The chemical modification of KCa channels by carbon monoxide in vascular smooth muscle cells.
The chemical modification of big conductance calcium-activated potassium (KCa) channels in rat tail artery smooth muscle cells by carbon monoxide (CO) was investigated using the cell-free single channel recording technique. Exposure of the internal surface of cell membranes to diethyl pyrocarbonate (DEPC) neither affected the characteristics of KCa channels nor modified the stimulatory effect of CO on KCa channels. However, when DEPC was applied to the external surface of cell membranes, the open probability of KCa channels was reduced. The pH and concentration dependence of the effect of DEPC indicated the specific modification of histidine residues. Kinetic analysis suggested that one externally located histidine residue was modified by DEPC. Treatment of the external surface of cell membranes with DEPC abolished the CO-induced increase in the open probability of KCa channels. Likewise, the presence of CO partially protected KCa channels from inhibition by DEPC. Moreover, photooxidation of the histidine residue located on the external membrane surface abolished the CO-induced activation of KCa channels. Our study demonstrates that the CO-induced increase in the open probability of KCa channels may rely specifically on the structure and topological locations of histidine residues.


Antagonism by DDT of the effect of valinomycin on a synthetic membrane.
The potassium conductance which is induced by 10(-6) molar valinomycin in a lecithin-decane membrane is reversed by 3 x 10(-6) molar DDT. Membranes not treated with valinomycin are not affected by DDT. This blockade of potassium conductance parallels the effect of DDT on axonic conduction. Dieldrin and lindane, whose physiological actions are in some ways like those of DDT, do not affect valinomycin-induced conductance of lecithin-decane membranes.


Role of organic osmolytes in water homoeostasis in skin.
The ability to conserve water is fundamental to terrestrial life. A number of organs such as the kidney and the bladder have important roles in the regulation of body water balance. The epidermis of skin is also fundamental to this process, and it is in a constant battle to prevent loss of water to the external, dry environment. Given this important role of the epidermis as a barrier to water loss, it is perhaps surprising that many of the cellular mechanisms by which human keratinocytes achieve cell volume homoeostasis, maintain epidermal hydration and adapt to biological effects from environmental stressors such as ultraviolet radiation are poorly understood. This article reviews what is known thus far and speculates about other potential mechanisms through which skin conducts water homoeostasis, with a particular emphasis on the putative role of organic osmolytes.


Response of two breeds of chickens to Ascaridia galli infections from two geographic sources.
Comparative resistance to different isolates of Ascaridia galli was investigated in a local chicken breed from Jordan (LC) and in the Lohmann LSL white chicken (LW) strain. In two trials, birds of LC and LW were inoculated orally at 1-day old with 250 embryonated A. galli eggs. In the first trial a German source of A. galli was used, whereas in the second trial, a Jordan source of A. galli was used. At week 7 of infection, infected LC birds harbored significantly (P<0.05) fewer worms and excreted less A. galli eggs than infected LW birds. A. galli isolated from Jordan were less infectious than A. galli from Germany. Results suggest that the variation in genetic background between LC and LW is involved in the resistance to A. galli infection. A. galli isolates from different geographic areas differ in their ability to infect different chicken genotypes.


Interferon lambda inhibits dengue virus replication in epithelial cells.
In viral disease, infection is controlled at the cellular level by type I interferon (IFN-I), but dengue virus (DENV) has the ability to inhibit this response. Type III interferon, also known as lambda IFN (IFN-III or IFN-λ), is a complementary pathway to the antiviral response by IFN-I. This work analyzed the IFN-λ (IFN-III) mediated antiviral response against DENV serotype 2 (DENV-2) infection. Dengue fever patients were sampled to determine their IFN-λ levels by ELISA. To study the IFN-λ response during DENV infection we selected the epithelial cell line C33-A, and we demonstrated that it is permissive to DENV-2 infection. The effect of IFN-λ on virus replication was determined in these cells, in parallel to the expression of IFN-stimulated genes (ISGs), and Suppressor of Cytokine Signaling (SOCS), genes measured by RT-qPCR. We found increased (~1.8 times) serological IFN-λ in dengue fever patients compared to healthy blood donors. IFN-λ inhibited DENV-2 replication in a dose-dependent manner in vitro. The reduction of viral titer corresponded with increased ISG mRNA levels (MX1 and OAS1), with the highest inhibition occurring at ISG's peak expression. Presence of IFN-negative regulators, SOCS1 and SOCS3, during DENV-2 infection was associated with reduced IFN-λ1 expression. Evidence described here suggests that IFN-λ is a good candidate inhibitor of viral replication in dengue infection. Mechanisms for the cellular and organismal interplay between DENV and IFN- λ need to be further studied as they could provide insights into strategies to treat this disease. Furthermore, we report a novel epithelial model to study dengue infection in vitro.


The Maine-Seattle back questionnaire: a 12-item disability questionnaire for evaluating patients with lumbar sciatica or stenosis: results of a derivation and validation cohort analysis.
Analysis of health-related quality of life data obtained from a prospective cohort study of patients with sciatica due to an intervertebral disc herniation or lumbar spinal stenosis. To derive and validate a shortened version of a previously validated 23-item modification of the Roland-Morris Disability Questionnaire. For patients with low back pain, improving health-related quality of life is often the main goal of therapy. The Roland-Morris Disability Questionnaire is one of the best validated and most frequently used back-specific functional status measures. A shortened version may permit more widespread use in clinical and research settings. Data from 507 patients with sciatica enrolled in the Maine Lumbar Spine Study were used to derive a shortened version of a 23-item modification of the original Roland-Morris Disability Questionnaire using qualitative and cluster analysis techniques. The internal consistency, construct validity, reproducibility, and responsiveness in detecting change over a 3-month period for a new 12-item scale was compared to the original 23-item scale. The 12-item scale was then validated in an independent cohort of 148 patients with lumbar spinal stenosis. Internal consistency was very good but modestly lower for the 12-item instrument compared to the 23-item original scale. Reproducibility over a 3-month interval was good and did not differ between the 12-item and original scale. Findings from the validation cohort were similar or better than the derivation cohort. A high degree of construct validity with patient-reported symptoms was demonstrated for the 12-item and original scales. The responsiveness and interpretability of the 12-item scale over 3 months was excellent and comparable to the original scale. Responsiveness assessed in patients with lumbar spinal stenosis in the independent validation cohort showed consistent findings compared to patients with a disc herniation in the derivation cohort. This short, simple, self-administered 12-item back-specific functional status questionnaire performed extremely well in comparison with the original 23-item scale. If validated in additional study populations, this new questionnaire may be useful in the clinical setting as a way for providers to prospectively compare their outcomes of care to other patient populations, and to study treatment effectiveness.


Calcium-activated gene transfection from DNA/poly(amic acid-co-imide) complexes.
In this study, we synthesized a water-soluble poly(amic acid-co-imide) (PA-I) from ethylenediaminetetraacetic dianhydride (EDTA) and 2,2'-(ethylenedioxy)bis(ethylamine) that possesses comparable transfection efficiency to that of polyethylenimine (PEI), when prepared in combination with divalent calcium cations. The polycondensation of monomers afforded poly(amic acid) (PA) precursors, and subsequent thermal imidization resulted in the formation of PA-I. At a polymer/DNA ratio (indicated by the molar ratio of nitrogen in the polymer to phosphate in DNA) of 40, complete retardation of the DNA band was observed by gel electrophoresis, indicating the strong association of DNA with PA-I. A zeta potential of -22 mV was recorded for the PA-I polymer solution, and no apparent cytotoxicity was observed at concentrations up to 500 μg·mL(-1). In the presence of divalent Ca(2+), the transfection efficiency of PA-I was higher than that of PA, due to the formation of a copolymer/Ca(2+)/DNA polyplex and the reduction in negative charge due to thermal cyclization. Interestingly, a synergistic effect of Ca(2+) and the synthesized copolymer on DNA transfection was observed. The use of Ca(2+) or copolymer alone resulted in unsatisfactory delivery, whereas the formation of three-component polyplexes synergistically increased DNA transfection. Our findings demonstrated that a PA-I/Ca(2+)/DNA polyplex could serve as a promising candidate for gene delivery.


Generating squeezed vacuum field with nonzero orbital angular momentum with atomic ensembles.
We demonstrated that by using a pump field with nonzero orbital angular momentum (OAM) in the polarization self-rotation squeezing process it is possible to generate a squeezed vacuum optical field with the matching OAM. We found a similar level of maximum quantum noise reduction for a first-order Laguerre-Gaussian pump beam and a regular Gaussian pump beam, even though the optimal operational conditions differed in these two cases. Also, we investigated the effect of self-defocusing on the level of the vacuum squeezing by simultaneously monitoring the minimum quantum noise level and the output beam transverse profile at various pump laser powers and atomic densities and found no direct correlations between the increased beam size and the degree of measured squeezing.


Effects of soy isoflavones on endothelial function in healthy postmenopausal women.
To evaluate the effects of soy isoflavone administration on endothelial function in healthy postmenopausal women. Sixty naturally postmenopausal women were randomly assigned to receive isoflavone or placebo tablets for 6 months. Endothelium-dependent vasodilatation was measured by brachial reactivity technique along with levels of plasma soluble intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, P-selectin and soluble thrombomodulin, von Willebrand factor, and tissue plasminogen activator. Differences between endothelium-dependent and endothelium-independent vasodilatation were assessed by evaluating brachial reactivity parameters after reactive hyperemia and after sublingual administration of nitroglycerin; furthermore, in the active group, the effect of isoflavones was also evaluated during the intra-arterial infusion of N-monomethyl-L-arginine. Serum levels of lipids [high-density lipoprotein and low-density lipoprotein cholesterol, triglycerides and lipoprotein(a)] and hemostatic factors (prothrombin, fibrinogen, plasminogen activator inhibitor-1, and fibrin D-dimer) were also measured. To confirm the absorption of isoflavones, their blood concentrations were determined. Isoflavone treatment versus placebo was associated with a significant improvement in endothelium-dependent vasodilatation but had no impact on endothelial-independent arterial diameter and flow. Intra-arterial infusion of N-monomethyl-L-arginine inhibited the significant effect of isoflavones on endothelium-mediated vasodilatation. Furthermore, isoflavone group experienced statistically significant reductions in plasma concentrations of ICAM-1, VCAM-1, and E-selectin. Levels of soluble thrombomodulin, von Willebrand factor, tissue plasminogen activator, lipids, and hemostatic factors did not change significantly throughout the study in both groups. Our findings suggest a positive influence of soy isoflavones on endothelial function in healthy postmenopausal women as evidenced by an improvement in endothelium-dependent vasodilatation and a reduction in plasma adhesion molecule levels.


Regulatory assessment of brand changes in the commercial tobacco product market.
Regulatory oversight of tobacco product design has gained momentum in the US and internationally. Appropriate standards for assessing commercial brands and characterising product features must be considered a priority. An area of potential concern is in-market design changes adopted within a single commercial brand over time. Internal tobacco industry documents were identified and used to assess internal discussion of product guidelines and practices regarding in-market brand changes. Commercial tobacco products undergo a constant process of revision in-market, beginning at the most basic level of physical product characteristics and components, and including every aspect of design. These revisions commonly exceed guidelines for acceptable product variance adopted within the industry. While consumer and market testing is conducted to ensure that products remain acceptable to users, explicit marketing often may not accompany brand changes. In the absence of such marketing, it should not be assumed that a brand remains unchanged. For manufacturers, assessment of competitor brands includes identification and analysis of non-routine changes; that is, those changes likely to significantly alter the character of a given brand. Regulators must adopt a similar practice in determining standards for product evaluation in the face of ongoing commercial product revision.


Evidence that the Kennedy and polyamine pathways are dysregulated in human brain in cases of dementia with Lewy bodies.
Disruptions of brain metabolism are considered integral to the pathogenesis of dementia, but thus far little is known of how dementia with Lewy bodies (DLB) impacts the brain metabolome. DLB is less well known than other neurodegenerative diseases such as Alzheimer's and Parkinson's disease which is perhaps why it is under-investigated. This exploratory study aimed to address current knowledge gaps in DLB research and search for potentially targetable biochemical pathways for therapeutics. It also aimed to better understand metabolic similarities and differences with other dementias. Combined metabolomic analyses of 1H NMR and tandem mass spectrometry of neocortical post-mortem brain tissue (Brodmann region 7) from autopsy confirmed cases of DLB (n = 15) were compared with age/gender-matched, non-cognitively impaired healthy controls (n = 30). Following correction for multiple comparisons, only 2 metabolites from a total of 219 measured compounds significantly differed. Putrescine was suppressed (55.4%) in DLB and O-phosphocholine was elevated (52.5%). We identified a panel of 5 metabolites (PC aa C38:4, O-Phosphocholine, putrescine, 4-Aminobutyrate, and SM C16:0) capable of accurately discriminating between DLB and control subjects. Deep Learning (DL) provided the best predictive model following 10-fold cross validation (AUROC (95% CI) = 0.80 (0.60-1.0)) with sensitivity and specificity equal to 0.92 and 0.88, respectively. Altered brain levels of putrescine and O-phosphocholine indicate that the Kennedy pathway and polyamine metabolism are perturbed in DLB. These are accompanied by a consistent underlying trend of lipid dysregulation. As yet it is unclear whether these are a cause or consequence of DLB onset.


Educational attainment and cardiovascular disease in the United States: A quasi-experimental instrumental variables analysis.
There is ongoing debate about whether education or socioeconomic status (SES) should be inputs into cardiovascular disease (CVD) prediction algorithms and clinical risk adjustment models. It is also unclear whether intervening on education will affect CVD, in part because there is controversy regarding whether education is a determinant of CVD or merely correlated due to confounding or reverse causation. We took advantage of a natural experiment to estimate the population-level effects of educational attainment on CVD and related risk factors. We took advantage of variation in United States state-level compulsory schooling laws (CSLs), a natural experiment that was associated with geographic and temporal differences in the minimum number of years that children were required to attend school. We linked census data on educational attainment (N = approximately 5.4 million) during childhood with outcomes in adulthood, using cohort data from the 1992-2012 waves of the Health and Retirement Study (HRS; N = 30,853) and serial cross-sectional data from 1971-2012 waves of the National Health and Nutrition Examination Survey (NHANES; N = 44,732). We examined self-reported CVD outcomes and related risk factors, as well as relevant serum biomarkers. Using instrumental variables (IV) analysis, we found that increased educational attainment was associated with reduced smoking (HRS β -0.036, 95%CI: -0.06, -0.02, p < 0.01; NHANES β -0.032, 95%CI: -0.05, -0.02, p < 0.01), depression (HRS β -0.049, 95%CI: -0.07, -0.03, p < 0.01), triglycerides (NHANES β -0.039, 95%CI: -0.06, -0.01, p < 0.01), and heart disease (HRS β -0.025, 95%CI: -0.04, -0.002, p = 0.01), and improvements in high-density lipoprotein (HDL) cholesterol (HRS β 1.50, 95%CI: 0.34, 2.49, p < 0.01; NHANES β 0.86, 95%CI: 0.32, 1.48, p < 0.01), but increased BMI (HRS β 0.20, 95%CI: 0.002, 0.40, p = 0.05; NHANES β 0.13, 95%CI: 0.01, 0.32, p = 0.05) and total cholesterol (HRS β 2.73, 95%CI: 0.09, 4.97, p = 0.03). While most findings were cross-validated across both data sets, they were not robust to the inclusion of state fixed effects. Limitations included residual confounding, use of self-reported outcomes for some analyses, and possibly limited generalizability to more recent cohorts. This study provides rigorous population-level estimates of the association of educational attainment with CVD. These findings may guide future implementation of interventions to address the social determinants of CVD and strengthen the argument for including educational attainment in prediction algorithms and primary prevention guidelines for CVD.


Genomic BCR-ABL1 breakpoint characterization by a multi-strategy approach for "personalized monitoring" of residual disease in chronic myeloid leukemia patients.
For monitoring minimal residual disease (MRD) in chronic myeloid leukemia (CML) the most recommended method is quantitative RT-PCR (RT-qPCR) for measuring BCR-ABL1 transcripts. Several studies reported that a DNA-based assay enhances the sensitivity of detection of the BCR-ABL1 genomic rearrangement, even if its characterization results difficult. We developed a DNA-based method for detecting and quantifying residual BCR-ABL1 positive leukemic stem cells in CML patients. We propose two alternative approaches: the first one is a fluorescence in situ hybridization (FISH)-based step followed by Sanger sequencing; the second one employs MinION, a single molecule sequencer based on nanopore technology. Finally, after defining the BCR-ABL1 genomic junction, we performed the target CML patient-specific quantification, using droplet digital PCR (ddPCR). FISH and MinION steps, respectively, together with ddPCR analysis, greatly reduce the complexity that has impeded the use of "personalized monitoring" of CML in clinical practice. Our report suggests a feasible pipeline, in terms of costs and reproducibility, aimed at characterizing and quantifying the genomic BCR-ABL1 rearrangement during MRD monitoring in CML patients.


Effective donor abilities of E-t-Bu and EPh (E = O, S, Se, Te) to a high valent transition metal.
Amido rotation in the chromium(vi), d(0)-system NCr(NPr(i)2)2X is under investigation as a method for the parameterization of ligands for their donor properties toward high valent metals. In this study, two new series were prepared and studied based on chalcogenide ligands, X = EBu(t) and EPh and where E = O, S, Se, Te; the OPh and SPh compounds were previously reported. The ligand donor parameters for these ligands correlate with the Cr-E-C angles in these chalcogenide series. In addition, it was found that NBO calculated overlaps and DFT calculated bond dissociation enthalpies correlate within X = halide-, EBu(t)- and EPh-series. All of the new complexes were characterized by X-ray diffraction.


Seizure Treatment in Children Transported to Tertiary Care: Recommendation Adherence and Outcomes.
Convulsive seizures account for 15% of pediatric air transports. We evaluated seizure treatment received in community hospital emergency departments among transported patients for adherence to recommended management. This study was a retrospective cohort study of children transported for an acute seizure to a tertiary pediatric hospital from 2010 to 2013. Seizure treatment was evaluated for adherence to recommended management. The primary outcome was intubation. Among 126 events, 61% did not receive recommended acute treatment. The most common deviation from recommended care was administration of >2 benzodiazepine doses. Lack of adherence to recommended care was associated with a greater than twofold increased risk of intubation (relative risk 2.4; 95% confidence interval, 1.4-4.13) and 1.5-fold increased risk of admission to the ICU (relative risk 1.65; 95% confidence interval, 1.24-2.16). Duration of ventilation was commonly <24 hours (87%) for patients who did or did not receive recommended acute seizure care. Among events treated initially with a benzodiazepine, only 32% received a recommended weight-based dosage, and underdosing was most common. Adherence to evidence-based recommended acute seizure treatment during initial care of pediatric patients using medical air transportation was poor. Intubation was more common when patients did not receive recommended acute seizure care. Educational efforts with a sustained quality focus should be directed to increase adherence to appropriate pediatric seizure treatment of children in community emergency departments.


Health status and socio-economic factors associated with health facility utilization in rural and urban areas in Zambia.
With regards to equity, the objective for health care systems is "equal access for equal needs". We examined associations of predisposing, enabling and need factors with health facility utilization in areas with high HIV prevalence and few people being aware of their HIV status. The data is from a population-based survey among adults aged 15years or older conducted in 2003. The current study is based on a subset of this data of adults 15-49 years with a valid HIV test result. A modified Health behaviour model guided our analytical approach. We report unadjusted and adjusted odds ratios and their 95% confidence intervals from logistic regression analyses. Totals of 1042 males and 1547 females in urban areas, and 822 males and 1055 females in rural areas were included in the study. Overall, 53.1% of urban and 56.8% of rural respondents utilized health facilities past 12 months. In urban areas, significantly more females than males utilized health facilities (OR=1.4 (95% CI [1.1, 1.6]). Higher educational attainment (10+ years of schooling) was associated with utilization of health facilities in both urban (OR=1.7, 95% CI [1.3, 2.1]) and rural (OR=1.4, 95% CI [1.0, 2.0]) areas compared to respondents who attained up to 7 years of schooling. Respondents who self-rated their health status as very poor/ poor/fair were twice more likely to utilize health facilities compared to those who rated their health as good/excellent. Respondents who reported illnesses were about three times more likely to utilize health facilities compared to those who did not report the illnesses. In urban areas, respondents who had mental distress were 1.7 times more likely to utilize health facilities compare to those who had no mental distress. Compared to respondents who were HIV negative, respondents who were HIV positive were 1.3 times more likely to utilize health facilities. The health care needs were the factors most strongly associated with health care seeking. After accounting for need differentials, health care seeking differed modestly by urban and rural residence, was somewhat skewed towards women, and increased substantially with socioeconomic position.


Mutant K-ras oncogenes in colon cancers Do not predict Patient's chemotherapy response or survival
One half of human colon cancers bear mutant c-K-ras oncogenes. Mutant K-ras oncogenes are associated with shortened survival in non-small cell lung cancers, and, in cell line models, with resistance to cis-platinum and to ionizing radiation. This study examines whether mutant K-ras alleles in colon cancer alter patients' response to chemotherapy or survival. We studied 37 patients who received chemotherapy with 5-fluorouracil and leucovorin, Exon 1 of the c-K-ras gene was PCR amplified from DNA extracted from paraffin-embedded tumor blocks. The presence of mutant or wild-type c-K-ras alleles was determined by dideoxy sequencing of the PCR-amplified c-K-ras DNA. c-K-ras mutations at codons 12 or 13 were present in 19 and absent in 18 cases. Responses to chemotherapy were equally likely in patients with either wild-type or mutant c-K-ras, occurring in 28% of patients with wild-type ras and 32% of patients with mutant ras (P = 0.8). Survival was also indistinguishable among both groups. Median survival from diagnosis was 35 months for ras wild-type patients and 31 months for ras mutant patients (P = 0.96). Median survival from starting chemotherapy was 14 months for ras wild-type patients and 17 months for ras mutant patients (P = 0.26). Patients with colon cancers bearing either wild-type or mutant c-K-ras alleles are indistinguishable in overall survival and are equally likely to respond to 5-fluorouracil-based chemotherapy.


Lineage relationship between prostate adenocarcinoma and small cell carcinoma.
Prostate cancer displays different morphologies which, in turn, affect patient outcome. This fact prompted questions about the lineage relationship between differentiated, more treatable prostate adenocarcinoma and poorly differentiated, less treatable non-adenocarcinoma including small cell carcinoma, and the molecular mechanism underlying prostate cancer differentiation. Newly available non-adenocarcinoma/small cell carcinoma PDX LuCaP lines were analyzed for expression of stem cell transcription factors (scTF) LIN28A, NANOG, POU5F1, SOX2, which are responsible for reprogramming or de-differentiation. cDNA of these genes were cloned from small cell carcinoma LuCaP 145.1 into expression vectors to determine if they could function in reprogramming. Expression of scTF was detected in small cell carcinoma LuCaP 93, 145.1, 145.2, and non-adenocarcinoma LuCaP 173.1, 173.2A. Transfection of scTF from LuCaP 145.1 altered the gene expression of prostate non-small cell carcinoma cells, as well as fibroblasts. The resultant cells grew in stem-like colonies. Of note was a 10-fold lower expression of B2M in the transfected cells. Low B2M was also characteristic of LuCaP 145.1. Conversely, B2M was increased when stem cells were induced to differentiate. This work suggested a pathway in the emergence of non-adenocarcinoma/small cell carcinoma from adenocarcinoma through activation of scTF genes that produced cancer de-differentiation.


Introduction and methods of the evidence-based guidelines for the diagnosis and management of autism spectrum disorder by the Italian National Institute of Health.
Autism Spectrum Disorder (ASD) is a neuro-developmental disorder that affects communication and behavior with a prevalence of approximately 1% worldwide. Health outcomes of interventions for ASD are largely Participant Reported Outcomes (PROs). Specific guidelines can help support the best care for people with ASD to optimize these health outcomes but they have to adhere to standards for their development to be trustworthy. The goal of this article is to describe the new methodological standards of the Italian National Institute of Health and novel aspects of this guideline development process. This article will serve as a reference standard for future guideline development in the Italian setting. We applied the new standards of the Italian National Institute of Health to the two guidelines on diagnosis and management of children/adolescents and adults with ASD, with a focus on the scoping, panel composition, management of conflict of interest, generation and prioritization of research questions, early stakeholders' involvement, and PROs. Recommendations are based on the Grading of Recommendations Assessment, Development and Evaluation (GRADE) Evidence-to-Decision frameworks. Following a public application process, the ISS established two multidisciplinary panels including people with ASD and/or their caregivers. Seventy-nine research questions were identified as potentially relevant for the guideline on children and adolescents with ASD and 31 for the one on adults with ASD. Questions deemed to have the highest priority were selected for inclusion in the guidelines. Other stakeholders valued their early involvement in the process which will largely focus on PROs. The panels then successfully piloted the development of recommendations using the methodological standards and process set by the ISS with a focus on PROs. In this article, we describe the development of practice guidelines that focus on PROs for the diagnosis and management of ASD based on novel methods for question prioritization and stakeholder involvement. The recommendations allow for the adoption or adaptation to international settings.


The enactment of national health insurance: a Boolean analysis of twenty advanced industrial countries.
The scholarly literature on health care politics has generated a series of hypotheses to explain U.S. exceptionalism in health policy and to explain the adoption of national health insurance (NHI) more generally. Various cultural, institutional, and political conditions are held to make the establishment of some form of national health insurance policy more (or less) likely to occur. The literature is dominated by national and comparative case studies that illustrate the theoretical logic of these hypotheses but do not provide a framework for examining the hypotheses cross-nationally. This article is an initial attempt to address that void by using Boolean analysis to examine systematically several of the major propositions that emerge from the case study literature on the larger universe of twenty advanced industrial democracies. This comparative analysis offers considerable support for the veto points hypothesis while still finding each of the factors examined to be relevant in certain scenarios. We conclude with a discussion of the implications of these findings for future research and for advocates of national health insurance in the United States.


Dynamic intravoxel incoherent motion imaging of skeletal muscle at rest and after exercise.
The purpose of this work was to demonstrate the feasibility of intravoxel incoherent motion imaging (IVIM) for non-invasive quantification of perfusion and diffusion effects in skeletal muscle at rest and following exercise. After IRB approval, eight healthy volunteers underwent diffusion-weighted MRI of the forearm at 3 T and eight different b values between 0 and 500 s/mm(2) with a temporal resolution of 57 s per dataset. Dynamic images were acquired before and after a standardized handgrip exercise. Diffusion (D) and pseudodiffusion (D*) coefficients as well as the perfusion fraction (FP ) were measured in regions of interest in the flexor digitorum superficialis and profundus (FDS/FDP), brachioradialis, and extensor carpi radialis longus and brevis muscles by using a multi-step bi-exponential analysis in MATLAB. Parametrical maps were calculated voxel-wise. Differences in D, D*, and FP between muscle groups and between time points were calculated using a repeated measures analysis of variance with post hoc Bonferroni tests. Mean values and standard deviations at rest were the following: D*, 28.5 ± 11.4 × 10(-3) mm(2) /s; FP , 0.03 ± 0.01; D, 1.45 ± 0.09 × 10(-3) mm(2) /s. Changes of IVIM parameters were clearly visible on the parametrical maps. In the FDS/FDP, D* increased by 289 ± 236% (p < 0.029), FP by 138 ± 58% (p < 0.01), and D by 17 ± 9% (p < 0.01). A significant increase of IVIM parameters could also be detected in the brachioradialis muscle, which however was significantly lower than in the FDS/FDP. After 20 min, all parameters were still significantly elevated in the FDS/FDP but not in the brachioradialis muscle compared with the resting state. The IVIM approach allows simultaneous quantification of muscle perfusion and diffusion effects at rest and following exercise. It may thus provide a useful alternative to other non-invasive methods such as arterial spin labeling. Possible fields of interest for this technique include perfusion-related muscle diseases, such as peripheral arterial occlusive disease.


BMI is an important driver of β-cell loss in type 1 diabetes upon diagnosis in 10 to 18-year-old children.
Body weight-related insulin resistance probably plays a role in progression to type 1 diabetes, but has an uncertain impact following diagnosis. In this study, we investigated whether BMI measured at diagnosis was an independent predictor of C-peptide decline 1-year post-diagnosis. Multicentre longitudinal study carried out at diagnosis and up to 1-year follow-up. Data on C-peptide were collected from seven diabetes centres in Europe. Patients were grouped according to age at diagnosis (<5 years, n=126; >5 years <10 years, n=295; >10 years <18 years, n=421; >18 years, n=410). Linear regression was used to investigate whether BMI was an independent predictor of change in fasting C-peptide over 1 year. Models were additionally adjusted for baseline insulin dose and HbA1c. In individuals diagnosed between 0 and 5 years, 5 and 10 years and those diagnosed >18 years, we found no association between BMI and C-peptide decline. In patients aged 10-18 years, higher BMI at baseline was associated with a greater decline in fasting C-peptide over 1 year with a decrease (β 95% CI; P value) of 0.025 (0.010, 0.041) nM/kg per m(2) higher baseline BMI (P=0.001). This association remained significant after adjusting for gender and differences in HbA1c and insulin dose (β=0.026, 95% CI=0.0097, 0.042; P=0.002). These observations indicate that increased body weight and increased insulin demand are associated with more rapid disease progression after diagnosis of type 1 diabetes in an age group 10-18 years. This should be considered in studies of β-cell function in type 1 diabetes.


Changes in the abundance of grassland species in monocultures versus mixtures and their relation to biodiversity effects.
Numerous studies have reported positive effects of species richness on plant community productivity. Such biodiversity effects are usually quantified by comparing the performance of plant mixtures with reference monocultures. However, several mechanisms, such as the lack of resource complementarity and facilitation or the accumulation of detrimental agents, suggest that monocultures are more likely than mixtures to deteriorate over time. Increasing biodiversity effects over time could therefore result from declining monocultures instead of reflecting increases in the functioning of mixtures. Commonly, the latter is assumed when positive trends in biodiversity effects occur. Here, we analysed the performance of 60 grassland species growing in monocultures and mixtures over 9 years in a biodiversity experiment to clarify whether their temporal biomass dynamics differed and whether a potential decline of monocultures contributed significantly to the positive net biodiversity effect observed. Surprisingly, individual species' populations produced, on average, significantly more biomass per unit area when growing in monoculture than when growing in mixture. Over time, productivity of species decreased at a rate that was, on average, slightly more negative in monocultures than in mixtures. The mean net biodiversity effect across all mixtures was continuously positive and ranged between 64-217 g per m(2). Short-term increases in the mean net biodiversity effect were only partly due to deteriorating monocultures and were strongly affected by particular species gaining dominance in mixtures in the respective years. We conclude that our species performed, on average, comparably in monocultures and mixtures; monoculture populations being slightly more productive than mixture populations but this trend decreased over time. This suggested that negative feedbacks had not yet affected monocultures strongly but could potentially become more evident in the future. Positive biodiversity effects on aboveground productivity were heavily driven by a small, but changing, set of species that behaved differently from the average species.


Extracellular signal-regulated kinase phosphorylates tumor necrosis factor alpha-converting enzyme at threonine 735: a potential role in regulated shedding.
The ectodomain of certain transmembrane proteins can be released by the action of cell surface proteases, termed secretases. Here we have investigated how mitogen-activated protein kinases (MAPKs) control the shedding of membrane proteins. We show that extracellular signal-regulated kinase (Erk) acts as an intermediate in protein kinase C-regulated TrkA cleavage. We report that the cytosolic tail of the tumor necrosis factor alpha-converting enzyme (TACE) is phosphorylated by Erk at threonine 735. In addition, we show that Erk and TACE associate. This association is favored by Erk activation and by the presence of threonine 735. In contrast to the Erk route, the p38 MAPK was able to stimulate TrkA cleavage in cells devoid of TACE activity, indicating that other proteases are also involved in TrkA shedding. These results demonstrate that secretases are able to discriminate between the different stimuli that trigger membrane protein ectodomain cleavage and indicate that phosphorylation by MAPKs may regulate the proteolytic function of membrane secretases.


Simple estimation of absolute free energies for biomolecules.
One reason that free energy difference calculations are notoriously difficult in molecular systems is due to insufficient conformational overlap, or similarity, between the two states or systems of interest. The degree of overlap is irrelevant, however, if the absolute free energy of each state can be computed. We present a method for calculating the absolute free energy that employs a simple construction of an exactly computable reference system which possesses high overlap with the state of interest. The approach requires only a physical ensemble of conformations generated via simulation and an auxiliary calculation of approximately equal central-processing-unit cost. Moreover, the calculations can converge to the correct free energy value even when the physical ensemble is incomplete or improperly distributed. As a "proof of principle," we use the approach to correctly predict free energies for test systems where the absolute values can be calculated exactly and also to predict the conformational equilibrium for leucine dipeptide in implicit solvent.


The significant role of mast cells in cancer.
Mast cells (MC) are a bone marrow-derived, long-lived, heterogeneous cellular population that function both as positive and negative regulators of immune responses. They are arguably the most productive chemical factory in the body and influence other cells through both soluble mediators and cell-to-cell interaction. MC are commonly seen in various tumors and have been attributed alternatively with tumor rejection or tumor promotion. Tumor-infiltrating MC are derived both from sentinel and recruited progenitor cells. MC can directly influence tumor cell proliferation and invasion but also help tumors indirectly by organizing its microenvironment and modulating immune responses to tumor cells. Best known for orchestrating inflammation and angiogenesis, the role of MC in shaping adaptive immune responses has become a focus of recent investigations. MC mobilize T cells and antigen-presenting dendritic cells. They function as intermediaries in regulatory T cells (Treg)-induced tolerance but can also modify or reverse Treg-suppressive properties. The central role of MC in the control of innate and adaptive immunity endows them with the ability to tune the nature of host responses to cancer and ultimately influence the outcome of disease and fate of the cancer patient.


The Petroleum Ether Extract from Hypericum perforatum Root Cultures Exhibits Potent Antiproliferative Activity in Human Keratinocytes and Fibroblasts.
The hyperforin content of Hypericum perforatum herb was repeatedly reported to be responsible for a multitude of pharmacological activities. Our recent report about the hyperforin accumulation in in vitro root cultures of H. perforatum provides an alternative perspective to achieve constant product quality and to serve the rapidly growing market. In this study, the antiproliferative effect of a petroleum ether extract from the in vitro root cultures was investigated. When normalized to 1 µM hyperforin content, the extract reduced the viability of human keratinocytes (HaCaT) and human dermal fibroblast monolayers to 33 and 36%, respectively, after 72 h of incubation. A cytotoxicity assay and live-dead cell staining confirmed that the extract lacked a cytotoxic effect and that the reduction in cell viability was mainly due to the antiproliferative activity. Application of the 1 µM hyperforin-normalized extract to a 3D artificial skin construct significantly reduced the proliferation of HaCaT in the presence of fibroblasts. This effect was proved by the reduction in thickness of the epidermal construct from 100 µm (control) to 17 µm (treated). Notably, 1 µM pure hyperforin lacked effectiveness in both monolayer cultures and 3D artificial skin constructs. Nor were fractions of the extract containing colupulone and xanthones active. The combination of these constituents also failed to reassemble the antiproliferative activity, which indicates a synergistic role of yet unidentified components present in the extract. Our findings may introduce H. perforatum root cultures as a novel lead system for the treatment of hypertrophic scars.


An all-optical single-step process for production of nanometric-sized fluorescent diamonds.
Nanodiamonds (NDs) containing negatively charged Nitrogen-Vacancy (NV) centers are promising materials for applications in photonics, quantum computing, and sensing of environmental parameters like temperature, strain and magnetic fields. However, the production of fluorescent NDs remains a technological challenge, requiring a complex multi-step process involving controlled introduction of substitutional nitrogen into the diamond lattice, annealing and fragmentation from macrocrystals to nanocrystals. Here, we report on a single-step, all-optical process for the production of nanometric-sized fluorescent diamonds based on laser ablation of a carbon substrate at low temperature (100 °C) under a nitrogen atmosphere. We demonstrate that this synthesis route yields fluorescent NDs with a concentration of native NV centers controlled by adjusting the experimental ablation conditions. Spin-polarization dependent optical-transitions are observed by optically detected magnetic resonance spectroscopy, thus providing strong evidence of the presence of negatively charged NV centers in the as-grown NDs. Finally, we propose a thermodynamic model able to describe the nucleation of NDs and the formation of NV centers in the present single-step optical process.


Sporulation in Bacillus brevis: studies on protease and protein turnover.
Bacillus brevis ATCC 9999 sporulated without the production of extracellular protease when grown and allowed to sporulate on nutrient agar plates containing either azo-albumin, casein, or gelatin. Intracellular protease could not be detected in extracts of cells grown in nutrient broth-salts media. However, approximately one-fifth of the level of protease activity in extracts of B. subtilis SB 19 was found in extracts of B. brevis ATCC 9999 grown in minimal media. When protein turnover studies were performed by chasing previously incorporated (14)C-l-phenylalanine with a large excess of unlabeled phenylalanine, no protein turnover could be detected if the excess phenylalanine was added at T(0) and only a small amount of turnover (1 to 1.6% per hr) was detected if the excess phenylalanine was added at T(2.5-2.7). This compares with a measured turnover rate of 8.5% per hr in B. subtilis SB 19 under similar conditions.


How stable are stages of change for nutrition behaviors in the Netherlands?
This paper describes the stability of the stages of change concept of the Transtheoretical Model for three different nutrition behaviors (fat, fruit and vegetable intake) among adult individuals who are unexposed to planned interventions. Secondary analyses were conducted on data collected in control groups (n = 386 and n = 739) of two intervention studies in the Netherlands. Data on dietary intakes and stages of change was collected at baseline and follow up, either 1 year (study 1) or 3 months (study 2) post-baseline. Higher levels of agreement between baseline and follow-up measures of stage of change were found for pre-contemplation and maintenance than for the other stages. However, many forward as well as backward stage transitions occurred between baseline and follow-up also among respondents in pre-contemplation and maintenance at baseline. The results indicate that many stage transitions may occur in individuals, also when they are not exposed to planned interventions. An additional explanation for the stage instability may be low reliability of the staging algorithm. The results imply that if classification into stages of change is used to tailor interventions, these interventions may be tailored to the wrong stage, at least with longer time intervals between stage assessment and intervention. Further research is needed to assess 'spontaneous' stage-transitions in shorter time intervals.


Activity of isoflavones and berberine on vasomotor symptoms and lipid profile in menopausal women.
The aim of this study was to evaluate the efficacy of a food supplement combination based on isoflavones and berberine (ISB) in the treatment of menopausal symptoms and dyslipidaemia. Isoflavones are extracted from soy and absorbed in the body after being activated by lactobacillus. Berberine, extracted from the plant Berberis aristata, lowers plasma cholesterol and triglycerides (TG) by increasing low-density lipoprotein (LDL) receptors and reducing hepatic synthesis of TG. One hundred twenty women with a mean age of 54.8 ± 0.6 years were enrolled and randomized to treatment with ISB (estromineral lipid [EL] = 60 cases) or calcium and vitamin D(3) (CaD = 60 cases). Menopausal symptoms, plasma cholesterol, and TG were evaluated at baseline, and after 4 and 12 weeks. EL treatment significantly lowered plasma total cholesterol (-13.5% ± 0.7 vs -0.2% ± 0.5), LDL cholesterol (-12.4% ± 1.5 vs + 0.8 % ± 0.7) and TG (-18.9% ± 2.5 vs -1.3% ± 1.2) and improved menopausal symptoms compared with CaD treatment. Safety parameters were unchanged during the study. The combination of berberine and isoflavones was effective in lowering cardiovascular (CV) risk factors in menopausal women with moderate dyslipidaemia and in improving their quality of life.


Lactate and T (2) measurements of synovial aspirates at 1.5 T: differentiation of septic from non-septic arthritis.
The aim of this study was to differentiate septic from non-septic arthritis by measuring lactate concentration with (1)H magnetic resonance spectroscopy (HMRS) and by estimating total protein content with the assessment of T (2) values. In 30 patients with acute arthritis, synovial fluid was aspirated. Lactate concentrations were analyzed with single voxel HMRS at 1.5 T. T (2) relaxation times were mapped with a multi-spin echo sequence. All samples underwent microbiological testing and routine laboratory analysis to quantify lactate concentration and total protein content. Values obtained in septic and non-septic arthritis were compared with a Mann-Whitney U test. Synovial fluid from patients with septic arthritis (n = 10) had higher concentrations of lactate (11.4 +/- 4.0 mmol/L) and higher total protein content (51.8 +/- 10.7 g/L) than fluid obtained in non-septic arthritis (n = 20; 5.2 +/- 1.1 mmol/L and 40.4 +/- 6.9 g/L, respectively, p < 0.001 and <0.01, respectively). Measured lactate concentrations and T (2) relaxation times (as an indicator of total protein content) were moderately correlated to laboratory-confirmed lactate concentration (r (2) = 0.71) and total protein content (r (2) = 0.73). Markedly increased lactate concentrations (>6 mmol/L) in combination with low T (2) values (<550 ms) identify septic arthritis with a sensitivity of 70% and a specificity of 89%. Spectroscopic measurements of lactate concentration in combination with the estimation of protein content using T (2) may be of value in the differentiation of septic from non-septic arthritis.


Burden of increasing opioid use in the treatment of migraine: Results from the Migraine in America Symptoms and Treatment Study.
We sought to assess factors associated with the frequency of self-reported prescription opioid use in persons with migraine, including demographic variables, comorbidities, headache characteristics, and patterns of consultation. Despite the dose-dependent effect of opioids on migraine progression and the association with negative outcomes, migraine treatment often includes opioids. The Migraine in America Symptoms and Treatment Study focuses on individuals with migraine who receive prescription acute medications, including those receiving and those not receiving opioids. This web-based panel survey identified people in the United States with migraine using a validated screener. This analysis stratified people with migraine into 4 groups based on days of monthly opioid use: non-opioid users, ≤3 days, 4-9 days, and ≥10 days per month. Of 15,133 respondents with migraine, 4701 (31%) reported acute prescription medication use for headache/migraine in the previous 3 months (mean age 45 years, 71.6% [3367/4701] female), of whom 32.5% (1528/4701) reported opioid use. About one-third of respondents with primary care or neurology consults in the prior 6 months reported receiving an opioid, and more than half of respondents (209/391, 53.5%) with a pain clinic consultation did so. Models compared those using opioids ≤3 days/month (879/4701, 18.7%), 4-9 days/month (304/4701, 6.5%), ≥10 days/month (345/4701, 7.3%) to non-opioid users (3173/4701, 67.5%). Compared to non-opioid users, infrequent users (≤3 days/month) were more likely to be male and less likely to have chronic migraine or to screen positive for anxiety and depression; and frequent opioid users (the 4-9 days/month and the ≥10 days/month groups) were more likely to be male, to smoke, to be obese, to report greater pain interference, to have moderate to severe disability, to have symptoms of anxiety and depression, to use fewer triptans and nonsteroidal anti-inflammatory drugs, and to have poor acute treatment optimization. Among prescription medication users, this cross-sectional analysis shows that increasing use of prescription opioids is associated with male gender, chronic migraine, more severe disability, anxiety and depression, poor acute treatment optimization, and treatment in a pain clinic.


Generational distribution of a Candida glabrata population: Resilient old cells prevail, while younger cells dominate in the vulnerable host.
Similar to other yeasts, the human pathogen Candida glabrata ages when it undergoes asymmetric, finite cell divisions, which determines its replicative lifespan. We sought to investigate if and how aging changes resilience of C. glabrata populations in the host environment. Our data demonstrate that old C. glabrata are more resistant to hydrogen peroxide and neutrophil killing, whereas young cells adhere better to epithelial cell layers. Consequently, virulence of old compared to younger C. glabrata cells is enhanced in the Galleria mellonella infection model. Electron microscopy images of old C. glabrata cells indicate a marked increase in cell wall thickness. Comparison of transcriptomes of old and young C. glabrata cells reveals differential regulation of ergosterol and Hog pathway associated genes as well as adhesion proteins, and suggests that aging is accompanied by remodeling of the fungal cell wall. Biochemical analysis supports this conclusion as older cells exhibit a qualitatively different lipid composition, leading to the observed increased emergence of fluconazole resistance when grown in the presence of fluconazole selection pressure. Older C. glabrata cells accumulate during murine and human infection, which is statistically unlikely without very strong selection. Therefore, we tested the hypothesis that neutrophils constitute the predominant selection pressure in vivo. When we altered experimentally the selection pressure by antibody-mediated removal of neutrophils, we observed a significantly younger pathogen population in mice. Mathematical modeling confirmed that differential selection of older cells is sufficient to cause the observed demographic shift in the fungal population. Hence our data support the concept that pathogenesis is affected by the generational age distribution of the infecting C. glabrata population in a host. We conclude that replicative aging constitutes an emerging trait, which is selected by the host and may even play an unanticipated role in the transition from a commensal to a pathogen state.


Carotid intima-media thickness in surgical menopause: women who received HRT versus who did not.
The effects of hormone replacement therapy (HRT) in the natural menopausal period have been extensively studied. However, these effects have almost none been studied in purely surgical menopause. The aim of this study was to measure intima-media thickness (IMT) of carotid arteries bilaterally in two groups of surgical menopausal women who received HRT versus who did not. A B-mode ultrasound unit was used for the measurements of the IMTs of carotid arteries in two groups. Measurements of Group 1 (n=65, untreated group) were compared with those of Group 2 (n=70, treated group), in 2-years of follow-up. Patients in Group 2 received daily doses of 0.625 mg of oral conjugated estrogen preparates. Serum estradiol levels, lipid profiles, and blood pressures were measured, pre and postoperatively. For the statistical analyses in terms of differences of IMTs between two groups, general factorial analysis of variation was used. Among preoperative values of low-density lipoprotein (LDL), high-density lipoprotein (HDL), cholesterol, systolic and diastolic blood pressures, estradiol, and age, only the estradiol values showed significant difference between both groups. The statistical results concerning the postoperative IMT differences for both groups showed that there was a statistically significant difference when comparing both groups, showing an increase in IMT in Group 1. In surgical menopausal women, the direction of the HRT effect is in agreement with evidence from earlier studies on the effects of HRT in natural menopausal women.


Biocompatibility of a novel cyanoacrylate based tissue adhesive: cytotoxicity and biochemical property evaluation.
Cyanoacrylate (CA) is most widely used as a medical and commercial tissue adhesive because of easier wound closure, good cosmetic results and little discomfort. But, CA-based tissue adhesives have some limitations including the release of cytotoxic chemicals during biodegradation. In previous study, we made prepolymerized allyl 2-CA (PACA) based tissue adhesive, resulting in longer chain structure. In this study, we investigated a biocompatibility of PACA as alternative tissue adhesive for medical application, comparing with that of Dermabond® as commercial tissue adhesive. The biocompatibility of PACA was evaluated for short-term (24 hr) and long-term (3 and 7 days) using conventional cytotoxicity (WST, neutral red, LIVE/DEAD and TUNEL) assays, hematoxylin-eosin (H&E) and Masson trichrome (MT) staining. Besides we examined the biochemical changes in cells and DNA induced by PACA and Dermabond® utilizing Raman spectroscopy which could observe the denaturation and conformational changes in protein, as well as disintegration of the DNA/RNA by cell death. In particular, we analyzed Raman spectrum using the multivariate statistical methods including principal component analysis (PCA) and support vector machine (SVM). As a result, PACA and Dermabond® tissue adhesive treated cells and tissues showed no difference of the cell viability values, histological analysis and Raman spectral intensity. Also, the classification analysis by means of PCA-SVM classifier could not discriminate the difference between the PACA and Dermabond® treated cells and DNA. Therefore we suggest that novel PACA might be useful as potential tissue adhesive with effective biocompatibility.


Stem cell and gene therapeutic strategies for the treatment of multiple sclerosis.
Multiple sclerosis is a disease of the central nervous system that predmoninantly affects young adults. The pathogenic mechanisms are complex, however numerous studies indicate that the disease is initiated by an autoimmune attack on protein targets present in the central nervous system. Given that a dysfunctional immune system perpetuates the pathophysiological mechanisms that characterize this inflammatory disorder, several therapeutic approaches that target immune cells or their secreted mediators have been generated and are currently used clinically. Although these strategies have been partially beneficial to a proportion of patients, current therapies are not particularly effective at preventing disease progression. As such there is a large and unmet need for the development of more effective treatments. Owing to a number of promising results obtained in mouse models of multiple sclerosis, cell therapies implementing hematopoietic, mesenchymal and neural stem cells may provide practical vehicles for in situ immunomodulation, neuroprotection and regeneration. In concert with these approaches, gene therapy strategies are being investigated to restore antigen-specific tolerance or to deliver anti-inflammatory molecules. Furthermore targeted delivery of glial or neurotropic factors, which counteract the activity of inhibitory molecules within the neurodegenerative component of the lesion, is also being pursued. It is conceivable that these experimental approaches alone, or in combination with emerging and current treatments, may establish a rational protocol for the treatment of multiple sclerosis and potentially other autoimmune disorders.


Occupational lifting of heavy loads and preterm birth: a study within the Danish National Birth Cohort.
To examine the association between occupational lifting during pregnancy and preterm birth. The risk of preterm birth was estimated for total burden lifted per day and number of medium and heavy loads lifted per day. In a study population of 62 803 pregnant women enrolled to the Danish National Birth Cohort from 1996 to 2002, the association between self-reported occupational lifting in the first part of pregnancy and preterm birth was analysed using logistic regression models with adjustment for age, parity, cervical cone biopsy, assisted reproduction and smoking. Associations between lifting and extremely (before 28 weeks), very (28-32 weeks) and moderately (33-37 weeks) preterm birth were analysed using Cox regression models. We found a dose-response relation between total daily burden lifted and preterm birth with an OR of 1.50 (95% CI 1.03 to 2.19) with loads over 1000 kg/day. No threshold value was found. The associations were strongest for extremely and very preterm birth with HRs (95% CIs) of 4.3 (1.4 to 13.8) and 1.7 (0.7 to 4.0), respectively. Lifting heavy loads (>20 kg) more than10 times/day was associated with preterm birth up to an OR of 2.03 (95% CI 1.14 to 3.62). In a society with social welfare and a highly regulated working environment, occupational lifting was associated with an increased risk of preterm birth.


Chilli Anthracnose: The Epidemiology and Management.
Indian cuisine is renowned and celebrated throughout the world for its spicy treat to the tongue. The flavor and aroma of the food generated due to the use of spices creates an indelible experience. Among the commonly utilized spices to stimulate the taste buds in Indian food, whole or powdered chilli constitutes an inevitable position. Besides being a vital ingredient of of Indian food, chilli occupy an important position as an economic commodity, a major share in Indian economy. Chilli also has uncountable benefits to human health. Fresh green chilli fruits contain more Vitamin C than found in citrus fruits, while red chilli fruits have more Vitamin A content than as found in carrots. The active component of the spice, Capsaicin possesses the antioxidant, anti-mutagenic, anti-carcinogenic and immunosuppressive activities having ability to inhibit bacterial growth and platelet aggregation. Though introduced by the Portuguese in the Seventeenth century, India has been one of the major producers and exporters of this crop. During 2010-2011, India was the leading exporter and producer of chilli in the world, but recently due to a decline in chilli production, it stands at third position in terms of its production. The decline in chilli production has been attributed to the diseases linked with crop like anthracnose or fruit rot causing the major share of crop loss. The disease causes severe damage to both mature fruits in the field as well as during their storage under favorable conditions, which amplifies the loss in yield and overall production of the crop. This review gives an account of the loss in production and yield procured in chili cultivation due to anthracnose disease in Indian sub-continent, with emphasis given to the sustainable management strategies against the conventionally recommended control for the disease. Also, the review highlights the various pathogenic species of Colletotrichum spp, the causal agent of the disease, associated with the host crop in the country. The information in the review will prove of immense importance for the groups targeting the problem, for giving a collective information on various aspects of the epidemiology and management of the disease.


The Impact of Li Grain Size on Coulombic Efficiency in Li Batteries.
One of the most promising means to increase the energy density of state-of-the-art lithium Li-ion batteries is to replace the graphite anode with a Li metal anode. While the direct use of Li metal may be highly advantageous, at present its practical application is limited by issues related to dendrite growth and low Coulombic efficiency, CE. Here operando electrochemical scanning transmission electron microscopy (STEM) is used to directly image the deposition/stripping of Li at the anode-electrolyte interface in a Li-based battery. A non-aqueous electrolyte containing small amounts of H2O as an additive results in remarkably different deposition/stripping properties as compared to the "dry" electrolyte when operated under identical electrochemical conditions. The electrolyte with the additive deposits more Li during the first cycle, with the grain sizes of the Li deposits being significantly larger and more variable. The stripping of the Li upon discharge is also more complete, i.e., there is a higher cycling CE. This suggests that larger grain sizes are indicative of better performance by leading to more uniform Li deposition and an overall decrease in the formation of Li dendrites and side reactions with electrolyte components, thus potentially paving the way for the direct use of Li metal in battery technologies.


Developing and implementing a standardized process for global trigger tool application across a large health system.
To complement voluntary adverse event reporting, which may detect only specific categories of harms and may represent merely a fraction of actual adverse events, the Adventist Health System (AHS) began using the Institute for Healthcare Improvement (IHI) Global Trigger Tool (GTT) to more accurately gauge the number, types, and severity levels of adverse events and developed a centralized process to do so uniformly. AHS began using the GTT in 2009 in 25 of its 42 hospitals that used a common electronic medical record (EMR). The common EMR and centralized record review enables AHS to apply the GTT uniformly and provides consistency of data collected. AHS sends quarterly reports to participating facilities to communicate findings and provides case studies illustrating the most egregious harms. Case study recipients are encouraged to further examine patient records, explore events leading to harm, and share the information with process/quality improvement committees, medical executive committees, and boards of directors to identify opportunities for quality improvement. AHS staffing and record review processes have evolved since 2009. A GTT review of 17,295 patient records indicated that adverse events clustered as medication-related glycemic events; medication-related delirium, confusion, or oversedation related to analgesics, sedatives, and muscle relaxants; pressure ulcers; medication-related bleeding; and medication-related skin/mucosal reaction/itching. The AHS process demonstrates how a large health system uses the GTT to detect harms. Since 2009 AHS has improved and streamlined its reporting, data entry and review processes. AHS used major harms findings to initiate systemwide collaborative improvement projects for glycemic management and pressure ulcers.


Determining health care rights from behind a veil of ignorance.
Should our society establish positive rights to health care that each citizen could claim, as many health policy analysts believe? Or should it provide only background rules of contract and property law and leave the provision of health care to the free market, as Richard Epstein advocates in Mortal Peril? In this article, Professor Korobkin argues that this question should be addressed from the Rawlsian "veil of ignorance" perspective. That is, the question should be answered by asking what kind of society would individuals agree to form if they had no knowledge of their individual skills or endowments; if they did not know whether they were rich or poor, healthy or sick, weak or strong. Professor Korobkin contends that individuals behind such a veil of ignorance would balance their inherent risk aversion (which favors a safety net of "rights") against the inefficient incentives created by rights regimes that would reduce net social wealth (which favors a free market). Whether they would choose to establish rights to health care or not is ultimately an empirical question that turns on how inefficient any particular right would be. The question thus requires a case-by-case analysis of proposed rights. The article then considers the policy issues of (1) community rating of private health insurance and (2) the mandated provision of emergency medical care. It concludes that in these cases the inefficient incentives created by establishing rights are probably smaller and/or controllable enough to lead individuals behind the veil of ignorance to favor a regime of positive rights.


Human Milk Oligosaccharides and Associations With Immune-Mediated Disease and Infection in Childhood: A Systematic Review.
Complex sugars found in breastmilk, human milk oligosaccharides (HMOs), may assist in early-life immune programming and prevention against infectious diseases. This study aimed to systematically review the associations between maternal levels of HMOs and development of immune-mediated or infectious diseases in the offspring. PubMed and EMBASE databases were searched (last search on 22 February 2018) according to a predetermined search strategy. Original studies published in English examining the effect of HMOs on immune-mediated and infectious disease were eligible for inclusion. Of 847 identified records, 10 articles from 6 original studies were included, with study quality ranging from low to high. Of three studies to examine allergic disease outcomes, one reported a protective effect against cow's milk allergy (CMA) by 18 months of age associated with lower lacto-N-fucopentaose (LNFP) III concentrations (OR: 6.7, 95% CI 2.0-22). Another study found higher relative abundance of fucosyloligosaccharides was associated with reduced diarrhea incidence by 2 years, due to (i) stable toxin-E. coli infection (p = 0.04) and (ii) "all causes" (p = 0.042). Higher LNFP-II concentrations were associated with (i) reduced cases of gastroenteritis and respiratory tract infections at 6 weeks (p = 0.004, p = 0.010) and 12 weeks (p = 0.038, p = 0.038) and (ii) reduced HIV transmission (OR: 0.45; 95% CI: 0.21-0.97) and mortality risk among HIV-exposed, uninfected infants (HR: 0.33; 95% CI: 0.14-0.74) by 24 months. Due to heterogeneity of the outcomes reported, pooling of results was not possible. There was limited evidence that low concentrations of LNFP-III are associated with CMA and that higher fucosyloligosaccharide levels protect infants against infectious disease. Further research is needed.


Single-molecule spectroscopy of protein folding in a chaperonin cage.
Molecular chaperones are known to be essential for avoiding protein aggregation in vivo, but it is still unclear how they affect protein folding mechanisms. We use single-molecule Förster resonance energy transfer to follow the folding of a protein inside the GroEL/GroES chaperonin cavity over a time range from milliseconds to hours. Our results show that confinement in the chaperonin decelerates the folding of the C-terminal domain in the substrate protein rhodanese, but leaves the folding rate of the N-terminal domain unaffected. Microfluidic mixing experiments indicate that strong interactions of the substrate with the cavity walls impede the folding process, but the folding hierarchy is preserved. Our results imply that no universal chaperonin mechanism exists. Rather, a competition between intra- and intermolecular interactions determines the folding rates and mechanisms of a substrate inside the GroEL/GroES cage.


The accuracy of Doppler ultrasound measurement of pressure gradients across irregular, dual, and tunnellike obstructions to blood flow.
The accuracy of Doppler-estimated pressure gradients in the setting of irregular, multiple, and tunnellike stenoses was investigated. An in vitro model of the left ventricular outflow tract was designed to allow pulsatile flow of red cells in saline across valve orifices from 0.01 to 2.5 cm2. Simultaneous pressure gradients were estimated by both Doppler and direct-pressure manometer techniques. Gradients obtained by the two methods correlated well for valve areas in the range of clinical stenoses at pressure gradients of 10 to 150 mm Hg (r = .97 to .99). Model valves were constructed with a large orifice (0.75 to 1.25 cm2) placed beside a small orifice (0.02 to 0.25 cm2) in the same outflow tract. A distinct jet was recorded when the Doppler transducer was aligned with each orifice. Doppler-estimated gradients for each pair of large and small orifices were identical and correlated well with those measured by manometer (r = .97 to .99). Irregularly shaped orifices also provided good correlation between the two methods (r = .98 to .99). Pulsatile flow was generated through long tunnellike obstructions with cross-sectional areas varying from 0.06 to 1.25 cm2. Tunnel length varied from 0.1 to 4 cm. Tunnel areas above 0.25 cm2 gave good Doppler-to-manometer correspondence at all tunnel lengths. Doppler underestimated manometer-determined values in the 0.25 cm2 tunnel by 8% at 3 cm and by 15% at 4 cm. In the 0.06 cm2 tunnel, Doppler underestimated manometer gradients by 12%, 15%, 32%, and 42% at lengths of 1, 2, 3, and 4 cm, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Dendritic cell activation by an E. coli-derived monophosphoryl lipid A enhances the efficacy of PD-1 blockade.
Cancer immunotherapy is a powerful approach for cancer treatment, but its clinical effects rely on the tumor's immune conditions. In particular, low response rates to PD-1 blockades are highly correlated with impaired T cell priming. Here, we demonstrate that E. coli-derived monophosphoryl lipid A (EcML) activates dendritic cells in a toll-like receptor-4 (TLR-4)-dependent manner and increases the sensitivity of cancer cells to anti-PD-1 immunotherapy. EcML is a mixture of 4'-monophosphoryl lipids A (MPLAs) produced directly by an engineered Escherichia coli strain; it has a unique congener composition that differentiates it from the well-established MPLA adjuvants, 3-O-desacyl-4'-monophosphoryl lipid A and glucopyranosyl lipid A. Given that active dendritic cells initiate adaptive immune responses, we investigated the anti-tumor activity of an aqueous formulation of EcML. Upon sensing EcML via TLR-4, dendritic cells matured into powerful antigen-presenting cells that could stimulate naïve T cells. EcML reduced tumor growth in the B16F10 mouse model via dendritic cell activation and potentiated PD-1 blockade therapy in the B16F10-OVA melanoma model. These data identify EcML as a promising TLR-4 agonist that can induce anti-tumor immune responses and potentiate PD-1 blockade therapy against tumors.


Enteropathogenic Escherichia coli inhibits ileal sodium-dependent bile acid transporter ASBT.
Apical sodium-dependent bile acid transporter (ASBT) is responsible for the absorption of bile acids from the intestine. A decrease in ASBT function and expression has been implicated in diarrhea associated with intestinal inflammation. Whether infection with pathogenic microorganisms such as the enteropathogenic Escherichia coli (EPEC) affect ASBT activity is not known. EPEC is a food-borne enteric pathogen that translocates bacterial effector molecules via type three secretion system (TTSS) into host cells and is a major cause of infantile diarrhea. We investigated the effects of EPEC infection on ileal ASBT function utilizing human intestinal Caco2 cells and HEK-293 cells stably transfected with ASBT-V5 fusion protein (2BT cells). ASBT activity was significantly inhibited following 60 min infection with EPEC but not with nonpathogenic E. coli. Mutations in bacterial escN, espA, espB, and espD, the genes encoding for the elements of bacterial TTSS, ablated EPEC inhibitory effect on ASBT function. Furthermore, mutation in the bacterial BFP gene encoding for bundle-forming pili abrogated the inhibition of ASBT by EPEC, indicating the essential role for bacterial aggregation and the early attachment. The inhibition by EPEC was associated with a significant decrease in the V(max) of the transporter and a reduction in the level of ASBT on the plasma membrane. The inhibition of ASBT by EPEC was blocked in the presence of protein tyrosine phosphatase inhibitors. Our studies provide novel evidence for the alterations in the activity of ASBT by EPEC infection and suggest a possible effect for EPEC in influencing intestinal bile acid homeostasis.


Honey is potentially effective in the treatment of atopic dermatitis: Clinical and mechanistic studies.
As manuka honey (MH) exhibits immunoregulatory and anti-staphylococcal activities, we aimed to investigate if it could be effective in the treatment of atopic dermatitis (AD). Adult volunteers with bilateral AD lesions were asked to apply MH on one site overnight for seven consecutive days and leave the contralateral site untreated as possible. Three Item Severity score was used to evaluate the response. Skin swabs were obtained from both sites before and after treatment to investigate the presence of staphylococci and enterotoxin production. In addition, the ability of MH and its methanolic and hexane extracts to down regulate IL4-induced CCL26 protein release from HaCaT cells was evaluated by enzyme linked immunosorbent assay. Also, the ability of MH to modulate calcium ionophore-induced mast cell degranulation was assessed by enzyme immunoassay. In 14 patients, AD lesions significantly improved post MH treatment versus pre-treatment as compared to control lesions. No significant changes in the skin staphylococci were observed after day 7, irrespective of honey treatment. Consistent with the clinical observation, MH significantly down regulated IL4-induced CCL26 release from HaCaT cells in a dose-dependent manner. This effect was partially lost, though remained significant, when methanolic and hexane extracts of MH were utilized. In addition, mast cell degranulation was significantly inhibited following treatment with MH. MH is potentially effective in the treatment of AD lesions based on both clinical and cellular studies through different mechanisms. This needs to be confirmed by randomized and controlled clinical trials.


Relationship Between Physical Activity and Overweight and Obesity in Children: Findings From the 2012 National Health and Nutrition Examination Survey National Youth Fitness Survey.
This study examined the relationship between childhood obesity and overweight and functional activity and its enjoyment. A cross-sectional design was used to analyze data from the 2012 National Health and Nutrition Examination Survey National Youth Fitness Survey. Multivariate logistic regression models were used. Data for 1,640 children ages 3-15 yr were retrieved. Physical activity was negatively associated with risk of obesity (odds ratio [OR] = 0.93; 95% confidence interval [CI] [0.87, 0.98]). Although children who were obese and overweight were more likely to have functional limitations (ORs = 1.58-1.61), their enjoyment of physical activity participation was not significantly different from that of the healthy-weight group. Physical activity lowered the risk of obesity. Children who were obese had functional limitations compared with healthy-weight children, but both groups enjoyed physical activity equally. Future studies are needed to determine barriers to participation among these children in recreation and sporting activities.


Multiphyletic origins of methylotrophy in Alphaproteobacteria, exemplified by comparative genomics of Lake Washington isolates.
We sequenced the genomes of 19 methylotrophic isolates from Lake Washington, which belong to nine genera within eight families of the Alphaproteobacteria, two of the families being the newly proposed families. Comparative genomic analysis with a focus on methylotrophy metabolism classifies these strains into heterotrophic and obligately or facultatively autotrophic methylotrophs. The most persistent metabolic modules enabling methylotrophy within this group are the N-methylglutamate pathway, the two types of methanol dehydrogenase (MxaFI and XoxF), the tetrahydromethanopterin pathway for formaldehyde oxidation, the serine cycle and the ethylmalonyl-CoA pathway. At the same time, a great potential for metabolic flexibility within this group is uncovered, with different combinations of these modules present. Phylogenetic analysis of key methylotrophy functions reveals that the serine cycle must have evolved independently in at least four lineages of Alphaproteobacteria and that all methylotrophy modules seem to be prone to lateral transfers as well as deletions.


Spontaneous wormlike micelles formed in a single-tailed zwitterionic surface-active ionic liquid aqueous solution.
Wormlike micelles were successfully fabricated by the self-assembly of a single-tailed zwitterionic surface-active ionic liquid (SAIL), 3-(1-hexadecyl-3-imidazolio) propanesulfonate β-naphthalene sulfonate (C16IPS-Nsa), in aqueous solutions without any additives. With increasing zwitterionic SAIL concentration, spontaneous transition from micelles to wormlike micelles and then a hexagonal phase occurs. Interestingly, the wormlike micelles are closed, stretched and directionally arranged, which was rarely found in previous research studies. This kind of wormlike micelle is useful to serve as novel soft template in the synthesis of functional materials. 1H NMR illustrated that the introduced π-π stacking and hydrophobic interactions originated from the hydrophobic aromatic counterions are responsible for the structural transformation. Density functional theory (DFT) simulated the optimum configuration of C16IPS-Nsa and calculated the interaction energy of C16IPS-Nsa-H2O is 39.6 kJ mol-1. This work paves the way for regulating the self-assembly structures of amphiphiles through changing specific weak interactions.


Are new medical students' specialty preferences gendered? Related motivational factors at a Dutch medical school.
Female students currently outnumber male students in most medical schools. Some medical specialties are highly gender segregated. Therefore, it is interesting to know whether medical students have early specialization preferences based on their gender. Consequently, we like to know importance stipulated to motivational factors. Our study investigates new medical students' early specialization preferences and motivational factors. New students at a Dutch medical school (n = 657) filled in a questionnaire about specialty preferences (response rate = 94%; 69.5% female, 30.5% male). The students chose out of internal medicine, psychiatry, neurology, pediatrics, surgery, gynecology and family medicine, "other" or "I don't know." Finally, they valued ten motivational factors. Forty percent of the medical students reported no specialty preference yet. Taken together, female medical students preferred pediatrics and wished to combine work and care, whereas male students opted for surgery and valued career opportunities. Gender-driven professional preferences in new medical students should be noticed in order to use competencies. Changes in specialty preferences and motivational factors in pre- and post graduates should further assess the role of medical education.


E. adenophorum Induces Cell Cycle and Apoptosis of Renal Cells through Mitochondrial Pathway and Caspase Activation in Saanen Goat.
The cytotoxicity effects of E. adenophorum on cell cycle and apoptosis of renal cells in Saanen goat was evaluated by TUNEL, DAPI, AO/EB staining, DNA fragmentation assay, Caspase activity, Western-blot, qRT-PCR and flow cytometry analysis. 16 saanen goats randomly divided into four groups were fed on 0%, 40%, 60% and 80% E. adenophorum diets. The Results showed that E. adenophorum induced typical apoptotic features of renal cells. E. adenophorum significantly suppressed renal cells viability, caused cell cycle activity arrest and induced typical apoptotic features in a dose-dependent manner. However, the protein levels of Fas/FasL, Bid and caspase-8 did not appear significant changes in the process of E. adenophorum-induced apoptosis. Moreover, E. adenophorum administration slightly decreased Bcl-2 expression, promoted Bax translocation to mitochondria, triggered the release of Cyt c from mitochondria into cytosol and activated caspase-9, -3, and cleaved PARP. The mitochondrial p53 translocation was significantly activated, accompanied by a significant increase in the loss of ΔΨm, Cyt c release and caspase-9 activation. Above all, these data suggest that E. adenophorum induces renal cells apoptosis via the activation of mitochondria-mediated apoptosis pathway in renal cells. These findings may provide new insights to understand the mechanisms involved in E. adenophorum-caused cytotoxicity of renal cells.


Review: Pathogenesis of Helicobacter pylori infection.
The original strategies developed by Helicobacter pylori to persistently colonise its host and to deregulate its cellular functions make this bacterium an outstanding model to study host-pathogen interaction and the mechanisms responsible for bacterial-induced carcinogenesis. During the last year, significant results were obtained on the role of bacterial factors essential for gastric colonisation such as spiral shape maintenance, orientation through chemotaxis and the formation of bacteria clonal population islands inside the gastric glands. Particularities of the H pylori cell surface, a structure important for immune escape, were demonstrated. New insights in the bacterial stress response revealed the importance of DNA methylation-mediated regulation. Further findings were reported on H pylori components that mediate natural transformation and mechanisms of bacterial DNA horizontal transfer which maintain a high level of H pylori genetic variability. Within-host evolution was found to be niche-specific and probably associated with physiological differences between the antral and oxyntic gastric mucosa. In addition, with the progress of CryoEM, high-resolution structures of the major virulence factors, VacA and CagT4SS, were obtained. The use of gastric organoid models fostered research revealing, preferential accumulation of bacteria at the site of injury during infection. Several studies further characterised the role of CagA in the oncogenic properties of H pylori, identifying the activation of novel CagA-dependent pathways, leading to the promotion of genetic instabilities, epithelial-to-mesenchymal transition and finally carcinogenesis. Recent studies also highlight that microRNA-mediated regulation and epigenetic modifications, through DNA methylation, are key events in the H pylori-induced tumorigenesis process.


Internet-based early intervention to prevent posttraumatic stress disorder in injury patients: randomized controlled trial.
Posttraumatic stress disorder (PTSD) develops in 10-20% of injury patients. We developed a novel, self-guided Internet-based intervention (called Trauma TIPS) based on techniques from cognitive behavioral therapy (CBT) to prevent the onset of PTSD symptoms. To determine whether Trauma TIPS is effective in preventing the onset of PTSD symptoms in injury patients. Adult, level 1 trauma center patients were randomly assigned to receive the fully automated Trauma TIPS Internet intervention (n=151) or to receive no early intervention (n=149). Trauma TIPS consisted of psychoeducation, in vivo exposure, and stress management techniques. Both groups were free to use care as usual (nonprotocolized talks with hospital staff). PTSD symptom severity was assessed at 1, 3, 6, and 12 months post injury with a clinical interview (Clinician-Administered PTSD Scale) by blinded trained interviewers and self-report instrument (Impact of Event Scale-Revised). Secondary outcomes were acute anxiety and arousal (assessed online), self-reported depressive and anxiety symptoms (Hospital Anxiety and Depression Scale), and mental health care utilization. Intervention usage was documented. The mean number of intervention logins was 1.7, SD 2.5, median 1, interquartile range (IQR) 1-2. Thirty-four patients in the intervention group did not log in (22.5%), 63 (41.7%) logged in once, and 54 (35.8%) logged in multiple times (mean 3.6, SD 3.5, median 3, IQR 2-4). On clinician-assessed and self-reported PTSD symptoms, both the intervention and control group showed a significant decrease over time (P<.001) without significant differences in trend. PTSD at 12 months was diagnosed in 4.7% of controls and 4.4% of intervention group patients. There were no group differences on anxiety or depressive symptoms over time. Post hoc analyses using latent growth mixture modeling showed a significant decrease in PTSD symptoms in a subgroup of patients with severe initial symptoms (n=20) (P<.001). Our results do not support the efficacy of the Trauma TIPS Internet-based early intervention in the prevention of PTSD symptoms for an unselected population of injury patients. Moreover, uptake was relatively low since one-fifth of individuals did not log in to the intervention. Future research should therefore focus on innovative strategies to increase intervention usage, for example, adding gameplay, embedding it in a blended care context, and targeting high-risk individuals who are more likely to benefit from the intervention. International Standard Randomized Controlled Trial Number (ISRCTN): 57754429; http://www.controlled-trials.com/ISRCTN57754429 (Archived by WebCite at http://webcitation.org/6FeJtJJyD).


Mediator subunit MED31 is required for radial patterning of Arabidopsis roots.
Stem cell specification in multicellular organisms relies on the precise spatiotemporal control of RNA polymerase II (Pol II)-dependent gene transcription, in which the evolutionarily conserved Mediator coactivator complex plays an essential role. In Arabidopsis thaliana, SHORTROOT (SHR) and SCARECROW (SCR) orchestrate a transcriptional program that determines the fate and asymmetrical divisions of stem cells generating the root ground tissue. The mechanism by which SHR/SCR relays context-specific regulatory signals to the Pol II general transcription machinery is unknown. Here, we report the role of Mediator in controlling the spatiotemporal transcriptional output of SHR/SCR during asymmetrical division of stem cells and ground tissue patterning. The Mediator subunit MED31 interacted with SCR but not SHR. Reduction of MED31 disrupted the spatiotemporal activation of CYCLIND6;1 (CYCD6;1), leading to defective asymmetrical division of stem cells generating ground tissue. MED31 was recruited to the promoter of CYCD6;1 in an SCR-dependent manner. MED31 was involved in the formation of a dynamic MED31/SCR/SHR ternary complex through the interface protein SCR. We demonstrate that the relative protein abundance of MED31 and SHR in different cell types regulates the dynamic formation of the ternary complex, which provides a tunable switch to strictly control the spatiotemporal transcriptional output. This study provides valuable clues to understand the mechanism by which master transcriptional regulators control organ patterning.


Prevalence and risk factors associated with smoking among school children, Southern Brazil.
To estimate the prevalence of smoking among students and associated factors. Secondary data from the Vigescola Survey, conducted in the cities of Curitiba, Florianópolis and Porto Alegre (Southern Brazil) between 2002 and 2004, were used. Sample comprised 3,690 school children, aged between 13 and 15 years, and enrolled in the 7th and 8th grades of primary school and 1st grade of high school, in public and private schools. Weighted proportions and odds ratio (OR) were estimated and multiple logistic regression was used to analyze results. Smoking prevalence rates were 10.7% (95% CI: 10.2;11.3) in Florianópolis, 12.6% (95% CI: 12.4;12.9) in Curitiba and 17.7% (95% CI: 17.4;18.0) in Porto Alegre. Risk factors associated with smoking among schoolchildren in Curitiba were: female sex (OR=1.49), smoking father (OR=1.59), smoking friends (OR=3.46), exposure to secondhand tobacco smoke outside the home (OR=3.26), and having some object with cigarette brand logos (OR=3.29). In Florianópolis, variables associated with smoking were: female schoolchildren (OR=1.26), having smoking friends (OR=9.31), exposure to secondhand tobacco smoke at home (OR=2.03) and outside the home (OR=1.45) and having seen advertisements on posters (OR=1.82). In Porto Alegre, variables associated with tobacco use among school children were: female sex (OR=1.57), aged between 14 years (OR=1.77) and 15 years (OR=2.89), smoking friends (OR=9.12), exposure to secondhand tobacco smoke at home (OR=1.87) and outside the home (OR=1.77) and having some object with cigarette brand logos (OR=2.83). Smoking prevalence among school children aged between 13 and 15 years is high. Factors significantly associated with it and common to the three capitals were as follows: having smoking friends and being exposed to environmental smoke outside the home.


Global pediatric craniopharyngioma management modalities and outcomes.
Pediatric craniopharyngioma is a complex pathology, with optimal management involving a multidisciplinary approach and thoughtful care coordination. To date, no studies have compared various treatment modalities and outcomes described in different global regions. We conducted a comprehensive systematic review to compare demographics, clinical presentation, treatment approach and outcomes of children diagnosed with craniopharyngioma globally. A systematic review was conducted in accordance with the Preferred Reporting Item for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Search terms included "craniopharyngioma" and country-specific terms. Inclusion criteria included full-text studies published between 2000-2022, primarily examining pediatric patients 18-years old or younger diagnosed with craniopharyngioma, and reporting management and outcomes of interest. Data extracted included country of origin, demographical data, initial presentation and treatment modality, and outcomes. Descriptive statistics and between-group comparisons based on country of origin were performed. Of 797 search results, 35 articles were included, mostly originating from high-income countries (HIC) (n = 25, 71.4%). No studies originated from low-income countries (LIC). When comparing HIC to middle-income countries (MIC), no differences in patient demographics were observed. No differences in symptomatology at initial presentation, tumor type, surgical approach or extent of surgical resection were observed. HIC patients undergoing intracystic therapy were more likely to receive bleomycin (n = 48, 85.7%), while the majority of MIC patients received interferon therapy (n = 10, 62.5%). All MIC patients undergoing radiation therapy underwent photon therapy (n = 102). No statistically significant differences were observed in postoperative complications or mean follow-up duration between HIC and MIC (78.1 ± 32.2 vs. 58.5 ± 32.1 months, p = 0.241). Pediatric craniopharyngioma presents and is managed similarly across the globe. However, no studies originating from LICs and resource-poor regions examine presentation and management to date, representing a significant knowledge gap that must be addressed to complete the global picture of pediatric craniopharyngioma burden and management.


Colobomatous macrophthalmia with microcornea syndrome maps to the 2p23-p16 region.
Colobomatous macrophthalmia with microcornea syndrome (OMIM 602499) is a rare, autosomal dominant malformation characterized by microcornea, uveal coloboma, axial enlargement of the globe, and myopia. Using what is currently the largest described pedigree and candidate localization approach, we first excluded the candidate genes PAX2, PAX3, PAX6, and PAX9. Subsequently, the chromosome 14q24 region containing the CHX10, SIX1, and SIX4 genes were also excluded. Positive LOD scores were obtained with the DNA markers selected from the 2p23-p16 region. A maximum pairwise LOD score of 3.61 (Theta = 0) was noted with the DNA marker D2S1788. Haplotype analysis positioned the locus between DNA markers D2S2263 and D2S1352 within a 22 Mb physical interval. This region contains major candidate genes, such as SIX2, SIX3, and CYP1B1; however, mutation analysis did not identify a causative mutation in these genes. Macrophthalmia, colobomatous, with microcornea (MACOM) is proposed as the gene symbol for this malformation linked to 2p23-p16.


Participatory technology.
The hunger to participate that exists today in various segments of the American public is in part a response to what some people perceive as an unresponsiveness of institutions and processes to the felt needs of the individual and of society. It is also, in part, an expression of a desire for a redistribution of power in American public life. Technology is one of the major determinants of the nature of public as well as private life in contemporary society. Participatory technology is an attempt on the part of diverse individuals and groups to influence technological processes through participation in existing or new public processes by which technology is or can be developed , controlled, and implemented. Like other processes of direct citizen participation in governmental decision making, it raises many questions about the adequacy of existing theories and practices of representative government. These questions cannot be answered on an a priori basis. Members of the educational, scientific, technical, and governmental communities should analyze these questions in an effort to develop answers that are appropriate to the particular situations for which they are responsible and with which they are concerned.


A novel osteoporosis model with ascorbic acid deficiency in Akr1A1 gene knockout mice.
The AKR1A1 protein is a member of the aldo-keto reductase superfamily that is responsible for the conversion of D-glucuronate to L-gulonate in the ascorbic acid (vitamin C) synthesis pathway. In a pCAG-eGFP transgenic mouse line that was produced by pronuclear microinjection, the integration of the transgene resulted in a 30-kb genomic DNA deletion, including the Akr1A1 gene, and thus caused the knockout (KO) of the Akr1A1 gene and targeting of the eGFP gene. The Akr1A1 KO mice (Akr1A1eGFP/eGFP) exhibited insufficient serum ascorbic acid levels, abnormal bone development and osteoporosis. Using micro-CT analysis, the results showed that the microarchitecture of the 12-week-old Akr1A1eGFP/eGFP mouse femur was shorter in length and exhibited less cortical bone thickness, enlargement of the bone marrow cavity and a complete loss of the trabecular bone in the distal femur. The femoral head and neck of the proximal femur also showed a severe loss of bone mass. Based on the decreased levels of serum osteocalcin and osteoblast activity in the Akr1A1eGFP/eGFP mice, the osteoporosis might be caused by impaired bone formation. In addition, administration of ascorbic acid to the Akr1A1eGFP/eGFP mice significantly prevented the condition of osteoporotic femurs and increased bone formation. Therefore, through ascorbic acid administration, the Akr1A1 KO mice exhibited controllable osteoporosis and may serve as a novel model for osteoporotic research.