Gamma-ray emission from the shell of supernova remnant W44 revealed by the Fermi LAT.
Recent observations of supernova remnants (SNRs) hint that they accelerate cosmic rays to energies close to ~10(15) electron volts. However, the nature of the particles that produce the emission remains ambiguous. We report observations of SNR W44 with the Fermi Large Area Telescope at energies between 2 x 10(8) electron volts and 3 x10(11) electron volts. The detection of a source with a morphology corresponding to the SNR shell implies that the emission is produced by particles accelerated there. The gamma-ray spectrum is well modeled with emission from protons and nuclei. Its steepening above approximately 10(9) electron volts provides a probe with which to study how particle acceleration responds to environmental effects such as shock propagation in dense clouds and how accelerated particles are released into interstellar space.


Clinical and molecular characteristics of carnitineacylcarnitine translocase deficiency with c.270delC and a novel c.408C>A variant.
Carnitine-acylcarnitine translocase deficiency (CACTD) is a rare, autosomal recessive, and highly lethal fatty acid oxidation (FAO) disorder caused by defective acylcarnitine transport across the mitochondrial membrane. CACTD is characterized by severe episodes of hypoglycemia and hyperammonemia, seizures, cardiomyopathy, liver dysfunction, severe neurological damage, and muscle weakness. Herein, we described the clinical features, biochemical, and molecular findings of three patients with CACTD, presented with poor feeding, hypoglycemia, liver dysfunctions, and hyperammonemia, but died despite intensive treatment. All cases had similar signs and symptoms like poor feeding and respiratory failure associated with liver dysfunction. Urinary organic acid profiles in the presence of hypoglycemia and hyperammonemia led us to the possible diagnosis of one of fatty acid β-oxidation defects. Results of the molecular analyses were compatible with CACTD. In addition to known mutation (c.270delC;p.Phe91Leufs*38) we detected a novel one (c.408C > A;p.Cys136*). All three cases died despite a very intensive therapy. Based on our experience with these three cases, it can be said that CACTD has a relatively poor prognosis, molecular studies are of most importance in suspected cases for the final diagnosis and such studies might be of help while giving genetic counselling and guidance to parents for future pregnancies.


Pathophysiology and treatment of alien hand syndrome.
Alien hand syndrome (AHS) is a disorder of involuntary, yet purposeful, hand movements that may be accompanied by agnosia, aphasia, weakness, or sensory loss. We herein review the most reported cases, current understanding of the pathophysiology, and treatments. We performed a PubMed search in July of 2014 using the phrases "alien hand syndrome," "alien hand syndrome pathophysiology," "alien hand syndrome treatment," and "anarchic hand syndrome." The search yielded 141 papers (reviews, case reports, case series, and clinical studies), of which we reviewed 109. Non-English reports without English abstracts were excluded. Accumulating evidence indicates that there are three AHS variants: frontal, callosal, and posterior. Patients may demonstrate symptoms of multiple types; there is a lack of correlation between phenomenology and neuroimaging findings. Most pathologic and functional imaging studies suggest network disruption causing loss of inhibition as the likely cause. Successful interventions include botulinum toxin injections, clonazepam, visuospatial coaching techniques, distracting the affected hand, and cognitive behavioral therapy. The available literature suggests that overlap between AHS subtypes is common. The evidence for effective treatments remains anecdotal, and, given the rarity of AHS, the possibility of performing randomized, placebo-controlled trials seems unlikely. As with many other interventions for movement disorders, identifying the specific functional impairments caused by AHS may provide the best guidance towards individualized supportive care.


Study of protein targets for covalent modification by the antitumoral and anti-inflammatory prostaglandin PGA1: focus on vimentin.
Prostaglandins with cyclopentenone structure (cyPG) display potent antiproliferative actions that have elicited their study as potential anticancer agents. Several natural and synthetic analogs of the cyPG prostaglandin A(1) (PGA(1)) have proven antitumoral efficacy in cancer cell lines and animal models. In addition, PGA(1) has been used as an inhibitor of transcription factor NF-kappaB-mediated processes, including inflammatory gene expression and viral replication. An important determinant for these effects is the ability of cyPG to form Michael adducts with free thiol groups. The chemical nature of this interaction implies that PGA(1) could covalently modify cysteine residues in a large number of cellular proteins potentially involved in its beneficial effects. However, only a few targets of PGA(1) have been identified. In previous work, we have observed that a biotinylated analog of PGA(1) that retains the cyclopentenone moiety (PGA(1)-B) binds to multiple targets in fibroblasts. Here, we have addressed the identification of these targets through a proteomic approach. Cell fractionation followed by avidin affinity chromatography yielded a fraction enriched in proteins modified by PGA(1)-B. Analysis of this fraction by SDS-PAGE and LC-MS/MS allowed the identification of the chaperone Hsp90, elongation and initiation factors for protein synthesis and cytoskeletal proteins including actin, tubulin and vimentin. Furthermore, we have characterized the modification of vimentin both in vitro and in intact cells. Our observations indicate that cysteine 328 is the main site for PGA(1) addition. These results may contribute to a better understanding of the mechanism of action of PGA(1) and the potential of cyPG-based therapeutic strategies.


Potential of the natural products against leishmaniasis in Old World - a review of in-vitro studies.
Leishmaniasis is a vector-borne disease among the 10 most Neglected Tropical Diseases with diverse clinical manifestations caused by protozoan parasites of the Leishmania genus. Around 80% of leishmaniasis cases are found in the Old World affecting populations mainly in low and middle-income countries. Its control relies mostly on chemotherapy which still presents many drawbacks. Natural products may offer an inexhaustible source of chemical diversity with therapeutic potential. Despite the lack of knowledge on traditional products with activity against Leishmania parasites, many reports describe the search for natural extracts and compounds with antileishmanial properties against promastigote and amastigote parasite forms. This review summarizes the research of 74 publications of the last decade (2008-2018) focused on the identification of endemic plant-derived products that are active against Old World Leishmania parasites responsible for cutaneous and visceral leishmaniasis. The present review combines data on antileishmanial activity of 423 plants species, belonging to 94 different families, including a large range of crude extracts which lead to the isolation of 86 active compounds. Most studied plants came from Asia and most promising plant families for antileishmanial activity were Asteraceae and Lamiaceae. From the chemical point of view, terpenoids were the most frequently isolated natural products. These studies suggest that natural products isolated from Old World flora are a rich source of new chemical scaffolds for future leishmaniasis treatment as well as for other Neglected Tropical Diseases warranting further investigation.


A systematic review of PET and PET/CT in oncology: a way to personalize cancer treatment in a cost-effective manner?
A number of diagnostic tests are required for the detection and management of cancer. Most imaging modalities such as computerized tomography (CT) are anatomical. However, positron emission tomography (PET) is a functional diagnostic imaging technique using compounds labelled with positron-emitting radioisotopes to measure cell metabolism. It has been a useful tool in studying soft tissues such as the brain, cardiovascular system, and cancer. The aim of this systematic review is to critically summarize the health economic evidence of oncologic PET in the literature. Eight electronic databases were searched from 2005 until February 2010 to identify economic evaluation studies not included in previous Health Technology Assessment (HTA) reports. Only full health economic evaluations in English, French, or German were considered for inclusion. Economic evaluations were appraised using published quality criteria for assessing the quality of decision-analytic models. Given the variety of methods used in the health economic evaluations, the economic evidence has been summarized in qualitative form. From this new search, 14 publications were identified that met the inclusion criteria. All publications were decision-analytic models and evaluated PET using Fluorodeoxyglucose F18 (FDG-PET). Eight publications were cost-effectiveness analyses; six were cost-utility analyses. The studies were from Australia, Belgium, Canada, France, Italy, Taiwan, Japan, the Netherlands, the United Kingdom, and the United States. In the base case analyses of these studies, cost-effectiveness results ranged from dominated to dominant. The methodology of the economic evaluations was of varying quality. Cost-effectiveness was primarily influenced by the cost of PET, the specificity of PET, and the risk of malignancy. Owing to improved care and less exposure to ineffective treatments, personalized medicine using PET may be cost-effective. However, the strongest evidence for the cost-effectiveness of PET is still in the staging of non-small cell lung cancer. Management decisions relating to the assessment of treatment response or radiotherapy treatment planning require further research to show the impact of PET on patient management and its cost-effectiveness. Because of the potential for increased patient throughput and the possible greater accuracy, the cost-effectiveness of PET/CT may be superior to that of PET. Only four studies of the cost-effectiveness of PET/CT were found in this review, and this is clearly an area for future research.


Association between insomnia symptoms and mortality: a prospective study of U.S. men.
Insomnia complaints are common in older adults and may be associated with mortality risk. However, evidence regarding this association is mixed. Thus, we prospectively examined whether men with insomnia symptoms had an increased risk of mortality during 6 years of follow-up. A prospective cohort study of 23,447 US men participating in the Health Professionals Follow-Up Study and free of cancer, reported on insomnia symptoms in 2004, were followed through 2010. Deaths were identified from state vital statistic records, the National Death Index, family reports, and the postal system. We documented 2025 deaths during 6 years of follow-up (2004-2010). The multivariable-adjusted hazard ratios of total mortality were 1.25 (95% confidence interval [CI], 1.04-1.50) for difficulty initiating sleep, 1.09 (95% CI, 0.97-1.24) for difficulty maintaining sleep, 1.04 (95% CI, 0.88-1.22) for early-morning awakenings, and 1.24 (95% CI, 1.05-1.46) for nonrestorative sleep, comparing men with those symptoms most of the time with men without those symptoms, after adjusting for age, lifestyle factors, and presence of common chronic conditions. Men with difficulty initiating sleep and nonrestorative sleep most of the time had a 55% (hazard ratio, 1.55; 95% CI, 1.19-2.04; P-trend=0.01) and 32% (hazard ratio, 1.32; 95% CI, 1.02-1.72; P-trend=0.002) increased risk of cardiovascular disease mortality, respectively, relative to men without those symptoms. Some insomnia symptoms, especially difficulty initiating asleep and nonrestorative sleep, are associated with a modestly higher risk of mortality.


Quantitative effects on cardiovascular risk factors and coronary heart disease risk of replacing partially hydrogenated vegetable oils with other fats and oils.
Reduced consumption of trans-fatty acids (TFA) is desirable to lower coronary heart disease (CHD) risk. In practice, partially hydrogenated vegetable oils (PHVO) that contain both TFAs and other fatty acids are the unit of replacement and could be replaced with diverse alternative fats and oils. We performed quantitative estimates of CHD effects if a person's PHVO consumption were to be replaced with alternative fats and oils based on (1) randomized dietary trials and (2) prospective observational studies. We performed meta-analyses of (1) the effects of TFAs on blood lipids and lipoproteins in controlled dietary trials and (2) associations of habitual TFA consumption with CHD outcomes in prospective cohort studies. On the basis of these results and corresponding findings for saturated fatty acids (SFA), cis-monounsaturated fatty acids (MUFA) and cis-polyunsaturated fatty acids (PUFA), we calculated the effects on CHD risk for replacing 7.5% of energy from three different PHVO formulations (containing 20, 35 or 45% TFAs) with butter, lard, palm or vegetable oils. In controlled trials, each 1% energy replacement of TFAs with SFAs, MUFAs or PUFAs, respectively, decreased the total cholesterol (TC)/high-density lipoprotein cholesterol (HDL-C) ratio by 0.31, 0.54 and 0.67; the apolipoprotein (Apo)-B/ApoAI ratio by 0.007, 0.010 and 0.011; and lipoprotein (Lp)(a) by 3.76, 1.39 and 1.11 mg/l (P<0.05 for each). We also included possible effects on C-reactive protein (CRP) of TFAs vs other fats from one trial. On the basis of these risk factor changes in controlled trials, CHD risk would be variably decreased by different fats and oils replacing 7.5% of energy from 20% TFA PHVO (CHD risk reduction: -2.7% (butter) to -9.9% (canola)); 35% TFA PHVO (-11.9% (butter) to -16.0% (canola)); or 45% TFA PHVO (-17.6% (butter) to -19.8% (canola)). In prospective cohort studies, each 2% energy replacement of TFAs with SFAs, MUFAs or PUFAs would lower CHD risk by 17% (95% confidence interval (CI)=7-25%), 21% (95% CI=12-30%) or 24% (95% CI=15-33%), respectively. On the basis of these associations in observational studies, CHD risk would be variably decreased by different fats and oils replacing 7.5% of energy from 20% TFA PHVO (CHD risk reduction: +0.5% (butter) to -21.8% (soybean)); 35% TFA PHVO (-14.4% (butter) to -33.4% (soybean)); or 45% TFA PHVO (-22.4% (butter) to -39.6% (soybean)). The demonstrated effects on TC/HDL-C, ApoB/ApoAI, Lp(a), and CRP in randomized feeding trials together accounted for approximately 65-80% and approximately 50% of the estimated risk reduction for replacing PHVO with animal fats and vegetable oils, respectively, that would be calculated from prospective cohort studies. Effects on CHD risk of removing PHVO from a person's diet vary depending on the TFA content of the PHVO and the fatty acid composition of the replacement fat or oil, with direct implications for reformulation of individual food products. Accounting for the summed effects of TFAs on multiple CHD risk factors provides more accurate estimates of potential risk reduction than considering each risk factor in isolation, and approaches the estimated risk reduction derived from prospective cohort studies.


Effect of mobile phone exposure on apoptotic glial cells and status of oxidative stress in rat brain.
The aim of this study was to investigate the effects of mobile phone exposure on glial cells in brain. The study carried out on 31 Wistar Albino adult male rats. The rat heads in a carousel exposed to 900 MHz microwave. For the study group (n:14), rats exposed to the radiation 2 h per day (7 days in a week) for 10 months. For the sham group (n:7), rats were placed into the carousel and the same procedure was applied except that the generator was turned off. For the cage control (n:10), nothing applied to rats in this group. In this study, rats were euthanized after 10 months of exposure periods and brains were removed. Brain tissues were immunohistochemically stained for the active (cleaved) caspase-3, which is a well-known apoptosis marker, and p53. The expression of the proteins was evaluated by a semi-quantitative scoring system. However, total antioxidative capacity (TAC), catalase, total oxidant status (TOS), and oxidative stress index were measured in rat brain. Final score for apoptosis in the exposed group was significantly lower than the sham (p < 0.001) and the cage control groups (p < 0.01). p53 was not significantly changed by the exposure (p > 0.05). The total antioxidant capacity and catalase in the experimental group was found higher than that in the sham group (p < 0.001, p < 0.05). In terms of the TOS and oxidative stress index, there was no statistically significant difference between exposure and sham groups (p > 0.05). In conclusion, the final score for apoptosis, total antioxidant capacity and catalase in rat brain might be altered by 900 MHz radiation produced by a generator to represent exposure of global systems for mobile communication (GSM) cellular phones.


Upregulation of TGF-beta, FOXP3, and CD4+CD25+ regulatory T cells correlates with more rapid parasite growth in human malaria infection.
Understanding the regulation of immune responses is central for control of autoimmune and infectious disease. In murine models of autoimmunity and chronic inflammatory disease, potent regulatory T lymphocytes have recently been characterized. Despite an explosion of interest in these cells, their relevance to human disease has been uncertain. In a longitudinal study of malaria sporozoite infection via the natural route, we provide evidence that regulatory T cells have modifying effects on blood-stage infection in vivo in humans. Cells with the characteristics of regulatory T cells are rapidly induced following blood-stage infection and are associated with a burst of TGF-beta production, decreased proinflammatory cytokine production, and decreased antigen-specific immune responses. Both the production of TGF-beta and the presence of CD4+CD25+FOXP3+ regulatory T cells are associated with higher rates of parasite growth in vivo. P. falciparum-mediated induction of regulatory T cells may represent a parasite-specific virulence factor.


Determinants of primary care physicians' referral pattern: a structural equation model approach.
This study examines patient referrals by primary care physicians (PCP) with nurse practitioners and physician assistants (NP-PA) in their medical practices as compared to practices without them. The study uses data from the Robert Wood Johnson Foundation community tracking study (CTS) Physician Survey, Round I (1996-97) and II (1998-99). Structural equations with binary dependent variables were used to examine the links among managed care, the use of NP-PA, the complexity of patient's conditions, and the number of referrals. PCP's with NP-PA were found to have a greater likelihood of treating patients with complex conditions instead of referring them to specialists. Managed care related variables (i.e., large group practice/HMO, the percentage of patients for whom PCPs acted as gatekeepers, and the percentage of managed care revenue from capitated/prepaid contracts) affected PCP' patient referrals, but only through the increased use of NP-PA. PCP's with NP-PA were also found to provide appropriate care to the complex patients. These findings indicate that NP-PA enable PCP to concentrate on patients with more complex conditions thus reducing the number of referrals. In addition, NP-PA is found to affect the gatekeeper role of PCPs.


Variable selection and prediction in biased samples with censored outcomes.
With the increasing availability of large prospective disease registries, scientists studying the course of chronic conditions often have access to multiple data sources, with each source generated based on its own entry conditions. The different entry conditions of the various registries may be explicitly based on the response process of interest, in which case the statistical analysis must recognize the unique truncation schemes. Moreover, intermittent assessment of individuals in the registries can lead to interval-censored times of interest. We consider the problem of selecting important prognostic biomarkers from a large set of candidates when the event times of interest are truncated and right- or interval-censored. Methods for penalized regression are adapted to handle truncation via a Turnbull-type complete data likelihood. An expectation-maximization algorithm is described which is empirically shown to perform well. Inverse probability weights are used to adjust for the selection bias when assessing predictive accuracy based on individuals whose event status is known at a time of interest. Application to the motivating study of the development of psoriatic arthritis in patients with psoriasis in both the psoriasis cohort and the psoriatic arthritis cohort illustrates the procedure.


Hormone-related risk factors and postmenopausal breast cancer among nulliparous versus parous women: An aggregated study.
Nulliparity is an established breast cancer risk factor, particularly when compared with parity at young ages. The authors aggregated data from 4 US prospective studies (1979-2006) including 32,641 nulliparous (1,612 breast cancers) and 204,270 parous (8,180 breast cancers) women to examine the hypothesis that nulliparity may increase susceptibility to established postmenopausal breast cancer risk factors. The aggregated hazard ratio for nulliparous versus all parous women = 1.27 (95% confidence interval: 1.21, 1.34), and that for nulliparous versus women <25 years of age at first birth = 1.38 (95% confidence interval: 1.30, 1.46). Among nulliparous women, the hazard ratios for current menopausal hormone therapy use (vs. never use), body mass index ≥30 kg/m(2) (vs. <25 kg/m(2)), and weekly consumption of ≥7 alcoholic drinks (vs. none) ranged from 1.3 to 1.6. The hazard ratios did not differ by parity. In a model including all women, the joint association for each of these factors and nulliparity combined compared with first birth before age 25 years was an approximately 2-fold increased breast cancer risk. Although the baseline risk is higher for nulliparous women compared with parous women, these results suggest that the associations between hormone-related factors and breast cancer do not differ by parity.


Cloning, characterization, and functional studies of a human 40-kDa catecholamine-regulated protein: implications in central nervous system disorders.
Catecholamine-regulated proteins (CRPs) have been shown to bind dopamine and other structurally related catecholamines; in particular, the 40-kDa CRP (CRP40) protein has been previously cloned and functionally characterized. To determine putative human homologs, BLAST analysis using the bovine CRP40 sequence identified a human established sequence tag (EST) with significant homology (accession #BQ224193). Using this EST, we cloned a recombinant human brain CRP40-like protein, which possessed chaperone activity. Radiolabeled dopamine binding studies with recombinant human CRP40 protein demonstrated the ability of this protein to bind dopamine with low affinity and high capacity. The full-length human CRP40 nucleotide sequence was elucidated (accession #DQ480334) with RNA ligase-mediated rapid amplification of complementary DNA ends polymerase chain reaction, while Northern blot hybridization suggested that human CRP40 is an alternative splice variant of the 70-kDa mitochondrial heat shock protein, mortalin. Human SH-SY5Y neuroblastoma cells treated with the antipsychotic drug, haloperidol, exhibited a significant increase in CRP40 messenger RNA expression compared to untreated control cells, while other dopamine agonists/antagonists also altered CRP40 expression and immunolocalization. In conclusion, these results show that we have cloned a splice variant of mortalin with a novel catecholamine binding function and that this chaperone-like protein may be neuroprotective in dopamine-related central nervous system disorders.


ABVD versus modified stanford V versus MOPPEBVCAD with optional and limited radiotherapy in intermediate- and advanced-stage Hodgkin's lymphoma: final results of a multicenter randomized trial by the Intergruppo Italiano Linfomi.
In this multicenter, prospective, randomized clinical trial on advanced Hodgkin's lymphoma (HL), the efficacy and toxicity of two chemotherapy regimens, doxorubicin, vinblastine, mechlorethamine, vincristine, bleomycin, etoposide, and prednisone (Stanford V) and mechlorethamine, vincristine, procarbazine, prednisone, epidoxirubicin, bleomycin, vinblastine, lomustine, doxorubicin, and vindesine (MOPPEBVCAD), were compared with doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) as standard therapy to select which regimen would best support a reduced radiotherapy program, which was limited to < or = two sites of either previous bulky or partially remitting disease (a modification of the original Stanford program). Three hundred fifty-five patients with stage IIB, III, or IV HL were randomly assigned. Three hundred thirty-four patients were assessable for the study and received six cycles of ABVD (n = 122), three cycles of Stanford V (n = 107), or six cycles of MOPPEBVCAD (n = 106); radiotherapy was administered to 76, 71, and 50 patients in these three arms, respectively. The complete response rates for ABVD, Stanford V, and MOPPEBVCAD were 89%, 76% and 94%, respectively; 5-year failure-free survival (FFS) and progression-free survival rates were 78%, 54%, 81% and 85%, 73%, and 94%, respectively (P < .01 for comparison of Stanford V with the other two regimens). Corresponding 5-year overall survival rates were 90%, 82%, and 89% for ABVD, Stanford V, and MOPPEBVCAD, respectively. Stanford V was more myelotoxic than ABVD but less myelotoxic than MOPPEBVCAD, which had larger reductions in the prescribed drug doses. When associated with conditioned and limited (not adjuvant) radiotherapy, ABVD and MOPPEBVCAD were superior to Stanford V chemotherapy in terms of response rate and FFS and progression-free survival. Patients were irradiated less often after MOPPEBVCAD, but this regimen was more toxic. ABVD is still the best choice when it is combined with optional, limited irradiation.


The relationship between mechanical work and energy expenditure of locomotion in horses.
Three-dimensional motion capture and metabolic assessment were performed on four standardbred horses while walking, trotting and galloping on a motorized treadmill at different speeds. The mechanical work was partitioned into the internal work (W(INT)), due to the speed changes of body segments with respect to the body centre of mass, and the external work (W(EXT)), due to the position and speed changes of the body centre of mass with respect to the environment. The estimated total mechanical work (W(TOT)=W(INT)+W(EXT)) increased with speed, while metabolic work (C) remained rather constant. As a consequence, the 'apparent efficiency' (eff(APP)=W(TOT)/C) increased from 10 % (walking) to over 100 % (galloping), setting the highest value to date for terrestrial locomotion. The contribution of elastic structures in the horse's limbs was evaluated by calculating the elastic energy stored and released during a single bounce (W(EL,BOUNCE)), which was approximately 1.23 J kg(-)(1) for trotting and up to 6 J kg(-)(1) for galloping. When taking into account the elastic energy stored by the spine bending and released as W(INT), as suggested in the literature for galloping, W(EL,BOUNCE) was reduced by 0.88 J kg(-)(1). Indirect evidence indicates that force, in addition to mechanical work, is also a determinant of the metabolic energy expenditure in horse locomotion.


Effect of biosynthetic human growth hormone on insulin action in individual tissues of the rat in vivo.
Excessive endogenous production or exogenous administration of human growth hormone (hGH) causes insulin resistance at both the hepatic and extrahepatic levels. However, which extrahepatic tissues are involved have not been defined. We have examined the diabetogenic action of authentic biosynthetic hGH on whole body glucose disposal, hepatic glucose output, and glucose metabolism in individual peripheral tissues. The use of a highly purified preparation of the hormone allowed us to examine the isolated effects of 22K hGH. The euglycemic hyperinsulinemic (approximately 100 mU/L) clamp plus 3H-2-deoxyglucose technique was used to quantitate the effects of hGH on insulin action in vivo. Administration of biosynthetic hGH at a dose of 10 IU/kg/24 h for 48 hours in male Wistar rats (approximately 340 g) produced a highly significant decrease in the steady state clamp glucose infusion rate (GIR) when compared with controls (8.1 +/- 0.6 v 18.7 +/- 0.7 mg/kg/min, P less than .001), reduced insulin-mediated suppression of hepatic glucose output (Ra) (3.9 +/- 0.6 v 0.7 +/- 0.3 mg/kg/min, P less than .05) and a decreased clamp glucose disposal rate (Rd) (12.0 +/- 0.4 v 18.10 +/- 1.1 mg/kg/min, P less than .001). There was a significant decrease in insulin-mediated glucose uptake as indicated by tissue accumulation of [3H]-2-deoxyglucose phosphorylation in diaphragm and hindlimb muscles. Insulin action was more substantially reduced in muscles (approximately 50%) than in adipose tissues (approximately 20%). These studies confirm that the diabetogenic action of hGH in the rat is due to a combination of inhibition of insulin suppression of hepatic glucose output and inhibition of the uptake and subsequent utilization of glucose in skeletal muscles.


Combining reduced glutathione and ascorbic acid has supplementary beneficial effects on boar sperm cryotolerance.
The main aim of this work was to evaluate how supplementing freezing and thawing media with reduced glutathione (GSH) and l-ascorbic acid (AA) affected the quality parameters of frozen-thawed boar spermatozoa. With this purpose, semen samples of 12 ejaculates coming from 12 boars were used. Each ejaculate was split into seven aliquots to which 5 mM of GSH and 100 μM of AA were added separately or together at two different steps of freeze-thawing. Various sperm parameters (levels of free cysteine residues in sperm nucleoproteins, sperm viability, acrosome membrane integrity, intracellular peroxide and superoxide levels [ROS], and total and progressive motility) were evaluated before freezing and at 30 and 240 minutes after thawing. Both GSH and AA significantly improved boar sperm cryotolerance when they were separately added to freezing and thawing media. However, the highest improvement was recorded when both freezing and thawing media were supplemented with 5 mM of GSH plus 100 μM of AA. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels and had no impact on superoxide levels. According to our results, we can conclude that supplementation of freezing and thawing media with both GSH and AA has a combined, beneficial effect on frozen-thawed boar sperm, which is greater than that obtained with the separate addition of either GSH or AA.


Contextualising the link between adolescents' use of digital technology and their mental health: a multi-country study of time spent online and life satisfaction.
Evidence on whether the amount of time children spend online affects their mental health is mixed. There may be both benefits and risks. Yet, almost all published research on this topic is from high-income countries. This paper presents new findings across four countries of varying wealth. We analyse data gathered through the Global Kids Online project from nationally representative samples of Internet-using children aged 9 to 17 years in Bulgaria (n = 1,000), Chile (n = 1,000), Ghana (n = 2,060) and the Philippines (n = 1,873). Data was gathered on Internet usage on week and weekend days. Measures of absolute (comparable across countries) and relative (compared to other children within countries) time use were constructed. Mental health was measured by Cantril's ladder (life satisfaction). The analysis also considers the relative explanatory power on variations in mental health of children's relationships with family and friends. Analysis controlled for age, gender and family socioeconomic status. In Bulgaria and Chile, higher-frequency Internet use is weakly associated with lower life satisfaction. In Ghana and the Philippines, no such pattern was observed. There was no evidence that the relationship between frequency of Internet use and life satisfaction differed by gender. In all four countries, the quality of children's close relationships showed a much stronger relationship with their life satisfaction than did time spent on the Internet. Time spent on the Internet does not appear to be strongly linked to children's life satisfaction, and results from one country should not be assumed to transfer to another. Improving the quality of children's close relationships offers a more fruitful area for intervention than restricting their time online. Future research could consider a wider range of countries and links between the nature, rather than quantity, of Internet usage and mental health.


Arsenic tolerance in Arabidopsis is mediated by two ABCC-type phytochelatin transporters.
Arsenic is an extremely toxic metalloid causing serious health problems. In Southeast Asia, aquifers providing drinking and agricultural water for tens of millions of people are contaminated with arsenic. To reduce nutritional arsenic intake through the consumption of contaminated plants, identification of the mechanisms for arsenic accumulation and detoxification in plants is a prerequisite. Phytochelatins (PCs) are glutathione-derived peptides that chelate heavy metals and metalloids such as arsenic, thereby functioning as the first step in their detoxification. Plant vacuoles act as final detoxification stores for heavy metals and arsenic. The essential PC-metal(loid) transporters that sequester toxic metal(loid)s in plant vacuoles have long been sought but remain unidentified in plants. Here we show that in the absence of two ABCC-type transporters, AtABCC1 and AtABCC2, Arabidopsis thaliana is extremely sensitive to arsenic and arsenic-based herbicides. Heterologous expression of these ABCC transporters in phytochelatin-producing Saccharomyces cerevisiae enhanced arsenic tolerance and accumulation. Furthermore, membrane vesicles isolated from these yeasts exhibited a pronounced arsenite [As(III)]-PC(2) transport activity. Vacuoles isolated from atabcc1 atabcc2 double knockout plants exhibited a very low residual As(III)-PC(2) transport activity, and interestingly, less PC was produced in mutant plants when exposed to arsenic. Overexpression of AtPCS1 and AtABCC1 resulted in plants exhibiting increased arsenic tolerance. Our findings demonstrate that AtABCC1 and AtABCC2 are the long-sought and major vacuolar PC transporters. Modulation of vacuolar PC transporters in other plants may allow engineering of plants suited either for phytoremediation or reduced accumulation of arsenic in edible organs.


Impacts of pr-10a overexpression at the molecular and the phenotypic level.
Biotechnological approaches using genetic modifications such as homologous gene overexpression can be used to decode gene functions under well-defined circumstances. However, only the recording of the resulting phenotypes allows inferences about the impact of the modification on the organisms' evolutionary, ecological or economic performance. We here compare a potato wild-type cell line with two genetically engineered cell cultures homologously overexpressing Pathogenesis Related Protein 10a (pr-10a). A detailed analysis of the relative gene-expression patterns of pr-10a and its regulators sebf and pti4 over time provides insights into the molecular response of heterotrophic cells to distinct osmotic and salt-stress conditions. Furthermore, this system serves as an exemplar for the tracing of respiration kinetics as a faster and more sensitive alternative to the laborious and time-consuming recording of growth curves. The utility and characteristics of the resulting data type and the requirements for its appropriate analysis are figured out. It is demonstrated how this novel type of phenotypic information together with the gene-expression-data provides valuable insights into the effect of genetic modifications on the behaviour of cells on both the molecular and the macroscopic level.


Migration of epoxidized soybean oil (ESBO) and phthalates from twist closures into food and enforcement of the overall migration limit.
Nineteen samples of food in glass jars with twist closures were collected by the national food inspectors at Danish food producers and a few importers, focusing on fatty food, such as vegetables in oil, herring in dressing or pickle, soft spreadable cheese, cream, dressings, peanut butter, sauces and infant food. The composition of the plasticizers in the gaskets was analysed by gas chromatography with flame ionization detection (GC-FID) and gas chromatography-mass spectrometry (GC-MS). Epoxidized soybean oil (ESBO) and phthalates were determined in the homogenized food samples. ESBO was the principal plasticizer in five of the gaskets; in 14 it was phthalates. ESBO was found in seven of the food samples at concentrations from 6 to 100 mg kg(-1). The highest levels (91-100 mg kg(-1)) were in oily foods such as garlic, chilli or olives in oil. Phthalates, i.e. di-iso-decylphthalate (DIDP) and di-iso-nonylphthalates (DINP), were found in seven samples at 6-173 mg kg(-1). The highest concentrations (99-173 mg kg(-1)) were in products of garlic and tomatoes in oil and in fatty food products such as sauce béarnaise and peanut butter. For five of the samples the overall migration from unused lids to the official fatty food simulant olive oil was determined and compared with the legal limit of 60 mg kg(-1). The results ranged from 76 to 519 mg kg(-1) and as a consequence the products were withdrawn from the market.


Antigenic drift in visna: virus variation during long-term infection of Icelandic sheep.
A group of 20 Icelandic sheep were infected intracerebrally with visna virus strain 1514, and 209 virus isolates were obtained from the blood, cerebrospinal fluid, and central nervous system (CNS) over a period of 7 years, during which eight animals developed clinical signs of visna necessitating sacrifice. (i) Using type-specific antisera, it was found that 12 (16%) of 76 isolates tested escaped neutralization. These 12 variant viruses were distributed randomly among animals and over time, and did not replace the infecting strain even though all sheep developed homotypic antibody within 3 months of infection. The one exception was sheep no. 1557 (an animal without clinical visna), where the last six isolates were variants. (ii) A total of 35 blood and CNS isolates from seven of these sheep (including five with clinical visna) were tested against serial samples of their own sera. Autologous antisera neutralized all isolates tested with the exception of isolates from sheep 1557. None of the isolates obtained at sacrifice from the five sheep with clinical visna escaped neutralization with autologous antisera. These data suggest that although variant viruses are encountered at considerable frequency during long-term infection of Icelandic sheep, the variants usually do not replace the infecting strain. Antigenic drift does not appear to be essential for virus persistence or for the development of clinically evident CNS lesions.


Charging effects on bonding and catalyzed oxidation of CO on Au8 clusters on MgO.
Gold octamers (Au8) bound to oxygen-vacancy F-center defects on Mg(001) are the smallest clusters to catalyze the low-temperature oxidation of CO to CO2, whereas clusters deposited on close-to-perfect magnesia surfaces remain chemically inert. Charging of the supported clusters plays a key role in promoting their chemical activity. Infrared measurements of the stretch vibration of CO adsorbed on mass-selected gold octamers soft-landed on MgO(001) with coadsorbed O2 show a red shift on an F-center-rich surface with respect to the perfect surface. The experiments agree with quantum ab initio calculations that predict that a red shift of the C-O vibration should arise via electron back-donation to the CO antibonding orbital.


CMR in Evaluating Valvular Heart Disease: Diagnosis, Severity, and Outcomes.
Cardiac magnetic resonance (CMR) is a versatile imaging tool that brings much to the assessment of valvular heart disease. Although it is best known for myocardial imaging (even in valve disease), it provides excellent assessment of all 4 heart valves, with some distinct advantages, including a free choice of image planes and accurate flow and volumetric quantification. These allow the severity of each valve lesion to be characterized, in addition to optimal visualization of the surrounding outflow tracts and vessels, to deliver a comprehensive package. It can assess each valve lesion separately (in multiple valve disease) and is not affected by hemodynamic status. The accurate quantitation of regurgitant lesions and the ability to characterize myocardial changes also provides an ability to predict future clinical outcomes in asymptomatic patients. This review outlines how CMR can be used in cardiac valve disease to compliment echocardiography and enhance the patient assessment. It covers the main CMR methods used, their strengths and limitations, and the optimal way to apply them to evaluate valve disease.


The cis-regulatory logic of Hedgehog gradient responses: key roles for gli binding affinity, competition, and cooperativity.
Gradients of diffusible signaling proteins control precise spatial patterns of gene expression in the developing embryo. Here, we use quantitative expression measurements and thermodynamic modeling to uncover the cis-regulatory logic underlying spatially restricted gene expression in a Hedgehog (Hh) gradient in Drosophila. When Hh signaling is low, the Hh effector Gli, known as Cubitus interruptus (Ci) in Drosophila, acts as a transcriptional repressor; when Hh signaling is high, Gli acts as a transcriptional activator. Counterintuitively and in contrast to previous models of Gli-regulated gene expression, we found that low-affinity binding sites for Ci were required for proper spatial expression of the Hh target gene decapentaplegic (dpp) in regions of low Hh signal. Three low-affinity Ci sites enabled expression of dpp in response to low signal; increasing the affinity of these sites restricted dpp expression to regions of maximal signaling. A model incorporating cooperative repression by Ci correctly predicted the in vivo expression of a reporter gene controlled by a single Ci site. Our work clarifies how transcriptional activators and repressors, competing for common binding sites, can transmit positional information to the genome. It also provides an explanation for the widespread presence of conserved, nonconsensus Gli binding sites in Hh target genes.


Effect of replacing glucose with lipid on the energy metabolism of newborn infants.
1. Indirect calorimetry and primed constant infusion of [U-13C]glucose were combined in 28 appropriate-for-gestational age newborn, parenterally fed infants, in order to measure glucose utilization and glucose oxidation and to estimate lipogenesis from glucose. 2. The infants were randomly allocated to either a group receiving glucose as the non-protein energy source or a group having one-quarter of the glucose energy replaced by intravenous fat. The energy intake (370 kJ day-1 kg-1) and protein intake (3.4 g day-1 kg-1) were similar in both groups. 3. Energy expenditure (P less than 0.005), non-protein carbon dioxide production (P less than 0.005) and non-protein oxygen consumption (P less than 0.05) were lower in the lipid-supplemented group. 4. The significant excess of glucose utilization over oxidation (P less than 0.001) can be accounted for by lipid synthesis from glucose. 5. Fat synthesis from glucose was higher in the glucose/amino acid group (P less than 0.02), but total fat storage was higher in the lipid-supplemented group (P less than 0.02). Nitrogen balance was similar in both groups. 6. As lipogenesis from glucose is an energy- and oxygen-consuming and a carbon dioxide-producing process, the data suggest that the differences between the glucose-only group and the lipid-supplemented group are due to different rates of lipogenesis from glucose.


No rapid audiovisual recalibration in adults on the autism spectrum.
Autism spectrum disorders (ASD) are characterized by difficulties in social cognition, but are also associated with atypicalities in sensory and perceptual processing. Several groups have reported that autistic individuals show reduced integration of socially relevant audiovisual signals, which may contribute to the higher-order social and cognitive difficulties observed in autism. Here we use a newly devised technique to study instantaneous adaptation to audiovisual asynchrony in autism. Autistic and typical participants were presented with sequences of brief visual and auditory stimuli, varying in asynchrony over a wide range, from 512 ms auditory-lead to 512 ms auditory-lag, and judged whether they seemed to be synchronous. Typical adults showed strong adaptation effects, with trials proceeded by an auditory-lead needing more auditory-lead to seem simultaneous, and vice versa. However, autistic observers showed little or no adaptation, although their simultaneity curves were as narrow as the typical adults. This result supports recent Bayesian models that predict reduced adaptation effects in autism. As rapid audiovisual recalibration may be fundamental for the optimisation of speech comprehension, recalibration problems could render language processing more difficult in autistic individuals, hindering social communication.


Direct compression of chitosan: process and formulation factors to improve powder flow and tablet performance.
Chitosan is a polymer derived from chitin that is widely available at relatively low cost, but due to compression challenges it has limited application for the production of direct compression tablets. The aim of this study was to use certain process and formulation variables to improve manufacturing of tablets containing chitosan as bulking agent. Chitosan particle size and flow properties were determined, which included bulk density, tapped density, compressibility and moisture uptake. The effect of process variables (i.e. compression force, punch depth, percentage compaction in a novel double fill compression process) and formulation variables (i.e. type of glidant, citric acid, pectin, coating with Eudragit S®) on chitosan tablet performance (i.e. mass variation, tensile strength, dissolution) was investigated. Moisture content of the chitosan powder, particle size and the inclusion of glidants had a pronounced effect on its flow ability. Varying the percentage compaction during the first cycle of a double fill compression process produced chitosan tablets with more acceptable tensile strength and dissolution rate properties. The inclusion of citric acid and pectin into the formulation significantly decreased the dissolution rate of isoniazid from the tablets due to gel formation. Direct compression of chitosan powder into tablets can be significantly improved by the investigated process and formulation variables as well as applying a double fill compression process.


Prevalence of odontogenic lesions among the Kanpur population: an institutional study.
The frequency and incidence of odontogenic cysts and tumors are controversial and depends in the geographic location. These lesions have great clinical and histopathological diversity, and reflect a need for epidemiological profile to be considered when these lesions are studied. The purpose of this study was to determine the prevalence of odontogenic cysts and tumors over a period of 10 years and to compare with other data reported around the world. Data for the study were obtained from the archives of the Department of Oral and Maxillofacial Pathology, diagnosed as the cases of Odontogenic cysts and tumors histopathologically, reported from January 2007 to March 2016. Case records of the patients that fit the histological classification of the World Health Organization (2005) were included in the study and the variables analyzed were: Age, sex, anatomical location, and histological type. In a total of 1319 biopsies reported, 112 were odontogenic in nature. Of these 112 odontogenic lesions reported, Odontogenic Cysts accounted for 54.4% and odontogenic tumors of 45.53%. Of them 61.60% of the patients were males. Common age of occurrence was 21-50 years and mandibular posterior was the most common site of occurrence. Most common odontogenic cyst and tumor reported was dentigerous cyst and ameloblastoma respectively. This study provides epidemiological information on odontogenic cyst and tumors at an institutional level. The relative frequency of these cysts and tumors can be analyzed at a global level to understand their prevalence, incidence, biological behaviour, and distribution.


Differences in Cortical Gray Matter Atrophy of Paraplegia and Tetraplegia after Complete Spinal Cord Injury.
Anatomical studies of spinal cord injury (SCI) using magnetic resonance imaging (MRI) report diverging observations, from "no changes" to "tissue atrophy in motor and non-motor regions." These discrepancies among studies can be attributed to heterogeneity in extent, level, and post-injury duration observed within the SCI population. But, no studies have investigated structural changes associated with different levels of injury (paraplegia vs. tetraplegia). High-resolution MRI images were processed using a voxel-based morphometry technique to compare regional gray matter volume (GMV) between 16 complete paraplegia and 7 complete tetraplegia SCI subjects scanned within 2 years of injury when compared to 22 age-matched healthy controls using one-way analysis of covariance (ANCOVA). A post-hoc analysis using a region of interest-based approach was utilized to quantify GMV differences between healthy controls and subgroups of SCI. A voxel-wise one-sample t-test was also performed to evaluate the mean effect of post-injury duration on GMV of the SCI group. ANCOVA resulted in altered GMV in inferior frontal gyrus, bilateral mid orbital gyrus extending to rectal gyrus, and anterior cingulate cortex. Post-hoc analysis, in general, indicated GM atrophy after SCI, but tetraplegia showed a greater decrease in GMV when compared to paraplegia and healthy controls. Further, the GMV of the middle frontal gyrus, superior frontal gyrus, inferior frontal gyrus, insula, mid-orbital gyrus, and middle temporal gyrus was positively correlated with post-injury duration in both paraplegia and tetraplegia groups. GM atrophy after SCI is affected by level of cord injury, with higher levels of injury resulting in greater loss of GMV. Magnitude of GMV loss in the frontal cortex after SCI also appears to be dynamic within the first 2 years of injury. Understanding the effect of injury level and injury duration on structural changes after SCI can help to better understand the mechanisms leading to positive and negative clinical outcome in SCI patients.


Double-digest RAD-sequencing: do pre- and post-sequencing protocol parameters impact biological results?
Next-generation sequencing technologies have opened a new era of research in population genetics. Following these new sequencing opportunities, the use of restriction enzyme-based genotyping techniques, such as restriction site-associated DNA sequencing (RAD-seq) or double-digest RAD-sequencing (ddRAD-seq), has dramatically increased in the last decade. From DNA sampling to SNP calling, the laboratory and bioinformatic parameters of enzyme-based techniques have been investigated in the literature. However, the impact of those parameters on downstream analyses and biological results remains less documented. In this study, we investigated the effects of sevral pre- and post-sequencing settings on ddRAD-seq results for two biological systems: a complex of butterfly species (Coenonympha sp.) and several populations of common beech (Fagus sylvatica). Our results suggest that pre-sequencing parameters (i.e., DNA quantity, number of PCR cycles during library preparation) have a significant impact on the number of recovered reads and SNPs, on the number of unique alleles and on individual heterozygosity. In the same way, we found that post-sequencing settings (i.e., clustering and minimum coverage thresholds) influenced loci reconstruction (e.g., number of loci, mean coverage) and SNP calling (e.g., number of SNPs; heterozygosity) but had only a marginal impact on downstream analyses (e.g., measure of genetic differentiation, estimation of individual admixture, and demographic inferences). In addition, replication analyses confirmed the reproducibility of the ddRAD-seq procedure. Overall, this study assesses the degree of sensitivity of ddRAD-seq data to pre- and post-sequencing protocols, and illustrates its robustness when studying population genetics.


An aggressive systemic mastocytosis preceded by ovarian dysgerminoma.
Aggressive systemic mastocytosis (ASM) is a rare malignant disease characterized by disordered mast cell accumulation in various organs. We here describe a female ASM patient with a previous history of ovarian dysgerminoma. Molecular cytogenomic analyses were performed to elucidate an etiological link between the ASM and dysgerminoma of the patient. This patient was affected by ovarian dysgerminoma which was treated by chemotherapy and surgical resection. Having subsequently been in complete remission for 2 years, she developed symptoms of ASM. A somatic D816A mutation in the KIT gene was detected in her bone marrow, which facilitated the diagnosis of ASM. Unexpectedly, this KIT D816A variant was also detected in the prior ovarian dysgerminoma sample. Whole-exome sequencing allowed us to identify a somatic nonsense mutation of the TP53 gene in the bone marrow, but not in the dysgerminoma. Microarray analysis of the patient's bone marrow revealed a copy-number-neutral loss of heterozygosity at the TP53 locus, suggestive of the homozygous nonsense mutation in the TP53 gene. In addition, the loss of heterozygosity at the TP53 locus was also detected in the dysgerminoma. These results indicated that either the mast cells causing the ASM in this case had originated from the preceding ovarian dysgerminoma as a clonal evolution of a residual tumor cell, which acquired the TP53 mutation, or that both tumors developed from a common cancer stem cell carrying the KIT D816A variation.


The effect of declining exposure on T cell-mediated immunity to Plasmodium falciparum - an epidemiological "natural experiment".
Naturally acquired immunity to malaria may be lost with lack of exposure. Recent heterogeneous reductions in transmission in parts of Africa mean that large populations of previously protected people may lose their immunity while remaining at risk of infection. Using two ethnically similar long-term cohorts of children with historically similar levels of exposure to Plasmodium falciparum who now experience very different levels of exposure, we assessed the effect of decreased parasite exposure on antimalarial immunity. Peripheral blood mononuclear cells (PBMCs) from children in each cohort were stimulated with P. falciparum and their P. falciparum-specific proliferative and cytokine responses were compared. We demonstrate that, while P. falciparum-specific CD4+ T cells are maintained in the absence of exposure, the proliferative capacity of these cells is altered considerably. P. falciparum-specific CD4+ T cells isolated from children previously exposed, but now living in an area of minimal exposure ("historically exposed") proliferate significantly more upon stimulation than cells isolated from children continually exposed to the parasite. Similarly, PBMCs from historically exposed children expressed higher levels of pro-inflammatory cytokines and lower levels of anti-inflammatory cytokines after stimulation with P. falciparum. Notably, we found a significant positive association between duration since last febrile episode and P. falciparum-specific CD4+ T cell proliferation, with more recent febrile episodes associated with lower proliferation. Considered in the context of existing knowledge, these data suggest a model explaining how immunity is lost in absence of continuing exposure to P. falciparum.


Thoracoscopic lung biopsy under regional anesthesia for interstitial lung disease.
Interstitial lung disease (ILD) management guidelines support lung biopsy-guided therapy. However, the high mortality associated with thoracoscopic lung biopsy using general anesthesia (GA) in patients with ILD has deterred physicians from offering this procedure and adopt a diagnostic approach based on high-resolution CT. Here we report that thoracoscopy under regional anesthesia could be a safer alternative for lung biopsy and effectively guide ILD treatment. This was a single-center retrospective review of prospectively maintained database and consisted of patients who underwent thoracoscopic lung biopsy between March 2016 and March 2018. Patients were divided into two groups: (A) GA, and (B) regional anesthesia using monitored anesthesia care (MAC) and thoracic epidural anesthesia (TEA). During the study period, 44 patients underwent thoracoscopic lung biopsy. Of these, 15 underwent MAC/TEA. There were no significant differences between the two groups with regard to pulmonary function test and clinicodemographic profile. However, operative time and hospital stay were shorter in MAC/TEA group (32.5±18.5 min vs 50.8±18.4; p=0.004, 1.0±1.3 days vs 10.0±34.7 days; p<0.001, respectively). Eight patients in the GA group, but none in the MAC/TEA group, experienced worsening of ILD after lung biopsy (p=0.03). Additionally, one patient in the GA group died due to acute ILD worsening. No cases of MAC/TEA group had to be converted to GA. In all cases a pathological diagnosis could be made. Thoracoscopy using regional anesthesia might be a safer alternative to lung biopsy in patients with ILD.


Chromosome rearrangements shape the diversification of secondary metabolism in the cyclosporin producing fungus Tolypocladium inflatum.
Genes involved in production of secondary metabolites (SMs) in fungi are exceptionally diverse. Even strains of the same species may exhibit differences in metabolite production, a finding that has important implications for drug discovery. Unlike in other eukaryotes, genes producing SMs are often clustered and co-expressed in fungal genomes, but the genetic mechanisms involved in the creation and maintenance of these secondary metabolite biosynthetic gene clusters (SMBGCs) remains poorly understood. In order to address the role of genome architecture and chromosome scale structural variation in generating diversity of SMBGCs, we generated chromosome scale assemblies of six geographically diverse isolates of the insect pathogenic fungus Tolypocladium inflatum, producer of the multi-billion dollar lifesaving immunosuppressant drug cyclosporin, and utilized a Hi-C chromosome conformation capture approach to address the role of genome architecture and structural variation in generating intraspecific diversity in SMBGCs. Our results demonstrate that the exchange of DNA between heterologous chromosomes plays an important role in generating novelty in SMBGCs in fungi. In particular, we demonstrate movement of a polyketide synthase (PKS) and several adjacent genes by translocation to a new chromosome and genomic context, potentially generating a novel PKS cluster. We also provide evidence for inter-chromosomal recombination between nonribosomal peptide synthetases located within subtelomeres and uncover a polymorphic cluster present in only two strains that is closely related to the cluster responsible for biosynthesis of the mycotoxin aflatoxin (AF), a highly carcinogenic compound that is a major public health concern worldwide. In contrast, the cyclosporin cluster, located internally on chromosomes, was conserved across strains, suggesting selective maintenance of this important virulence factor for infection of insects. This research places the evolution of SMBGCs within the context of whole genome evolution and suggests a role for recombination between chromosomes in generating novel SMBGCs in the medicinal fungus Tolypocladium inflatum.


Bisphenol A and child and youth behaviour: Canadian Health Measures Survey 2007 to 2011.
Bisphenol A (BPA) is a synthetic industrial chemical commonly used in consumer products. Results from the Canadian Heath Measures Survey (CHMS) indicate that more than 90% of children and youth aged 6 to 19 have detectable levels of urinary BPA. Childhood concentration levels of BPA have been linked with negative behavioural outcomes. The data are from the first two cycles (2007 to 2009 and 2009 to 2011) of the CHMS, which collected biomonitoring indicators via spot blood and urine samples. Behavioural outcomes--hyperactivity/inattention, emotional symptoms, conduct problems, peer problems, and prosocial behavior--were assessed with Goodman's Strengths and Difficulties Questionnaire. Geometric mean urinary BPA concentration was examined overall and by demographic and socioeconomic correlates. Six multiple logistic regression analyses were conducted to investigate associations between childhood BPA concentrations and risk status for each outcome. Children aged 6 to 8 had higher BPA concentrations than did older children and youth. Concentrations were significantly higher among children and youth exposed to second-hand smoke every day or almost every day and those in low or lower-middle income households. Higher BPA concentrations were associated with increased odds of hyperactivity among girls and lower prosocial behaviour among boys. These findings suggest an association between urinary BPA concentration and children's behavioural outcomes.


Social learning correlates of exercise self-efficacy: early experiences with physical activity.
Self-efficacy, the degree of confidence an individual has for performing a given behavior, is a central concept in social learning theory. Self-efficacy is a strong correlate of current exercise and a consistent predictor of future exercise behavior. This study was designed to explore the correlates of exercise self-efficacy in order to determine whether early experiences with exercise and sports fosters exercise self-efficacy in adulthood. This study explored historical and contemporaneous correlates of exercise self-efficacy using a path analytic approach and a randomly selected sample of adults. Historical experiences with exercise and sports were classified as more or less organized and as proximal or distal in time. The activities studied included those reported during elementary (ages 6-14), high school (15-18), and the period after high school. The historical activities, scaled using factor analysis, were found to have no significant direct association with self-efficacy and only marginal indirect associations with self-efficacy mediated by contemporaneous social learning variables. Contemporaneous variables drawn from social learning theory were found to have the largest direct and indirect associations with self-efficacy specifically related to vigorous exercise. Directions for experimental analysis and health promotion policy implications are discussed.


Correlation of serum anti-helicobacter pylori immunoglobulin a (IgA) with histological parameters of chronic gastritis in ibadan, Nigeria.
The seroprevalence of anti-H. pylori IgA antibodies has been reported to vary among populations and in relation to strains of Helicobacter pylori bacterium. However, there has been conflicting reports on the association between IgA serological status and the histological variables of chronic gastritis. This study was therefore conducted to clarify this relationship. Using an ELISA based commercial kit, anti-H. pylori IgA antibody tests were performed on 65 dyspeptic patients and 65 age- and sex-matched controls. The gastric biopsies of these patients were also examined histologically for the degrees of inflammation, activity, intestinal metaplasia and atrophy. The CagA status of the patients had been determined previously. There was an anti-H. pylori IgA antibody prevalence of 67.7% in dyspeptics and 56.9% in non-dyspeptic individuals. No correlations were observed between serum H. pylori IgA antibody and the graded parameters of chronic gastritis in dyspeptic patients, although twice more patients with mild gastric inflammation were found among IgA positive than among IgA negative patients. However, a statistically significant relationship was established between serum IgA positivity and the CagA status of the patients (p = 0.028). The seroprevalence of anti-H. pylori IgA antibody is high in our environment. Serum IgA status may be associated with milder degrees of gastritis in our patients but a larger cohort of patients is needed to confirm this. There seems to be a good agreement between serum IgA and CagA statuses among dyspeptic patients.


Disclosing complementary and alternative medicine use in the medical encounter: a qualitative study in women with breast cancer.
Despite recent findings that patients who use complementary and alternative medicine (CAM) typically choose not to mention this to their physicians, little is known about the reasons for this lack of communication. Understanding the reasons for nondisclosure of CAM use is critical to improving physician-patient communication and patient care. We are conducting a 5-year prospective cohort study consisting of 4 interview cycles. The multiethnic, population-based sample consists of 86 San Francisco residents with recently diagnosed breast cancer (response rate = 87%). Findings are determined using qualitative analysis of transcribed interviews. At initial contact, 72% of the participants were using at least 1 CAM therapy for breast cancer. Six months later, 65% of participants were using CAM. Of the women being treated by an alternative practitioner, 54% disclosed their CAM use to their physicians. Conversely, 94% discussed details of their biomedical treatments with their alternative practitioner. Reasons for not disclosing CAM use included anticipating the physician's disinterest, negative response, or unwillingness or inability to contribute useful information; the perception that the CAM therapies used were irrelevant to the biomedical treatment course; and the patients' views regarding the appropriate coordination of disparate healing strategies. Discussions of patients' CAM use are more poorly integrated into the medical encounter than discussions of biomedical treatment are with alternative practitioners. Patients' disclosure is often cautiously modulated, even by those who would welcome an open discussion with their physicians. Specific suggestions for discussing CAM use with patients are presented.


Interview with John Kaas.
Jon Kaas received a Ph.D. in Psychology with Irving Diamond at Duke University and completed a postdoctoral fellowship with Clinton Woolsey in neurophysiology at the University of Wisconsin, where he also collaborated with Ray Guillery. After 4 years as an Assistant Professor in Neurophysiology at Wisconsin, he moved to the Department of Psychology at Vanderbilt University where he is currently Centennial Professor. Awards include the Earl Sutherland Prize, Javits Neuroscience Investigator Award, Krieg Cortical Discoverer Award, and American Psychological Association Distinguished Scientific Contribution Award.


Brain-computer communication and slow cortical potentials.
A thought translation device (TTD) has been designed to enable direct brain-computer communication using self-regulation of slow cortical potentials (SCPs). However, accuracy of SCP control reveals high intersubject variability. To guarantee the highest possible communication speed, some important aspects of training SCPs are discussed. A baseline correction of SCPs can increase performance. Multichannel recordings show that SCPs are of highest amplitude around the vertex electrode used for feedback, but in some subjects more global distributions were observed. A new method for control of eye movement is presented. Sequential effects of trial-to-trial interaction may also cause difficulties for the user. Finally, psychophysiological factors determining SCP communication are discussed.


Association between circulating hemostatic measures and dementia or cognitive impairment: systematic review and meta-analyzes.
Hemostasis and thrombosis may be important contributors to cognitive decline and dementia. Certain blood markers may assist in diagnosis or management. To collate evidence for the association of circulating hemostatic variables and dementia or cognitive impairment. A systematic review of studies describing blood markers of hemostatic function and cognition/dementia. Abstracts were reviewed by two independent assessors and studies selected based on pre-specified criteria. We described methodological quality and performed meta-analyzes where data allowed. From 7103 titles, 485 abstracts and included 21 studies (n = 32,773) were assessed. In two longitudinal studies, the incident of vascular dementia risk was greater for higher D-dimer [hazard ratio (HR): 1.50, 95% confidence interval (CI): 1.15-1.96]. For case-control data, we calculated standardized mean differences (SMD) and 95% CI. Higher levels of: factor (F)VII (SMD: 0.93; 95% CI: 0.60-1.26), fibrinogen (SMD: 1.53; 95% CI: 1.17-1.87), prothrombin fragment 1 and 2 (SMD: 0.64; 95% CI: 0.32-0.96), plasminogen activator inhibitor (SMD: 0.68; 95% CI: 0.26-1.10), D-dimer (SMD: 2.00; 95% CI: 1.59-2.40) and von Willebrand factor (VWF) (SMD: 1.68; 95% CI: 1.30-2.06) showed modest but significant associations with vascular dementia. For patients with any dementia diagnosis, associations were with higher D-dimer (SMD: 0.36; 95% CI: 0.15-0.56) and VWF (SMD: 0.31; 95% CI: 0.11-0.51). For specific cognitive domains, significant (P < 0.001) positive correlations were fibrinogen and speed of processing (0.76; 95% CI: 0.67-0.84), verbal memory (0.69; 95% CI: 0.59-0.79) and non-verbal reasoning (0.57; 95% CI: 0.49-0.65). The present results suggest a modest association between hemostasis and vascular dementia including increased levels of thrombin generation markers (D-dimer and prothrombin fragment 1 + 2) and endothelial dysfunction (VWF and plasminogen activator inhibitor). Associations are weaker for specific cognitive tests and when all dementias are combined.


Optimal Curiosity-Driven Modular Incremental Slow Feature Analysis.
Consider a self-motivated artificial agent who is exploring a complex environment. Part of the complexity is due to the raw high-dimensional sensory input streams, which the agent needs to make sense of. Such inputs can be compactly encoded through a variety of means; one of these is slow feature analysis (SFA). Slow features encode spatiotemporal regularities, which are information-rich explanatory factors (latent variables) underlying the high-dimensional input streams. In our previous work, we have shown how slow features can be learned incrementally, while the agent explores its world, and modularly, such that different sets of features are learned for different parts of the environment (since a single set of regularities does not explain everything). In what order should the agent explore the different parts of the environment? Following Schmidhuber's theory of artificial curiosity, the agent should always concentrate on the area where it can learn the easiest-to-learn set of features that it has not already learned. We formalize this learning problem and theoretically show that, using our model, called curiosity-driven modular incremental slow feature analysis, the agent on average will learn slow feature representations in order of increasing learning difficulty, under certain mild conditions. We provide experimental results to support the theoretical analysis.


Gender differences in health: results from SHARE, ELSA and HRS.
We examine gender differences in health at ages 50 years and older in 11 European countries, England and the USA. We use the Survey of Health, Ageing and Retirement (SHARE) for 11 Continental European countries; the English Longitudinal Study of Ageing (ELSA) and the Health and Retirement Study (HRS) for the USA to examine gender differences in health behaviours, functioning problems, disability, disease prevalence and self-rated health. Women in all countries are more likely than men to have disabling, non-lethal conditions including functioning problems [odds ratio (OR) indicating the effect of female is 1.57-2.43], IADL difficulties (OR 1.45-2.94), arthritis (OR 1.46-2.90) and depressive symptoms (OR 1.45-3.35). On the other hand, self-reported heart disease is more common among men (OR indicating effect of female ranges from 0.43 to 0.86). These differences are not eliminated by controlling for smoking behaviour and weight. Self-reported hypertension (OR 0.72-1.53) is generally more common among women; stroke and diabetes do not show consistent sex differences. While subjective assessment of health is poorer among women, this is not true when indicators of functioning, disability and diseases are controlled. There is remarkable consistency in direction of gender differences in health across these 13 countries. The size of the differences is affected in many cases by the similarity in behaviours of men and women.


Therapeutic response of multidrug-resistant Plasmodium falciparum and P. vivax to chloroquine and sulfadoxine-pyrimethamine in southern Papua, Indonesia.
To determine the level of antimalarial drug resistance in southern Papua, Indonesia, we assessed the therapeutic efficacy of chloroquine plus sulfadoxine-pyrimethamine (CQ+SP) for Plasmodium falciparum infections as well as CQ monotherapy for P. vivax infections. Patients with P. falciparum failing therapy were re-treated with unsupervised quinine+/-doxycycline therapy and those with P. vivax with either unsupervised quinine+/-doxycycline or amodiaquine. In total, 143 patients were enrolled in the study (103 treated with CQ+SP and 40 with CQ). Early treatment failures occurred in four patients (4%) with P. falciparum and six patients (15%) with P. vivax. The failure rate by Day 28 for P. vivax was 65% (95% CI 49-81). After PCR correction for re-infections, the Day 42 recrudescence rate for P. falciparum infections was 48% (95% CI 31-65). Re-treatment with unsupervised quinine+/-doxycycline resulted in further recurrence of malaria in 48% (95% CI 31-65) of P. falciparum infections and 70% (95% CI 37-100) of P. vivax infections. Eleven patients with recurrent P. vivax were re-treated with amodiaquine; there were no early or late treatment failures. In southern Papua, a high prevalence of drug resistance of P. falciparum and P. vivax exists both to first- and second-line therapies. Preliminary data indicate that amodiaquine retains superior efficacy compared with CQ for CQ-resistant P. vivax.


Complement activated granulocytes can cause autologous tissue destruction in man.
Activation of polymorphonuclear granulocytes (PMNs) by C5a is thought to be important in the pathogenesis of multiple organ failure during sepsis and after trauma. In our experiment exposure of human PMNs to autologous zymosan activated plasma (ZAP) leads to a rapid increase in chemiluminescence. Heating the ZAP at 56( degrees )C for 30 min did not alter the changes, while untreated plasma induced only baseline activity. The respiratory burst could be completely abolished by decomplementation and preincubation with rabbit antihuman C5a antibodies. Observation of human omentum using electron microscopy showed intravascular aggregation of PMNs, with capillary thrombosis and diapedesis of the cells through endothelial junctions 90 s after exposure to ZAP. PMNs caused disruption of connections between the mesothelial cells. After 4 min the mesothelium was completely destroyed, and connective tissue and fat cells exposed. Native plasma and minimum essential medium did not induce any morphological changes. These data support the concept that C5a activated PMNs can cause endothelial and mesothelial damage in man. Even though a causal relationship between anaphylatoxins and organ failure cannot be proved by these experiments C5a seems to be an important mediator in the pathogenesis of changes induced by severe sepsis and trauma in man.


Challenges of assessing the clinical efficacy of asexual blood-stage Plasmodium falciparum malaria vaccines.
In the absence of any highly effective vaccine candidate against Plasmodium falciparum malaria, it remains imperative for the field to pursue all avenues that may lead to the successful development of such a formulation. The development of a subunit vaccine targeting the asexual blood-stage of Plasmodium falciparum malaria infection has proven particularly challenging with only limited success to date in clinical trials. However, only a fraction of potential blood-stage vaccine antigens have been evaluated as targets, and a number of new promising candidate antigen formulations and delivery platforms are approaching clinical development. It is therefore essential that reliable and sensitive methods of detecting, or ruling out, even modest efficacy of blood-stage vaccines in small clinical trials be established. In this article we evaluate the challenges facing blood-stage vaccine developers, assess the appropriateness and limitations of various in vivo approaches for efficacy assessment and suggest future directions for the field.


Evaluation of Cytotoxicity of Silorane and Methacrylate based Dental Composites using Human Gingival Fibroblasts.
The effects of leached substances from the restorative dental materials may induce local and systemic adverse effects. Thus the biological and toxic properties of the restorative dental materials must be compatible with the oral tissues or with general health. Therefore, the need for biocompatible restorative dental material implies the necessity of toxicity testing. It was the purpose of this investigation to determine and compare the possible toxic effect of silorane based composite (Filtek P90) on human gingival fibroblast (HGF) in vitro using cytotoxicity measuring parameters (MTT assay) in comparison with its methacrylate counterpart (Z100) for their viability, proliferation rate. Fresh healthy biopsy specimens of human gingival tissue of patients were obtained. For HGF, cells were cultured in Dulbecco's modified Eagle medium and grown to sub confluent monolayers. After attaining confluence, cells were treated with different doses of the Filtek P90 or Z 100 for different time point. HGF cells were observed for their proliferation, viability by MTT assay. The results of the cytotoxicity assay showed that, the percentage of viable cells was very good in the first 24h and marginally decreased in the next 48h period in all groups. However, the proliferation rate was never below 84% in all the groups, at any given concentration. Filtek P90 and Z100 treated cells exhibited insignificant decrease in the cell proliferation both in 24h and 48h exposure when compared to significant decrease in the cell survival rate in the positive control (Mitomycin C 250 μg/ml).) Comparison of the toxicity between Filtek P90 and Z100 in 24h & 48h separately showed that there was no significant difference (p<0.05) between these two composites in 24h and 48h' time period at all concentrations of the composites. To conclude, the new silorane based restorative composite showed comparable cytotoxic characteristics to clinically successful dimethacrylate composites suggesting the non-toxic nature in the oral environment and hence contributing to clinical success of these new restorative materials.


eHUGS: Enhanced Hierarchical Unbiased Graph Shrinkage for Efficient Groupwise Registration.
Effective and efficient spatial normalization of a large population of brain images is critical for many clinical and research studies, but it is technically very challenging. A commonly used approach is to choose a certain image as the template and then align all other images in the population to this template by applying pairwise registration. To avoid the potential bias induced by the inappropriate template selection, groupwise registration methods have been proposed to simultaneously register all images to a latent common space. However, current groupwise registration methods do not make full use of image distribution information for more accurate registration. In this paper, we present a novel groupwise registration method that harnesses the image distribution information by capturing the image distribution manifold using a hierarchical graph with its nodes representing the individual images. More specifically, a low-level graph describes the image distribution in each subgroup, and a high-level graph encodes the relationship between representative images of subgroups. Given the graph representation, we can register all images to the common space by dynamically shrinking the graph on the image manifold. The topology of the entire image distribution is always maintained during graph shrinkage. Evaluations on two datasets, one for 80 elderly individuals and one for 285 infants, indicate that our method can yield promising results.


Looking into the shadow: the eugenics argument in debates on reproductive technologies and practices.
Eugenics is often referred to in debates on the ethics of reproductive technologies and practices, in relation to the creation of moral boundaries between acceptable and unacceptable technologies, and acceptable and unacceptable uses of these technologies. Historians have argued that twentieth century eugenics cannot be reduced to a uniform set of practices, and that no simple lessons can be drawn from this complex history. Some authors stress the similarities between past eugenics and present reproductive technologies and practices (what I define throughout the paper as 'the continuity view') in order to condemn the latter. Others focus on the differences between past and present practices (what I define throughout the paper as 'the discontinuity view') in order to defend contemporary reproductive technologies. In this paper, I explore the meanings of the word 'eugenics' and the relationship between its past and present uses in terms of contemporary debates on reproductive technologies and practices. I argue that moral disagreement about present technologies originate in divergent views of condemnable and justifiable features of the past.


Validation of parent-reported physical activity and sedentary time by accelerometry in young children.
It is unknown if young children's parent-reported physical activity and sedentary time are correlated with direct measures. The study objectives were to compare parent-reported physical and sedentary activity versus directly measured accelerometer data in early childhood. From 2013 to 2014, 117 healthy children less than 6 years of age were recruited to wear Actical accelerometers for 7 days. Accelerometer data and questionnaires were available on 87 children (74%). Average daily physical activity was defined as the sum of activity ≥100 counts per minute, and sedentary time as the sum of activity <100 counts per minute during waking hours. Parents reported daily physical activity (unstructured free play in and out of school, and organized activities) and selected sedentary behaviors (screen time, stroller time, time in motor vehicle). Spearman correlation coefficients and Bland-Altman plots were used to assess the validity of parent-reported measures compared to accelerometer data. Total physical activity was significantly greater when measured by accelerometer than parent-report; the median difference was 131 min/day (p < 0.001). Parent-reported child physical activity was weak to moderately correlated with directly measured total physical activity (r = 0.39, 95% CI 0.19, 0.56). The correlations between types of physical activity (unstructured free play in and outside of school/daycare, and organized structured activity) and accelerometer were r = 0.30 (95% CI 0.09, 0.49); r = 0.42 (95% CI 0.23, 0.58); r = 0.26 (95% CI 0.05, 0.46), respectively. There was no correlation between parent-reported and accelerometer-measured total sedentary time in children (r = 0.10, 95% CI -0.12, 0.33). When the results were stratified by age group (<18, 18-47, and 48-70 months of age) no statistically significant correlations were observed and some inverse associations were observed. The correlation between parent-report of young children's physical activity and accelerometer-measured activity was weak to moderate depending on type of activity and age group. Parent-report of children's sedentary time was not correlated with accelerometer-measured sedentary time. Additional validation studies are needed to determine if parent-reported measures of physical activity and sedentary time are valid among children less than 6 years of age and across these young age groups.


Effect of 1,25(OH)2D3 on bone morphogenetic protein-3 mRNA expression.
Bone morphogenetic proteins (BMPs) are members to the transforming growth factor-beta superfamily. They induce ectopic bone formation in rat and are pleiotropic initiators of inducible osteogenic precursor cells. A lot of reports have studied the presence of BMPs and their effects on bone marker expression in many different cell lines, however none describe the regulation of BMP3 by different factors and expression conditions. When a human bone marrow stromal cell (HBMSC) culture was treated simultaneously with 1,25(OH)2D3 (10(-8) M) and BMP3 (2.5 ng/ml), the total osteocalcin content in the cell layer and in the culture medium was higher than when the culture was treated with either factor alone (162%). To elucidate this synergistic activity, Northern blot analysis was done to study the effect of 1,25(OH)2D3 on BMP3 mRNA expression. Several human cell lines (MNNG, U-2OS, MG-63, KHOS, TE85, HOS) and HBMSC were treated by 1,25(OH)2D3 (10(-8) M for 24 h). Purified mRNA from treated and untreated cells were denatured using glyoxal and dimethylsulfoxide, and were fractionated on a 1% agarose gel. After electrophoresis, RNA were blotted onto a nylon membrane and incubated with 32P-labeled BMP3 and GAPDH riboprobes. Northern blot analysis revealed that, the BMP3 mRNA level was increased in a few cell lines (MG-63, HBMSC, HOS) after the addition of 1,25(OH)2D3 when compared to the untreated cells (127%+/-1; 130.5%+/-19.5; 207%+/-14). An higher stimulation was observed in HBMSC primary culture when compared to differentiated HBMSC. In view of these results, we now investigate the following hypothesis: does the BMP3 promoter exhibit the vitamin D receptor response like the osteocalcin gene?


Ursolic Acid Induces Apoptosis of Prostate Cancer Cells via the PI3K/Akt/mTOR Pathway.
Ursolic acid (UA), a pentacyclic triterpenoid, is known to exert antitumor activity in breast, lung, liver and colon cancers. Nonetheless, the underlying mechanism of ursolic acid in prostate cancer cells still remains unclear. In the present study, we report the chemotherapeutic effects of ursolic acid as assessed using in vitro and in vivo models. Treatment of human prostate cancer cells (LNCaP and PC-3) with UA inhibited the proliferation and induced apoptosis in both cell lines as characterized by the increased Annexin V-binding. The induction of apoptosis by UA was associated with a decrease in the levels of Bcl-2, Bcl-xl, survivin, and activated caspase-3. Treatment with UA also inhibited the expression of phosphatidylinositol-3-kinase (PI3K), phosphorylation of Akt and mTOR signaling proteins. Further, administration of UA significantly inhibited the growth of LNCaP prostate tumor xenografts in athymic nude mice, which was associated with inhibition of cell proliferation, induction of apoptosis of tumor cells and decreased expression of PI3K downstream factors, such as p-Akt and p-mTOR in tumor xenograft tissues. Our study demonstrates that UA not only inhibits cell growth but also induces apoptosis through modulation of the PI3K/Akt/mTOR pathway in human prostate cancer cells. We suggest that UA may be a new chemotherapeutic candidate against prostate cancer.


GDF-5 can act as a context-dependent BMP-2 antagonist.
Bone morphogenetic protein (BMP)-2 and growth and differentiation factor (GDF)-5 are two related transforming growth factor (TGF)-β family members with important functions in embryonic development and tissue homeostasis. BMP-2 is best known for its osteoinductive properties whereas GDF-5-as evident from its alternative name, cartilage derived morphogenetic protein 1-plays an important role in the formation of cartilage. In spite of these differences both factors signal by binding to the same subset of BMP receptors, raising the question how these different functionalities are generated. The largest difference in receptor binding is observed in the interaction with the type I receptor BMPR-IA. GDF-5, in contrast to BMP-2, shows preferential binding to the isoform BMPR-IB, which is abrogated by a single amino acid (A57R) substitution. The resulting variant, GDF-5 R57A, represents a "BMP-2 mimic" with respect to BMP receptor binding. In this study we thus wanted to analyze whether the two growth factors can induce distinct signals via an identically composed receptor. Unexpectedly and dependent on the cellular context, GDF-5 R57A showed clear differences in its activity compared to BMP-2. In ATDC-5 cells, both ligands induced alkaline phosphatase (ALP) expression with similar potency. But in C2C12 cells, the BMP-2 mimic GDF-5 R57A (and also wild-type GDF-5) clearly antagonized BMP-2-mediated ALP expression, despite signaling in both cell lines occurring solely via BMPR-IA. The BMP-2- antagonizing properties of GDF-5 and GDF-5 R57A could also be observed in vivo when implanting BMP-2 and either one of the two GDF-5 ligands simultaneously at heterotopic sites. Although comparison of the crystal structures of the GDF-5 R57A:BMPR-IAEC- and BMP-2:BMPR-IAEC complex revealed small ligand-specific differences, these cannot account for the different signaling characteristics because the complexes seem identical in both differently reacting cell lines. We thus predict an additional component, most likely a not yet identified GDF-5-specific co-receptor, which alters the output of the signaling complexes. Hence the presence or absence of this component then switches GDF-5's signaling capabilities to act either similar to BMP-2 or as a BMP-2 antagonist. These findings might shed new light on the role of GDF-5, e.g., in cartilage maintenance and/or limb development in that it might act as an inhibitor of signaling events initiated by other BMPs.


Nephrotoxic effects of common and emerging drugs of abuse.
The kidneys can be injured in diverse ways by many drugs, both legal and illegal. Novel associations and descriptions of nephrotoxic effects of common and emerging drugs of abuse have appeared over the past several years. Anabolic androgenic steroids, illicitly used by athletes and others for decades to increase muscle mass and decrease body fat, are emerging as podocyte toxins given recent descriptions of severe forms of FSGS in long-term abusers. Synthetic cannabinoids, a new group of compounds with marijuana-like effects, recently became popular as recreational drugs and have been associated with an atypical form of AKI. 3,4-Methylenedioxymethamphetamine, commonly known as ecstasy, is a widely used synthetic recreational drug with mood-enhancing properties and a constellation of toxicities that can result in death. These toxic effects include hyperthermia, hypotonic hyponatremia due to its arginine vasopressin secretagogue-like effects, rhabdomyolysis, and cardiovascular collapse. Cocaine, a serotonin-norepinephrine-dopamine reuptake inhibitor that serves as an illegal stimulant, appetite suppressant, and anesthetic, also causes vasoconstriction and rhabdomyolysis. Recent adulteration of much of the world's supply of cocaine with levamisole, an antihelminthic agent with attributes similar to but distinct from those of cocaine, appears to have spawned a new type of ANCA-associated systemic vasculitis. This review discusses the nephrotoxic effects of these common and emerging drugs of abuse, of which both community and health care providers should become aware given their widespread abuse. Future investigation into pathogenetic mechanisms associated with these drugs is critical and may provide a window into ways to lessen and even prevent the nephrotoxic effects of these drugs of abuse and perhaps allow a deeper understanding of the nephrotoxicities themselves.


Implications of lemuriform extinctions for the Malagasy flora.
Madagascar's lemurs display a diverse array of feeding strategies with complex relationships to seed dispersal mechanisms in Malagasy plants. Although these relationships have been explored previously on a case-by-case basis, we present here the first comprehensive analysis of lemuriform feeding, to our knowledge, and its hypothesized effects on seed dispersal and the long-term survival of Malagasy plant lineages. We used a molecular phylogenetic framework to examine the mode and tempo of diet evolution, and to quantify the associated morphological space occupied by Madagascar's lemurs, both extinct and extant. Using statistical models and morphometric analyses, we demonstrate that the extinction of large-bodied lemurs resulted in a significant reduction in functional morphological space associated with seed dispersal ability. These reductions carry potentially far-reaching consequences for Malagasy ecosystems, and we highlight large-seeded Malagasy plants that appear to be without extant animal dispersers. We also identify living lemurs that are endangered yet occupy unique and essential dispersal niches defined by our morphometric analyses.


A deep-learning pipeline for the diagnosis and discrimination of viral, non-viral and COVID-19 pneumonia from chest X-ray images.
Common lung diseases are first diagnosed using chest X-rays. Here, we show that a fully automated deep-learning pipeline for the standardization of chest X-ray images, for the visualization of lesions and for disease diagnosis can identify viral pneumonia caused by coronavirus disease 2019 (COVID-19) and assess its severity, and can also discriminate between viral pneumonia caused by COVID-19 and other types of pneumonia. The deep-learning system was developed using a heterogeneous multicentre dataset of 145,202 images, and tested retrospectively and prospectively with thousands of additional images across four patient cohorts and multiple countries. The system generalized across settings, discriminating between viral pneumonia, other types of pneumonia and the absence of disease with areas under the receiver operating characteristic curve (AUCs) of 0.94-0.98; between severe and non-severe COVID-19 with an AUC of 0.87; and between COVID-19 pneumonia and other viral or non-viral pneumonia with AUCs of 0.87-0.97. In an independent set of 440 chest X-rays, the system performed comparably to senior radiologists and improved the performance of junior radiologists. Automated deep-learning systems for the assessment of pneumonia could facilitate early intervention and provide support for clinical decision-making.


The physical and practical problems experienced by cancer survivors: a rapid review and synthesis of the literature.
A rapid and comprehensive review to identify what is known and not known about the physical and practical problems faced by adult cancer survivors. A systematic literature review process was used. This focused on published reviews to enable a fast but rigorous identification of both the gaps and well-researched areas within survivorship. The search identified 5121 reviews, of which 42 were screened and 9 met the quality and inclusion criteria. The majority of papers focused on physical well being (n = 6) with the remaining papers focusing on practical well being (employment and finance). The quality of the reviews varied (ranging from weak to good). Gaps identified include sexual function, lower-limb lymphoedema, peripheral neuropathy, bladder and GI problems, hormonal sequelae, older cancer survivors, work impact of cancer and context-specific unmet supportive care needs. The review found a lack of standardised nomenclature for survivorship and methodological limitations. Four main gaps in knowledge relating to the practical and physical problems associated with cancer survivorship have been identified. These are key symptoms, unmet supportive care needs, employment and older cancer survivors, and should be addressed by future research and systematic literature reviews. Work is also needed to address the nomenclature of survivorship and to improve the methodology of research into cancer survivors (including standardised measures, theoretical frameworks, longitudinal design, inclusion of older survivors and age-matched controls for comparison). The review highlighted the need for better research within the identified areas in order to improve the experiences of cancer survivors.