Selective activation of the sodium-independent, cyclosporin A-sensitive calcium pore of cardiac mitochondria by doxorubicin.
Various potentially cytopathic actions of doxorubicin have been implicated in the development of cardiotoxicity. However, inhibition of mitochondrial respiration and disruption of calcium homeostasis are emerging as possible primary determinants of toxicity. The objective of this investigation was to identify the specific ion channel by which doxorubicin interferes with calcium homeostasis in isolated rat heart mitochondria. Calcium accumulation by tightly coupled mitochondria was determined spectrophotometrically using arsenazo III and membrane potential was monitored fluorometrically with rhodamine 123. Incubation of cardiac mitochondria with doxorubicin caused a decrease in net accumulation and a delayed spontaneous release of calcium. There was no discernable dissipation of membrane potential and no detectable effect of the drug on the electrophoretic uniport. Diltiazem had no effect while cyclosporin A completely inhibited doxorubicin-induced calcium release. Addition of cyclosporin A prior to calcium prevented the doxorubicin-induced inhibition of calcium accumulation and the release of calcium from cardiac mitochondria. These data suggest that doxorubicin has a specific action on the mitochondrial ADP/ATP antiport, independent of the electrophoretic uniport and the sodium-dependent antiport, which may initiate calcium cycling and the consequential inhibition of oxidative phosphorylation.


Parenchyma Abundance in Wood of Evergreen Trees Varies Independently of Nutrients.
The abundance of living cells in wood-mainly as interconnected axial and ray parenchyma networks-varies widely between species. However, the functional significance of this variation and its role in plant ecological strategies is poorly understood, as is the extent to which different parenchyma fractions are favored in relation to soil nutrients and hydraulic functions. We analyzed wood tissue fractions of 16 Australian angiosperm species sampled from two nearby areas with similar climate but very different soil nutrient profiles and investigated structure-function links with soil and tissue nutrient concentrations and other plant traits. We expected the variation in parenchyma fractions to influence nutrient concentrations in wood xylem, and to find species with lower parenchyma fractions and accordingly lower nutrient requirements on lower-nutrient soils. Surprisingly, both axial and ray parenchyma fractions were mostly unrelated to tissue and soil nutrient concentrations, except for nitrogen concentration in stem sapwood. Species from low nutrient soils showed higher fractional P translocation from both leaves and sapwood, but little patterning with respect to tissue nitrogen. While species from high and low nutrient soils clearly clustered along the soil-fertility axis, their tissue composition varied independently from plant functional traits related to construction costs and hydraulic anatomy. Our findings imply that there is considerable variation among species in the nutrient concentrations within different parenchyma tissues. The anatomical composition of wood tissue seems unrelated to plant nutrient requirements. Even though xylem parenchyma is involved in metabolic functions such as nutrient translocation and storage, parenchyma abundance on its own does not directly explain variation in these functions, even in co-occurring species. While parenchyma is highly abundant in wood of angiosperm trees, we are still lacking a convincing ecological interpretation of its variability and role in whole-tree nutrient budgets.


Levels of Visual Stress in Proficient Readers: Effects of Spectral Filtering of Fluorescent Lighting on Reading Discomfort.
Visual stress (VS) affects reading in 5-12% of the general population and 31-36% of children with reading disorders. Symptoms include print distortions and visual discomfort when reading, and are exacerbated by fluorescent lighting. Prior research has indicated that VS can also affect proficient readers. We therefore examined levels of visual discomfort in a group of expert readers (n = 24) under both standard and spectrally-filtered fluorescent lighting. Participants rated their awareness of six symptoms of VS under each lighting condition. Under the standard condition, 4(16.7%) of the group recorded moderate to high levels of VS. Differences in symptom levels and reading speed between conditions were analysed using the Wilcoxon Signed Rank Test. Under the filter condition, the group reported less discomfort regarding all six symptoms of VS surveyed. The differences were significant with respect to three of the symptoms (p = .029 - p < .001), with a medium effect size in all of them (r = .31 - r = .46) and total score (p = .007; r = .39). Variations in reading proficiency included significantly fewer self-corrections (p = .019) and total errors (p = .004). Here we present evidence that VS-type symptoms of reading discomfort are not confined to populations with reading difficulties and may also occur in proficient readers, and that simple adaptations to fluorescent lighting may alleviate such symptoms.


Micronucleus analysis in peripheral blood lymphocytes from melanoma patients treated with dacarbazine.
Dacarbazine is an antitumor drug used with considerable success in the chemotherapy of a number of human neoplasias, particularly advanced disseminated melanoma. Dacarbazine is mutagenic in prokaryotic and eukaryotic cells, but no effect in vivo have been evaluated. Peripheral blood lymphocytes from patients with metastatic melanoma undergoing dacarbazine chemotherapy every 21 days for a total of 7 cycles, were analyzed for the presence of micronuclei with the CREST antikinetochore antibody technique. Cytogenetic analysis on blood samples collected just before and 2 hours after the therapy was carried out at 48, 72 and 96 hours following lymphocyte stimulation. A significant increase in micronucleus frequency was found at both 72 and 96 hours after therapy. For the only two patients analyzed after more than one cycle, a decrease in micronuclei was observed after the third and the fourth therapy. Moreover, the CREST antibody technique showed that the frequency of micronuclei containing whole chromosomes (CREST+) was significantly higher after therapy at 72 and 96 hours. As the frequency of micronuclei containing acentric chromosome fragments (CREST-) was not significantly increased after therapy, either at 72 or 96 hours after lymphocyte stimulation, we suppose that DTIC mainly acted as an aneugenic agent. The lack of a significant micronucleus increase at 48 hours could suggest that this culture time is too short for providing cultures with a sufficient large number of diving cells. In conclusion, our results have shown that dacarbazine induced chromosome loss in lymphocytes from patients treated with this drug.


Postnatal exposure to benzene alters the neurobehavioral functioning of rats when tested during adulthood.
Male and female rats of the Fischer Strain were dosed with 550 mg/kg of benzene or corn oil vehicle on Days 9, 11 and 13 postpartum. The spontaneous motor activity of benzene exposed rats (males and females) was found to be elevated when tested at 100-130 days of age. When challenged with various doses of d-amphetamine (0.3-3 mg/kg), the benzene exposed rats were found to be less sensitive to the motor activity increasing effects of the drug. In a subsequent test to measure specific components of exploratory activity in an operant chamber (nose-poking, wall rearing, lever touching), female rats exposed to benzene were found to emit fewer rearing responses. Body weights and performance in a battery of tests to assess neurobehavioral toxicity (Days 45, 60 and 100 postpartum) were not affected by postnatal exposure to benzene. These data indicate that postnatal exposure to benzene can produce significant alterations in the motor activity of rats when tested during adulthood and the type of effect depended on the procedure used. Changes in the sensitivity of benzene exposed animals to d-amphetamine suggest long-term alterations in catecholaminergic function.


Risk of Introduction of Bovine Tuberculosis (TB) Into TB-Free Herds in Southern Bahia, Brazil, Associated With Movement of Live Cattle.
Bovine tuberculosis (TB) is a zoonotic disease, endemic in Brazil, with an impact on public health and trade. TB causes direct and indirect financial losses to infected farms and regions. Our study aimed at quantifying the risk of introduction of TB into TB-free herds of southern Bahia, Brazil, via movement of live cattle from other regions of the State. Results suggest that the annual risk of introducing TB into free farms of southern Bahia, either through legal or illegal trade of live cattle, is very low, varying, on average, between 0.001 and 0.006 depending on the region of origin of the animals. Noteworthy, illegal movements accounted for 90% of the risk demonstrating the importance of compliance with official regulations. These results are useful to inform both veterinary authorities and farmers in making decisions related to the regionalization of the control for TB in the country, with the ultimate goal of eliminating this major zoonotic disease from an important dairy region of Brazil.


Feasibility of implementing a mobile phone-based telemonitoring programme to support pregnant women at high-risk for pre-eclampsia in Karachi, Pakistan: protocol for a mixed-methods study.
High maternal mortality from pre-eclampsia/eclampsia results from lack of early identification and management of pregnant women at high-risk for pre-eclampsia. A potential tool to support pregnant women at high-risk for pre-eclampsia is telemonitoring. There is limited evidence on the use and effectiveness of telemonitoring for pregnant women in low-income and middle-income countries (LMICs) which limits the understanding of the process and mechanisms through which the intervention works in LMICs. This study will explore the feasibility of implementing a mobile phone-based telemonitoring programme for pregnant women at high-risk for pre-eclampsia in Karachi, Pakistan. A convergent mixed-methods study will be conducted at the Jinnah Postgraduate Medical Center (JPMC) in Karachi, Pakistan. This study will recruit 50 pregnant women at high-risk for pre-eclampsia to assess clinical feasibility across the five foci of Bowen's framework including acceptability, demand, implementation, practicality and limited-efficacy testing. Data sources will include semi-structured interviews with the enrolled women, caregivers and clinicians, as well as quantitative data from paper medical records, research logs and server data. The quantitative and qualitative data will be analysed separately and then integrated at the interpretation and reporting levels to advance our understanding of the telemonitoring programme's feasibility across the five areas of Bowen's framework. Ethics approvals have been obtained from JPMC, the National Bioethics Committee of Pakistan, University Health Network, Aga Khan University and the University of Toronto. The study results will be disseminated to the scientific community through publications and conference presentations. Findings of the study will provide evidence on the feasibility of using a telemonitoring programme where pregnant women at high-risk for pre-eclampsia in Pakistan will take their own blood pressure readings at home. Lessons learnt in this feasibility trial will be used to determine the appropriateness of a future effectiveness trial. NCT05662696.


Beta transforming growth factors (TGFbeta) at the porcine conceptus-maternal interface. Part II: uterine TGFbeta bioactivity and expression of immunoreactive TGFbetas (TGFbeta1, TGFbeta2, and TGFbeta3) and their receptors (type I and type II).
Porcine uterine tissues were collected from Days 10 to 14 of gestation (peri-implantation period) or corresponding days of the estrous cycle. Results indicated a marked increase in beta transforming growth factors (TGFbeta1, TGFbeta2, and TGFbeta3) and TGFbeta receptor (type I and type II) immunostaining in uterine luminal epithelium (ULE) between Days 10 and 14 of gestation, but there was no increase in ULE immunostaining on the corresponding days of the estrous cycle. Uterine glands and stroma were intensely immunopositive in pregnant gilts for TGFbeta isoforms and their receptors, but immunostaining was weak to undetectable in cycling gilts. No differences were detected in myometrium, in which immunostaining was moderate in both cycling and pregnant gilts. Additionally, TGFbeta2 and TGFbeta receptor (type I and type II) immunostaining was detected in uterine monocyte/macrophage-like cells. Western blotting detected the presence of all three TGFbeta isoforms in uterine luminal flushings. The CCL64 cell TGFbeta bioassay detected bioactive TGFbetas++ in uterine luminal flushings on Days 12, 13, an 14 of gestation. These results strongly indicate that uterine expression of TGFbetas and their receptors is pregnancy specific and that bioactive TGFbetas are present at the conceptus-maternal interface in the peri-implantation period in pigs. Thus TGFbetas are likely to be involved in autocrine-paracrine interactions between the maternal uterus and the conceptus.


Colonic fermentation of indigestible carbohydrates contributes to the second-meal effect.
Low postprandial blood glucose is associated with low risk of metabolic diseases. A meal's ability to diminish the glucose response to carbohydrates eaten during the following meal is known as the "second-meal effect" (SME). The reduced glycemia elicited by low-glycemic-index (LGI) foods consumed during the first meal has been suggested as the main mechanism for SME. However, LGI foods often increase colonic fermentation because of the presence of fiber and resistant starch. The objective was to study the SME of greater fermentation of high-glycemic-index (HGI) and LGI carbohydrates eaten during a previous meal. Ten healthy volunteers ate 3 breakfast test meals consisting of sponge cakes made with rapidly digestible, nonfermentable amylopectin starch plus cellulose (HGI meal), amylopectin starch plus the fermentable disaccharide lactulose (HGI-Lac meal), or slowly digestible, partly fermentable amylose starch plus cellulose (LGI meal). Five hours later, subjects were fed the same standard lunch containing 93 g available carbohydrates. Blood was collected for measurement of glucose, insulin, and nonesterified fatty acids (NEFAs). Breath hydrogen was measured as a marker of colonic fermentation. Postlunch gastric emptying was measured by using ultrasonography. Both the HGI-Lac and LGI meals improved glucose tolerance at lunch. In the case of the HGI-Lac meal, this effect was concomitant with low NEFA concentrations and delayed gastric emptying. Fermentable carbohydrates, independent of their effect on a food's glycemic index, have the potential to regulate postprandial responses to a second meal by reducing NEFA competition for glucose disposal and, to a minor extent, by affecting intestinal motility.


Sulfide dehydrogenase from the hyperthermophilic archaeon Pyrococcus furiosus: a new multifunctional enzyme involved in the reduction of elemental sulfur.
Pyrococcus furiosus is an anaerobic archaeon that grows optimally at 100 degrees C by the fermentation of carbohydrates yielding acetate, CO2, and H2 as the primary products. If elemental sulfur (S0) or polysulfide is added to the growth medium, H2S is also produced. The cytoplasmic hydrogenase of P. furiosus, which is responsible for H2 production with ferredoxin as the electron donor, has been shown to also catalyze the reduction of polysulfide to H2S (K. Ma, R. N. Schicho, R. M. Kelly, and M. W. W. Adams, Proc. Natl. Acad. Sci. USA 90:5341-5344, 1993). From the cytoplasm of this organism, we have now purified an enzyme, sulfide dehydrogenase (SuDH), which catalyzes the reduction of polysulfide to H2S with NADPH as the electron donor. SuDH is a heterodimer with subunits of 52,000 and 29,000 Da. SuDH contains flavin and approximately 11 iron and 6 acid-labile sulfide atoms per mol, but no other metals were detected. Analysis of the enzyme by electron paramagnetic resonance spectroscopy indicated the presence of four iron-sulfur centers, one of which was specifically reduced by NADPH. SuDH has a half-life at 95 degrees C of about 12 h and shows a 50% increase in activity after 12 h at 82 degrees C. The pure enzyme has a specific activity of 7 mumol of H2S produced.min-1.mg of protein-1 at 80 degrees C with polysulfide (1.2 mM) and NADPH (0.4 mM) as substrates. The apparent Km values were 1.25 mM and 11 microM, respectively. NADH was not utilized as an electron donor for polysulfide reduction. P. furiosus rubredoxin (K(m) = 1.6 microM) also functioned as an electron acceptor for SuDH, and SuDH catalyzed the reduction of NADP with reduced P. furiosus ferredoxin (K(m) = 0.7 microM) as an electron donor. The multiple activities of SuDH and its proposed role in the metabolism of S(o) and polysulfide are discussed.


Linking Behavioral and Computational Approaches to Better Understand Variant Vowel Pronunciations in Developing Readers.
The overarching goal of the new Florida State University/Haskins Laboratory/University of Connecticut Learning Disability (LD) Hub project is to align computational and behavioral theories of individual word reading development more closely with the challenges of learning to read a quasi-regular orthography (i.e., English) for both typically developing (TD) children and, more specifically, children with dyslexia. Our LD Hub adopts an integrated approach to better understand the neurocognitive bases of individual differences in word reading development by specifically examining the experiential (exogenous) and child-specific (endogenous) factors that determine acquisition of orthographic-phonological knowledge at different subword granularities using behavioral and computational modeling. Findings are intended to enrich understanding of the processes that influence individual differences in word reading development in TD and dyslexic children and significantly inform issues of practice (e.g., curriculum, instruction, diagnosis, and intervention). Here, we briefly provide the rationale for the Hub and present findings from the initial behavioral and computational modeling studies.


Mammalian EAK-7 activates alternative mTOR signaling to regulate cell proliferation and migration.
Nematode EAK-7 (enhancer-of-akt-1-7) regulates dauer formation and controls life span; however, the function of the human ortholog mammalian EAK-7 (mEAK-7) is unknown. We report that mEAK-7 activates an alternative mechanistic/mammalian target of rapamycin (mTOR) signaling pathway in human cells, in which mEAK-7 interacts with mTOR at the lysosome to facilitate S6K2 activation and 4E-BP1 repression. Despite interacting with mTOR and mammalian lethal with SEC13 protein 8 (mLST8), mEAK-7 does not interact with other mTOR complex 1 (mTORC1) or mTOR complex 2 (mTORC2) components; however, it is essential for mTOR signaling at the lysosome. This phenomenon is distinguished by S6 and 4E-BP1 activity in response to nutrient stimulation. Conventional S6K1 phosphorylation is uncoupled from S6 phosphorylation in response to mEAK-7 knockdown. mEAK-7 recruits mTOR to the lysosome, a crucial compartment for mTOR activation. Loss of mEAK-7 results in a marked decrease in lysosomal localization of mTOR, whereas overexpression of mEAK-7 results in enhanced lysosomal localization of mTOR. Deletion of the carboxyl terminus of mEAK-7 significantly decreases mTOR interaction. mEAK-7 knockdown decreases cell proliferation and migration, whereas overexpression of mEAK-7 enhances these cellular effects. Constitutively activated S6K rescues mTOR signaling in mEAK-7-knocked down cells. Thus, mEAK-7 activates an alternative mTOR signaling pathway through S6K2 and 4E-BP1 to regulate cell proliferation and migration.


Frontal fibrosing alopecia treatment options.
Frontal fibrosing alopecia (FFA) is a rare dermatologic disease that causes scarring and hair loss and is increasing in prevalence worldwide. FFA patients typically present with hair loss in the frontal scalp region and eyebrows which may be associated with sensations of itching or burning. FFA is a clinically distinct variant of lichen planopilaris (LPP) that affects predominantly postmenopausal women, although men and premenopausal women may also be affected. Early diagnosis and prompt treatment are necessary to prevent definitive scarring and permanent hair loss. Data from retrospective studies indicate that 5-alpha-reductase inhibitors (5aRIs) are effective in stabilizing the disease. In our clinical experience, we have seen optimal results treating FFA patients with oral finasteride in conjunction with hydroxychloroquine, topical calcineurin inhibitors (tacrolimus) and excimer laser in patients with signs of active inflammation.


Soil examination for a forensic trace evidence laboratory--Part 1: Spectroscopic techniques.
In the past, forensic soil examination was a routine aspect of trace evidence examination in forensic science. However, in Australia, the apparent need for soil examinations has diminished and with it the capability of forensic science laboratories to carry out soil examination has been eroded. In recent years, due to soil examinations contributing to some high profile investigations, interest in soil examinations has been renewed. Routine soil examinations conducted in a forensic science laboratory by trace evidence scientists can be facilitated if the examinations are conducted using the instrumentation routinely used by these examiners. Spectroscopic techniques such as visible microspectrophotometry (MSP) and Attenuated Total Reflectance (ATR) Fourier Transform Infrared spectroscopy (FTIR) are routinely used by trace evidence analysts for the colour and compositional analysis, respectively, of forensic items, including paints, fibres, inks and toners, tapes, adhesives and other miscellaneous examinations. This article presents an examination of the feasibility of using MSP and ATR-FTIR as a first step in the forensic comparison of soils with particular reference to Australian soil samples. This initial study demonstrates MSP and ATR-FTIR can effectively be used as a screening test for the discrimination of "forensic-sized" soil samples prior to submission for more detailed analyses by a soil expert.


Hemolytic properties of Ca(2+)-treated human erythrocytes under hydrostatic pressure.
The effect of intracellular Ca2+ on high pressure-induced hemolysis of human erythrocytes was examined. Red cells were incubated with Ca2+ (0.01-1 mM) in the presence of ionophore A23187. The Ca(2+)-loaded cells were subjected to a pressure of 200 mPa. Treatment with 0.1 mM Ca2+ had the greatest suppressive effect on the hemolysis. On removal of intracellular Ca2+, red cells showed a morphological change from echinocytes to normal discocytes but the hemolysis remained unaltered. Measurement of intracellular K+ and viscosity demonstrated that the suppressive effect of Ca2+ on the hemolysis is irreversible and is largely associated with the increase of intracellular viscosity induced by K+ efflux.


Identifying neural signatures of tobacco retail outlet exposure: Preliminary validation of a "community neuroscience" paradigm.
An extensive epidemiological literature indicates that increased exposure to tobacco retail outlets (TROs) places never smokers at greater risk for smoking uptake and current smokers at greater risk for increased consumption and smoking relapse. Yet research into the mechanisms underlying this effect has been limited. This preliminary study represents the first effort to examine the neurobiological consequences of exposure to personally relevant TROs among both smokers (n = 17) and nonsmokers (n = 17). Individuals carried a global positioning system (GPS) tracker for 2 weeks. Traces were used to identify TROs and control outlets that fell inside and outside their ideographically defined activity space. Participants underwent functional MRI (fMRI) scanning during which they were presented with images of these storefronts, along with similar store images from a different county and rated their familiarity with these stores. The main effect of activity space was additive with a Smoking status × Store type interaction, resulting in smokers exhibiting greater neural activation to TROs falling inside activity space within the parahippocampus, precuneus, medial prefrontal cortex, and dorsal anterior insula. A similar pattern was observed for familiarity ratings. Together, these preliminary findings suggest that the otherwise distinct neural systems involved in self-orientation/self-relevance and smoking motivation may act in concert and underlie TRO influence on smoking behavior. This study also offers a novel methodological framework for evaluating the influence of community features on neural activity that can be readily adapted to study other health behaviors.


Intercorrelations among general arousability, emerging and current sexual desire, and severity of sexual dysfunction in women.
Intercorrelations among general trait arousability, emerging sexual desire, current sexual desire, lack of sexual desire perceived as a problem, and sexual dysfunction were assessed in 65 women currently involved in marital or de facto heterosexual relationships. The Stimulus Screening Test was used to measure general arousability and the Assessment of Sexual Function and Dysfunction Questionnaire was used to measure emerging sexual desire, current sexual desire lack of sexual desire, and sexual dysfunction. Analysis showed that arousability was positively associated with current levels of sexual desire which was negatively associated with sexual dysfunction. There was a curvilinear relationship between arousability and sexual dysfunction, with women experiencing both high and low levels of arousability being more likely to score higher on measures of sexual dysfunction than those experiencing medium levels of arousal. The possible implications of these results are discussed.


[A questionable guideline on introduction of solid food to breast-fed infants].
In 1993, the Norwegian National Nutrition Council recommended that solid food be introduced to breast-fed infants gradually between the age of four and six months, and not later than six months. A critical appraisal of the literature reveals that the alleged inadequacy of the volume of breast milk after 4-6 months is an artifact caused by the inevitable fall of milk yield induced by supplementation and unnatural time scheduling of breast-feeding. The council has overlooked evidence showing that voluntary exclusive breast feeding on demand provides enough energy and nutrients to most infants for at least nine months. There is no documented need for iron-rich solid food before the age of 7-9 months. Mothers who wish to breast-feed exclusively after the age of six months should therefore be encouraged to do so. Solid food should not be emphatically recommended until the infant is nine months old.


[Creutzfeldt-Jakob disease and transfusional risk: the point in 1998].
As both iatrogenic and experimental transmission of Creutzfeldt-Jakob disease (CJD) have been demonstrated, potential blood transfusion-related CJD through blood component contamination is not unlikely. Recent studies also suggest that B lymphocytes would play a crucial role in the disease pathogenesis. However, to date, there is no epidemiological evidence suggesting that classical transmission of CJD would involve B lymphocytes. To reduce the theoretical risk of such transmission and as no biological marker of the disease is currently available, at-risk blood donors are excluded and leukocyte depletion has been introduced. Furthermore, the recent discovery of a new variant of the disease (nwCJD) which is caused by the bovine agent and has typical tropism for lymphoreticular tissues should lead to close surveillance.


Direct Oral Anticoagulants Combined with Antiplatelet Therapy in the Treatment of Coronary Heart Disease: An Updated Meta-analysis.
Direct oral anticoagulants (DOACs) combined with antiplatelet therapy for acute coronary syndrome (ACS) may reduce ischemic events, but there is no consensus on bleeding risk. Moreover, the effect of DOACs on stable coronary artery disease (CAD) needs to be elucidated. We conducted a meta-analysis to summarize the efficacy and safety of DOACs combined with antiplatelet therapy in the treatment of stable CAD and ACS. We searched PubMed, Web of Science, and the Cochrane Central Register of Controlled Trials, then performed a systematic review of all 17 randomized controlled trials. For patients with stable CAD, DOACs combined with antiplatelet therapy significantly reduced the rate of major adverse cardiovascular events (MACE) (risk ratio; 95% confidence interval: 0.88; 0.81-0.95) and ischemic stroke (0.62; 0.50-0.77), with a relatively low risk of major bleeding (1.72; 1.42-2.07). For patients with ACS, the combination of DOACs reduced the risk of MACE (0.91; 0.85-0.97), myocardial infarction (MI) (0.90; 0.83-0.98), and ischemic stroke (0.75; 0.58-0.97), accompanied by increased non-fatal bleeding events and intracranial hemorrhage (3.42; 1.76-6.65). Results were similar when restricting the analysis to phase III studies except for the rate of stroke in patients with ACS. Combination therapy reduced the incidence of MI in ACS patients, but the risk of bleeding from intracranial hemorrhaging outweighs the benefit of MACE driven by MI. That is due to combination therapy having no positive impact on mortality; thus, the benefit-risk balance may be more favorable in patients with stable CAD.


Acid suppression therapy does not predispose to Clostridium difficile infection: the case of the potential bias.
An adverse effect of acid-suppression medications on the occurrence of Clostridium difficile infection (CDI) has been a common finding of many, but not all studies. We hypothesized that association between acid-suppression medications and CDI is due to the residual confounding in comparison between patients with infection to those without, predominantly from non-tested and less sick subjects. We aimed to evaluate the effect of acid suppression therapy on incidence of CDI by comparing patients with CDI to two control groups: not tested patients and patients suspected of having CDI, but with a negative test. We conducted a case-control study of adult patients hospitalized in internal medicine department of tertiary teaching hospital between 2005-2010 for at least three days. Controls from each of two groups (negative for CDI and non-tested) were individually matched (1:1) to cases by primary diagnosis, Charlson comorbidity index, year of hospitalization and gender. Primary outcomes were diagnoses of International Classification of Diseases (ICD-9)-coded CDI occurring 72 hours or more after admission. Patients with CDI were similar to controls with a negative test, while controls without CDI testing had lower clinical severity. In multivariable analysis, treatment by acid suppression medications was associated with CDI compared to those who were not tested (OR = 1.88, p-value = 0.032). Conversely, use of acid suppression medications in those who tested negative for the infection was not associated with CDI risk as compared to the cases (OR = 0.66; p = 0.059). These findings suggest that the reported epidemiologic associations between use of acid suppression medications and CDI risk may be spurious. The control group choice has an important impact on the results. Clinical differences between the patients with CDI and those not tested and not suspected of having the infection may explain the different conclusions regarding the acid suppression effect on CDI risk.


The mouse carbonic anhydrase I gene contains two tissue-specific promoters.
We report the isolation and characterization of the mouse carbonic anhydrase I (CAI) gene. Direct RNA sequence analysis of the 5' nontranslated regions of CAI mRNA from mouse colon and mouse erythroleukemia cells demonstrated tissue specificity in the lengths and sequences of CAI transcripts. Analysis of several mouse CAI genomic clones showed that the transcripts arose from a single CAI gene with two tissue-specific promoters and eight exons. CAI transcripts in the colon were found to initiate just upstream of the erythroid exon 2 of the CAI gene region sequence. Erythroid transcripts originated from a novel promoter upstream of exon 1, which was located more than 10 but less than 250 kilobases upstream of exon 2. Erythroid exon 1 contained only a nontranslated sequence, which was spliced to exon 2 via a cryptic splice acceptor site located in the region that encoded the colon mRNA 5' nontranslated sequence. The remaining exon-intron junctions were conserved in comparison with those of the CAII and CAIII genes.


Significant Amplification of Instantaneous Extreme Precipitation With Convective Self-Aggregation.
This work explores the effect of convective self-aggregation on extreme rainfall intensities through an analysis at several stages of the cloud lifecycle. In addition to increases in 3-hourly extremes consistent with previous studies, we find that instantaneous rainrates increase significantly (+30%). We mainly focus on instantaneous extremes and, using a recent framework, relate their increase to increased precipitation efficiency: the local increase in relative humidity drives larger accretion efficiency and lower re-evaporation. An in-depth analysis based on an adapted scaling for precipitation extremes reveals that the dynamic contribution decreases (-25%) while the thermodynamic is slightly enhanced (+5%) with convective self-aggregation, leading to lower condensation rates. When the atmosphere is more organized into a moist convecting region and a dry convection-free region, deep convective updrafts are surrounded by a warmer environment which reduces convective instability and thus the dynamic contribution. The moister boundary-layer explains the positive thermodynamic contribution. The microphysic contribution is increased by +50% with aggregation. The latter is partly due to reduced evaporation of rain falling through a moister near-cloud environment, but also to the associated larger accretion efficiency. Thus, a potential change in convective organization regimes in a warming climate could lead to an evolution of tropical precipitation extremes significantly different than that expected from thermodynamical considerations. The relevance of self-aggregation to the real tropics is still debated. Improved fundamental understanding of self-aggregation, its sensitivity to warming and connection to precipitation extremes, is hence crucial to achieve accurate rainfall projections in a warming climate.


Diminished antiproteinuric effect of the angiotensin receptor blocker losartan during high potassium intake in patients with CKD.
Angiotensin II type 1 receptor blockers (ARBs) lower blood pressure (BP) and proteinuria and reduce renal disease progression in many-but not all-patients. Reduction of dietary sodium intake improves these effects of ARBs. Dietary potassium intake affects BP and proteinuria. We set out to address the effect of potassium intake on BP and proteinuria response to losartan in non-diabetic proteinuric chronic kidney disease (CKD) patients. We performed a post hoc analysis of a placebo-controlled interventional cross-over study in 33 non-diabetic proteinuric patients (baseline mean arterial pressure and proteinuria: 105 mmHg and 3.8 g/day, respectively). Patients were treated for 6 weeks with placebo, losartan and losartan/hydrochlorothiazide (HCT), combined with a habitual (∼200 mmol/day) and low-sodium (LS) diet (<100 mmol/day), in randomized order. To analyse the effects of potassium intake, we categorized patients based on median split of 24-h urinary potassium excretion, reflecting potassium intake. Mean potassium intake was stable during all six treatment periods. Losartan and losartan/HCT lowered BP and proteinuria in all treatment groups. Patients with high potassium intake showed no difference in the BP effects compared with patients with low potassium intake. The antiproteinuric response to losartan monotherapy and losartan combined with HCT during the habitual sodium diet was significantly diminished in patients with high potassium intake (20% versus 41%, P = 0.011; and 48% versus 64%, P = 0.036). These differences in antiproteinuric response abolished when shifting to the LS diet. In proteinuric CKD patients, the proteinuria, but not BP-lowering response to losartan during a habitual high-sodium diet was hampered during high potassium intake. Differences disappeared after sodium status change by LS diet.


Oxidative mitigation of aquatic methane emissions in large Amazonian rivers.
The flux of methane (CH4 ) from inland waters to the atmosphere has a profound impact on global atmospheric greenhouse gas (GHG) levels, and yet, strikingly little is known about the dynamics controlling sources and sinks of CH4 in the aquatic setting. Here, we examine the cycling and flux of CH4 in six large rivers in the Amazon basin, including the Amazon River. Based on stable isotopic mass balances of CH4 , inputs and outputs to the water column were estimated. We determined that ecosystem methane oxidation (MOX) reduced the diffusive flux of CH4 by approximately 28-96% and varied depending on hydrologic regime and general geochemical characteristics of tributaries of the Amazon River. For example, the relative amount of MOX was maximal during high water in black and white water rivers and minimal in clear water rivers during low water. The abundance of genetic markers for methane-oxidizing bacteria (pmoA) was positively correlated with enhanced signals of oxidation, providing independent support for the detected MOX patterns. The results indicate that MOX in large Amazonian rivers can consume from 0.45 to 2.07 Tg CH4 yr(-1) , representing up to 7% of the estimated global soil sink. Nevertheless, climate change and changes in hydrology, for example, due to construction of dams, can alter this balance, influencing CH4 emissions to atmosphere.


Quantitative measures of myo-IP6 in soil using solution 31P NMR spectroscopy and spectral deconvolution fitting including a broad signal.
Inositol phosphates, particularly myo-inositol hexakisphosphate (myo-IP6), are an important pool of soil organic phosphorus (P) in terrestrial ecosystems. To measure concentrations of myo-IP6 in alkaline soil extracts, solution 31P nuclear magnetic resonance (NMR) spectroscopy is commonly used. However, overlap of the NMR peaks of myo-IP6 with several other peaks in the phosphomonoester region requires spectral deconvolution fitting (SDF) to partition the signals and quantify myo-IP6. At present, two main SDF approaches are in use; the first fits a Lorentzian/Gaussian lineshape to the myo-IP6 peaks directly to the baseline without an underlying broad signal, and the second fits a Lorentzian/Gaussian lineshape to the myo-IP6 peaks simultaneously with an underlying broad peak. The aim of this study was to compare the recovery of added myo-IP6 to soil extracts using both SDF procedures for six soil samples of diverse origin and differing concentrations of organic P (112 to 1505 mg P per kgsoil). The average recovery of total added myo-IP6 was 95% (SD 5) and 122% (SD 32) using SDF with and without an underlying broad signal, respectively. The recovery of individual peaks of myo-IP6 differed, most notably, the C5 phosphate peak of myo-IP6 was overestimated by up to 213% when a broad peak was not included in SDF. Based on the SDF procedure that includes a broad peak, concentrations of myo-IP6 ranged from 0.6 to 90.4 mg P per kgsoil, which comprised 1-23% of total phosphomonoesters. Our results demonstrate that the SDF procedure with an underlying broad signal is essential for the accurate quantification of myo-IP6 in soil extracts.


Infliximab for induction and maintenance therapy for ulcerative colitis.
Infliximab, a chimeric monoclonal antibody directed against tumor necrosis factor alpha, is an established treatment for Crohn's disease but not ulcerative colitis. Two randomized, double-blind, placebo-controlled studies--the Active Ulcerative Colitis Trials 1 and 2 (ACT 1 and ACT 2, respectively)--evaluated the efficacy of infliximab for induction and maintenance therapy in adults with ulcerative colitis. In each study, 364 patients with moderate-to-severe active ulcerative colitis despite treatment with concurrent medications received placebo or infliximab (5 mg or 10 mg per kilogram of body weight) intravenously at weeks 0, 2, and 6 and then every eight weeks through week 46 (in ACT 1) or week 22 (in ACT 2). Patients were followed for 54 weeks in ACT 1 and 30 weeks in ACT 2. In ACT 1, 69 percent of patients who received 5 mg of infliximab and 61 percent of those who received 10 mg had a clinical response at week 8, as compared with 37 percent of those who received placebo (P<0.001 for both comparisons with placebo). A response was defined as a decrease in the Mayo score of at least 3 points and at least 30 percent, with an accompanying decrease in the subscore for rectal bleeding of at least 1 point or an absolute rectal-bleeding subscore of 0 or 1. In ACT 2, 64 percent of patients who received 5 mg of infliximab and 69 percent of those who received 10 mg had a clinical response at week 8, as compared with 29 percent of those who received placebo (P<0.001 for both comparisons with placebo). In both studies, patients who received infliximab were more likely to have a clinical response at week 30 (P< or =0.002 for all comparisons). In ACT 1, more patients who received 5 mg or 10 mg of infliximab had a clinical response at week 54 (45 percent and 44 percent, respectively) than did those who received placebo (20 percent, P<0.001 for both comparisons). Patients with moderate-to-severe active ulcerative colitis treated with infliximab at weeks 0, 2, and 6 and every eight weeks thereafter were more likely to have a clinical response at weeks 8, 30, and 54 than were those receiving placebo. (ClinicalTrials.gov numbers, NCT00036439 and NCT00096655.)


Ethical and Methodological Challenges in Research With Hard-to-Reach Groups: Examples From Research on Family Caregivers for Migrant Older Adults Living With Dementia.
Family caregivers of migrants with dementia constitute a population group that is hard to reach for research participation due to factors such as shame about the disease and past experiences of discrimination. In this article, research-ethical challenges associated with participant recruitment and qualitative data collection among relatives of migrants with dementia are discussed. Over a period of 8 years, 3 studies were conducted to investigate the experiences of family caregivers for persons with dementia of Turkish descent in Germany. Across these studies, a total of 32 family caregivers were interviewed. In this article, based on the "Principles of Biomedical Ethics" according to Beauchamp and Childress (2009), research-ethical conflicts associated with sampling methods and the presence of third parties during qualitative interviews are discussed. The potential risks emanating from sampling strategies and the presence of third parties during interviews regarding the voluntary nature of study participation are examined. Additionally, this article formulates recommendations for ensuring truly voluntary participation and protecting both the participants (family caregivers) and relatives with dementia from harm. These practical recommendations aim to help future researchers to avoid ethical pitfalls and represent a roadmap for making necessary methodological decisions.


Effect of Hemifacial Spasm on Intraocular Pressure Measurement.
To evaluate the effect of hemifacial spasm (HFS) on intraocular pressure (IOP) measurement. Twenty-four consecutive patients with HFS and 25 age- and gender-matched randomly selected eyes of healthy volunteers underwent corneal pachymetry and IOP measurements using Goldmann applanation tonometer (GAT) and noncontact tonometer (NCT). IOP measurements were performed before (during HFS) and 2 weeks after Botox injections in HFS patients and in healthy volunteers without Botox injections. There was no statistical difference between involved eye side and uninvolved eye side of HFS patients in measured central corneal thickness. Similarly, no difference was found between involved eye side of HFS patients and controls. There were no statistically significant differences comparing IOP values before treatment and levels measured at 2 weeks of Botox injections, either with GAT (p = 0.33, 0.11) or NCT (p = 0.80, 0.43) devices in the involved eyes and uninvolved eyes of patients with HFS, respectively. There were also no significant differences in these parameters (GAT (p = 0.63) and NCT (p = 0.54)) in controls. Contractions in facial muscles may not lead to significant increase in IOP in HFS patients. This result may help clinical decision making in the treatment of glaucoma patients with HFS. This trial is registered with NCT03390803.


Smoking is associated with advanced coronary atherosclerosis in youth.
Smoking is linked to atherosclerosis and coronary heart disease (CHD) in older adults. However, evidence that smoking affects coronary atherosclerosis in young people is incomplete. The Pathobiological Determinants of Atherosclerosis in Youth (PDAY) Study collected arteries, blood, and other tissues from persons 15 to 34 years of age dying of external causes and autopsied in forensic laboratories. Lesions in the proximal left anterior descending coronary arteries (LAD) from 1127 subjects were graded microscopically according to the American Heart Association criteria. Among individuals with advanced lesions (Grade 4 or 5), smokers had a greater prevalence of Grade 5 lesions than non-smokers (odds ratio 9.61, 95% confidence interval 2.34-39.57), a difference suggesting that smoking accelerates the transition from Grade 4 to Grade 5 lesions. This association occurred among both men and women, and among persons with and without other CHD risk factors. The difference in qualities of advanced lesions suggests that smoking possibly accelerates the transition from Grade 4 to Grade 5 lesions by promoting thrombosis and accretion on the intimal surface of the plaque.


Predictive modeling of signaling crosstalk during C. elegans vulval development.
Caenorhabditis elegans vulval development provides an important paradigm for studying the process of cell fate determination and pattern formation during animal development. Although many genes controlling vulval cell fate specification have been identified, how they orchestrate themselves to generate a robust and invariant pattern of cell fates is not yet completely understood. Here, we have developed a dynamic computational model incorporating the current mechanistic understanding of gene interactions during this patterning process. A key feature of our model is the inclusion of multiple modes of crosstalk between the epidermal growth factor receptor (EGFR) and LIN-12/Notch signaling pathways, which together determine the fates of the six vulval precursor cells (VPCs). Computational analysis, using the model-checking technique, provides new biological insights into the regulatory network governing VPC fate specification and predicts novel negative feedback loops. In addition, our analysis shows that most mutations affecting vulval development lead to stable fate patterns in spite of variations in synchronicity between VPCs. Computational searches for the basis of this robustness show that a sequential activation of the EGFR-mediated inductive signaling and LIN-12 / Notch-mediated lateral signaling pathways is key to achieve a stable cell fate pattern. We demonstrate experimentally a time-delay between the activation of the inductive and lateral signaling pathways in wild-type animals and the loss of sequential signaling in mutants showing unstable fate patterns; thus, validating two key predictions provided by our modeling work. The insights gained by our modeling study further substantiate the usefulness of executing and analyzing mechanistic models to investigate complex biological behaviors.


Effects of level of dietary intake and physical form of protein supplement on the digestibilities of different dietary carbohydrates between mouth and abomasum of young steers.
Steers fitted with simple rumen and abomasal cannulas were given isoenergetic diets of rolled barley and chopped straw, pelleted together with some tapioca alone (B) or with some tapioca replaced by coarse soyabean meal (M) or finely ground soyabean flour (F). The diets were given at two levels to support 0.5 (L) and 1.0 (H) kg/d live weight gain. Chromic oxide and PEG were given as digesta flow markers. Mouth to abomasum digestibilities of different dietary sugars at the low level of intake (LB) were 0.65, 0.68, 0.59, 0.56 and 0.94 for arabinose, galactose, xylose, cellulose-glucose and starch-glucose respectively. Corresponding values at the higher level of intake (HB) were 0.55, 0.66, 0.55, 0.44 and 0.93 respectively. Supplementation with either soya bean meal or flour had no significant effects on the mouth to abomasum of dietary carbohydrate digestibilities at either level of feeding.


The behavior change technique taxonomy (v1) of 93 hierarchically clustered techniques: building an international consensus for the reporting of behavior change interventions.
CONSORT guidelines call for precise reporting of behavior change interventions: we need rigorous methods of characterizing active content of interventions with precision and specificity. The objective of this study is to develop an extensive, consensually agreed hierarchically structured taxonomy of techniques [behavior change techniques (BCTs)] used in behavior change interventions. In a Delphi-type exercise, 14 experts rated labels and definitions of 124 BCTs from six published classification systems. Another 18 experts grouped BCTs according to similarity of active ingredients in an open-sort task. Inter-rater agreement amongst six researchers coding 85 intervention descriptions by BCTs was assessed. This resulted in 93 BCTs clustered into 16 groups. Of the 26 BCTs occurring at least five times, 23 had adjusted kappas of 0.60 or above. "BCT taxonomy v1," an extensive taxonomy of 93 consensually agreed, distinct BCTs, offers a step change as a method for specifying interventions, but we anticipate further development and evaluation based on international, interdisciplinary consensus.


Ameliorative effects of morel mushroom (Morchella esculenta) against Cadmium-induced reproductive toxicity in adult male rats.
Cadmium (Cd) is one of the major toxicants, which affects human health through occupational and environmental exposure. In the current study, we evaluated the protective effects of morel mushrooms against Cd-induced reproductive damages in rats. For this purpose, 30 male rats were divided into 6 groups (n=5/group), the first group served as the control group, second group was treated with an intraperitoneal (i.p) injection of 1 mg/kg/day of Cd. Third and fourth groups were co-treated with 1 mg/kg/day of Cd (i.p) and 10 and 20 mg/kg/day of morel mushroom extract (orally) respectively. The final 2 groups received oral gavage of 10 and 20 mg/kg/day of morel mushroom extract alone. After treatment for 17 days, the animals were euthanized, and testes and epididymis were dissected out. One testis and epididymis of each animal were processed for histology, while the other testis and epididymis were used for daily sperm production (DSP) and comet assay. Our results showed that Cd and morel mushrooms have no effect on animal weight, but Cd significantly decreases the DSP count and damages the heritable DNA which is reversed in co-treatment groups. Similarly, the histopathological results of testes and epididymis show that morel mushrooms control the damage to these tissues. Whereas the morel mushroom extract alone could enhance the production of testosterone. These results conclude that morel mushrooms not only control the damage done by Cd, but it could also be used as a protection mechanism for heritable DNA damage.


Shyness programme: longer term benefits, cost-effectiveness, and acceptability.
In two randomized controlled trials Titov et al. demonstrated significant benefit from an Internet- and email-based treatment programme for social phobia: the Shyness programme. Data are presented about the longer term outcomes (6 months after treatment), cost-effectiveness relative to face-to-face treatment, and the acceptability of the programme to participants. Participants completed outcome and acceptability questionnaires at 6 months after treatment. Repeated measures analyses of variance were calculated using an intention-to-treat design. Cost-effectiveness in years lived with disability averted were calculated based on between-group effect sizes. A total of 59% of treatment group participants completed the 6 month follow-up questionnaires. Between post-treatment and 6 month follow up participants continued to make improvements in symptoms of social phobia, while maintaining improvements in mood, psychological distress, and disability. At 6 month follow up the mean within-group effect size (Cohen's d) for the two social phobia measures increased from 1.2 to 1.4. Cost-effectiveness in years lived with disability (YLD) averted was calculated as one-quarter that of face-to-face group treatment, or $AUD1495 for one YLD gained, compared to $AUD5686/YLD gained. Participants rated the Internet treatment to be as effective and helpful as face-to-face treatment. The present results confirm the reliability of the short-term findings reported in the first two Shyness programmes. The procedure appears to be very cost-effective, and acceptable to participants. These data provide further support for the development of Internet-based virtual clinics for common mental disorders.


Evaluation of the association between Addiction Severity Index and depression with adherence to anti-retroviral therapy among HIV infected patients.
Adequate adherence to anti-retroviral therapy is required to achieve viral suppression and desirable treatment outcomes among HIV patients. The aim of this study was to examine the associations between adherence and severity of substance use as well as adherence and severity of depressive symptoms among Iranian HIV patients. In a prospective study, HIV patients with current substance use were assessed for adherence level via self report and pill count methods, severity of depressive symptoms (Beck Depression Inventory- II) and substance use (Addiction Severity Index) during a three months follow up after initiating antiretroviral therapy. The adherence level, severity of depressive symptoms and substance use were assessed one month, two months and three months after initiation of anti-retroviral therapy. Addiction Severity Index (ASI) composite scores were calculated for each domain and the associations between ASI domains and adherence as well as severity of depressive symptoms and adherence were assessed. Twenty six HIV patients with current substance use disorder completed the study. At the end of the first month, adherence to therapy via pill count and self-report were 80%±31.9% and 85.12%±32%, respectively. At the end of the second month, adherence to therapy via pill count and self report were 87%±32% and 93.94%±23% respectively. At the end of the third month, the measured adherence via pill count and self report were 85%±33.7% and 90.1%±25.7% respectively. Adherence was higher among married patients and those who used reminder systems. Composite scores of the medical status and psychiatric status were related to higher adherence after first month. Substance use was inversely associated with adherence at the second follow up (r=-0.4, p=0.04). Also, severity of depressive symptoms was not related to adherence level. The repeated measurement analysis showed a significant decrease in psychiatric status domain of the ASI composite score after three months of initiating therapy (p=0.02). Preventive measures should aim treatment of substance use among HIV patients in order to increase adherence level. Also, conducting psychological evaluations is necessary considering the high prevalence of depression among Iranian HIV patients.


Income non-reporting: implications for health inequalities research.
To determine whether, in the context of a face to face interview, socioeconomic groups differ in their propensity to provide details about the amount of their personal income, and to discuss the likely consequences of any differences for studies that use income based measures of socioeconomic position. The study used data from the 1995 Australian Health Survey. The sample was selected using a stratified multi-stage area design that covered urban and rural areas across all States and Territories and included non-institutionalised residents of private and non-private dwellings. The response rate was 91.5% for selected dwellings and 97.0% for persons within dwellings. Data were collected using face to face interviews. Income response, the dependent measure, was binary coded (0 if income was reported and 1 for refusals, "don't knows" and insufficient information). Socioeconomic position was measured using employment status, occupation, education and main income source. The socioeconomic characteristics of income non-reporters were initially examined using sex specific age adjusted proportions with 95% confidence intervals. Multivariate analysis was performed using logistic regression. Persons aged 15-64 (n = 33,434) who were reportedly in receipt of an income from one or more sources during the data collection reference period. The overall rate of income non-response was 9.8%. Propensity to not report income increased with age (15-29 years 5.8%, 30-49 10.6%, 50-64 13.8%). No gender differences were found (men 10.2%, women 9.3%). Income non-response was not strongly nor consistently related to education or occupation for men, although there was a suggested association among these variables for women, with highly educated women and those in professional occupations being less likely to report their income. Strong associations were evident between income non-response, labour force status and main income source. Rates were highest among the employed and those in receipt of an income from their own business or partnership, and lowest among the unemployed and those in receipt of a government pension or benefit (which excluded the unemployed). Given that differences in income non-reporting were small to moderate across levels of the education and occupation variables, and that propensity to not report income was greater among higher socioeconomic groups, estimates of the relation between income and health are unlikely to be affected by socioeconomic variability in income non-response. Probability estimates from a logistic regression suggested that higher rates of income non-reporting among employed persons who received their income from a business or partnership were not attributable to socio-economic factors. Rather, it is proposed that these higher rates were attributable to recall effects, or concerns about having one's income information disclosed to taxation authorities. Future studies need to replicate this analysis to determine whether the results can be inferred to other survey and data collection contexts. The analysis should also be extended to include an examination of the relation between socio-economic position and accuracy of income reporting. Little is known about this issue, yet it represents a potential source of bias that may have important implications for studies that investigate the association between income and health.


A nationwide school fruit and vegetable policy and childhood and adolescent overweight: A quasi-natural experimental study.
School free fruit and vegetable (FFV) policies are used to promote healthy dietary habits and tackle obesity; however, our understanding of their effects on weight outcomes is limited. We assess the effect of a nationwide FFV policy on childhood and adolescent weight status and explore heterogeneity by sex and socioeconomic position. This study used a quasi-natural experimental design. Between 2007 and 2014, Norwegian combined schools (grades 1-10, age 6 to 16 years) were obligated to provide FFVs while elementary schools (grades 1-7) were not. We used 4 nationwide studies (n = 11,215 children) from the Norwegian Growth Cohort with longitudinal or cross-sectional anthropometric data up to age 8.5 and 13 years to capture variation in FFV exposure. Outcomes were body mass index standard deviation score (BMISDS), overweight and obesity (OW/OB), waist circumference (WC), and weight to height ratio (WtHR) at age 8.5 years, and BMISDS and OW/OB at age 13 years. Analyses included longitudinal models of the pre- and post-exposure trajectories to estimate the policy effect. The participation rate in each cohort was >80%, and in most analyses <4% were excluded due to missing data. Estimates were adjusted for region, population density, and parental education. In pooled models additionally adjusted for pre-exposure BMISDS, there was little evidence of any benefit or unintended consequence from 1-2.5 years of exposure to the FFV policy on BMISDS, OW/OB, WC, or WtHR in either sex. For example, boys exposed to the FFV policy had a 0.05 higher BMISDS (95% CI: -0.04, 0.14), a 1.20-fold higher odds of OW/OB (95% CI: 0.86, 1.66) and a 0.3 cm bigger WC (95% CI: -0.3, 0.8); while exposed girls had a 0.04 higher BMISDS (95% CI: -0.04, 0.13), a 1.03 fold higher odds of OW/OB (95% CI: 0.75, 1.39), and a 0-cm difference in WC (95% CI: -0.6, 0.6). There was evidence of heterogeneity in the policy effect estimates at 8.5 years across cohorts and socioeconomic position; however, these results were inconsistent with other comparisons. Analysis at age 13 years, after 4 years of policy exposure, also showed little evidence of an effect on BMISDS or OW/OB. The main limitations of this study are the potential for residual confounding and exposure misclassification, despite efforts to minimize their impact on conclusions. In this study we observed little evidence that the Norwegian nationwide FFV policy had any notable beneficial effect or unintended consequence on weight status among Norwegian children and adolescents.


Patient and caregiver productivity loss and indirect costs associated with cardiovascular events in Portugal.
To present the Portuguese results of a multi-country cross-sectional survey aiming to estimate productivity loss in the first year after an acute coronary syndrome (ACS) or stroke. Patients previously hospitalized for ACS or stroke were enrolled during a routine cardiology/neurology visit 3-12 months after the index event and ≥4 weeks after returning to work. Productivity loss for the patient and the caregiver in the previous four weeks were reported by the patient using the validated iMTA Productivity Cost Questionnaire (iPCQ). Hours lost were converted into eight-hour work days and prorated to one year, combined with initial hospitalization and sick leave, and valued according to Portuguese labor costs. The analysis included 39 employed patients with ACS (mean age 51 years, 80% men, 95% with myocardial infarction, mean left ventricular ejection fraction 55%) and 31 with stroke (mean age 50 years, 80% men, all ischemic, 77% with modified Rankin Scale 0-1); 41% of ACS and 10% of stroke patients had a history of cardiovascular disease. Mean (SD) productivity loss for patients and caregivers was 47 (62) work days for ACS and 76 (101) work days for stroke. ACS patients lost 37 (39) and caregivers lost 10 (42) work days. Stroke patients and caregivers lost 65 (78) and 12 (38) work days, respectively. Total mean indirect cost per case was €5403 (€7095) and €8726 (€11558) for employed patients with ACS and stroke, respectively. The annual proportions of productive time lost by employed patients due to ACS and stroke in Portugal were 17% and 27%, respectively. Caregivers of these patients lost about 5% of their annual productive time.


High frequency of APOB gene mutations causing familial hypobetalipoproteinaemia in patients of Dutch and Spanish descent.
Familial hypobetalipoproteinaemia (FHBL) is an autosomal co-dominant hereditary disorder of lipoprotein metabolism characterised by decreased low density lipoprotein (LDL) cholesterol and apolipoprotein B (APOB) plasma levels. High levels of plasma APOB and LDL cholesterol are strong predictors for risk of cardiovascular disease (CVD), while individuals with low APOB and LDL cholesterol levels are thought to have lower than average risk for CVD, and in fact, heterozygous FHBL patients appear to be asymptomatic. Rather than identifying truncated APOB proteins in plasma fractions separated by gel electrophoresis, which will miss any mutations in proteins smaller than 30 kb, we analysed the APOB gene directly, using PCR. We identified nine different mutations, six of which are novel. Each mutation showed complete co-segregation with the FHBL phenotype in the families, and statistically significant differences between carriers and non-carriers were found for plasma total, LDL, and HDL cholesterol, triglycerides, and APOB levels, but not for APOA1 levels. All carriers of an APOB mutation were completely free from CVD. Prolonged low levels of LDL cholesterol and elevated levels of HDL cholesterol may reduce the progression of atherosclerotic disease, but this has not been unequivocally shown that this is indeed the case in individuals with FHBL, and is the subject of a current study.


[How to detect neuropathy in leprosy].
In leprosy, the early detection of peripheral nerve damage is essential for the prevention of disability. To date, there is no consensus on what is the best clinical test to reveal such abnormalities. In this prospective study we examined the effectiveness of five clinical tests to assess radial cutaneous nerve (RCN) damage (the most frequently involved). Light touch was assessed by two nylon threads (based on the Semmes-Weinstein monofilaments testing technique) bent on the skin at a pressure of 0.5 (N. 4 nylon) and 0.2 gram (N. 5 nylon). Pinprick and cooling sensations were examined by a needle and a drop of ether. The nerve thickness was assessed by palpation. Sensory findings were then compared to sensory nerve conduction values of the RCN and a sensitivity analysis was performed. The patient group consisted of 108 consecutive new leprosy sufferers (138 RCN) who attended the Institut de Léprologie Appliquée de Dakar during one year. Diagnosis and classification were based on Ridley and Jopling's criteria (clinical examination, skin smears and biopsy). Normal values were determined among 22 healthy subjects (44 RCN). The best tests in term of sensitivity were palpation (.60), N. 5 nylon (.65) and N. 5 + palpation (.79). Their positive predictive values were .84 (palpation), .94 (N. 5 nylon) and .83 (N. 5 + palpation). The best tests in term of area under the curve were palpation (.66), N. 5 nylon (.71) and N. 5 + palpation (.78). The results remain the same for the lepromatous or tuberculoid leprosy patients.(ABSTRACT TRUNCATED AT 250 WORDS)


A novel Apaf-1-independent putative caspase-2 activation complex.
Caspase activation is a key event in apoptosis execution. In stress-induced apoptosis, the mitochondrial pathway of caspase activation is believed to be of central importance. In this pathway, cytochrome c released from mitochondria facilitates the formation of an Apaf-1 apoptosome that recruits and activates caspase-9. Recent data indicate that in some cells caspase-9 may not be the initiator caspase in stress-mediated apoptosis because caspase-2 is required upstream of mitochondria for the release of cytochrome c and other apoptogenic factors. To determine how caspase-2 is activated, we have studied the formation of a complex that mediates caspase-2 activation. Using gel filtration analysis of cell lysates, we show that caspase-2 is spontaneously recruited to a large protein complex independent of cytochrome c and Apaf-1 and that recruitment of caspase-2 to this complex is sufficient to mediate its activation. Using substrate-binding assays, we also provide the first evidence that caspase-2 activation may occur without processing of the precursor molecule. Our data are consistent with a model where caspase-2 activation occurs by oligomerization, independent of the Apaf-1 apoptosome.


MR imaging of the pleura and chest wall.
Evaluation of pleural and chest-wall disease begins with a chest radiograph. In many cases, further evaluation of the chest wall and pleura requires cross-sectional imaging with CT scan or MR imaging. MR imaging is reserved for specific cases of tumors, infections, pleural effusions, and masses. The superior soft-tissue contrast of MR imaging is useful in evaluating the extent of infections and tumors involving the pleura and chest wall. The multiplanar capability of MR imaging aids in the evaluation of chest wall and pleural abnormalities, particularly in the apical regions.


Statin use in patients with diabetes and kidney disease: the Japanese experience.
Diabetes is a leading cause of chronic kidney disease (CKD) and end-stage renal disease (ESRD) in developed nations, including Japan and the United States. Japan has the unenviable distinction of having one of the world's highest rates of dialysis: in 2011, there were over 300,000 dialysis patients (2,383 per million people), with diabetic patients accounting for almost half of all incident cases. Concomitance of CKD and diabetes predicts a greater risk of cardiovascular disease (CVD) than either condition in isolation. Hence, appropriate management of modifiable cardiovascular (CV) risk factors, including dyslipidemia, is paramount in this high-risk group. The United States and Japan have distinct approaches to cholesterol management, with more stringent therapeutic targets for lipid control advocated in US guidelines. However, upward trends in cholesterol levels and coronary heart disease incidence in Japan may provide justification for more intensive CV risk factor management strategies by Japanese physicians to achieve maximum benefit. Attainment of recommended lipid goals in Japan is poor, particularly in patients with diabetes and/or CKD in whom CV risk factors are often undertreated. Statin therapy has been shown to be safe and effective in reducing CV risk in patients with diabetes and/or CKD stages 1-5. Moreover, statins may impart a renoprotective effect by preventing or delaying progressive loss of kidney function. This review summarizes evidence from studies in Western and Japanese populations to highlight the CV and renal benefits of lipid-lowering agents in CKD patients, including those with diabetes.


Venous Thromboembolism in Minimally Invasive Compared With Open Hysterectomy for Endometrial Cancer.
To evaluate whether minimally invasive surgery for endometrial cancer is independently associated with a decreased odds of venous thromboembolism compared with open surgery. We performed a secondary analysis cohort study of prospectively collected quality improvement data and examined patients undergoing hysterectomy for endometrial cancer from 2008 to 2013 recorded in the National Surgical Quality Improvement Program database. Patients undergoing minimally invasive (laparoscopic or robotic) surgery were compared with those undergoing open surgery with respect to 30-day postoperative venous thromboembolism. Demographic and procedure variables were examined as potential confounders. Data regarding receipt of perioperative venous thromboembolism prophylaxis were not available. Bivariable tests and logistic regression were used for analysis. Of 9,948 patients who underwent hysterectomy for the treatment of endometrial cancer, 61.9% underwent minimally invasive surgery and 38.1% underwent open surgery. Patients undergoing minimally invasive surgery had a lower venous thromboembolism incidence (0.7%, n=47) than patients undergoing open surgery (2.2%, n=80) (P<.001). In a multivariate model adjusting for age, body mass index, race, operative time, Charlson comorbidity score, and surgical complexity, minimally invasive surgery remained associated with decreased odds of venous thromboembolism (adjusted odds ratio 0.36, 95% confidence interval 0.24-0.53) compared with open surgery. Minimally invasive surgery for the treatment of endometrial cancer is independently associated with decreased odds of venous thromboembolism compared with open surgery.


Analysis of 1135 gut metagenomes identifies sex-specific resistome profiles.
Several gastrointestinal diseases show a sex imbalance, although the underlying (patho)physiological mechanisms behind this are not well understood. The gut microbiome may be involved in this process, forming a complex interaction with host immune system, sex hormones, medication and other environmental factors. Here we performed sex-specific analyses of fecal microbiota composition in 1135 individuals from a population-based cohort. The overall gut microbiome composition of females and males was significantly different (p = 0.001), with females showing a greater microbial diversity (p = 0.009). After correcting for the effects of intrinsic factors, smoking, diet and medications, female hormonal factors such as the use of oral contraceptives and undergoing an ovariectomy were associated with microbial species and pathways. Females had a higher richness of antibiotic-resistance genes, with the most notable being resistance to the lincosamide nucleotidyltransferase (LNU) gene family. The higher abundance of resistance genes is consistent with the greater prescription of the Macrolide-Lincosamide-Streptogramin classes of antibiotics to females. Furthermore, we observed an increased resistance to aminoglycosides in females with self-reported irritable bowel syndrome. These results throw light upon the effects of common medications that are differentially prescribed between sexes and highlight the importance of sex-specific analysis when studying the gut microbiome and resistome.


Isolation of mesenchymal stem cells from equine umbilical cord blood.
There are no published studies on stem cells from equine cord blood although commercial storage of equine cord blood for future autologous stem cell transplantations is available. Mesenchymal stem cells (MSC) have been isolated from fresh umbilical cord blood of humans collected non-invasively at the time of birth and from sheep cord blood collected invasively by a surgical intrauterine approach. Mesenchymal stem cells isolation percentage from frozen-thawed human cord blood is low and the future isolation percentage of MSCs from cryopreserved equine cord blood is therefore expectedly low. The hypothesis of this study was that equine MSCs could be isolated from fresh whole equine cord blood. Cord blood was collected from 7 foals immediately after foaling. The mononuclear cell fraction was isolated by Ficoll density centrifugation and cultured in a DMEM low glucose based media at 38.5 degrees C in humidified atmosphere containing 5% CO2. In 4 out of 7 samples colonies with MSC morphology were observed. Cellular morphology varied between monolayers of elongated spindle-shaped cells to layered cell clusters of cuboidal cells with shorter cytoplasmic extensions. Positive Alizarin Red and von Kossa staining as well as significant calcium deposition and alkaline phosphatase activity confirmed osteogenesis. Histology and positive Safranin O staining of matrix glycosaminoglycans illustrated chondrogenesis. Oil Red O staining of lipid droplets confirmed adipogenesis. We here report, for the first time, the isolation of mesenchymal-like stem cells from fresh equine cord blood and their differentiation into osteocytes, chondrocytes and adipocytes. This novel isolation of equine cord blood MSCs and their preliminary in vitro differentiation positions the horse as the ideal pre-clinical animal model for proof-of-principle studies of cord blood derived MSCs.


Amniotic fluid stem cells: future perspectives.
The existence of stem cells in human amniotic fluid was reported for the first time almost ten years ago. Since this discovery, the knowledge about these cells has increased dramatically. Today, amniotic fluid stem (AFS) cells are widely accepted as a new powerful tool for basic research as well as for the establishment of new stem-cell-based therapy concepts. It is possible to generate monoclonal genomically stable AFS cell lines harboring high proliferative potential without raising ethical issues. Many different groups have demonstrated that AFS cells can be differentiated into all three germ layer lineages, what is of relevance for both, the scientific and therapeutical usage of these cells. Of special importance for the latter is the fact that AFS cells are less tumorigenic than other pluripotent stem cell types. In this paper, we have summarized the current knowledge about this relatively young scientific field. Furthermore, we discuss the relevant future perspectives of this promising area of stem cell research focusing on the next important questions, which need to be answered.


Mediastinal goiter diagnosed by functional imaging.
A 63-year-old asymptomatic woman with cured Hodgkin diseases presented for restaging. The chest computed tomography showed a mass at the right side of the upper mediastinum. The benignity and the origin of the tissue were unknown. First, we performed a bronchoscopy-guided biopsy but without success. In the next step, we initiated radionuclide imaging with technetium-99m pertechnetate (Tc-99m) and radioiodine (I-123). Low uptake of Tc-99m and intense accumulation of I-123 after 2 and 24 h to the mediastinal mass suggested that the mass was a mediastinal goiter. Based on iodine uptake and the fact that our patient had no symptoms of tracheal compression, we decide to go for a radioiodine therapy.


Direct molecular regulation of the myogenic determination gene Myf5 by Pax3, with modulation by Six1/4 factors, is exemplified by the -111 kb-Myf5 enhancer.
The Myf5 gene plays an important role in myogenic determination during mouse embryo development. Multiple genomic regions of the Mrf4-Myf5 locus have been characterised as enhancer sequences responsible for the complex spatiotemporal expression of the Myf5 gene at the onset of myogenesis. These include an enhancer sequence, located at -111 kb upstream of the Myf5 transcription start site, which is responsible of Myf5 activation in ventral somitic domains (Ribas et al., 2011. Dev. Biol. 355, 372-380). We show that the -111 kb-Myf5 enhancer also directs transgene expression in some limb muscles, and is active at foetal as well as embryonic stages. We have carried out further characterisation of the regulation of this enhancer and show that the paired-box Pax3 transcription factor binds to it in vitro as in vivo, and that Pax binding sites are essential for its activity. This requirement is independent of the previously reported regulation by TEAD transcription factors. Six1/4 which, like Pax3, are important upstream regulators of myogenesis, also bind in vivo to sites in the -111 kb-Myf5 enhancer and modulate its activity. The -111 kb-Myf5 enhancer therefore shares common functional characteristics with another Myf5 regulatory sequence, the hypaxial and limb 145 bp-Myf5 enhancer, both being directly regulated in vivo by Pax3 and Six1/4 proteins. However, in the case of the -111 kb-Myf5 enhancer, Six has less effect and we conclude that Pax regulation plays a major role in controlling this aspect of the Myf5 gene expression at the onset of myogenesis in the embryo.


Comparison of Cytotoxic Activity of Bile on HepG2 and CCRF-CEM Cell Lines: An in Vitro Study.
The aim of this study was to examine the effect of crude bile on the human HepG2 and CCRF-CEM cell lines. Cells were exposed to different dilutions of bile. Antiproliferative effects were determined by the cytotoxic MTT assay. Cells undergoing apoptosis were identified by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Bile administration resulted in dose-dependent cytotoxicity in both HepG2 and CCRF-CEM cell lines. Incubated cells exhibited morphologic features of apoptosis. Bile has significant cytotoxic activity in HepG2 and CCRF-CEM cancer cells via induction of apoptosis. The mechanism of apoptosis needs to be further evaluated. It may have clinical utility in the treatment of cancer after in vivo confirmation of activity.


Effect of Maillard reaction on biochemical properties of peanut 7S globulin (Ara h 1) and its interaction with a human colon cancer cell line (Caco-2).
The purpose of this study was to determine the influence of Maillard reaction (MR, glycation) on biochemical and biological properties of the major peanut allergen Ara h 1. Three different time/temperature conditions of treatment were applied (37, 60, and 145 °C). The extent of MR was assessed by SDS-PAGE, loss of free amino groups, fluorescence intensity, content of bound sugar and fructosamine. The Caco-2 model system was applied to study effects of hydrolysed and non-hydrolysed Ara h 1 on proliferation and interleukin-8 (IL-8) secretion from Caco-2 cells. We demonstrated significant differences in the biochemical properties of Ara h 1 glycated at different time/temperature conditions. Glycation of Ara h 1 at 37 °C was shown to cause least biochemical changes, not limiting pepsin hydrolysis. Loss of free amino groups, increase of fluorescence and brown colour of Ara h 1 glycated at 60 and 145 °C indicated advanced and final stages of MR. Non-treated Ara h 1 inhibited Caco-2 cell proliferation and stimulated IL-8 secretion. This effect was less pronounced for glycated Ara h 1. Incubation of Caco-2 cells with non-hydrolysed Ara h 1, glycated at the temperature of 37 and 60 °C, did not stimulate IL-8 secretion. Each applied time/temperature-treatment combination caused different biochemical changes of Ara h 1, underlining diversity of formed MRPs. MR, independently of temperature/time conditions, reduced the pro-inflammatory properties of native Ara h 1, reflected in stimulation of IL-8 secretion from intestinal epithelial cells.


Validation of PCR-RFLP techniques for the evaluation of codon 72 of p53 and CYP1A1 gene's polymorphisms in relation with ovarian cancer in a Romanian population.
Epidemiological studies suggest that the onset and progression of ovarian cancer are associated with the presence of estrogens. CYP1A1 gene has two polymorphisms, which may affect the estrogens' metabolites and contribute to increased susceptibility to neoplastic transformation of ovarian cells. P53 is a tumor suppressor gene, which acts to preserve stability of human genome. Codon 72 polymorphism of p53 gene was correlated with susceptibility for ovarian cancer. The aim of our study was to validate the use of PCR-RFLP techniques for the evaluation of p53 codon 72 and CYP1A1 gene polymorphisms in relation with ovarian cancer in a Romanian population and to evaluate gene-environment interaction in this context. The case-control study included 42 subjects. The assessment of risk and protective factors was performed using a questionnaire. Polymorphisms of CYP1A1 and p53 genes were assessed using the validated PCR-RFLP techniques. The statistical analysis was performed using Epi Info 3.5.1 software. Frequency of Arg/Arg genotype of p53 gene was higher among cases (43%) compared with controls (33.3%), but the difference was not statistically significant (p=0.75). The presence of Ile/Val polymorphism of CYP1A1 gene was identified in 9.5% of the cases and the MspI polymorphism of CYP1A1 gene was not identified in our subjects. Validation of techniques consisted in the optimization of RFLP methods for p53 and CYP1A1 genes polymorphism analyzing that allowed highlighting the existence of codon 72 polymorphism of p53 gene and Ile/Val polymorphism of CYP1A1 gene in the population from this region.


Testosterone and reproductive effort in male primates.
Considerable evidence suggests that the steroid hormone testosterone mediates major life-history trade-offs in vertebrates, promoting mating effort at the expense of parenting effort or survival. Observations from a range of wild primates support the "Challenge Hypothesis," which posits that variation in male testosterone is more closely associated with aggressive mating competition than with reproductive physiology. In both seasonally and non-seasonally breeding species, males increase testosterone production primarily when competing for fecund females. In species where males compete to maintain long-term access to females, testosterone increases when males are threatened with losing access to females, rather than during mating periods. And when male status is linked to mating success, and dependent on aggression, high-ranking males normally maintain higher testosterone levels than subordinates, particularly when dominance hierarchies are unstable. Trade-offs between parenting effort and mating effort appear to be weak in most primates, because direct investment in the form of infant transport and provisioning is rare. Instead, infant protection is the primary form of paternal investment in the order. Testosterone does not inhibit this form of investment, which relies on male aggression. Testosterone has a wide range of effects in primates that plausibly function to support male competitive behavior. These include psychological effects related to dominance striving, analgesic effects, and effects on the development and maintenance of the armaments and adornments that males employ in mating competition.


Practical visualization of internal structure of white matter for image interpretation: staining a spin-echo T2-weighted image with three echo-planar diffusion-weighted images.
To our knowledge, no method satisfactory for clinical use has been developed to visualize white matter fiber tracts with diffusion-weighted MR imaging. The purpose of this study was to determine whether superposition of a spin-echo T2-weighted image and a color-coded image derived from three orthogonal diffusion-weighted images could show fiber tract architecture of the brain with an image quality appropriate for accurate reading with a computer monitor. MR images from 50 consecutive cases were reviewed. Three diffusion-weighted images per section were acquired with three orthogonal motion-probing gradients. These images were registered to a corresponding spin-echo T2-weighted image. A color-coded image was synthesized from three diffusion-weighted images by assigning red, green, or blue to each diffusion-weighted image and then adding a spin-echo T2-weighted image with a weighting factor. The ability of the superposed image to delineate the white matter pathways was evaluated on the basis of the known anatomy of these pathways and qualitatively compared with that of the spin-echo T2-weighted image. The main white matter fiber pathways, in particular the superior longitudinal fascicle, corpus callosum, tapetum, optic radiation, and internal capsule, were more clearly and easily identified on the superposed image than on the spin-echo T2-weighted image. The time required to produce the superposed image was approximately 40 minutes. Superposition of a spin-echo T2-weighted image and a color-coded image created from three orthogonal diffusion-weighted images showed structures of the brain that were not clearly visible on the spin-echo T2-weighted image alone. Such superposition presents images that are easy to interpret correctly.


Moderators of Effects of Internet-Delivered Exercise and Pain Coping Skills Training for People With Knee Osteoarthritis: Exploratory Analysis of the IMPACT Randomized Controlled Trial.
Internet-delivered exercise, education, and pain coping skills training is effective for people with knee osteoarthritis, yet it is not clear whether this treatment is better suited to particular subgroups of patients. The aim was to explore demographic and clinical moderators of the effect of an internet-delivered intervention on changes in pain and physical function in people with knee osteoarthritis. Exploratory analysis of data from 148 people with knee osteoarthritis who participated in a randomized controlled trial comparing internet-delivered exercise, education, and pain coping skills training to internet-delivered education alone. Primary outcomes were changes in knee pain while walking (11-point Numerical Rating Scale) and physical function (Western Ontario and McMaster Universities Osteoarthritis Index function subscale) at 3 and 9 months. Separate regression models were fit with moderator variables (age, gender, expectations of outcomes, self-efficacy [pain], education, employment status, pain catastrophizing, body mass index) and study group as covariates, including an interaction between the two. Participants in the intervention group who were currently employed had significantly greater reductions in pain at 3 months than similar participants in the control group (between-group difference: mean 2.38, 95% CI 1.52-3.23 Numerical Rating Scale units; interaction P=.02). Additionally, within the intervention group, pain at 3 months reduced by mean 0.53 (95% CI 0.28-0.78) Numerical Rating Scale units per unit increase in baseline self-efficacy for managing pain compared to mean 0.11 Numerical Rating Scale units (95% CI -0.13 to 0.35; interaction P=.02) for the control group. People who were employed and had higher self-efficacy at baseline were more likely to experience greater improvements in pain at 3 months after an internet-delivered exercise, education, and pain coping skills training program. There was no evidence of a difference in the effect across gender, educational level, expectation of treatment outcome, or across age, body mass index, or tendency to catastrophize pain. Findings support the effectiveness of internet-delivered care for a wide range of people with knee osteoarthritis, but future confirmatory research is needed. Australian New Zealand Clinical Trials Registry ACTRN12614000243617; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=365812&isReview=true (Archived by WebCite at http://www.webcitation.org/6z466oTPs).