apoptosis regulatory genes altered by histone deacetylase inhibitors in chronic lymphocytic leukemia cells.
Histone deacetylases (HDACs) play a key role in the regulation of acetylation status not only of histones but also of many other non-histone proteins involved in cell cycle regulation, differentiation or apoptosis. Therefore, histone deacetylase inhibitors (HDACi) have emerged as promising anticancer agents. Herein, we report the characterization of apoptosis in B-cell chronic lymphocytic leukemia (CLL) induced by two HDACi, Kendine 92 and SAHA. Both inhibitors induce dose-, time- and caspase-dependent apoptosis through the mitochondrial pathway. Interestingly, Kendine 92 and SAHA show a selective cytotoxicity for B lymphocytes and induce apoptosis in CLL cells with mutated or deleted TP53 as effectively as in tumor cells harboring wild-type TP53. The pattern of apoptosis-related gene and protein expression profile has been characterized. It has shown to be irrespective of TP53 status and highly similar between SAHA and Kendine 92 exposure. The balance between the increased BAD, BNIP3L, BNIP3, BIM, PUMA and AIF mRNA expression levels, and decreased expression of BCL-W, BCL-2, BFL-1, XIAP and FLIP indicates global changes in the apoptosis mRNA expression profile consistent with the apoptotic outcome. Protein expression analysis shows increased levels of NOXA, BIM and PUMA proteins upon Kendine 92 and SAHA treatment. Our results highlight the capability of these molecules to induce apoptosis not only in a selective manner but also in those cells frequently resistant to standard treatments. Thus, Kendine 92 is a novel HDACi with anticancer efficacy for non-proliferating CLL cells.


Germline Mutations in the BRIP1, BARD1, PALB2, and NBN Genes in Women With Ovarian Cancer.
Epithelial ovarian cancer (EOC) is the most lethal gynecological malignancy, responsible for 13 000 deaths per year in the United States. Risk prediction based on identifying germline mutations in ovarian cancer susceptibility genes could have a clinically significant impact on reducing disease mortality. Next generation sequencing was used to identify germline mutations in the coding regions of four candidate susceptibility genes-BRIP1, BARD1, PALB2 and NBN-in 3236 invasive EOC case patients and 3431 control patients of European origin, and in 2000 unaffected high-risk women from a clinical screening trial of ovarian cancer (UKFOCSS). For each gene, we estimated the prevalence and EOC risks and evaluated associations between germline variant status and clinical and epidemiological risk factor information. All statistical tests were two-sided. We found an increased frequency of deleterious mutations in BRIP1 in case patients (0.9%) and in the UKFOCSS participants (0.6%) compared with control patients (0.09%) (P = 1 x 10(-4) and 8 x 10(-4), respectively), but no differences for BARD1 (P = .39), NBN1 ( P = .61), or PALB2 (P = .08). There was also a difference in the frequency of rare missense variants in BRIP1 between case patients and control patients (P = 5.5 x 10(-4)). The relative risks associated with BRIP1 mutations were 11.22 for invasive EOC (95% confidence interval [CI] = 3.22 to 34.10, P = 1 x 10(-4)) and 14.09 for high-grade serous disease (95% CI = 4.04 to 45.02, P = 2 x 10(-5)). Segregation analysis in families estimated the average relative risks in BRIP1 mutation carriers compared with the general population to be 3.41 (95% CI = 2.12 to 5.54, P = 7×10(-7)). Deleterious germline mutations in BRIP1 are associated with a moderate increase in EOC risk. These data have clinical implications for risk prediction and prevention approaches for ovarian cancer and emphasize the critical need for risk estimates based on very large sample sizes before genes of moderate penetrance have clinical utility in cancer prevention.


Understanding the Limiting Factors of Solvent-Annealed Small-Molecule Bulk-Heterojunction Organic Solar Cells from a Chemical Perspective.
A detailed account of the limiting factors of solvent-annealed bulk-heterojunction small-molecule organic solar cells is given. This account is based on the extensive characterisation of solar cell devices made from a library of five diketopyrolopyrole (DPP) donor dyes. Their chemical structure is designed in such a way as to provide insights into the energetics of solar cell active layer micro-structure formation. Numerous chemical and physical properties of the active layers are assessed and inter-related such as light absorption, molecular packing in the solid state, crystal-forming properties in thin films, charge carrier mobility and charge carrier recombination kinetics. A myriad of characterisation techniques are used such as UV/Vis absorption spectroscopy, photoluminescence spectroscopy, XRD, AFM and photo-induced transient measurements, which provide information on the optical properties of the active layers, morphology and recombination kinetics. Consequently, a mechanism for the solvent-vapour-annealing-assisted formation of crystalline domains of donor molecules in the active layer is proposed, and the micro-structural features are related to the J-V characteristics of the devices. According to this model, the crystalline phase in which the donor crystallise in the active layer is the key determinant to direct the formation of the micro-structure.


Low-dose vs standard-dose unfractionated heparin for percutaneous coronary intervention in acute coronary syndromes treated with fondaparinux: the FUTURA/OASIS-8 randomized trial.
The optimal unfractionated heparin regimen for percutaneous coronary intervention (PCI) in patients with non-ST-segment elevation acute coronary syndromes treated with fondaparinux is uncertain. To compare the safety of 2 unfractionated heparin regimens during PCI in high-risk patients with non-ST-segment elevation acute coronary syndromes initially treated with fondaparinux. Double-blind randomized parallel-group trial in 179 hospitals in 18 countries involving 2026 patients undergoing PCI within 72 hours, nested within a cohort of 3235 high-risk patients with non-ST-segment elevation acute coronary syndromes initially treated with fondaparinux enrolled from February 2009 to March 2010. Patients received intravenously either low-dose unfractionated heparin, 50 U/kg, regardless of use of glycoprotein IIb/IIIa (GpIIb-IIIa) inhibitors or standard-dose unfractionated heparin, 85 U/kg (60 U/kg with GpIIb-IIIa inhibitors), adjusted by blinded activated clotting time (ACT). Composite of major bleeding, minor bleeding, or major vascular access-site complications up to 48 hours after PCI. Key secondary outcomes include composite of major bleeding at 48 hours with death, myocardial infarction, or target vessel revascularization within day 30. The primary outcome occurred in 4.7% of those in the low-dose group vs 5.8% in the standard-dose group (odds ratio [OR], 0.80; 95% confidence interval [CI], 0.54-1.19; P = .27). The rates of major bleeding were not different but the rates of minor bleeding were lower with 0.7% in the low-dose group vs 1.7% in the standard-dose group (OR, 0.40; 95% CI, 0.16-0.97; P = .04). For the key secondary outcome, the rates for low-dose group were 5.8% vs 3.9% in the standard-dose group (OR, 1.51; 95% CI, 1.00-2.28; P = .05) and for death, myocardial infarction, or target vessel revascularization it was 4.5% for the low-dose group vs 2.9% for the standard-dose group (OR, 1.58; 95% CI, 0.98-2.53; P = .06). Catheter thrombus rates were very low (0.5% in the low-dose group and 0.1% in the standard-dose group, P = .15). Low-dose compared with standard-dose unfractionated heparin did not reduce major peri-PCI bleeding and vascular access-site complications. clinicaltrials.gov Identifier: NCT00790907.


Hydrogenated polycyclic aromatic hydrocarbons (HnPAHs) as catalysts for hydrogenation reactions in the interstellar medium: a quantum chemical model.
The sticking of H atoms onto dust grains and large hydrocarbon molecules has received considerable attention because it is thought to govern the formation of H2 and other H-containing molecules in the interstellar medium. Using the density functional theory (DFT) approximation, we have investigated the capacity of neutral hydrogenated polycyclic aromatic hydrocarbons (HnPAH) to catalyze simple hydrogenation reactions by acting as a source of atomic hydrogen. In particular, the interaction of OH and CO with H1-anthracene (singly hydrogenated) and H14-anthracene (fully hydrogenated) to form H2O and HCO was modeled following the Eley-Rideal mechanism. In this process, a hydrogen atom is abstracted from the HnPAH molecule forming the corresponding hydrogenated compound. The results were compared to the most known case of the HnPAH-catalyzed formation of H2. It was observed that whereas H2 is formed by overcoming activation barriers of approximately 0.02 and 0.10 eV with H1-anthracene and H14-anthracene, respectively, H2O is produced in a barrierless fashion with both hydrocarbon molecules. The production of HCO was found to be a highly unfavorable process (with activation barriers of 0.73 eV and 3.13 eV for H1- and H14-anthracene, respectively). Complementary calculations performed using the rest of the Hn-anthracene molecules (from 2 to 13 extra H atoms) showed that in all the cases the reaction with OH is barrierless as well. This efficient mechanism could therefore be a possible route for water formation in the cold interstellar medium.


Internet-based learning in the health professions: a meta-analysis.
The increasing use of Internet-based learning in health professions education may be informed by a timely, comprehensive synthesis of evidence of effectiveness. To summarize the effect of Internet-based instruction for health professions learners compared with no intervention and with non-Internet interventions. Systematic search of MEDLINE, Scopus, CINAHL, EMBASE, ERIC, TimeLit, Web of Science, Dissertation Abstracts, and the University of Toronto Research and Development Resource Base from 1990 through 2007. Studies in any language quantifying the association of Internet-based instruction and educational outcomes for practicing and student physicians, nurses, pharmacists, dentists, and other health care professionals compared with a no-intervention or non-Internet control group or a preintervention assessment. Two reviewers independently evaluated study quality and abstracted information including characteristics of learners, learning setting, and intervention (including level of interactivity, practice exercises, online discussion, and duration). There were 201 eligible studies. Heterogeneity in results across studies was large (I(2) > or = 79%) in all analyses. Effect sizes were pooled using a random effects model. The pooled effect size in comparison to no intervention favored Internet-based interventions and was 1.00 (95% confidence interval [CI], 0.90-1.10; P < .001; n = 126 studies) for knowledge outcomes, 0.85 (95% CI, 0.49-1.20; P < .001; n = 16) for skills, and 0.82 (95% CI, 0.63-1.02; P < .001; n = 32) for learner behaviors and patient effects. Compared with non-Internet formats, the pooled effect sizes (positive numbers favoring Internet) were 0.10 (95% CI, -0.12 to 0.32; P = .37; n = 43) for satisfaction, 0.12 (95% CI, 0.003 to 0.24; P = .045; n = 63) for knowledge, 0.09 (95% CI, -0.26 to 0.44; P = .61; n = 12) for skills, and 0.51 (95% CI, -0.24 to 1.25; P = .18; n = 6) for behaviors or patient effects. No important treatment-subgroup interactions were identified. Internet-based learning is associated with large positive effects compared with no intervention. In contrast, effects compared with non-Internet instructional methods are heterogeneous and generally small, suggesting effectiveness similar to traditional methods. Future research should directly compare different Internet-based interventions.


A mathematical model of the growth of sea lice, Lepeophtheirus salmonis, populations on farmed Atlantic salmon, Salmo salar L., in Scotland and its use in the assessment of treatment strategies.
Sea lice are a persistent problem for farmed and wild salmonid populations. Control can be achieved through the use of veterinary medicines. A model was developed to describe the patterns of sea lice infection on salmon farms in Scotland and to predict the likely effect of various treatment strategies. This model takes into account development rates and mortality using compartments representing life history stages and external infection pressure. The national sea lice infection pattern was described using parameters representing stage survival, background infection levels and egg viability rates. The patterns observed across farms varied greatly and the model gave broad agreement to observed trends with different parameters being required in the model for sites using hydrogen peroxide and cypermethrin treatments. The parameter estimates suggest that the background infection pressure on sites where cypermethrin was administered was higher than for those using hydrogen peroxide. Both models had comparable magnitudes of sensitivity with survival from one stage to another being the most sensitive parameter, followed by feedback rates at which gravid females produce eggs, with background infection levels the least sensitive. The effect of different cypermethrin treatment strategies was assessed using the model. Increasing treatments in a production cycle gave more effective control. However, the model showed that timing of treatments is most important if sea lice are to be effectively controlled.


The MYC-WDR5 Nexus and Cancer.
The MYC oncogenes encode a family of transcription factors that feature prominently in cancer. MYC proteins are overexpressed or deregulated in a majority of malignancies and drive tumorigenesis by inducing widespread transcriptional reprogramming that promotes cell proliferation, metabolism, and genomic instability. The ability of MYC to regulate transcription depends on its dimerization with MAX, which creates a DNA-binding domain that recognizes specific sequences in the regulatory elements of MYC target genes. Recently, we discovered that recognition of target genes by MYC also depends on its interaction with WDR5, a WD40-repeat protein that exists as part of several chromatin-regulatory complexes. Here, we discuss how interaction of MYC with WDR5 could create an avidity-based chromatin recognition mechanism that allows MYC to select its target genes in response to both genetic and epigenetic determinants. We rationalize how the MYC-WDR5 interaction provides plasticity in target gene selection by MYC and speculate on the biochemical and genomic contexts in which this interaction occurs. Finally, we discuss how properties of the MYC-WDR5 interface make it an attractive point for discovery of small-molecule inhibitors of MYC function in cancer cells.


Vitamin D deficiency increases blood-brain barrier dysfunction after ischemic stroke in male rats.
Because vitamin D hormone deficiency (VDHdef) can worsen severity and outcome for ischemic stroke, we examined the role of VDH in maintaining blood-brain-barrier (BBB integrity) in a rat model of stroke. In most types of stroke, the BBB is markedly compromised, potentially leading to a cascade of injury processes and functional deficits, so we examined a number of biomarkers associated with BBB disruption to determine whether VDH deficiency would further compromise the BBB following a stroke. Male Wistar rats were randomly assigned to one of two diet cohorts, VDH-sufficient (VDHsuf) and VDHdef. The VDHsuf group was fed standard rat chow and the VDHdef group got a VDH-null version of the same diet for 8 weeks. Animals from both cohorts were subjected to transient middle cerebral artery occlusion (tMCAO) surgery, killed at 72 h post-stroke, and their brains evaluated for BBB permeability and injury severity using expression of immunoglobulin (IgG), matrix metalloproteinase-9 (MMP-9) activity and alteration of tight junction (TJ) proteins as markers of BBB disruption. We also evaluated modulation of glucose transporter-1 (GLUT1), osteopontin (OPN), β-catenin and vitamin D receptor (VDR) expression in VDHsuf and VDHdef subjects. At the time of MCAO, rats on the VDHdef diet had circulating VDH levels one-fourth that of rats fed control chow. IgG extravasation after MCAO, indicating more severe BBB injury, was significantly higher in the MCAO+VDHdef than the MCAO+VDHsuf rats. Following MCAO, expression of MMP-9, GLUT1, VDR and OPN increased and the TJ proteins occludin and claudin-5 decreased significantly in the VDHdef compared to the VDHsuf group. We also observed significantly lower expression of β-catenin in the MCAO group of both VDHsuf and VDHdef rats. Under these conditions, VDH deficiency itself can compromise the BBB. We think that low serum VDH levels are likely to complicate stroke severity and its chronic consequences.


Placebo response in Parkinson's disease: comparisons among 11 trials covering medical and surgical interventions.
Placebo-associated improvements have been previously documented in small series of Parkinson's disease (PD) patients. Using a strict definition of placebo-associated improvement, we examined rates and timing of placebo responses to identify patient- and study-based characteristics, predicting positive placebo response in several PD clinical trials. We collected individual patient data from the placebo groups of 11 medical and surgical treatment trials involving PD patients with differing PD severities and placebo-assignment likelihoods. We defined a positive placebo response as > or = 50% improvement in total Unified Parkinson's Disease Rating Scale motor (UPDRSm) score or a decrease by > or = 2 points on at least two UPDRSm items compared to baseline. We calculated positive placebo response rates at early (3-7 weeks), mid (8-18 weeks), and late (23-35 weeks) stages of follow-up. Odds ratios for patient- and study-based characteristics were obtained from a model fitted using generalized estimating equations. There were 858 patients on placebo who met inclusion criteria for analysis. Three studies involved patients without need of symptomatic treatment, two involved patients without motor fluctuations needing symptomatic treatment, and six (three medical and three surgical) involved patients with motor fluctuations. The overall placebo response rate was 16% (range: 0-55%). Patients with higher baseline UPDRSm scores and studies that focused on PD with motor fluctuations, surgical interventions, or those with a higher probability of placebo assignment showed increased odds of positive placebo response. Placebo responses were temporally distributed similarly during early, mid, and late phases of follow-up. Placebo-related improvements occur in most PD clinical trials and are similarly distributed across all 6 months of follow-up. Recognition of factors that impact placebo response rates should be incorporated into individual study designs for PD clinical trials.


Successful treatment of PR3-ANCA-positive interstitial pneumonia with a moderate dose of glucocorticoid and rituximab.
Antineutrophil cytoplasmic antibody (ANCA)-positive interstitial pneumonia (IP) is reported as IP that is ANCA-positive and does not involve organ damage associated with vasculitis other than the lungs. While the combination of glucocorticoid and rituximab is effective in ANCA-associated vasculitis, the treatment strategy for ANCA-positive IP has not been established. Here, we report the first case of successful treatment of proteinase 3 (PR3)-ANCA-positive IP with a moderate dose of glucocorticoid and rituximab. The patient was an 80-year-old male who presented with subacute dry cough and dyspnoea. Blood tests revealed elevated levels of C-reactive protein, Krebs von den Lungen 6 (KL-6), and PR3-ANCA. Chest computed tomography (CT) showed interstitial shadows and infiltrates around honeycomb cysts. 18F-fluorodeoxyglucose (FDG) positron emission tomography CT revealed an uptake of FDG in the IP area. After starting treatment with a moderate dose of prednisolone and rituximab, the patient's clinical symptoms disappeared, C-reactive protein and KL-6 turned to be normal, and infiltrates around the cysts of honeycomb lungs disappeared. Prednisolone was gradually decreased to 2 mg, and no relapse or adverse events were observed during the course of treatment. Our case suggests that early treatment with a moderate dose of glucocorticoid and rituximab is effective for PR3-ANCA-positive IP.


A histone methyltransferase modulates antigenic variation in African trypanosomes.
To evade the host immune system, several pathogens periodically change their cell-surface epitopes. In the African trypanosomes, antigenic variation is achieved by tightly regulating the expression of a multigene family encoding a large repertoire of variant surface glycoproteins (VSGs). Immune evasion relies on two important features: exposing a single type of VSG at the cell surface and periodically and very rapidly switching the expressed VSG. Transcriptional switching between resident telomeric VSG genes does not involve DNA rearrangements, and regulation is probably epigenetic. The histone methyltransferase DOT1B is a nonessential protein that trimethylates lysine 76 of histone H3 in Trypanosoma brucei. Here we report that transcriptionally silent telomeric VSGs become partially derepressed when DOT1B is deleted, whereas nontelomeric loci are unaffected. DOT1B also is involved in the kinetics of VSG switching: in DeltaDOT1B cells, the transcriptional switch is so slow that cells expressing two VSGs persist for several weeks, indicating that monoallelic transcription is compromised. We conclude that DOT1B is required to maintain strict VSG silencing and to ensure rapid transcriptional VSG switching, demonstrating that epigenetics plays an important role in regulating antigenic variation in T. brucei.


The Brain Connectivity Workshops: moving the frontiers of computational systems neuroscience.
Understanding the link between neurobiology and cognition requires that neuroscience moves beyond mere structure-function correlations. An explicit systems perspective is needed in which putative mechanisms of how brain function is constrained by brain structure are mathematically formalized and made accessible for experimental investigation. Such a systems approach critically rests on a better understanding of brain connectivity in its various forms. Since 2002, frontier topics of connectivity and neural system analysis have been discussed in a multidisciplinary annual meeting, the Brain Connectivity Workshop (BCW), bringing together experimentalists and theorists from various fields. This article summarizes some of the main discussions at the two most recent workshops, 2006 at Sendai, Japan, and 2007 at Barcelona, Spain: (i) investigation of cortical micro- and macrocircuits, (ii) models of neural dynamics at multiple scales, (iii) analysis of "resting state" networks, and (iv) linking anatomical to functional connectivity. Finally, we outline some central challenges and research trajectories in computational systems neuroscience for the next years.


Calculation of protein requirements; a comparison of calculations based on bodyweight and fat free mass.
In dietary practice, it is common to estimate protein requirements on actual bodyweight, but corrected bodyweight (in cases with BMI <20 kg/m2 and BMI ≥30 kg/m2) and fat free mass (FFM) are also used. Large differences on individual level are noticed in protein requirements using these different approaches. To continue this discussion, the answer is sought in a large population to the following question: Will choosing actual bodyweight, corrected bodyweight or FFM to calculate protein requirements result in clinically relevant differences? This retrospective database study, used data from healthy persons ≥55 years of age and in- and outpatients ≥18 years of age. FFM was measured by air displacement plethysmography technology or bioelectrical impedance analysis. Protein requirements were calculated as 1) 1.2 g (g) per kilogram (kg) actual bodyweight or 2) corrected bodyweight or 3) 1.5 g per kg FFM. To compare these three approaches, the approach in which protein requirement is based on FFM, was used as reference method. Bland-Altman plots with limits of agreement were used to determine differences, analyses were performed for both populations separately and stratified by BMI category and gender. In total 2291 subjects were included. In the population with relatively healthy persons (n = 506, ≥55 years of age) mean weight is 86.5 ± 18.2 kg, FFM is 51 ± 12 kg and in the population with adult in- and outpatients (n = 1785, ≥18 years of age) mean weight is 72.5 ± 18.4 kg, FFM is 51 ± 11 kg. Clinically relevant differences were found in protein requirement between actual bodyweight and FFM in most of the participants with overweight, obesity or severe obesity (78-100%). Using corrected bodyweight, an overestimation in 48-92% of the participants with underweight, healthy weight and overweight is found. Only in the Amsterdam UMC population, protein requirement is underestimated when using the approach of corrected bodyweight in participants with severe obesity. The three approaches in estimation of protein requirement show large differences. In the majority of the population protein requirement based on FFM is lower compared to actual or corrected bodyweight. Correction of bodyweight reduces the differences, but remain unacceptably large. It is yet unknown which method is the best for estimation of protein requirement. Since differences vary by gender due to differences in body composition, it seems more accurate to estimate protein requirement based on FFM. Therefore, we would like to advocate for more frequent measurement of FFM to determine protein requirements, especially when a deviating body composition is to be expected, for instance in elderly and persons with overweight, obesity or severe obesity.


Endometrial ablation for severe menorrhagia in a patient with hereditary hemorrhagic Telangiectasia. A case report.
Hereditary hemorrhagic telangiectasia is a rare, inherited disease characterized by abnormal visceral and superficial blood vessel anastomoses. These telangiectasias predispose the patient to a lifelong history of recurrent bleeding for the nasal and gastrointestinal mucosa. Less commonly involved organs include the liver, brain and lung. To date there is no cure for this disease. Management requires many palliative minor surgical procedures to stop actively bleeding sites. Major surgery is often contraindicated in these patients due do coexisting medical sequelae of their underlying disease. Menorrhagia was diagnosed in a 42 year-old multipara with known history of hereditary hemorrhagic telangiectasia. The bleeding was unresponsive to hormonal therapy. Substantial preexisting conditions, including profound anemia, history of multiple strokes, a seizure disorder and ventricular arrhythmias, precluded major surgical intervention, including hysterectomy. Serial injections of leuprolide acetate injections were followed by hysteroscopic "rollerball" electrocoagulation of the endometrium under regional anesthesia. On long-term follow up, the patient was cured of her menorrhagia. Endometrial ablation provides patients who have significant medical complications with an effective, minimally invasive alternative to hysterectomy for control of menorrhagia.


Psychopathology in great apes: concepts, treatment options and possible homologies to human psychiatric disorders.
Many captive great apes show gross behavioral abnormalities such as stereotypies, self-mutilation, inappropriate aggression, fear or withdrawal, which impede attempts to integrate these animals in existing or new social groups. These abnormal behaviors resemble symptoms associated with psychiatric disorders in humans such as depression, anxiety disorders, eating disorders, and post-traumatic stress disorder. Due to the outstanding importance of social interaction and the prolonged period of infantile and juvenile dependence, early separation of infants from their mothers and other adverse rearing conditions, solitary housing, and sensory deprivation are among the major albeit non-specific sources of psychopathology in apes. In contrast to the wealth of research in monkeys, psychopathology in apes has been under-studied, and only a few studies have examined how to alleviate abnormal behavior in captive apes. Recent studies have shown that the enrichment of living conditions and behavioral treatment (conditioning) may ameliorate some pathological features, and careful familiarization with novel physical and social environments can help re-socialize behaviorally disturbed animals, but usually not to the extent of successful mating and raising offspring. The possibility of psychopharmacological treatment of the most severe disturbed animal patients has only been reluctantly considered, but a few case reports have revealed encouraging results. This article proposes the need to expand research into ape psychopathology, which would require an essentially interdisciplinary approach of primatology and psychiatry, ultimately to the benefit of both.


Peripherally placed totally implantable venous-access port systems of the forearm: clinical experience in 763 consecutive patients.
The aim of this study is to evaluate the effectiveness and safety of percutaneously placed totally implantable venous-access ports (TIVAPs) of the forearm. Between January 2006 and October 2008, peripheral TIVAPs were implanted in 763 consecutive patients by ultrasound and fluoroscopic guidance. All catheters were implanted under local anesthesia and were tunneled subcutaneously. Indication, technical success, and complications were retrospectively analyzed according to Society of Interventional Radiology (SIR) criteria. Presence of antibiotic prophylaxis, periprocedurally administered drugs (e.g., sedation), and laboratory results at the time of implantation were analyzed. Maintenance during the service interval was evaluated. In total, 327,499 catheter-days were analyzed. Technical success rate was 99.3%. Reasons for initial failure of implantation were either unexpected thrombosis of the subclavian vein, expanding tumor mass of the mediastinum, or failure of peripheral venous access due to fragile vessels. Mean follow-up was 430 days. There were 115 complications observed (15.1%, 0.03 per 100 catheter-days), of which 33 (4.3%) were classified as early (within 30 days from implantation) and 82 (10.7%) as late. Catheter-related venous thrombosis was found in 65 (8.5%) of 763 (0.02 per 100 catheter-days) TIVAPs. Infections were observed in 41 (5.4%) of 763 (0.01 per 100 catheter-days) devices. Other complications observed included dislocation of the catheter tip (0.8%), occlusion (0.1%), or rupture (0.1%) of the port catheter. Dislocated catheters were corrected during a second interventional procedure. In conclusion, implantation of percutaneously placed peripheral TIVAPs shows a high technical success rate and low risk of early complications when ultrasound and fluoroscopic guidance are used. Late complications are observed three times as often as early complications.


Noise suppression in coherent imaging.
Several noise suppression techniques in coherent imaging systems are described. For holographic imaging the diffuse wave, periodic phase modulation, and multiple wave techniques are compared and the implementation of the last is considered. For lens-type imaging systems the use of multiple incoherent waves results in excellent noise suppression.


Gata2b is a restricted early regulator of hemogenic endothelium in the zebrafish embryo.
The adult blood system is established by hematopoietic stem cells (HSCs), which arise during development from an endothelial-to-hematopoietic transition of cells comprising the floor of the dorsal aorta. Expression of aortic runx1 has served as an early marker of HSC commitment in the zebrafish embryo, but recent studies have suggested that HSC specification begins during the convergence of posterior lateral plate mesoderm (PLM), well before aorta formation and runx1 transcription. Further understanding of the earliest stages of HSC specification necessitates an earlier marker of hemogenic endothelium. Studies in mice have suggested that GATA2 might function at early stages within hemogenic endothelium. Two orthologs of Gata2 exist in zebrafish: gata2a and gata2b. Here, we report that gata2b expression initiates during the convergence of PLM, becoming restricted to emerging HSCs. We observe Notch-dependent gata2b expression within the hemogenic subcompartment of the dorsal aorta that is in turn required to initiate runx1 expression. Our results indicate that Gata2b functions within hemogenic endothelium from an early stage, whereas Gata2a functions more broadly throughout the vascular system.


THE INFLUENCE OF ENVIRONMENT UPON THE BIOLOGICAL PROCESSES OF THE VARIOUS MEMBERS OF THE COLON GROUP OF BACILLI : AN EXPERIMENTAL STUDY.
Assuming the typical colon bacillus and the typical typhoid bacillus to represent the types of this group that present the greatest divergences in biological peculiarities, we conclude, as others also have done, that there is a series of closely related forms that may be regarded as intermediate or transitional and which serve to establish a biological relationship, either near or remote, between these two typical members. From our own studies we are inclined to regard the typical colon bacillus as the type of this group, for the reason that its functional equilibrium, as observed in the intestine, is so permanent a quality that it may readily be perpetuated under what is ordinarily regarded as favorable artificial circumstances, and that with the continuance of such conditions there is no conspicuous tendency on the part of this organism to deviate from what we regard as its norm; whereas, on the other hand, with all the other members of this group with which we have worked, there is not only a lack of uniformity in the adjustment of the functions, but such as exists is readily disturbed under artificial environment; though it must be borne in mind that even with the typical colon bacillus we have also shown functional modifications to be possible under particular conditions. When the members of the colon group are cultivated under circumstances favorable to the development of both the function of fermentation and that of proteolysis, fermentation invariably takes precedence and no evidence of proteolysis is manifested until after fermentation has ceased. The cultivation of all the members of the colon group under circumstances that favor the development of one function, viz. that of proteolysis, at the expense of another, viz. that of fermentation, results first in an apparent increase of vigor; but this is of temporary duration and is quickly followed by the decline and death of the cell. The result of this increased activity of the proteolytic function is the formation of much larger amounts of indol by typical colon cultures than has ever been obtained by us by any other method. By the method of experimentation through which we were enabled to accentuate the proteolytic activity of the typical colon bacillus, as caused by an increase of indol formation, we have also induced the function of indol formation not only in atypical colon bacilli that had been devoid of it, but in every specimen of typical typhoid bacilli to which we had access as well. We feel justified in regarding one of the differential tests between the typhoid and colon bacillus, namely that of indol formation on the part of the latter and the absence of this function from the former, as of questionable value, for the reason, as shown above, that by particular methods of cultivation indol production has been shown to accompany the development of a number of specimens that we have every reason to regard as genuine typhoid bacilli. As a result of our own experiments, together with the observations of others, there can be no doubt that the bacillus coli communis at times possesses pathogenic properties, and that by artificial methods of treatment it may often be brought from a condition of benignity to one of virulence. The spleen of a typhoid patient has always been regarded as the only trustworthy source from which to obtain the typical typhoid bacillus. While we believe this to be true, still our investigations show that other members of the colon group may also be present in this viscus; in fact, from such spleens we have isolated practically all of the varieties of this group with which we are acquainted. From our experience, the value of the serum test for the differentiation of typhoid and colon bacilli would seem to be questionable. We are inclined, however, to attribute the irregularities recorded above as due more to the method of application than to defects of the principles involved; for, as stated, by the use of dried blood, as in our experiments, it is not possible to make the test with constant and accurate, or even approximately accurate, dilutions of the serum. Our irregularities may be in part due to this defect. We therefore lay less stress upon this than upon the other features of our work.


Spontaneous clearance of chronic hepatitis C infection is associated with an internal ribosomal entry site IIId stem loop structure variant.
To investigate if any mutations in hepatitis C virus (HCV) internal ribosome entry site (IRES) can inhibit the translation of viral polyprotein. A 26-year-old male patient infected with HCV 10 years ago was followed up. After 9 years of chronic infection. The patient had managed to resolve the infection for a period of 9 months, after which the patient experienced a viral recurrence characterized by high viral load and diverse HCV quasispecies. The IRES structures of the viral strains that disappeared were comparable with those that are currently active using structural mutational analysis. A novo mutational position 254 combined with a rarely observed mutation at position 253 in the stem of the IIId subdomain were observed and the new conformation had an octa-apical loop (AGUGUUGG) and a shift in the 3 ` GU from the loop to the stem. These mutations were found to be highly deleterious, and they affected the direct binding of the IIId loop to the 40S ribosomal subunit with a subsequent inhibition of translation of viral polyprotein and clearance of the virus.


Preferential degradation of the terminal carbohydrate moiety of membrane glycoproteins in rat hepatoma cells and after transfer to the membranes of mouse fibroblasts.
Glycoproteins in the plasma membrane of rat hepatoma cells were labeled at their externally exposed tyrosine residues with 131I and at their galactose and sialic acid residues with 3H. The degradation of both isotopes in the total cell protein fraction, in glycoproteins purified by concanavalin A, and in glycoproteins separated on two-dimensional gels was determined. Similarly, the total cellular membrane glycoproteins were metabolically labeled with [35S]methionine and [3H]fucose. The fate of both incorporated labels was followed by lectin chromatography or by precipitation of the proteins with specific antibodies followed by electrophoretic gel separation. In both labeling experiments, the carbohydrate markers were lost from the ligand-recognized fraction with similar kinetics as from the total cell protein fraction. In some glycoprotein species which were separated by two-dimensional gel electrophoresis, the polypeptide portion exhibited up to a twofold slower rate of degradation relative to that of the carbohydrate moiety. This difference is most pronounced in carbohydrate-rich glycoproteins. To corroborate this finding, double-labeled membrane glycoproteins were incorporated into reconstituted phospholipid vesicles which were then transferred via fusion into the plasma membrane of mouse fibroblasts. Both the polypeptide and carbohydrate moieties of the transferred membrane glycoproteins were degraded with the same relative kinetics as in the original hepatoma cells. The rate of degradation is mostly a function of the structural properties of the membrane components as shown by the preservation of metabolically stable fucogangliosides of Reuber H-35 hepatoma cells transferred onto the fibroblasts. The technique of insertion of membrane components into the plasma membrane of another cell should assist in the elucidation of the exact route and mechanism of membrane protein destruction.


Unique crystal structure of a novel surfactant protein from the foam nest of the frog Leptodactylus vastus.
Breeding by releasing eggs into stable biofoams ("foam nests") is a peculiar reproduction mode within anurans, fish, and tunicates; not much is known regarding the biochemistry or molecular mechanisms involved. Lv-ranaspumin (Lv-RSN-1) is the predominant protein from the foam nest of the frog Leptodactylus vastus. This protein shows natural surfactant activity, which is assumed to be crucial for stabilizing foam nests. We elucidated the amino acid sequence of Lv-RSN-1 by de novo sequencing with mass-spectrometry and determined the high-resolution X-ray structure of the protein. It has a unique fold mainly composed of a bundle of 11 α-helices and two small antiparallel β-strands. Lv-RSN-1 has a surface rich in hydrophilic residues and a lipophilic cavity in the region of the antiparallel β-sheet. It possesses intrinsic surface-active properties, reducing the surface tension of water from 73 to 61 mN m(-1) (15 μg mL(-1)). Lv-RSN-1 belongs to a new class of surfactants proteins for which little has been reported regarding structure or function.


An experimental histopathologic study of surgical glove powders.
Tissue reactions to surgical glove powders, ie, talc (magnesium silicate) and Keoflo (low cross-linked cornstarch) were studied histologically from day 1 to 16 weeks. These materials were tested as a contaminant on the surface of surgical suture or in a pellet form implanted in abdominal muscle of rat. Use of Keoflo resulted in an intense acute inflammatory response, with a progressive decrease in the amount of starch with time after implant. By the fourth week, most of the starch had disappeared with minimal tissue damage and scar formation. Rats implanted with talc showed an initial mild to moderate acute inflammation followed by chronic inflammatory response and granuloma formation by the third day. From the fourth week on, granulomas showed talc crystals within the giant cells surrounded by histiocytes, lymphocytes, some collagen, and fibroblasts. This study indicates that talc molecules are not absorbed, whereas low cross-linked cornstarch is an absorbable substance; therefore the latter is a safe material for use as surgical glove powder.


Medical student awareness of the Patient-centered Medical Home.
The Patient-centered Medical Home (PCMH) has come to the forefront of primary care practice redesign and can potentially improve health care outcomes and reduce costs. There are several initiatives in medical schools to teach concepts of the PCMH to students, but it is unknown what knowledge and attitudes medical students currently possess. We report students' awareness and opinions at two medical schools without comprehensive PCMH curricula. A total of 1,408 first- through fourth-year students at both schools were invited to participate. We distributed an electronic survey to all students via institutional e-mail listserves. Descriptive statistics were used. A total of 359 students participated, for a response rate of 25.5%. Despite no comprehensive curricula, 40.9% students had still encountered the topic of the PCMH. Family medicine and primary care clerkships serve as the most frequent point of exposure for students. Although many students reported not understanding the overall PCMH concept, most cited "some" understanding when presented with individually defined PCMH principles with the one exception: Value-based Payment. A significant portion of first-, second-, and third-year students rated learning about the PCMH by graduation as "important," while most fourth-year students rated this as "somewhat important." Students performed well on one knowledge question about PCMH principles; however, 29.6% of respondents believed that primary care physicians function as gatekeepers in the PCMH model. Medical students appear to have limited exposure and knowledge of the PCMH concept, suggesting the need to develop curricula about the PCMH in medical schools.


Feasibility Study on Applying Radiophotoluminescent Glass Dosimeters for CyberKnife SRS Dose Verification.
CyberKnife is one of multiple modalities for stereotactic radiosurgery (SRS). Due to the nature of CyberKnife and the characteristics of SRS, dose evaluation of the CyberKnife procedure is critical. A radiophotoluminescent glass dosimeter was used to verify the dose accuracy for the CyberKnife procedure and validate a viable dose verification system for CyberKnife treatment. A radiophotoluminescent glass dosimeter, thermoluminescent dosimeter, and Kodak EDR2 film were used to measure the lateral dose profile and percent depth dose of CyberKnife. A Monte Carlo simulation for dose verification was performed using BEAMnrc to verify the measured results. This study also used a radiophotoluminescent glass dosimeter coupled with an anthropomorphic phantom to evaluate the accuracy of the dose given by CyberKnife. Measurements from the radiophotoluminescent glass dosimeter were compared with the results of a thermoluminescent dosimeter and EDR2 film, and the differences found were less than 5%. The radiophotoluminescent glass dosimeter has some advantages in terms of dose measurements over CyberKnife, such as repeatability, stability, and small effective size. These advantages make radiophotoluminescent glass dosimeters a potential candidate dosimeter for the CyberKnife procedure. This study concludes that radiophotoluminescent glass dosimeters are a promising and reliable dosimeter for CyberKnife dose verification with clinically acceptable accuracy within 5%.


Maximum likelihood estimation of reference centiles.
We propose the use of centile estimates which are based on the fitting of appropriate densities by maximum likelihood. In the case of cross-sectional centile estimation, we show that this approach will generally lead to more precise estimates than would result from the use of non-parametric centile estimates. When longitudinal data are available or a series of cross-sectional data at different time points, the maximum likelihood approach can be used to simultaneously fit densities to each cross-section, subject to constraints (for example, smoothness constraints) on the parameters. The variances of these centile estimates are readily obtained and missing values and unequally spaced records are easily accommodated. We illustrate the procedure by means of an application using the Johnson family of densities to a study of weight gain in pregnancy.


LOPDF: a framework for extracting and producing open data of scientific documents for smart digital libraries.
Results of scientific experiments and research work, either conducted by individuals or organizations, are published and shared with scientific community in different types of scientific publications such as books, chapters, journals, articles, reference works and reference works entries. One aspect of these documents is their contents and the other is metadata. Metadata of scientific documents could be used to increase mutual cooperation, find people with common interest and research work, and to find scientific documents in the matching domains. The major issue in getting these benefits from metadata of scientific publications is availability of these data in unstructured (or semi-structured) format so that it can not be used to ask smart queries that can help in computing and performing different types of analysis on scientific publications data. Also, acquisition and smart processing of publications data is a complicated as well as time and resource consuming task. To address this problem we have developed a generic framework named as Linked Open Publications Data Framework (LOPDF). The LOPDF framework can be used to crawl, process, extract and produce machine understandable data (i.e., LOD) about scientific publications from different publisher specific sources such as portals, XML export and websites. In this paper we present the architecture, process and algorithm that we developed to process textual publications data and to produce semantically enriched data as RDF datasets (i.e., open data). The resulting datasets can be used to make smart queries by making use of SPARQL protocol. We also present the quantitative as well as qualitative analysis of our resulting datasets which ultimately can be used to compute the research behavior of organizations in rapidly growing knowledge society. Finally, we present the potential usage of producing and processing such open data of scientific publications and how results of performing smart queries on resulting open datasets can be used to compute the impact and perform different types of analysis on scientific publications data.


A sagittal plane model of the knee and cruciate ligaments with application of a sensitivity analysis.
In this investigation the complex multi-bundle structure of the cruciate ligaments and their interaction with the tibiofemoral joint was modeled analytically by representing the different regions of the cruciates with ligament elements. A sensitivity analysis was then performed to describe the effect that variations of the model input parameters had on the model variables (outputs). The effect that the cruciate ligament bundles had in controlling joint kinematics was dependent on knee flexion angle, and the load applied to the tibiofemoral joint. For passive range of knee motion with the thigh in the horizontal plane (a common rehabilitation activity), all cruciate ligament bundles were strained with the joint positioned between 0 and 10 deg of knee flexion, between 10 and 50 deg only the anterior bundle of the posterior cruciate ligament A-PCL was strained, and from 50 to 90 deg both the anteromedial portion of the anterior cruciate ligament A-ACL and the A-PCL were strained. This finding indicates that a strain distribution about a transverse cross section of the cruciates exists, and demonstrates the importance of differentiating between the strained and unstrained (unloaded) states of these ligaments. The strain value of a cruciate ligament bundle was an indication of how the bundle controls joint kinematics, while the unstrained values describe how much the ligament bundle must deform before it becomes strained and a restraint to tibiofemoral joint motion. In response to anterior and posterior directed loads, applied parallel to the tibial plateau, the respective, ACL and PCL load values were larger in magnitude. The sensitivity of the model outputs to the input parameters was highly dependent on knee flexion angle. The geometrical input parameters of the model (including the ligament insertion site locations and articular surface geometry) had the most pronounced effect on the model output quantities, while the stiffness and initial strain conditions of the ligament bundles had less of an effect on the model outputs. When loaded, the strain values of the ligament bundles were sensitive to the ligament insertion site position. The greatest sensitivity of the model outputs was the femoral insertion of the ACL; supporting clinical impressions and previous experimental findings. Changes in the anterior-posterior dimension of the femoral articular surface did not produce a substantial effect on the model outputs, while changes in the proximal-distal dimension created a large effect; similar results were found for the tibial surface dimensions. These findings indicate that rigid body contact between the articular surfaces may not be a realistic assumption particularly with application to the prediction of tibiofemoral compressive loading and the force/strain values of the cruciate ligament elements. This also has important implications for the design and clinical application of total knee replacements (that function as rigid bodies), particularly those that spare the PCL.


Exposure to phthalates and breast cancer risk in northern Mexico.
Phthalates, ubiquitous environmental pollutants that may disturb the endocrine system, are used primarily as plasticizers of polyvinyl chloride and as additives in consumer and personal care products. In this study, we examined the association between urinary concentrations of nine phthalate metabolites and breast cancer (BC) in Mexican women. We age-matched 233 BC cases to 221 women residing in northern Mexico. Sociodemographic and reproductive characteristics were obtained by direct interviews. Phthalates were determined in urine samples (collected pretreatment from the cases) by isotope dilution/high-performance liquid chromatography coupled to tandem mass spectrometry. Phthalate metabolites were detected in at least 82% of women. The geometric mean concentrations of monoethyl phthalate (MEP) were higher in cases than in controls (169.58 vs. 106.78 microg/g creatinine). Controls showed significantly higher concentrations of mono-n-butyl phthalate, mono(2-ethyl-5-oxohexyl) phthalate, and mono(3-carboxypropyl) phthalate (MCPP) than did the cases. After adjusting for risk factors and other phthalates, MEP urinary concentrations were positively associated with BC [odds ratio (OR), highest vs. lowest tertile = 2.20; 95% confidence interval (CI), 1.33-3.63; p for trend < 0.01]. This association became stronger when estimated for premenopausal women (OR, highest vs. lowest tertile = 4.13; 95% CI, 1.60-10.70; p for trend < 0.01). In contrast, we observed significant negative associations for monobenzyl phthalate (MBzP) and MCPP. We show for the first time that exposure to diethyl phthalate, the parent compound of MEP, may be associated with increased risk of BC, whereas exposure to the parent phthalates of MBzP and MCPP might be negatively associated. These findings require confirmation.


Application of complex-lag distributions for estimation of arbitrary order phase derivatives in digital holographic interferometry.
This Letter proposes a method to estimate phase derivatives of arbitrary order in digital holographic interferometry. Based on the desired order, the generalized complex-lag distribution is computed from the reconstructed interference field. Subsequently, the phase derivative is estimated by tracing the peak of the distribution. Simulation and experimental results are presented to validate the method's potential.


Resource implications of preparing individual participant data from a clinical trial to share with external researchers.
Demands are increasingly being made for clinical trialists to actively share individual participant data (IPD) collected from clinical trials using responsible methods that protect the confidentiality and privacy of clinical trial participants. Clinical trialists, particularly those receiving public funding, are often concerned about the additional time and money that data-sharing activities will require, but few published empirical data are available to help inform these decisions. We sought to evaluate the activity and resources required to prepare anonymised IPD from a clinical trial in anticipation of a future data-sharing request. Data from two UK publicly funded clinical trials were used for this exercise: 2437 participants with epilepsy recruited from 90 hospital outpatient clinics in the SANAD trial and 146 children with neuro-developmental problems recruited from 18 hospitals in the MENDS trial. We calculated the time and resources required to prepare each anonymised dataset and assemble a data pack ready for sharing. The older SANAD trial (published 2007) required 50 hours of staff time with a total estimated associated cost of £3185 whilst the more recently completed MENDS trial (published 2012) required 39.5 hours of staff time with total estimated associated cost of £2540. Clinical trial researchers, funders and sponsors should consider appropriate resourcing and allow reasonable time for preparing IPD ready for subsequent sharing. This process would be most efficient if prospectively built into the standard operational design and conduct of a clinical trial. Further empirical examples exploring the resource requirements in other settings is recommended. SANAD: International Standard Randomised Controlled Trials Registry: ISRCTN38354748 . Registered on 25 April 2003. EU Clinical Trials Register Eudract 2006-004025-28 . Registered on 16 May 2007. International Standard Randomised Controlled Trials Registry: ISRCTN05534585 /MREC 07/MRE08/43. Registered on 26 January 2007.


Design considerations in developing a text messaging program aimed at smoking cessation.
Cell phone text messaging is gaining increasing recognition as an important tool that can be harnessed for prevention and intervention programs across a wide variety of health research applications. Despite the growing body of literature reporting positive outcomes, very little is available about the design decisions that scaffold the development of text messaging-based health interventions. What seems to be missing is documentation of the thought process of investigators in the initial stages of protocol and content development. This omission is of particular concern because many researchers seem to view text messaging as the intervention itself instead of simply a delivery mechanism. Certainly, aspects of this technology may increase participant engagement. Like other interventions, however, the content is a central driver of the behavior change. To address this noted gap in the literature, we discuss the protocol decisions and content development for SMS Turkey (or Cebiniz birakin diyor in Turkish), a smoking cessation text messaging program for adult smokers in Turkey. Content was developed in English and translated into Turkish. Efforts were made to ensure that the protocol and content were grounded in evidence-based smoking cessation theory, while also reflective of the cultural aspects of smoking and quitting in Turkey. Methodological considerations included whether to provide cell phones and whether to reimburse participants for texting costs; whether to include supplementary intervention resources (eg, personal contact); and whether to utilize unidirectional versus bidirectional messaging. Program design considerations included how messages were tailored to the quitting curve and one's smoking status after one's quit date, the number of messages participants received per day, and over what period of time the intervention lasted. The content and methods of effective smoking cessation quitline programs were a useful guide in developing SMS Turkey. Proposed guidelines in developing text messaging-based behavior change programs are offered.


Improving structure-based virtual screening by multivariate analysis of scoring data.
Three different multivariate statistical methods, PLS discriminant analysis, rule-based methods, and Bayesian classification, have been applied to multidimensional scoring data from four different target proteins: estrogen receptor alpha (ERalpha), matrix metalloprotease 3 (MMP3), factor Xa (fXa), and acetylcholine esterase (AChE). The purpose was to build classifiers able to discriminate between active and inactive compounds, given a structure-based virtual screen. Seven different scoring functions were used to generate the scoring matrices. The classifiers were compared to classical consensus scoring and single scoring functions. The classifiers show a superior performance, with rule-based methods being most effective. The precision of correctly predicting an active compound is about 90% for three of the targets and about 25% for acetylcholine esterase. On the basis of these results, a new two-stage approach is suggested for structure-based virtual screening where limited activity information is available.


When nurses become the "second" victim.
Well-intentioned, conscientious nurses make medication errors. The subsequent feelings of guilt, remorse, and loss of personal and professional self-esteem these nurses experience are well documented. In this paper, we analyze the concept of "second victim" within the context of medication administration errors. We also examine factors that contribute to nurses becoming second victims after making an error. Implications for nurses and nursing practice include nurses being given a greater degree of authority in designing the nursing work environment. Implications for nurses and nursing practice are presented. Further study is needed to more fully understand this phenomenon and to discover appropriate support mechanisms and interventions to minimize the harm to nurses.


Genes expressed during the differentiation of pancreatic AR42J cells into insulin-secreting cells.
Pancreatic AR42J cells have the feature of pluripotency of the common precursor cells of the pancreas. Dexamethasone (Dx) converts them to exocrine cells, whereas activin A (Act) converts them into endocrine cells expressing pancreatic polypeptide. A combination of Act and betacellulin (BTC) converts them further into insulin-secreting cells. The present study identifies some of the genes involved in the process of differentiation that is induced by these factors, using the mRNA differential display and screening of the cDNA expression array. The expression levels of 7 genes were increased by Act alone, and a combination of Act and BTC increased the expression of 25 more genes. Of these, 16 represented known genes or homologues of genes characterized previously. Nine of the identified genes were unrelated to any other sequences in the database. An inhibitor of the mitogen-activated protein kinase pathway, PD098059, which blocks the differentiation into insulin-secreting cells, inhibited the expression of 18 of the 25 genes, suggesting that the proteins encoded by these genes are associated with the differentiation into insulin-producing cells. These include known genes encoding extracellular signaling molecules, such as parathyroid hormone-related peptide, cytoskeletal proteins, and intracellular signaling molecules. Identification and characterization of these differentially expressed genes should help to clarify the molecular mechanism of differentiation of pancreatic cells and the gene products that enable the beta-cells to produce insulin.


Regulation of the complement membrane attack pathway.
Complement activation, triggered through classic, alternative, or lectin pathways of activation, leads to activation of a final common reaction sequence, the membrane attack pathway, in which five soluble plasma proteins assemble into a multimolecular complex that inserts into and through the membrane, creating a functional pore, the membrane attack complex. The active products of membrane attack are the anaphylactic and chemotactic fragment C5a and the membrane attack complex itself. These active products are important in immune defense but also carry the potential to damage host cells. Regulation of the membrane attack pathway is essential to protect host cells from damage at sites of C activation, and several protective systems have evolved to achieve this in vivo. Nevertheless, in disease these protective systems may be overwhelmed, and the availability of agents that specifically inhibit the membrane attack pathway might be of considerable therapeutic benefit. In this article I first describe the membrane attack pathway, its active products, and its role in disease. Second, I discuss systems operating to prevent damage to host cells by membrane attack, the regulatory proteins and activities that form the core of the article. Finally, I describe recent developments in the production of therapeutic agents with the capacity to inhibit membrane attack pathway.


Feedback topology and XOR-dynamics in Boolean networks with varying input structure.
We analyze a model of fixed in-degree random Boolean networks in which the fraction of input-receiving nodes is controlled by the parameter gamma. We investigate analytically and numerically the dynamics of graphs under a parallel XOR updating scheme. This scheme is interesting because it is accessible analytically and its phenomenology is at the same time under control and as rich as the one of general Boolean networks. We give analytical formulas for the dynamics on general graphs, showing that with a XOR-type evolution rule, dynamic features are direct consequences of the topological feedback structure, in analogy with the role of relevant components in Kauffman networks. Considering graphs with fixed in-degree, we characterize analytically and numerically the feedback regions using graph decimation algorithms (Leaf Removal). With varying gamma , this graph ensemble shows a phase transition that separates a treelike graph region from one in which feedback components emerge. Networks near the transition point have feedback components made of disjoint loops, in which each node has exactly one incoming and one outgoing link. Using this fact, we provide analytical estimates of the maximum period starting from topological considerations.


Development of active tuberculosis during treatment of head and neck carcinoma: a case series.
Patients with head and neck carcinoma are considered to be at high risk of developing tuberculosis, since the risk of morbidity due to tuberculosis in these patients is 2.86 to 16 times the risk in the general population. This case series describes an 83-year-old Japanese man, a 60-year-old Japanese man, and a 69-year-old Japanese man who developed active pulmonary tuberculosis while being treated for head and neck carcinoma. Two had previously developed tuberculosis and were treated for more than 50 years, but no symptoms or imaging findings suggested tuberculosis onset in the patients at initiation of treatment for head and neck carcinoma. Initially, local radiotherapy was performed for all three patients. Chemotherapy was continued for two patients who had pulmonary metastasis since initial consultation. For the other patient, surgery was performed for recurrence. In all three cases, active tuberculosis infection was observed during maintenance chemotherapy or immediately following surgery. Due to the high risk of developing tuberculosis, the possibility of prophylactic administration of anti-tuberculosis agents to patients with head and neck carcinoma should be investigated, although prophylactic administration is not a cost-effective option for all patients with head and neck carcinoma. However, if tuberculosis onset occurs, it leads to various problems; it has a major impact on not only patients with cancer but also various people in the social environment. In the future, it is essential to consider prophylactic administration in patients requiring long-term maintenance drug therapy, especially in those who are treated at out-patient chemotherapy clinics, where there are several patients with cancer with low disease resistance.


Serial determination of thyroxine concentrations in hyperthyroid cats.
Serum thyroxine (T4) concentrations of 10 hyperthyroid cats were measured at hourly intervals between 9 AM and 4 PM. In 5 cats, blood samples were obtained by jugular venipuncture; the remaining 5 cats had blood samples obtained from jugular catheters. Over the 7-hour period, a significant temporal (diurnal) variation was not identified in the serum T4 concentrations of the cats (P greater than 0.01). The lowest mean serum T4 concentration (9.1 micrograms/dl) was measured at 3 PM and was 14.2% less than the highest mean serum T4 concentration (10.6 micrograms/dl) measured at 9 AM. Though there were fluctuations in serum T4 concentrations during the 7-hour period, the differences were not systematic. The maximal variation in serum T4 concentrations over the 7-hour period averaged less than 21%. Despite the random variations during the 7-hour period, none of the measured serum T4 concentrations was in the normal range. Measurement of serum T4 concentration from randomly obtained blood samples was determined to be reliable for diagnosing feline hyperthyroidism.


Biocompatibility of (Ba,Ca)(Zr,Ti)O3 piezoelectric ceramics for bone replacement materials.
Total joint replacement implants are generally designed to physically mimic the biological environment to ensure compatibility with the host tissue. However, implant instability exposes patients to long recovery periods, high risk for revision surgeries, and high expenses. Introducing electrical stimulation to the implant site to accelerate healing is promising, but the cumbersome nature of wired devices is detrimental to the implant design. We propose a novel strategy to stimulate cells at the implant site by utilizing piezoelectric ceramics as electrical stimulation sources. The inherent ability of these materials to form electric surface potentials under mechanical load allows them to act as internal power sources. This characteristic is commonly exploited in non-biomedical applications such as transducers or sensors. We investigate calcium/zirconium-doped barium titanate (BCZT) ceramics in an in vitro environment to determine their potential as implant materials. BCZT exhibits low cytotoxicity with human osteoblast and endothelial cells as well as high piezoelectric responses. Microstructural adaptation was identified as a route for optimizing piezoelectric behavior. Our results show that BCZT is a promising system for biomedical applications. Its characteristic ability to autonomously generate electric surface potentials opens the possibility to functionalize existing bone replacement implant designs to improve implant ingrowth and long-term stability.


Recurrence and Frequency of Disturbance have Cumulative Effect on Methanotrophic Activity, Abundance, and Community Structure.
Alternate prolonged drought and heavy rainfall is predicted to intensify with global warming. Desiccation-rewetting events alter the soil quality and nutrient concentrations which drive microbial-mediated processes, including methane oxidation, a key biogeochemical process catalyzed by methanotrophic bacteria. Although aerobic methanotrophs showed remarkable resilience to a suite of physical disturbances induced as a single event, their resilience to recurring disturbances is less known. Here, using a rice field soil in a microcosm study, we determined whether recurrence and frequency of desiccation-rewetting impose an accumulating effect on the methanotrophic activity. The response of key aerobic methanotroph subgroups (type Ia, Ib, and II) were monitored using qPCR assays, and was supported by a t-RFLP analysis. The methanotrophic activity was resilient to recurring desiccation-rewetting, but increasing the frequency of the disturbance by twofold significantly decreased methane uptake rate. Both the qPCR and t-RFLP analyses were congruent, showing the dominance of type Ia/Ib methanotrophs prior to disturbance, and after disturbance, the recovering community was predominantly comprised of type Ia (Methylobacter) methanotrophs. Both type Ib and type II (Methylosinus/Methylocystis) methanotrophs were adversely affected by the disturbance, but type II methanotrophs showed recovery over time, indicating relatively higher resilience to the disturbance. This revealed distinct, yet unrecognized traits among the methanotroph community members. Our results show that recurring desiccation-rewetting before a recovery in community abundance had an accumulated effect, compromising methanotrophic activity. While methanotrophs may recover well following sporadic disturbances, their resilience may reach a 'tipping point' where activity no longer recovered if disturbance persists and increase in frequency.


Calcium-sensing receptor activation increases cell-cell adhesion and β-cell function.
The extracellular calcium-sensing receptor (CaR) is expressed in pancreatic β-cells where it is thought to facilitate cell-to-cell communication and augment insulin secretion. However, it is unknown how CaR activation improves β-cell function. Immunocytochemistry and western blotting confirmed the expression of CaR in MIN6 β-cell line. The calcimimetic R568 (1µM) was used to increase the affinity of the CaR and specifically activate the receptor at a physiologically appropriate extracellular calcium concentration. Incorporation of 5-bromo-2'-deoxyuridine (BrdU) was used to measure cell proliferation, whilst changes in non-nutrient-evoked cytosolic calcium were assessed using fura-2-microfluorimetry. AFM-single-cell-force spectroscopy related CaR-evoked changes in epithelial (E)-cadherin expression to improved functional tethering between coupled cells. Activation of the CaR over 48hr doubled the expression of E-cadherin (206±41%) and increased L-type voltage-dependent calcium channel expression by 70% compared to control. These changes produced a 30% increase in cell-cell tethering and elevated the basal-to-peak amplitude of ATP (50µM) and tolbutamide (100µM)-evoked changes in cytosolic calcium. Activation of the receptor also increased PD98059 (1-100µM) and SU1498 (1-100µM)-dependent β-cell proliferation. Our data suggest that activation of the CaR increases E-cadherin mediated functional tethering between β-cells and increases expression of L-type VDCC and secretagogue-evoked changes in [Ca(2+)](i). These findings could explain how local changes in calcium, co-released with insulin, activate the CaR on neighbouring cells to help ensure efficient and appropriate secretory function.


Polio control after certification: major issues outstanding.
Now that the global eradication of wild poliovirus is almost within sight, planning for the post-certification era is becoming a priority issue. It is agreed that a stockpile of appropriate polio vaccines will need to be established, and a surveillance and response capacity will need to be maintained, in order to protect the world against any possible future outbreaks attributable either to the persistence of wild poliovirus or vaccine-derived polioviruses (VDPVs) or to the unintentional or intentional release of poliovirus from a laboratory or vaccine store. Although it has been suggested that the stockpile should consist of monovalent oral poliovirus vaccine (mOPV), many questions remain concerning its nature, financing, management, and use--in particular, because of uncertainties over future national vaccination policies, and over the availability of different vaccines, after the certification of wild poliovirus eradication. There are further uncertainties concerning the possible role and efficacy of inactivated poliovirus vaccine (IPV) used either routinely or in outbreak control in low-hygiene settings, the potential for rapid geographical spread of polioviruses should an outbreak occur after certification, and the risks inherent in introducing additional oral polio vaccine (OPV) viruses into populations in which the vaccine coverage and prevalence of immunity have declined, and which may thus favour the spread of VDPVs. Given these important gaps in knowledge, no country should discontinue polio vaccination until a coordinated policy for the post-certification era has been developed and the recommended measures have been put in place.