Early lineage specification of long-lived germline precursors in the colonial ascidian Botryllus schlosseri.
In many taxa, germline precursors segregate from somatic lineages during embryonic development and are irreversibly committed to gametogenesis. However, in animals that can propagate asexually, germline precursors can originate in adults. Botryllus schlosseri is a colonial ascidian that grows by asexual reproduction, and on a weekly basis regenerates all somatic and germline tissues. Embryonic development in solitary ascidians is the classic example of determinative specification, and we are interested in both the origins and the persistence of stem cells responsible for asexual development in colonial ascidians. In this study, we characterized vasa as a putative marker of germline precursors. We found that maternally deposited vasa mRNA segregates early in development to a posterior lineage of cells, suggesting that germline formation is determinative in colonial ascidians. In adults, vasa expression was observed in the gonads, as well as in a population of mobile cells scattered throughout the open circulatory system, consistent with previous transplantation/reconstitution results. vasa expression was dynamic during asexual development in both fertile and infertile adults, and was also enriched in a population of stem cells. Germline precursors in juveniles could contribute to gamete formation immediately upon transplantation into fertile adults, thus vasa expression is correlated with the potential for gamete formation, which suggests that it is a marker for embryonically specified, long-lived germline progenitors. Transient vasa knockdown did not have obvious effects on germline or somatic development in adult colonies, although it did result in a profound heterochrony, suggesting that vasa might play a homeostatic role in asexual development.


Training health professionals in smoking cessation.
Cigarette smoking is one of the leading causes of preventable death world wide. There is good evidence that brief interventions from health professionals can increase smoking cessation attempts. A number of trials have examined whether skills training for health professionals can lead them to have greater success in helping their patients who smoke. To determine the effectiveness of training health care professionals in the delivery of smoking cessation interventions to their patients, and to assess the additional effects of training characteristics such as intervention content, delivery method and intensity. The Cochrane Tobacco Addiction Group's Specialised Register, electronic databases and the bibliographies of identified studies were searched and raw data was requested from study authors where needed. Searches were updated in March 2012. Randomized trials in which the intervention was training of health care professionals in smoking cessation. Trials were considered if they reported outcomes for patient smoking at least six months after the intervention. Process outcomes needed to be reported, however trials that reported effects only on process outcomes and not smoking behaviour were excluded. Information relating to the characteristics of each included study for interventions, participants, outcomes and methods were extracted by two independent reviewers. Studies were combined in a meta-analysis where possible and reported in narrative synthesis in text and table. Of seventeen included studies, thirteen found no evidence of an effect for continuous smoking abstinence following the intervention. Meta-analysis of 14 studies for point prevalence of smoking produced a statistically and clinically significant effect in favour of the intervention (OR 1.36, 95% CI 1.20 to 1.55, p= 0.004). Meta-analysis of eight studies that reported continuous abstinence was also statistically significant (OR 1.60, 95% CI 1.26 to 2.03, p= 0.03).Healthcare professionals who had received training were more likely to perform tasks of smoking cessation than untrained controls, including: asking patients to set a quit date (p< 0.0001), make follow-up appointments (p< 0.00001), counselling of smokers (p< 0.00001), provision of self-help material (p< 0.0001) and prescription of a quit date (p< 0.00001). No evidence of an effect was observed for the provision of nicotine gum/replacement therapy. Training health professionals to provide smoking cessation interventions had a measurable effect on the point prevalence of smoking, continuous abstinence and professional performance. The one exception was the provision of nicotine gum or replacement therapy, which did not differ between groups.


Are Skyline Plot-Based Demographic Estimates Overly Dependent on Smoothing Prior Assumptions?
In Bayesian phylogenetics, the coalescent process provides an informative framework for inferring changes in the effective size of a population from a phylogeny (or tree) of sequences sampled from that population. Popular coalescent inference approaches such as the Bayesian Skyline Plot, Skyride, and Skygrid all model these population size changes with a discontinuous, piecewise-constant function but then apply a smoothing prior to ensure that their posterior population size estimates transition gradually with time. These prior distributions implicitly encode extra population size information that is not available from the observed coalescent data or tree. Here, we present a novel statistic, $\Omega$, to quantify and disaggregate the relative contributions of the coalescent data and prior assumptions to the resulting posterior estimate precision. Our statistic also measures the additional mutual information introduced by such priors. Using $\Omega$ we show that, because it is surprisingly easy to overparametrize piecewise-constant population models, common smoothing priors can lead to overconfident and potentially misleading inference, even under robust experimental designs. We propose $\Omega$ as a useful tool for detecting when effective population size estimates are overly reliant on prior assumptions and for improving quantification of the uncertainty in those estimates.[Coalescent processes; effective population size; information theory; phylodynamics; prior assumptions; skyline plots.].


Serum HIV-1 RNA levels and time to development of AIDS in the Multicenter Hemophilia Cohort Study.
To determine if the long-term incidence of the acquired immunodeficiency syndrome (AIDS) is related to human immunodeficiency virus type 1 (HIV-1) RNA levels measured early in HIV-1 infection. Epidemiologic cohort study. Five hemophilia treatment centers in the United States. A total of 165 subjects with hemophilia and HIV-1 infection (age at HIV-1 seroconversion, 1-66 years) followed from 1979 to 1995. The HIV-1 RNA level was measured by polymerase chain reaction over a range of 200 to 1 million or more HIV-1 RNA copies/mL in archived serum specimens collected 12 to 36 months (median, 27 months) after the estimated date of HIV-1 seroconversion. Kaplan-Meier methods were used to examine the risk of AIDS and proportional hazards models were used to estimate relative hazards. The HIV-1 RNA values were similar in subjects younger than 17 years at seroconversion (median, 5214 copies/mL) and those 18 to 34 years old (median, 4693 copies/mL), but higher in those 35 years or older (median, 12069 copies/mL) (P = .02 compared with each younger group). At 10 years after seroconversion, the proportions of subjects with AIDS were 72% among subjects with 100,000 or more HIV-1 RNA copies/mL measured 12 to 36 months after HIV-1 seroconversion (n = 9), 52% among subjects with 10,000 to 99,999 copies/mL (n = 55), 22% among subjects with 1000 to 9,999 copies/mL (n = 82), and 0% among subjects with fewer than 1000 copies/mL (n = 19) (P < .001). The age-adjusted relative hazard for AIDS for subjects with 10,000 or more copies/mL was 14.3 (95% confidence interval, 1.9-105.6) compared with subjects with fewer than 1000 copies/mL. The HIV-1 RNA level during early chronic HIV-1 infection is a strong, age-independent predictor of clinical outcome; low levels define persons with a high probability of long-term AIDS-free survival.


Impact of representative payees on substance use by homeless persons with serious mental illness.
Assignment of representative payees, third parties responsible for managing clients' funds, has been proposed to counter potential use of public support payments for abused substances by people with severe mental illness and substance use disorders. This study examines substance use outcomes in a sample of homeless persons with serious mental illness and substance use disorders, some of whom were assigned representative payees. The subjects were participating in the Access to Community Care and Effective Services and Supports (ACCESS) program, a federally funded demonstration program on integrating service systems. Clients were assessed at baseline and three months after case management services were initiated. Factorial repeated-measures analysis of covariance was used to examine substance use among four client subgroups, two of which had payees and two of which did not. Clients in this sample (N = 1,348) showed significant improvement on all measures of substance use over the first three months in the program. Those with payees showed no greater improvement in substance abuse than those without payees, although they did have fewer days of homelessness. This study failed to find evidence that merely adding external money management services to existing services improves substance abuse outcomes among clients who had dual diagnoses and were homeless. Besides assigning a payee, structured behavioral interventions may be needed to produce additional clinical benefits.


Usefulness of psychosocial theory variables in explaining fat-related dietary behavior in Chinese Americans: association with degree of acculturation.
To determine the usefulness of variables from psychosocial models of health behavior in explaining fat-related dietary behavior among a sample of Chinese Americans. A survey questionnaire was administered to a convenience sample of Chinese Americans and analyzed for descriptive statistics and relationships among variables. Participants were 600 healthy individuals, ranging from 25 to 70 years of age, living in New York City. Demographic factors, degree of acculturation, food preferences, and 13 social psychological scales derived from the Theory of Planned Behavior, the Healthy Belief Model, and Social Cognitive Theory. Dependent measures assessed were intention to reduce dietary fat and behaviors related to the selection of reduced-fat diets. Descriptive statistics, Pearsons' correlation coefficients, t-tests, one-way analyses of variance, and multiple regression analyses were used. Attitude, overall health concern, and self-efficacy accounted for 58% of the variance in behavioral intention for the entire sample. Attitude, perceived barriers, and self-efficacy accounted for 19% of the variance in the prediction of dietary fat reduction behaviors. In general, a gradient was seen in the increased predictiveness of each regression model by degree of acculturation of the immigrants to American culture: predictiveness (R2) for behavior ranged from 15% for the least to 34% for the most acculturated. Acculturation was significantly related to declines in the influence of habit and of social norms. These effects were not seen by length of residency. Nutrition educators should assess the degree of acculturation of groups with whom they work and recognize that the degree of acculturation impacts the relative importance of various psychosocial variables in fat reduction behaviors.


Treatment of hyponatremia.
Hyponatremia is an electrolyte disorder that is defined by a serum sodium concentration of less than 136 mmol/L. Hyponatremia occurs at a high incidence. It is commonly associated with mild to moderate mental impairment. Hypoosmolar hyponatremia occurs in the setting of plasma volume deficiency ("hypovolemia", e. g. after gastrointestinal fluid loss), liver cirrhosis and cardiac failure ("hypervolemic" hyponatremia) and syndrome of inappropriate antidiuretic hormone secretion ("euvolemic" hyponatremia). Excessive antidiuretic hormone and continued fluid intake are the pathogenetic causes of these hyponatremias. Whereas hypovolemic hyponatremia is best corrected by isotonic saline, conventional proposals for euvolemic and hypervolemic hyponatremia consist of the following: fluid restriction, lithium carbonate, demeclocycline, urea and loop diuretic. None of these nonspecific treatments is entirely satisfactory. Recently a new class of pharmacological agents -orally available vasopressin antagonists, collectively called vaptans- have been described. A number of clinical trials using vaptans have been performed already. They showed vaptans to be effective, specific and safe in the treatment of euvolemic and hypervolemic hyponatremia.


Virus-associated organosulfur metabolism in human and environmental systems.
Viruses influence the fate of nutrients and human health by killing microorganisms and altering metabolic processes. Organosulfur metabolism and biologically derived hydrogen sulfide play dynamic roles in manifestation of diseases, infrastructure degradation, and essential biological processes. Although microbial organosulfur metabolism is well studied, the role of viruses in organosulfur metabolism is unknown. Here, we report the discovery of 39 gene families involved in organosulfur metabolism encoded by 3,749 viruses from diverse ecosystems, including human microbiomes. The viruses infect organisms from all three domains of life. Six gene families encode for enzymes that degrade organosulfur compounds into sulfide, whereas others manipulate organosulfur compounds and may influence sulfide production. We show that viral metabolic genes encode key enzymatic domains, are translated into protein, and are maintained after recombination, and sulfide provides a fitness advantage to viruses. Our results reveal viruses as drivers of organosulfur metabolism with important implications for human and environmental health.


ON THE PIGMENT OF THE NEGRO'S SKIN AND HAIR.
The pigmentary granules of the negro's skin and hair can be freed in several ways from the cells in which they are lodged and collected in any desired amount. As thus obtained, these granules are found to be insoluble in dilute alkalies, dilute hydrochloric acid (hot or cold), alcohol, or other organic solvents when applied in the order named. If, after they have been subjected to the action of dilute hydrochloric acid, they are again treated with dilute alkalies, they are found to give up their pigment, and, on the continued application of heat, the granules dissolve entirely in the alkaline solution, leaving only an insignificant residue. The pigmentary granules are composed of a colourless ground substance or substratum, a pigment, and much inorganic matter. Their inorganic constituents, as thus far determined, are calcium, magnesium, iron, and silicic, phosphoric, and sulphuric acids; and these constituents possibly play an important part in the deposition and fixation of the colouring matter in the granules. The pigment isolated from the granules, and sufficiently freed from adherent inorganic matter, contains only the merest trace of iron-so little, in fact, that we must think of it when entirely pure as free of iron. Heating the isolated pigment with barium hydrate at a temperature of 260 degrees C. entirely frees it from the closely adherent ground substance, and it is then found that the vapours of pyrrol are no longer emitted when it is subjected to dry distillation, and the odour of burnt feathers is no longer discerned, although nitrogen is still present. We can not conclude as the result of our work that the pigment is a derivative of haemoglobin; it seems to us more probable that it is ultimately derived from the proteids of the parenchymatous juices. The total quantity of soluble pigment in the skin of a negro of average size is found to weigh about 1 gramme; the weight of the pigmentary granules is about 3.3 grammes, if we are right in our assumption that they contain sixty-five per cent of water and five per cent of mineral constituents in their natural state in the epidermis. The pigments of the epidermis and hair of the negro are very likely identical. In the present state of our knowledge we can only say that it seems highly probable that the pigment of the negro's hair is not different from the dark pigment found in the hair of the white races, and we may infer that the pigment of the black skin differs only in amount and not in kind from that deposited in the skin of the white man.


Evaluation of an integrated anaerobic/aerobic SBR system for the treatment of wool dyeing effluents. Purification of wool dyeing effluent in a SBR.
This work examined the performance of a sequencing batch reactor treating dyeing effluents from a factory that processes mainly wool and wool/polyester blends. Different operational conditions were evaluated, namely the influence of the anaerobic and the subsequent aerobic phase on the organic load removal, as well as the effect of the length of the aeration period (from 8 to 12 h) on process efficiency. A comparison between a fill stage in fast and slow modes was carried out. Results indicate that the cycle 2 conditions (fast fill and 12 h aeration time) improved the overall efficiency (85+/-6% soluble COD and 95+/-4% BOD5 removal yields) with a predominant COD uptake occurring in the aerobic stage. Slow, linear COD removal was observed in the anaerobic phase, in contrast with an exponential COD decrease in the oxic phase. For SS a level under 100 mg/l was general achieved in the exit of the reactor. With respect to dye degradation, a noticeable decrease of the absorbance measured in the UV-visible range was observed. This could be explained by the reduction of the azo bonds of some of the present dyes in the anaerobic step, in which ORP values lower than -400 mV were attained. Some oxidation of anthraquinone sulphonate dyes and of the aromatic amines resulting from azo bond cleavage could also have been taken place, as well as bioelimination mechanisms such as dye sorption.


Effect of caloric restriction on the expression of heat shock protein 70 and the activation of heat shock transcription factor 1.
The regulation of heat shock protein 70 (hsp70) expression is an excellent example of a cellular mechanism that has evolved to protect all living organisms from various types of physiological stresses; therefore, the reported age-related alterations in the ability of cells to express hsp70 in response to stress could seriously compromise the ability of a senescent organism in respond to changes in its environment. Because caloric restriction (CR) is the only experimental manipulation known to retard aging and increase the survival of rodents, it was of interest to analyze the effect of CR on the age-related alteration in the induction of hsp70 expression in rat hepatocytes. The effect of CR on the nuclear transcription of hsp70 gene in rat hepatocytes in response to various levels of heat shock was determined, and it was found that the age-related decline in the transcription of hsp70 at all temperatures studied was reversed by CR. Because the heat shock transcription factor (HSF) mediates the heat-induced transcription of hsp70, the effect of CR on the induction of HSF binding activity by heat shock was studied and found to arise from HSF1, which has been shown to be involved in the induction of HSF binding activity in other cell types. The age-related decrease in the induction of HSF1 binding activity in rat hepatocytes was reversed by CR, and did not appear to be due to an accumulation of inhibitory molecules with age. Interestingly, the level of HSF1 protein was significantly higher in hepatocytes isolated from old rats fed ad libitum compared to hepatocytes obtained from rats fed the CR diet even though the levels of HSF1 binding activity were lower for hepatocytes isolated from the old rats fed ad libitum. The levels of the mRNA transcript for HSF1 was not significantly altered by age or CR. Thus, the changes in HSF1 binding activity with age and CR do not arise from changes in the level of HSF1 protein available for activation.


Strengthening System and Implementation Research Capacity for Child Mental Health and Family Well-being in Sub-Saharan Africa.
Children in Sub-Saharan Africa (SSA) experience high rates of mental health problems, and the region has limited access to mental health resources and research capacity to address the needs. Despite the success of numerous evidence-based interventions (EBIs) and emerging methodology from the field of implementation science for addressing child mental health needs, most EBIs and implementation science methodology have not been applied in SSA contexts. The SMART-Africa Center aims to address these child welfare, mental health, services, and EBI implementation research gaps by establishing a regional trans-disciplinary collaborative center and studying strategies to strengthening mental health system and implementation research capacity. Our paper describes the overall framework and strategies that SMART-Africa team developed to strengthen capacity in three SSA countries (Ghana, Kenya, and Uganda) while focusing on its contextualization for the Kenyan school-community mental health settings. Methods to document the progress and impacts are also described. The design of the system and research strengthening activities is guided by a SMART-Africa Capacity Building framework. Two areas of capacity are focused. Mental health system capacity focuses on building political wills, leadership, transdisciplinary partnership, and stakeholders' global competency in evidence child mental health policy, intervention, and service implementation research. Implementation research capacity building focuses on building researchers' implementation research competency by carrying out an EBI implementation research (using a Hybrid Type II effectiveness-implementation). For illustration purpose, we describe how the system strengthening strategies has been applied in Kenya, and how the mixed methods design applied to assess the value and impacts of the capacity building activities. Feedback data and evaluation data collection using qualitative and quantitative methods for both areas of capacity building are still ongoing. Data will be analyzed and compared across countries in 2020-2021. Our work has shown some feasibility of applying the theory-guided system strengthening model in improving child mental health service system and research capacity in one of the three SMART-Africa partnering countries. Our mental health landscape and resource mapping in Kenya also illustrated that capacity building in SSA countries involved complex dynamic, history, and some overlap efforts with multiple partnerships, and these are critical to consider in training activity and evaluation design.


Exploring Research-Methods Blogs in Psychology: Who Posts What About Whom, and With What Effect?
During the methods crisis in psychology and other sciences, much discussion developed online in forums such as blogs and other social media. Hence, this increasingly popular channel of scientific discussion itself needs to be explored to inform current controversies, record the historical moment, improve methods communication, and address equity issues. Who posts what about whom, and with what effect? Does a particular generation or gender contribute more than another? Do blogs focus narrowly on methods, or do they cover a range of issues? How do they discuss individual researchers, and how do readers respond? What are some impacts? Web-scraping and text-analysis techniques provide a snapshot characterizing 41 current research-methods blogs in psychology. Bloggers mostly represented psychology's traditional leaderships' demographic categories: primarily male, mid- to late career, associated with American institutions, White, and with established citation counts. As methods blogs, their posts mainly concern statistics, replication (particularly statistical power), and research findings. The few posts that mentioned individual researchers substantially focused on replication issues; they received more views, social-media impact, comments, and citations. Male individual researchers were mentioned much more often than female researchers. Further data can inform perspectives about these new channels of scientific communication, with the shared aim of improving scientific practices.


Scope of telemedicine in neonatology.
There is a widespread shortage of physicians worldwide, especially in rural areas. This shortage is more prevalent when it comes to subspecialty care, even in developed countries. One way to provide access to specialty care is using technology via telemedicine. Telemedicine has evolved over the last two decades, and its use is becoming widespread in developed countries. However, its use in the neonatal population is still limited and practiced only in some centers. It is now apparent that telemedicine can be successfully used in the neonatal population for screening premature infants for retinopathy of prematurity, congenital heart disease, bedside clinical rounds, neonatal resuscitation with the support of a tertiary care hospital, and family support. This avoids unnecessary transfer and appears to provide the same quality of care that the baby would have received at the tertiary care facility. This approach also improves family satisfaction, as the baby and the mother are kept together, and reduces the cost of care. This review focuses on the use of telemedicine in neonatal care, concentrating on the main areas where telemedicine has been shown to be successful and effective, including the status of telemedicine in China.


Detection and molecular characterisation of a diagnosis escape variant associated with occult hepatitis B virus in Brazil.
Many studies have identified mutations in the hepatitis B surface antigen (HBsAg) as important factors limiting the ability of commercial serological assays to detect this viral antigen. However, an association between mutations in the HBsAg gene and the occurrence of occult HBV infection (OBI) in patients has not been established. To detect hepatitis B virus (HBV) DNA in patients with anti-HBc as a unique serological marker, a previously published, cost-effective TaqMan-based real-time polymerase chain reaction (PCR) test with minor groove binding probes was adapted for use in this study. The current study also aimed to investigate HBsAg mutations and genotypes of HBV in OBI at the Viral Hepatitis Ambulatory Clinic in Rio de Janeiro to determine any possible association. Intra-assay and inter-assay reproducibility were determined, and the mean coefficient of variation values obtained were 2.07 and 3.5, respectively. Probit analysis indicated that the 95% detection level was 25 IU/mL. The prevalence of OBI was investigated in 35 serum samples with an 'anti-HBc alone' profile from individuals who attended our clinic between 2011 and 2013. HBV DNA was detected in only one sample, resulting in an OBI rate of 2.9%. Nucleotide sequencing of the pre-S/S region was performed to genotype and analyse mutations within the HBsAg gene of this HBV DNA. The HBV in the OBI case was classified as sub-genotype A1, and a sequence analysis of the small S gene revealed 12 mutations in the major hydrophilic region compared to the consensus A1 sequence. Most of these mutations occurred in amino acid residues that have been reported as clinically relevant because they have been implicated in vaccine escape and/or inability to detect HBsAg by commercial serological assays. Our study suggests the importance of specific HBsAg mutations, different from those in D, B, and C genotypes, in sub-genotype A1 HBV associated with OBI.


A signature-based machine learning model for distinguishing bipolar disorder and borderline personality disorder.
Mobile technologies offer new opportunities for prospective, high resolution monitoring of long-term health conditions. The opportunities seem of particular promise in psychiatry where diagnoses often rely on retrospective and subjective recall of mood states. However, deriving clinically meaningful information from the complex time series data these technologies present is challenging, and the current implications for patient care are uncertain. In this study, 130 participants with bipolar disorder (n = 48) or borderline personality disorder (n = 31) and healthy volunteers (n = 51) completed daily mood ratings using a bespoke smartphone app for up to 1 year. A signature-based learning method was used to capture the evolving interrelationships between the different elements of mood and exploit this information to classify participants' diagnosis and to predict subsequent mood. The three participant groups could be distinguished from one another on the basis of self-reported mood using the signature methodology. The methodology classified 75% of participants into the correct diagnostic group compared with 54% using standard approaches. Subsequent mood ratings were correctly predicted with >70% accuracy. Prediction of mood was most accurate in healthy volunteers (89-98%) compared to bipolar disorder (82-90%) and borderline personality disorder (70-78%). The signature method provided an effective approach to the analysis of mood data both in terms of diagnostic classification and prediction of future mood. It also highlighted the differing predictability and the overlap inherent within disorders. The three cohorts offered internally consistent but distinct patterns of mood interaction in their reporting which have the potential to enable more efficient and accurate diagnoses and thus earlier treatment.


Electron paramagnetic resonance and saturation transfer electron paramagnetic resonance studies on erythrocytes from goats with and without heritable myotonia.
Erythrocytes from myotonic goats, an animal model of heritable myotonia, and normal goats were studied using electron paramagnetic resonance (EPR) and saturation transfer electron paramagnetic resonance (ST-EPR) spin labeling techniques. Three fatty acid spin labels with the nitroxide moiety at progressively greater distances from the carboxyl group were used to monitor different regions within the erythrocyte membrane. Since spin labels have been shown to induce hemolytic and morphologic alterations in erythrocytes, conditions for minimizing these alterations were first defined by hemolysis studies and scanning electron microscopy. Using these defined conditions for our studies we observed no significant differences in any of the EPR or ST-EPR parameters for normal and myotonic goat erythrocytes with any of the fatty acid spin labels used. Our results do not support the theory that myotonia is the result of a generalized membrane defect characterized by increased membrane fluidity as determined by fatty acid spin labels.


The draft genome of MD-2 pineapple using hybrid error correction of long reads.
The introduction of the elite pineapple variety, MD-2, has caused a significant market shift in the pineapple industry. Better productivity, overall increased in fruit quality and taste, resilience to chilled storage and resistance to internal browning are among the key advantages of the MD-2 as compared with its previous predecessor, the Smooth Cayenne. Here, we present the genome sequence of the MD-2 pineapple (Ananas comosus (L.) Merr.) by using the hybrid sequencing technology from two highly reputable platforms, i.e. the PacBio long sequencing reads and the accurate Illumina short reads. Our draft genome achieved 99.6% genome coverage with 27,017 predicted protein-coding genes while 45.21% of the genome was identified as repetitive elements. Furthermore, differential expression of ripening RNASeq library of pineapple fruits revealed ethylene-related transcripts, believed to be involved in regulating the process of non-climacteric pineapple fruit ripening. The MD-2 pineapple draft genome serves as an example of how a complex heterozygous genome is amenable to whole genome sequencing by using a hybrid technology that is both economical and accurate. The genome will make genomic applications more feasible as a medium to understand complex biological processes specific to pineapple.


A stable ZnCo2O4 cocatalyst for photocatalytic CO2 reduction.
Spinel ZnCo2O4 nanostructures have shown great opportunities in energy related areas. However, the applications of ZnCo2O4 for the conversion of CO2 are much less reported. Herein, we present the use of mesoporous ZnCo2O4 nanorods as efficient and high stable cocatalysts for the photochemical reduction of CO2 under mild reaction conditions.


From first to second wave: follow-up of the prospective COVID-19 cohort (KoCo19) in Munich (Germany).
In the 2nd year of the COVID-19 pandemic, knowledge about the dynamics of the infection in the general population is still limited. Such information is essential for health planners, as many of those infected show no or only mild symptoms and thus, escape the surveillance system. We therefore aimed to describe the course of the pandemic in the Munich general population living in private households from April 2020 to January 2021. The KoCo19 baseline study took place from April to June 2020 including 5313 participants (age 14 years and above). From November 2020 to January 2021, we could again measure SARS-CoV-2 antibody status in 4433 of the baseline participants (response 83%). Participants were offered a self-sampling kit to take a capillary blood sample (dry blood spot; DBS). Blood was analysed using the Elecsys® Anti-SARS-CoV-2 assay (Roche). Questionnaire information on socio-demographics and potential risk factors assessed at baseline was available for all participants. In addition, follow-up information on health-risk taking behaviour and number of personal contacts outside the household (N = 2768) as well as leisure time activities (N = 1263) were collected in summer 2020. Weighted and adjusted (for specificity and sensitivity) SARS-CoV-2 sero-prevalence at follow-up was 3.6% (95% CI 2.9-4.3%) as compared to 1.8% (95% CI 1.3-3.4%) at baseline. 91% of those tested positive at baseline were also antibody-positive at follow-up. While sero-prevalence increased from early November 2020 to January 2021, no indication of geospatial clustering across the city of Munich was found, although cases clustered within households. Taking baseline result and time to follow-up into account, men and participants in the age group 20-34 years were at the highest risk of sero-positivity. In the sensitivity analyses, differences in health-risk taking behaviour, number of personal contacts and leisure time activities partly explained these differences. The number of citizens in Munich with SARS-CoV-2 antibodies was still below 5% during the 2nd wave of the pandemic. Antibodies remained present in the majority of SARS-CoV-2 sero-positive baseline participants. Besides age and sex, potentially confounded by differences in behaviour, no major risk factors could be identified. Non-pharmaceutical public health measures are thus still important.


TNF-α down-regulates sarcoplasmic reticulum Ca²⁺ ATPase expression and leads to left ventricular diastolic dysfunction through binding of NF-κB to promoter response element.
TNF-alpha (TNF-α) causes left ventricular diastolic dysfunction. Down-regulation of sarcoplasmic reticulum Ca(2+)-ATPase 2a protein (SERCA2a) expression is one of the major mechanisms underlying diastolic dysfunction. We investigated whether TNF-α modulates SERCA2a expression and alters cardiac diastolic function, and its detailed signalling pathway. We used both in vitro cellular cardiomyocyte model and in vivo rat model to address this issue. We found that TNF-α decreased the levels of both SERCA2a mRNA and protein in the cardiomyocytes, with corresponding impairment of diastolic calcium reuptake, a cellular phenotype of cardiac diastolic function. An ∼2 kb promoter of the SERCA2a gene (atp2a2) along with its serial deletions was cloned into the luciferase reporter system. TNF-α significantly decreased the promoter activity, and truncation of the SERCA2a gene promoter with the putative nuclear factor kappa-B (NF-κB) response element abolished TNF-α-induced SERCA2a gene suppression. Chromatin immunoprecipitation and gel retardation also confirmed the binding of NF-κB to this putative-binding site. TNF-α increased the phosphorylation of IKK and the degradation of IκB, resulted in NF-κB nuclear translocation, and decreased SERCA2a gene promoter activity. This process was attenuated by NF-κB blockers and simvastatin. In the in vivo rat model, lipopolysaccharide treatment significantly elevated the serum TNF-α level, as well as phosphorylation of IKK, resulting in a decrease in myocardial SERCA2a expression, diastolic calcium reuptake, and diastolic dysfunction. Oral treatment with simvastatin led to an increase in SERCA2a expression, alleviation, and prevention of the diastolic dysfunction. TNF-α suppresses SERCA2a gene expression via the IKK/IκB/NF-κB pathway and binding of NF-κB to the SERCA2a gene promoter, and its effect is blocked by simvastatin, demonstrating the potential therapeutic effect of statins in treating inflammation-related diastolic dysfunction.


Risk model of cardiovascular surgery in 845 Marfan patients using the Japan adult cardiovascular surgery database.
The aim of this study was to evaluate the short-term operative results of patients with Marfan syndrome who underwent thoracic or abdominal aortic surgery in a 4-year period in Japan. Data were collected from the Japan Cardiovascular Surgery Database (JCVSD). We retrospectively analyzed the data of 845 patients with Marfan syndrome who underwent cardiovascular surgery between January 2008 and January 2011. Logistic regression was used to generate risk models. The early mortality rate was 4.4% (37/845). Odds ratios (OR), 95% confidence intervals (CI), and P values for structures and processes in the mortality prediction model were as follows: renal insufficiency (OR, 11.37; CI, 3.7234.66; P < 0.001); respiratory disorder (OR, 11.12; CI, 3.20-38.67; P < 0.001); aortic dissection (OR, 13.02; CI, 2.8060.60; P = 0.001); pseudoaneurysm (OR, 11.23; CI, 1.38-91.66; P = 0.024); thoracoabdominal aneurysm (OR, 2.67; CI, 1.22-5.84; P = 0.014); and aorticrupure (OR, 4.23; CI, 1.26-14.23; P = 0.002). The mortality prediction model had a Cindex of 0.82 and a Hosmer-Lemeshow P value of 0.56. In conclusion, this study demonstrated that renal insufficiency and respiratory disorder had great impact on the operative mortality of Marfan patients undergoing cardiovascular surgery. Because patients with aortic dissection or aortic rupture showed high operative mortality, close follow-up to avoid emergency operation is mandatory to improve the operative results. Achieving good results from surgery of the thoracoabdominal aorta was quite challenging, also in Marfan patients.


Scholarly Output in Peruvian National Dentistry according to Gender Disparity: A 10-Year Bibliometric Study.
To evaluate Peruvian scientific publications in dentistry according to sex disparity (2011-2020). This was a retrospective bibliometric study. The unit of analysis was made up of Peruvian dentistry publications indexed in the Scopus database during the last 10 years. Records with metadata (410) corresponding to the period 2011-2020 were downloaded and standardized and refined by analyzing the metadata. The search strategy was developed based on the individual profiles of each Peruvian institution that has a dental school or college. It was evaluated according to the AF-ID of each institution in the Scopus database. In addition, the information provided by the Scopus SciVal tool was used. Finally, publications, impact, and collaboration indicators were used, such as total number per document, per author, average of citations, h-index, collaboration rate, number of institutions, the Source Normalized Impact per Paper indicator, the CiteScore, and the Scopus Field-Weighted Citation Impact. The greatest increase was evident in 2018, with 2019 and 2020 being the maximum peak of scientific publication growth. However, sustained growth has not been evidenced in relation to the female sex. The analysis of coauthorship by the authors revealed four large clusters, of which the first three were represented by male researchers, such as Arriola-Guillen L., Mayta-Tovalino F., and Mendoza-Azpur G., and one by a female, Guerrero María E. Evaluating the national scientific publication in dentistry according to the CiteScore, it was found that most of the publications (145) from Peru were published in Q4 journals, although 90 manuscripts were published in Q1 journals. The Peruvian national dental publication in the last 10 years was mainly supported by male dentists, which invites us to reflect on the need to equalize opportunities so that female researchers can also reduce these gaps.


Continuity and utilization of health and community care in elderly patients with heart failure before and after hospitalization.
The period after hospitalization due to deteriorated heart failure (HF) is characterized as a time of high generalized risk. The transition from hospital to home is often problematic due to insufficient coordination of care, leading to a fragmentation of care rather than a seamless continuum of care. The aim was to describe health and community care utilization prior to and 30 days after hospitalization, and the continuity of care in patients hospitalized due to de novo or deteriorated HF from the patients' perspective and from a medical chart review. This was a cross-sectional study with consecutive inclusion of patients hospitalized at a county hospital in Sweden due to deteriorated HF during 2014. Data were collected by structured telephone interviews and medical chart review and analyzed with the Spearman's rank correlation coefficient and Chi square. A P value of 0.05 was considered significant. A total of 121 patients were included in the study, mean age 82.5 (±6.8) and 49% were women. Half of the patients had not visited any health care facility during the month prior to the index hospital admission, and 79% of the patients visited the emergency room (ER) without a referral. Among these elderly patients, a total of 40% received assistance at home prior to hospitalization and 52% after discharge. A total of 86% received written discharge information, one third felt insecure after hospitalization and lacked knowledge of which health care provider to consult with and contact in the event of deterioration or complications. Health care utilization increased significantly after hospitalization. Most patients had not visited any health care facility within 30 days before hospitalization. Health care utilization increased significantly after hospitalization. Flaws in the continuity of care were found; even though most patients received written information at discharge, one third of the patients lacked knowledge about which health care provider to contact in the event of deterioration and felt insecure at home after discharge.


Direct activation of the ventral striatum in anticipation of aversive stimuli.
The brain "reward" system, centered on the limbic ventral striatum, plays a critical role in the response to pleasure and pain. The ventral striatum is activated in animal and human studies during anticipation of appetitive/pleasurable events, but its role in aversive/painful events is less clear. Here we present data from three human fMRI studies based on aversive conditioning using unpleasant cutaneous electrical stimulation and show that the ventral striatum is reliably activated. This activation is observed during anticipation and is not a consequence of relief after the aversive event. Further, the ventral striatum is activated in anticipation regardless of whether there is an opportunity to avoid the aversive stimulus or not. Our data suggest that the ventral striatum, a crucial element of the brain "reward" system, is directly activated in anticipation of aversive stimuli.


Accumulation of lipochitin oligosaccharides and NodD-activating compounds in an efficient plant--Rhizobium nodulation assay.
During legume plant--Rhizobium spp. interactions, leading to the formation of nitrogen-fixing root nodules, the two major determinants of host plant-specificity are plant-produced nod gene inducers (NodD protein activating compounds) and bacterial lipochitin oligosaccharides (LCOs or Nod factors). In a time course, we describe the accumulation of LCOs in an efficient nodulation assay with Vicia sativa subsp. nigra and Rhizobium leguminosarum, in connection with the presence of NodD-activating compounds in the exudate of V. sativa roots. Relatively small amounts of both LCOs and NodD-activating compounds were found to be required for initiation of nodulation during the first days after inoculation. A strong increase in the amount of NodRlv-V[18:4,Ac] LCOs preceded root infection and nodule primordium formation. In contrast to the situation with non-nodulating rhizobia and nonmitogenic LCOs, the amount of NodD-activating compounds in the culture medium remained small after addition of nodulating rhizobia or mitogenic LCOs. Furthermore, addition of nodulating rhizobia or mitogenic LCOs resulted in nearly complete inhibition of root hair formation and elongation, whereas nonmitogenic LCOs stimulated root hair growth. Retention of NodD-activating compounds in the root may inhibit root hair growth.


The Earning and Spending Habits of Male Sex Workers in Lima, Peru.
Over the past decade, data have identified male sex work as a potentially viable economic decision; despite this, male sex workers (MSWs) continue to be perceived as group with access to few assets and resources. Using data from a pilot skills- building intervention for MSWs in Lima, Peru, an analysis of the economic characteristics of 209 MSWs is presented. The majority reported livable incomes with median earnings of US$250 per month, 83% earning above the urban poverty line. Interestingly, non-sex work was also an important source of income, especially for the high-earning MSWs. Spending data revealed that a large portion of income went to necessities (55%), luxuries (11%), and gifts (11%), with less toward savings (5%) and studies (1%). Such data on MSWs' earnings and spending, which suggest that a lack of overall income is not the MSW's main impediment to escaping poverty, could direct future poverty alleviation and health improvement programs in this key population.


Consumer Attitudes Toward Animal Welfare-Friendly Products and Willingness to Pay: Exploration of Mexican Market Segments.
The study aim was to identify consumer segmentation based on nonhuman animal welfare (AW) attitudes and their relationship with demographic features and willingness to pay (WTP) for welfare-friendly products (WFP) in Mexico. Personal interviews were conducted with 843 Mexican consumers who stated they purchased most of the animal products in their home. Respondents were selected using a quota sampling method with age, gender, education, and origin as quota control variables. The multivariate analysis suggested there were three clusters or consumer profiles labeled "skeptical," "concerned," and "ethical," which helped explain the association between AW attitudes, some demographic variables, and WTP for WFP. This study is one of the first to address consumer profiling in Latin America, and the findings could have implications for the commercialization of WFP. Hence, customers should receive information to consider welfare innovations when deciding to purchase animal products. The growth of the WFP food market establishes an element of a far more multifaceted phenomenon of sustainable consumption and support of a new paradigm called responsible marketing in emerging markets such as Mexico.


Time- and dose-dependent effects of roundup on human embryonic and placental cells.
Roundup is the major herbicide used worldwide, in particular on genetically modified plants that have been designed to tolerate it. We have tested the toxicity and endocrine disruption potential of Roundup (Bioforce on human embryonic 293 and placental-derived JEG3 cells, but also on normal human placenta and equine testis. The cell lines have proven to be suitable to estimate hormonal activity and toxicity of pollutants. The median lethal dose (LD(50)) of Roundup with embryonic cells is 0.3% within 1 h in serum-free medium, and it decreases to reach 0.06% (containing among other compounds 1.27 mM glyphosate) after 72 h in the presence of serum. In these conditions, the embryonic cells appear to be 2-4 times more sensitive than the placental ones. In all instances, Roundup (generally used in agriculture at 1-2%, i.e., with 21-42 mM glyphosate) is more efficient than its active ingredient, glyphosate, suggesting a synergistic effect provoked by the adjuvants present in Roundup. We demonstrated that serum-free cultures, even on a short-term basis (1 h), reveal the xenobiotic impacts that are visible 1-2 days later in serum. We also document at lower non-overtly toxic doses, from 0.01% (with 210 microM glyphosate) in 24 h, that Roundup is an aromatase disruptor. The direct inhibition is temperature-dependent and is confirmed in different tissues and species (cell lines from placenta or embryonic kidney, equine testicular, or human fresh placental extracts). Furthermore, glyphosate acts directly as a partial inactivator on microsomal aromatase, independently of its acidity, and in a dose-dependent manner. The cytotoxic, and potentially endocrine-disrupting effects of Roundup are thus amplified with time. Taken together, these data suggest that Roundup exposure may affect human reproduction and fetal development in case of contamination. Chemical mixtures in formulations appear to be underestimated regarding their toxic or hormonal impact.


A benzimidazolyl terpyridine-Fe2+ system and its recognition driven molecular model of a traffic light.
A new benzimidazolyl terpyridyl ligand (BIT) selectively detects Fe2+, F- and CN- ions in acetonitrile. The Fe2+ complex [Fe(H-BIT)2]·(ClO4)4 (1) displays the fast and selective detection of 2,4,6-trinitrophenol (TNP) with strong binding (log β = 8.51 ± 0.24) and a low limit of detection (1.4 × 10-7 M) among several nitroaromatics. The color of complex 1 changes from purple to red, yellow, and green upon the separate addition of 2,4,6-trinitrophenol and tetrabutylammonium salts of fluoride and cyanide, respectively. The overall color changes thus display a model of a traffic light. The structures of BIT, 1 and [Fe(BIT)2]·(TNP)2 (2) have been well characterized using spectroscopic techniques and single crystal X-ray crystallography.


Wireless Instantaneous Neurotransmitter Concentration System-based amperometric detection of dopamine, adenosine, and glutamate for intraoperative neurochemical monitoring.
In a companion study, the authors describe the development of a new instrument named the Wireless Instantaneous Neurotransmitter Concentration System (WINCS), which couples digital telemetry with fast-scan cyclic voltammetry (FSCV) to measure extracellular concentrations of dopamine. In the present study, the authors describe the extended capability of the WINCS to use fixed potential amperometry (FPA) to measure extracellular concentrations of dopamine, as well as glutamate and adenosine. Compared with other electrochemical techniques such as FSCV or high-speed chronoamperometry, FPA offers superior temporal resolution and, in combination with enzyme-linked biosensors, the potential to monitor nonelectroactive analytes in real time. The WINCS design incorporated a transimpedance amplifier with associated analog circuitry for FPA; a microprocessor; a Bluetooth transceiver; and a single, battery-powered, multilayer, printed circuit board. The WINCS was tested with 3 distinct recording electrodes: 1) a carbon-fiber microelectrode (CFM) to measure dopamine; 2) a glutamate oxidase enzyme-linked electrode to measure glutamate; and 3) a multiple enzyme-linked electrode (adenosine deaminase, nucleoside phosphorylase, and xanthine oxidase) to measure adenosine. Proof-of-principle analyses included noise assessments and in vitro and in vivo measurements that were compared with similar analyses by using a commercial hardwired electrochemical system (EA161 Picostat, eDAQ; Pty Ltd). In urethane-anesthetized rats, dopamine release was monitored in the striatum following deep brain stimulation (DBS) of ascending dopaminergic fibers in the medial forebrain bundle (MFB). In separate rat experiments, DBS-evoked adenosine release was monitored in the ventrolateral thalamus. To test the WINCS in an operating room setting resembling human neurosurgery, cortical glutamate release in response to motor cortex stimulation (MCS) was monitored using a large-mammal animal model, the pig. The WINCS, which is designed in compliance with FDA-recognized consensus standards for medical electrical device safety, successfully measured dopamine, glutamate, and adenosine, both in vitro and in vivo. The WINCS detected striatal dopamine release at the implanted CFM during DBS of the MFB. The DBS-evoked adenosine release in the rat thalamus and MCS-evoked glutamate release in the pig cortex were also successfully measured. Overall, in vitro and in vivo testing demonstrated signals comparable to a commercial hardwired electrochemical system for FPA. By incorporating FPA, the chemical repertoire of WINCS-measurable neurotransmitters is expanded to include glutamate and other nonelectroactive species for which the evolving field of enzyme-linked biosensors exists. Because many neurotransmitters are not electrochemically active, FPA in combination with enzyme-linked microelectrodes represents a powerful intraoperative tool for rapid and selective neurochemical sampling in important anatomical targets during functional neurosurgery.


Synthesis, anticancer and antibacterial activity of salinomycin N-benzyl amides.
A series of 12 novel monosubstituted N-benzyl amides of salinomycin (SAL) was synthesized for the first time and characterized by NMR and FT-IR spectroscopic methods. Molecular structures of three salinomycin derivatives in the solid state were determined using single crystal X-ray method. All compounds obtained were screened for their antiproliferative activity against various human cancer cell lines as well as against the most problematic bacteria strains such as methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis (MRSE), and Mycobacterium tuberculosis. Novel salinomycin derivatives exhibited potent anticancer activity against drug-resistant cell lines. Additionally, two N-benzyl amides of salinomycin revealed interesting antibacterial activity. The most active were N-benzyl amides of SAL substituted at -ortho position and the least anticancer active derivatives were those substituted at the -para position.


Alterations in the common fragile site gene Parkin in ovarian and other cancers.
The cloning and characterization of the common fragile site (CFS) FRA6E (6q26) identified Parkin, the gene involved in the pathogenesis of many cases of juvenile, early-onset and, rarely, late-onset Parkinson's disease, as the third large gene to be localized within a large CFS. Initial analyses of Parkin indicated that in addition to playing a role in Parkinson's disease, it might also be involved in the development and/or progression of ovarian cancer. These analyses also indicated striking similarities among the large CFS-locus genes: fragile histidine triad gene (FHIT; 3p14.2), WW domain-containing oxidoreductase gene (WWOX; 16q23), and Parkin (6q26). Analyses of FHIT and WWOX in a variety of different cancer types have identified the presence of alternative transcripts with whole exon deletions. Interestingly, various whole exon duplications and deletions have been identified for Parkin in juvenile and early-onset Parkinson's patients. Therefore, we performed mutational/exon rearrangement analysis of Parkin in ovarian cancer cell lines and primary tumors. Four (66.7%) cell lines and four (18.2%) primary tumors were identified as being heterozygous for the duplication or deletion of a Parkin exon. Additionally, three of 23 (13.0%) nonovarian tumor-derived cell lines were also identified as having a duplication or deletion of one or more Parkin exons. Analysis of Parkin protein expression with antibodies revealed that most of the ovarian cancer cell lines and primary tumors had diminished or absent Parkin expression. While functional analyses have not yet been performed for Parkin, these data suggest that like FHIT and WWOX, Parkin may represent a tumor suppressor gene.


A Systematic Review of Methods and Procedures Used in Ecological Momentary Assessments of Diet and Physical Activity Research in Youth: An Adapted STROBE Checklist for Reporting EMA Studies (CREMAS).
Ecological momentary assessment (EMA) is a method of collecting real-time data based on careful timing, repeated measures, and observations that take place in a participant's typical environment. Due to methodological advantages and rapid advancement in mobile technologies in recent years, more studies have adopted EMA in addressing topics of nutrition and physical activity in youth. The aim of this systematic review is to describe EMA methodology that has been used in studies addressing nutrition and physical activity in youth and provide a comprehensive checklist for reporting EMA studies. Thirteen studies were reviewed and analyzed for the following 5 areas of EMA methodology: (1) sampling and measures, (2) schedule, (3) technology and administration, (4) prompting strategy, and (5) response and compliance. Results of this review showed a wide variability in the design and reporting of EMA studies in nutrition and physical activity among youth. The majority of studies (69%) monitored their participants during one period of time, although the monitoring period ranged from 4 to 14 days, and EMA surveys ranged from 2 to 68 times per day. More than half (54%) of the studies employed some type of electronic technology. Most (85%) of the studies used interval-contingent prompting strategy. For studies that utilized electronic devices with interval-contingent prompting strategy, none reported the actual number of EMA prompts received by participants out of the intended number of prompts. About half (46%) of the studies failed to report information about EMA compliance rates. For those who reported, compliance rates ranged from 44-96%, with an average of 71%. Findings from this review suggest that in order to identify best practices for EMA methodology in nutrition and physical activity research among youth, more standardized EMA reporting is needed. Missing the key information about EMA design features and participant compliance might lead to misinterpretation of results. Future nutrition and physical activity EMA studies need to be more rigorous and thorough in descriptions of methodology and results. A reporting checklist was developed with the goal of enhancing reliability, efficacy, and overall interpretation of the findings for future studies that use EMAs.


Comparison of microleakage in Class II cavities restored with silorane-based and methacrylate-based composite resins using different restorative techniques over time.
Despite the growing tendency toward tooth-colored restorations in dentistry, polymerization shrinkage and subsequent marginal microleakage remains a problem. The aim of this in vitro study was to compare microleakage between silorane-based and methacrylate-based composite resins at different time intervals and with different restorative techniques. In this in vitro study, 108 sound extracted human molar teeth were used. Mesial and distal proximal class II boxes with dimensions of 1.5 mm depth and 4 mm width were prepared. The gingival margins of all cavities were 1 mm below the cement enamel junction. The teeth were randomly divided into three groups based on test materials. In the first group, the teeth were restored by a nanocomposite (Filtek Z350XT, 3MESPE) and SE Bond adhesive (Kuraray, Japan), in the second group, the teeth were restored with a silorane-based (Filtek P90, 3MESPE) and Filtek P90 Adhesive (3M ESPE, USA) and in the third group, the teeth were restored with a microhybrid posterior composite resin (Filtek P60, 3MESPE) and SE Bond adhesive (Kuraray, Japan). Half of the proximal cavities in each of these three groups were restored in two horizontal layers and the other half in four horizontal layers. After a period of aging (24-h, 3-month and 6-month) in water and then application of 500 thermal cycles, the teeth were immersed for 24-h in 0.5% fuchsin and evaluated under a stereomicroscope at ×36 magnification to evaluate leakage in gingival margin. Data was statistically analyzed using Kruskal-Wallis and Mann-Whitney U-tests. P ≤ 0.05 was considered as significant. In Z350XT statistically significant differences were observed in microleakage in comparison of 24-h and 6-month intervals (P = 0.01) that was higher in 6-month. Comparison of microleakage in P90 and P60 composite resins was also statistically significant and was less in P90. Microleakage was not significantly different between P90 and Z350XT at 24-h. However, this difference was significant at 3-month and 6-month intervals. Differences in microleakage of P60 and Z-350XT composite resins were not statistically significant in all intervals (P = 0.38). P90 showed the lowest microleakage during storage in water. Z350XT had microleakage similar to P90 within 24-h, but after 6-month of storage in water, it showed the highest microleakage among all the groups. The number of layers (2 layers vs. 4 layers) did not result in any differences in microleakage scores of the composite resins (P = 0.42). Water storage times did not result in any significant effect on microleakage of P90 and P60.


Mapping iterative medical imaging algorithm on cell accelerator.
Algebraic reconstruction techniques require about half the number of projections as that of Fourier backprojection methods, which makes these methods safer in terms of required radiation dose. Algebraic reconstruction technique (ART) and its variant OS-SART (ordered subset simultaneous ART) are techniques that provide faster convergence with comparatively good image quality. However, the prohibitively long processing time of these techniques prevents their adoption in commercial CT machines. Parallel computing is one solution to this problem. With the advent of heterogeneous multicore architectures that exploit data parallel applications, medical imaging algorithms such as OS-SART can be studied to produce increased performance. In this paper, we map OS-SART on cell broadband engine (Cell BE). We effectively use the architectural features of Cell BE to provide an efficient mapping. The Cell BE consists of one powerPC processor element (PPE) and eight SIMD coprocessors known as synergetic processor elements (SPEs). The limited memory storage on each of the SPEs makes the mapping challenging. Therefore, we present optimization techniques to efficiently map the algorithm on the Cell BE for improved performance over CPU version. We compare the performance of our proposed algorithm on Cell BE to that of Sun Fire ×4600, a shared memory machine. The Cell BE is five times faster than AMD Opteron dual-core processor. The speedup of the algorithm on Cell BE increases with the increase in the number of SPEs. We also experiment with various parameters, such as number of subsets, number of processing elements, and number of DMA transfers between main memory and local memory, that impact the performance of the algorithm.


Multiplex touchdown PCR for rapid typing of the opportunistic pathogen Propionibacterium acnes.
The opportunistic human pathogen Propionibacterium acnes is composed of a number of distinct phylogroups, designated types IA1, IA2, IB, IC, II, and III, which vary in their production of putative virulence factors, their inflammatory potential, and their biochemical, aggregative, and morphological characteristics. Although multilocus sequence typing (MLST) currently represents the gold standard for unambiguous phylogroup classification and individual strain identification, it is a labor-intensive and time-consuming technique. As a consequence, we developed a multiplex touchdown PCR assay that in a single reaction can confirm the species identity and phylogeny of an isolate based on its pattern of reaction with six primer sets that target the 16S rRNA gene (all isolates), ATPase (types IA1, IA2, and IC), sodA (types IA2 and IB), atpD (type II), and recA (type III) housekeeping genes, as well as a Fic family toxin gene (type IC). When applied to 312 P. acnes isolates previously characterized by MLST and representing types IA1 (n=145), IA2 (n=20), IB (n=65), IC (n=7), II (n=45), and III (n=30), the multiplex displayed 100% sensitivity and 100% specificity for detecting isolates within each targeted phylogroup. No cross-reactivity with isolates from other bacterial species was observed. This multiplex assay will provide researchers with a rapid, high-throughput, and technically undemanding typing method for epidemiological and phylogenetic investigations. It will facilitate studies investigating the association of lineages with various infections and clinical conditions, and it will serve as a prescreening tool to maximize the number of genetically diverse isolates selected for downstream higher-resolution sequence-based analyses.


Livelihood opportunities amongst adults with and without disabilities in Cameroon and India: A case control study.
Proven links between disability and poverty suggest that development programmes and policies that are not disability-inclusive will leave persons with disabilities behind. Despite this, there is limited quantitative evidence on livelihood opportunities amongst adults with disabilities in Low and Middle Income Countries. This study adds to the limited evidence base, contributing data from one African and one Asian Setting. We undertook a population-based case-control study of adults (18+) with and without disabilities in North-West Cameroon and in Telangana State, India. We found that adults with disabilities were five times less likely to be working compared to age-sex matched controls in both settings. Amongst adults with disabilities, current age, marital status and disability type were key predictors of working. Inclusive programmes are therefore needed to provide adequate opportunities to participate in livelihood prospects for adults with disabilities in Cameroon and India, on an equal basis as others. These findings are of crucial importance at this stage of the Sustainable Development Agenda, to ensure that the mandate of inclusive development is achieved.


The role of cholesterol in the activity of pneumolysin, a bacterial protein toxin.
The mechanism via which pneumolysin (PLY), a toxin and major virulence factor of the bacterium Streptococcus pneumoniae, binds to its putative receptor, cholesterol, is still poorly understood. We present results from a series of biophysical studies that shed light on the interaction of PLY with cholesterol in solution and in lipid bilayers. PLY lyses cells whose walls contain cholesterol. Using standard hemolytic assays we have demonstrated that the hemolytic activity of PLY is inhibited by cholesterol, partially by ergosterol but not by lanosterol and that the functional stoichiometry of the cholesterol-PLY complex is 1:1. Tryptophan (Trp) fluorescence data recorded during PLY-cholesterol titration studies confirm this ratio, reveal a significant blue shift in the Trp fluorescence peak with increasing cholesterol concentrations indicative of increasing nonpolarity in the Trp environment, consistent with cholesterol binding by the tryptophans, and provide a measure of the affinity of cholesterol binding: K(d) = 400 +/- 100 nM. Finally, we have performed specular neutron reflectivity studies to observe the effect of PLY upon lipid bilayer structure.


Higher-risk myelodysplastic syndromes with del(5q): is sequential azacitidine-lenalidomide combination the way to go?
High-risk myelodysplastic syndromes (HR-MDS) and acute myeloid leukemia (AML) with deletions of long arm of chromosome 5 (del[5q]) are characterized by rapid progression and poor survival. The majority of these patients are elderly with comorbidities, therefore limiting the use of intensive therapies which, even if used, unfortunately yield low responses. While azacitidine prolongs survival in patients with HR-MDS by a median of 9.5 months, responses only occur in less than half of the patients, and azacitidine therapy is not curative, with most patients relapsing within 2 years. Therefore, strategies to improve outcomes in these patients are needed. Azacitidine and lenalidomide both have meaningful single-agent clinical activity in HR-MDS and AML with del(5q). Early-phase trials in HR-MDS without del(5q) suggest increased activity with a concurrent azacitidine-lenalidomide combination. In this article, we review the results of a Phase I trial of a sequential azacitidine-lenalidomide combination approach in patients with HR-MDS and AML with del(5q) abnormality.


Refluxed Esterification of Fullerene-Conjugated P25 TiO2 Promotes Free Radical Scavenging Capacity and Facilitates Antiaging Potentials in Human Cells.
Titanium dioxide nanomaterials have good capability to prevent human cells from damage under UV irradiation. However, some studies indicated that the nanoscale of titanium dioxide could potentially cause harmful effects such as free radical generation under UV irradiation and thereby accelerate the progress of cell aging. Fullerenes can scavenge large amounts of free radicals due to the fact that fullerenes contain enormous amount of π electrons with low lying lowest unoccupied molecular orbital, but its adverse properties, such as the poor solubility in water, restricted the applicability. In this study, we employed water-soluble carboxylic acid fullerenes (C60-COOH and C70-COOH) as the free radical scavenger and modify onto the surface of titanium dioxide by refluxed esterification (P25/C60-COOH or C70-COOH) technique. The conformation and properties of these nanomaterials were characterized by techniques and equipment such as X-ray diffraction, energy dispersive spectroscopy analysis, scanning electron microscopy, thermal gravimetric analysis, high-resolution transmission electron microscopy, and Fourier transform infrared spectroscopy. We also introduced methylene blue and rhodamine B as indicators to evaluate and demonstrate the scavenging capacity of these nanomaterials. Moreover, we examined the biocompatibility and UV protection capacity of our P25/fullerene composites in human 293T cells, and applied luciferase activity assay to investigate the possible underlying cell protection mechanisms exhibited by these nanomaterials. Our data indicate that both P25/C60-COOH and P25/C70-COOH could protect human cells against UV exposure. P25/C70-COOH exhibits great anti-inflammation capacity, whereas P25/C60-COOH exhibits great anti-oxidative stress and anti-DNA damage capacity. Our results suggest that most of our P25/fullerene composite materials have the ability to reduce free radicals and exhibit high biomedical potential in anti-inflammation, anti-oxidant, and anti-aging applications.


Corrupted coordination of epigenetic modifications leads to diverging chromatin states and transcriptional heterogeneity in CLL.
Cancer evolution is fueled by epigenetic as well as genetic diversity. In chronic lymphocytic leukemia (CLL), intra-tumoral DNA methylation (DNAme) heterogeneity empowers evolution. Here, to comprehensively study the epigenetic dimension of cancer evolution, we integrate DNAme analysis with histone modification mapping and single cell analyses of RNA expression and DNAme in 22 primary CLL and 13 healthy donor B lymphocyte samples. Our data reveal corrupted coherence across different layers of the CLL epigenome. This manifests in decreased mutual information across epigenetic modifications and gene expression attributed to cell-to-cell heterogeneity. Disrupted epigenetic-transcriptional coordination in CLL is also reflected in the dysregulation of the transcriptional output as a function of the combinatorial chromatin states, including incomplete Polycomb-mediated gene silencing. Notably, we observe unexpected co-mapping of typically mutually exclusive activating and repressing histone modifications, suggestive of intra-tumoral epigenetic diversity. Thus, CLL epigenetic diversification leads to decreased coordination across layers of epigenetic information, likely reflecting an admixture of cells with diverging cellular identities.


Harmonic mode locking of a Nd:BEL laser using a 20-GHz dielectric resonator/optical modulator.
A 20-GHz dielectric-resonator/optical modulator is developed and used as an FM mode locker at the 84th harmonic of a conventional 238-MHz diode-pumped Nd:BEL laser cavity. Depending on the mode-locker drive frequency, two distinct regimes of mode locking were observed: 2.9-ps pulses at a repetition rate of 238 MHz and 3.9-ps pulses at a repetition rate of 20 GHz. These are to our knowledge the shortest pulses ever reported for active mode locking of a Nd laser.


Several Grassland Soil Nematode Species Are Insensitive to RNA-Mediated Interference.
Phenotypic analysis of defects caused by RNA mediated interference (RNAi) in Caenorhabditis elegans has proven to be a powerful tool for determining gene function. In this study we investigated the effectiveness of RNAi in four non-model grassland soil nematodes, Oscheius sp FVV-2., Rhabditis sp, Mesorhabditis sp., and Acrobeloides sp. In contrast to reference experiments performed using C. elegans and Caenorhabditis briggsae, feeding bacteria expressing dsRNA and injecting dsRNA into the gonad did not produce the expected RNAi knockdown phenotypes in any of the grassland nematodes. Quantitative reverse-transcribed PCR (qRT-PCR) assays did not detect a statistically significant reduction in the mRNA levels of endogenous genes targeted by RNAi in Oscheius sp., and Mesorhabditis sp. From these studies we conclude that due to low effectiveness and inconsistent reproducibility, RNAi knockdown phenotypes in non-Caenorhabditis nematodes should be interpreted cautiously.


Enhanced sampling in the well-tempered ensemble.
We introduce the well-tempered ensemble (WTE) which is the biased ensemble sampled by well-tempered metadynamics when the energy is used as collective variable. WTE can be designed so as to have approximately the same average energy as the canonical ensemble but much larger fluctuations. These two properties lead to an extremely fast exploration of phase space. An even greater efficiency is obtained when WTE is combined with parallel tempering. Unbiased Boltzmann averages are computed on the fly by a recently developed reweighting method [M. Bonomi, J. Comput. Chem. 30, 1615 (2009)]. We apply WTE and its parallel tempering variant to the 2d Ising model and to a Gō model of HIV protease, demonstrating in these two representative cases that convergence is accelerated by orders of magnitude.


Vasopressin for cardiac arrest: a systematic review and meta-analysis.
The current guidelines for cardiopulmonary resuscitation recommend vasopressin as an alternative to epinephrine for the treatment of adult shock-refractory ventricular fibrillation. The objective of this study was to determine the effectiveness of vasopressin in the treatment of cardiac arrest. We performed a systematic review and meta-analysis of 1519 patients with cardiac arrest from 5 randomized controlled trials that compared vasopressin and epinephrine. Two reviewers conducted a systematic search of electronic databases, complemented by hand searches, to identify randomized trials. Reviewers evaluated the quality of the trials, extracted data, and derived pooled estimates using a random-effects model. There were no statistically significant differences between the vasopressin and epinephrine groups in failure of return of spontaneous circulation (risk ratio [RR], 0.81; 95% confidence interval [CI], 0.58-1.12), death before hospital admission (RR, 0.72; 95% CI, 0.38-1.39), death within 24 hours (RR, 0.74; 95% CI, 0.38-1.43), death before hospital discharge (RR, 0.96; 95% CI, 0.87-1.05), or combination of number of deaths and neurologically impaired survivors (RR, 1.00; 95% CI, 0.94-1.07). Subgroup analysis based on initial cardiac rhythm showed no statistically significant difference in the rate of death before hospital discharge between the vasopressin and epinephrine groups in any of the 3 subgroups: ventricular fibrillation or ventricular tachycardia (RR, 0.97; 95% CI, 0.79-1.19), pulseless electrical activity (RR, 1.02; 95% CI, 0.95-1.10), or asystole (RR, 0.97; 95% CI, 0.94-1.00). There is no clear advantage of vasopressin over epinephrine in the treatment of cardiac arrest. Guidelines for Advanced Cardiac Life Support should not recommend vasopressin in resuscitation protocols until more solid human data on its superiority are available.


Electrical stimulation of excitable tissue: design of efficacious and safe protocols.
The physical basis for electrical stimulation of excitable tissue, as used by electrophysiological researchers and clinicians in functional electrical stimulation, is presented with emphasis on the fundamental mechanisms of charge injection at the electrode/tissue interface. Faradaic and non-Faradaic charge transfer mechanisms are presented and contrasted. An electrical model of the electrode/tissue interface is given. The physical basis for the origin of electrode potentials is given. Various methods of controlling charge delivery during pulsing are presented. Electrochemical reversibility is discussed. Commonly used electrode materials and stimulation protocols are reviewed in terms of stimulation efficacy and safety. Principles of stimulation of excitable tissue are reviewed with emphasis on efficacy and safety. Mechanisms of damage to tissue and the electrode are reviewed.


Acute effects of glucagon-like peptide-1, GLP-19-36 amide, and exenatide on mesenteric blood flow, cardiovascular parameters, and biomarkers in healthy volunteers.
Glucagon-like peptide-1 (GLP-1, GLP-17-36amide) and its sister peptide glucagon-like peptide 2 (GLP-2) influence numerous intestinal functions and GLP-2 greatly increases intestinal blood flow. We hypothesized that GLP-1 also stimulates intestinal blood flow and that this would impact on the overall digestive and cardiovascular effects of the hormone. To investigate the influence of GLP-1 receptor agonism on mesenteric and renal blood flow and cardiovascular parameters, we carried out a double-blinded randomized clinical trial. A total of eight healthy volunteers received high physiological subcutaneous injections of GLP-1, GLP-19-36 amide (bioactive metabolite), exenatide (stable GLP-1 agonist), or saline on four separate days. Blood flow in mesenteric, celiac, and renal arteries was measured by Doppler ultrasound. Blood pressure, heart rate, cardiac output, and stroke volume were measured continuously using an integrated system. Plasma was analyzed for glucose, GLP-1 (intact and total), exenatide and Pancreatic polypeptide (PP), and serum for insulin and C-peptide. Neither GLP-1, GLP-19-36 amide, exenatide nor saline elicited any changes in blood flow parameters in the mesenteric or renal arteries. GLP-1 significantly increased heart rate (two-way ANOVA, injection [P = 0.0162], time [P = 0.0038], and injection × time [P = 0.082]; Tukey post hoc GLP-1 vs. saline and GLP-19-36amide [P < 0.011]), and tended to increase cardiac output and decrease stroke volume compared to GLP-19-36 amide and saline. Blood pressures were not affected. As expected, glucose levels fell and insulin secretion increased after infusion of both GLP-1 and exenatide.


Expectant management of severe pre-eclampsia in the mid-trimester.
To determine maternal and perinatal outcomes with expectant management of severe pre-eclampsia in the mid-trimester, using a defined entry point. Prospective case series. Thirty-nine women admitted from 24 to 27 week's gestation with severe pre-eclampsia, whose pregnancies were otherwise stable, were managed expectantly with careful clinical and biochemical monitoring of maternal and foetal status, together with careful blood pressure control, in a high-care obstetric ward. The aim was to safely prolong the pregnancies and thereby improve perinatal outcome. Gestation was prolonged by a median of 12 (range 3--47) days, with greater periods gained at earlier gestations. The overall perinatal loss was 26% and the neonatal loss 17%. The rates of significant maternal complications were low. Expectant management of selected women with severe pre-eclampsia from 24 to 27 weeks' gestation in a tertiary care unit is acceptably safe and improves perinatal outcome.


Iron and restless legs syndrome: treatment, genetics and pathophysiology.
In this article, we review the original findings from MRI and autopsy studies that demonstrated brain iron status is insufficient in individuals with restless legs syndrome (RLS). The concept of deficient brain iron status is supported by proteomic studies from cerebrospinal fluid (CSF) and from the clinical findings where intervention with iron, either dietary or intravenous, can improve RLS symptoms. Therefore, we include a section on peripheral iron status and how peripheral status may influence both the RLS symptoms and treatment strategy. Given the impact of iron in RLS, we have evaluated genetic data to determine if genes are directly involved in iron regulatory pathways. The result was negative. In fact, even the HFE mutation C282Y could not be shown to have a protective effect. Lastly, a consistent finding in conditions of low iron is increased expression of proteins in the hypoxia pathway. Although there is lack of clinical data that RLS patients are hypoxic, there are intriguing observations that environmental hypoxic conditions worsen RLS symptoms; in this chapter we review very compelling data for activation of hypoxic pathways in the brain in RLS patients. In general, the data in RLS point to a pathophysiology that involves decreased acquisition of iron by cells in the brain. Whether the decreased ability is genetically driven, activation of pathways (eg, hypoxia) that are designed to limit cellular uptake is unknown at this time; however, the data strongly support a functional rather than structural defect in RLS, suggesting that an effective treatment is possible.


Prediction of 10-year vascular risk in patients with diabetes: the AD-ON risk score.
To formulate a combined cardiovascular risk score in diabetes that could be useful both to physicians and healthcare funders. Data were derived from the Action in Diabetes and Vascular Disease: Preterax and Diamicron Modified Release Controlled Evaluation Observational (ADVANCE-ON) study, a randomized controlled trial (mean duration 5 years) with a post-randomization follow-up (mean 4.9 years), that included 11 140 high-risk patients with diabetes. The outcome analysed was the occurrence of either fatal or non-fatal macrovascular or renal disease. A Cox regression model was used to determine weightings in the risk score. The resultant score was recalibrated to each of three major global regions, as covered by the ADVANCE-ON study. Over a median of 9.9 years, 1145 patients experienced at least one component of the combined outcome event. The resultant score, the AD-ON risk score, incorporated 13 demographic or clinical variables. Its discrimination was modest [c-statistic = 0.668 (95% confidence interval 0.651, 0.685)] but its calibration was excellent (predicted and observed risks coincided well, within disparate global regions). In terms of the integrated discrimination improvement index, its performance was marginally superior, over a 10-year risk horizon, to existing risk scores in clinical use, from a restricted version of the same data, for macrovascular and renal disease separately. The AD-ON risk score has advantages over the existing vascular risk scores in diabetes that used data from the original ADVANCE trial, which treat macrovascular and renal diseases separately. These advantages include its simplicity of use and global application.


Genomic epidemiology of group B streptococci spanning 10 years in an Irish maternity hospital, 2008-2017.
The genomic epidemiology of group b streptococcal (GBS) isolates from the Rotunda maternity hospital, Dublin, 2008-2017, was investigated. Whole genome sequences of isolates (invasive, n = 114; non-invasive, n = 76) from infants and women were analysed using the PubMLST database (https://pubmlst.org/sagalactiae/). Serotypes III (36%), Ia (18%), V (17%), II (11%) and Ib, (9%) and sequence types (ST) 17 (23%), ST-23 (14%), ST-1 (12%) and ST-19 (7%) were most common. Core genome MLST (cgMLST) differentiated isolates of the same ST, grouped STs into five lineages congruent with known clonal complexes and identified known mother-baby pairs and suspected linked infant cases. Clonal complex (CC) 17 accounted for 40% and 22% of infant and maternal invasive cases, respectively and 21% of non-invasive isolates. CC23 and CC19 were associated with maternal disease (30%) and carriage (24%), respectively. Erythromycin (26%) and clindamycin (18%) resistance increased over the study period and was associated with presence of the erm(B) gene (55%), CC1 (33%) and CC19 (24%). A multi-resistant integrative conjugative element incorporated in the PI-1 locus was detected in CC17, an ST-12 and ST-23 isolate confirming the global dissemination of this element. All isolates possessed one or more pilus islands. Genes encoding other potential protective proteins including Sip, C5a peptidase and Srr1 were present in 100%, 99.5% and 65.8% of isolates, respectively. The srr2 gene was unique to CC17. The PubMLST.org website provides a valuable framework for genomic GBS surveillance to inform on local and global GBS epidemiology, preventive and control measures.


Tumour necrosis factor alpha gene polymorphisms in women with recurrent pregnancy loss: a meta-analysis.
Tumour necrosis factor alpha (TNF-α) concentration is associated with recurrent pregnancy loss (RPL). Moreover, TNF-α promoter polymorphisms affect cellular TNF-α concentrations. Although several studies on TNF-α polymorphisms have been reported, these studies only define specific loci and result in conflicting conclusions. The meta-analysis in this study was performed to examine the relationship between various TNF-α polymorphisms and RPL. We searched for articles, using MEDLINE and Embase, and performed meta-analysis of 21 studies involving 3437 cases and 4016 controls. The results demonstrated that the -308G/A polymorphism is positively associated with RPL, particularly in cases involving three or more miscarriages. For -1031T/C, -863C/A, and -376G/A polymorphisms, recessive and homozygote models revealed significant associations with RPL. However, -857C/T, -238G/A, and +488G/A polymorphisms showed no association with RPL. A subgroup analysis with respect to ethnicity demonstrated that the -308G/A and -238G/A polymorphisms are associated with RPL in Asian and Middle Eastern populations, respectively. This meta-analysis showed the associations between TNF-α polymorphisms and RPL. However, further studies of genetic polymorphisms in TNF-α other than -308G/A and -238G/A are required.


Carbohydrate-lectin recognition of sequence-defined heteromultivalent glycooligomers.
Multivalency as a key principle in nature has been successfully adopted for the design and synthesis of artificial glycoligands by attaching multiple copies of monosaccharides to a synthetic scaffold. Besides their potential in various applied areas, e.g. as antiviral drugs, for the vaccine development and as novel biosensors, such glycomimetics also allow for a deeper understanding of the fundamental aspects of multivalent binding of both artificial and natural ligands. However, most glycomimetics so far neglect the purposeful arranged heterogeneity of their natural counterparts, thus limiting more detailed insights into the design and synthesis of novel glycomimetics. Therefore, this work presents the synthesis of monodisperse glycooligomers carrying different sugar ligands at well-defined positions along the backbone using for the first time sequential click chemistry and stepwise assembly of functional building blocks on solid support. This approach allows for straightforward access to sequence-defined, multivalent glycooligomers with full control over number, spacing, position, and type of sugar ligand. We demonstrate the synthesis of a set of heteromultivalent oligomers presenting mannose, galactose, and glucose residues. All heteromultivalent structures show surprisingly high affinities toward Concanavalin A lectin receptor in comparison to their homomultivalent analogues presenting the same number of binding ligands. Detailed studies of the ligand/receptor interaction using STD-NMR and 2fFCS indeed indicate a change in binding mechanism for trivalent glycooligomers presenting mannose or combinations of mannose and galactose residues. We find that galactose residues do not participate in the binding to the receptor, but they promote steric shielding of the heteromultivalent glycoligands and thus result in an overall increase in affinity. Furthermore, the introduction of nonbinding ligands seems to suppress receptor clustering of multivalent ligands. Overall these results support the importance of heteromultivalency specifically for the design of novel glycoligands and help to promote a fundamental understanding of multivalent binding modes.


Towards an anti-fibrotic therapy for scleroderma: targeting myofibroblast differentiation and recruitment.
In response to normal tissue injury, fibroblasts migrate into the wound where they synthesize and remodel new extracellular matrix. The fibroblast responsible for this process is called the myofibroblast, which expresses the highly contractile protein alpha-smooth muscle actin (alpha-SMA). In normal tissue repair, the myofibroblast disappears. Conversely, abnormal myofibroblast persistence is a key feature of fibrotic dieases, including scleroderma (systemic sclerosis, SSc). Myofibroblasts can be derived from differentiation of local resident fibroblasts or by recruitment of microvascular pericytes. Controlling myofibroblast differentiation and persistence is crucial for developing anti-fibrotic therapies targeting SSc. Insights have been recently generated into how the proteins transforming growth factor beta (TGFbeta), endothelin-1 (ET-1), connective tissue growth factor (CCN2/CTGF) and platelet derived growth factor (PDGF) contribute to myofibroblast differentiation and pericyte recruitment in general and to the persistent myofibroblast phenotype of lesional SSc fibroblast, specifically. This minireview summarizes recent findings pertinent to the origin of myofibroblasts in SSc and how this knowledge might be used to control the fibrosis in this disease. TGFbeta, ET-1, CCN2 and PDGF are likely to cooperate in driving tissue repair and fibrogenic responses in fibroblasts. TGFbeta, ET-1 and CCN2 appear to contribute to myofibroblast differentiation; PDGF appears to be involved with pericyte recruitment. Thus, different therapeutic strategies may exist for targeting the multisystem fibrotic disorder SSc.


Beyond frequency: who is most bothered by vasomotor symptoms?
Most menopausal women report vasomotor symptoms (hot flashes, night sweats). However, not all women with vasomotor symptoms, including frequent symptoms, are bothered by them. The primary aim was to identify correlates of vasomotor symptom bother beyond symptom frequency. The Study of Women's Health Across the Nation participants reporting vasomotor symptoms at annual visit 7 comprised the sample (N = 1,042). Assessments included hot flash and night sweats frequency (number per week) and bother (1, not at all- 4, very much). Negative affect (index of depressive symptoms, anxiety, perceived stress, negative mood), symptom sensitivity, sleep problems, and vasomotor symptom duration (number of years) were examined cross-sectionally in relation to bother in ordinal logistic regression models with symptom frequency and covariates. Hot flashes and night sweats were considered separately. In multivariable models controlling for hot flash frequency, negative affect (odds ratio [OR] = 1.27, 95% CI: 1.08-1.51), symptom sensitivity (OR = 1.18, 95% CI: 1.03-1.37), sleep problems (OR = 1.38, 95% CI: 1.04-1.85), poorer health (OR = 1.24, 95% CI: 1.03-1.48), duration of hot flashes (OR = 1.14, 95% CI: 1.06-1.23), younger age (OR = 0.94, 95% CI: 0.89-0.99), and African American race (vs white, OR = 1.59, 95% CI: 1.12-2.26) were associated with hot flash bother. After controlling for night sweats frequency and covariates, sleep problems (OR = 1.84, 95% CI:1.33-2.55) and night sweats duration (OR = 1.10, 95% CI: 1.02-1.20) were associated with night sweats bother. Beyond frequency, factors associated with bothersome hot flashes include mood, symptom sensitivity, symptom duration, sleep problems, age, and race. Correlates of bothersome night sweats include sleep problems and symptom duration. In addition to reducing frequency, interventions for vasomotor symptoms might consider addressing modifiable factors related to symptom bother.


Analysis of Predictive Equations for Estimating Resting Energy Expenditure in a Large Cohort of Morbidly Obese Patients.
The treatment of obesity requires creating an energy deficit through caloric restriction and physical activity. Energy needs are estimated assessing the resting energy expenditure (REE) that in the clinical practice is estimated using predictive equations. In the present cross sectional study, we compared, in a large cohort of morbidly obese patients, the accuracy of REE predictive equations recommended by current obesity guidelines [Harris-Benedict, WHO/FAO/ONU and Mifflin-St Jeor (MJ)] and/or developed for obese patients (Muller, Muller BC, Lazzer, Lazzer BC), focusing on the effect of comorbidities on the accuracy of the equations. Data on REE measured by indirect calorimetry and body composition were collected in 4,247 obese patients (69% women, mean age 48 ± 19 years, mean BMI 44 ± 7 Kg/m2) admitted to the Istituto Auxologico Italiano from 1999 to 2014. The performance of the equations was assessed in the whole cohort, in 4 groups with 0, 1, 2, or ≥ 3 comorbidities and in a subgroup of 1,598 patients with 1 comorbidity (47.1% hypertension, 16.7% psychiatric disorders, 13.3% binge eating disorders, 6.1% endocrine disorders, 6.4% type 2 diabetes, 3.5% sleep apnoea, 3.1% dyslipidemia, 2.5% coronary disease). In the whole cohort of obese patients, as well as in each stratum of comorbidity number, the MJ equation had the highest performance for agreement measures and bias. The MJ equation had the best performance in obese patients with ≥3 comorbidities (accuracy of 61.1%, bias of -89.87) and in patients with type 2 diabetes and sleep apnoea (accuracy/bias 69%/-19.17 and 66%/-21.67 respectively), who also have the highest levels of measured REE. In conclusion, MJ equation should be preferred to other equations to estimate the energy needs of Caucasian morbidly obese patients when measurement of the REE cannot be performed. As even MJ equation does not precisely predict REE, it should be better to plan the diet intervention by measuring rather than estimating REE. Future studies focusing on the clinical differences that determine the high inter-individual variability of the precision of the REE predictive equations (e.g., on the organ-tissue metabolic rate), could help to develop predictive equations with a better performance.


Volumetric optoacoustic tomography enables non-invasive in vivo characterization of impaired heart function in hypoxic conditions.
Exposure to chronic hypoxia results in pulmonary hypertension characterized by increased vascular resistance and pulmonary vascular remodeling, changes in functional parameters of the pulmonary vasculature, and right ventricular hypertrophy, which can eventually lead to right heart failure. The underlying mechanisms of hypoxia-induced pulmonary hypertension have still not been fully elucidated while no curative treatment is currently available. Commonly employed pre-clinical analytic methods are largely limited to invasive studies interfering with cardiac tissue or otherwise ex vivo functional studies and histopathology. In this work, we suggest volumetric optoacoustic tomography (VOT) for non-invasive assessment of heart function in response to chronic hypoxia. Mice exposed for 3 consecutive weeks to normoxia or chronic hypoxia were imaged in vivo with heart perfusion tracked by VOT using indocyanide green contrast agent at high temporal (100 Hz) and spatial (200 µm) resolutions in 3D. Unequivocal difference in the pulmonary transit time was revealed between the hypoxic and normoxic conditions concomitant with the presence of pulmonary vascular remodeling within hypoxic models. Furthermore, a beat-to-beat analysis of the volumetric image data enabled identifying and characterizing arrhythmic events in mice exposed to chronic hypoxia. The newly introduced non-invasive methodology for analysis of impaired pulmonary vasculature and heart function under chronic hypoxic exposure provides important inputs into development of early diagnosis and treatment strategies in pulmonary hypertension.