Spectral contrast imaging microscopy.
We introduce a new technique, spectral contrast imaging microscopy (SCIM), for super-resolution microscopic imaging. Based on a novel contrast mechanism that encodes each local spatial frequency with a corresponding optical wavelength, SCIM provides a real-time high-resolution spectral contrast microscopic image with superior contrast. We show that two microscopic objects, separated by a distance smaller than the diffraction limit of the optical system, can be spatially resolved in the SCIM image as different colors. Results with numerical simulation and experiments using a high-resolution United States Air Force target are presented. The ability of SCIM for imaging biological cells is also demonstrated.


Assessment of risks of pulmonary infection during 12 months following immunosuppressive treatment for active connective tissue diseases: a large-scale prospective cohort study.
Pulmonary infections (PI) are leading causes of death in patients with connective tissue diseases (CTD). The PREVENT study (Pulmonary infections in patients REceiving immunosuppressiVE treatmeNT for CTD) assessed risk of PI in patients with active CTD in the contemporary era of advanced immunosuppressive therapy. In patients who started corticosteroids (n = 763), conventional immunosuppressants or biologics for active CTD were enrolled. Clinical and laboratory data, usage of drugs, and occurrence of PI were collected for 12 months. Baseline risk factors were investigated using Cox regression analysis. A nested case-control (NCC) study was performed with 1:2 matched case-control pairs to assess the risk for each drug category. During the observation period, 32 patients died (4.2%) and 66 patients were lost to followup (8.6%). Patients with PI (n = 61, 8%) had a significantly worse accumulated survival rate than patients without (p < 0.01). Cox hazard regression analysis using baseline data showed that these factors were significantly associated with PI: age ≥ 65 years (HR 3.87, 95% CI 2.22-6.74), ≥ 20 pack-years of smoking (2.63, 1.37-5.04), higher serum creatinine level (1.21, 1.05-1.41 per 1.0 mg/dl increase), and maximum prednisolone (PSL) dose during the first 2 weeks of treatment (2.81, 1.35-5.86 per 1.0 mg/kg/day increase). Logistic regression analysis by an NCC study revealed that maximum PSL dose within 14 days before PI (OR 4.82, 95% CI 1.36-17.01 per 1.0 mg/dl increase; 2.57, 1.28-5.16 if ≥ 0.5 mg/kg/day) was significantly associated with the events, while other immunosuppressants were not. Physicians should be aware of the higher risks for corticosteroids of PI than other immunosuppressants and assess these risk factors before immunosuppressive treatment, to prevent PI.


Coevolutionary networks with homophily and heterophily.
We have investigated a simple coevolutionary network model incorporating three processes-changes of opinions, homophily, and heterophily. In this model, each node holds one of G opinions and changes its opinion, as in the voter model. Homophily is the tendency for connections to form between individuals of the same opinions and heterophily is the opposite effect. If there is no heterophily, this model corresponds to the Holme and Newman model [Phys. Rev. E 74, 056108 (2006)]. We show that the behavior of this model without heterophily can be understood in terms of a mean field approximation. We also find that this model with heterophily exhibits topologically complicated behaviors such as the small-world property.


Is vagal stimulation or inhibition benefit on the regulation of the stomach brain axis in obesity?
Objective: Possible effects of the vagus inhibition and stimulation on the hypothalamic nuclei, myenteric plexes and the vagus nerve were investigated.Methods: The female rats divided to the inhibition (INH), stimulation (STI) and, sham (SHAM) groups were fed with high fat diet (including 40% of energy from animal fat). After nine weeks, the rats were allowed to recover for 4 weeks in INH group. In STI group, the left vagus nerve stimulated (30 Hz/500 msn/30 sec.) starting 2nd post operative day for 5 minutes during 4 weeks. Healthy female rats used as control (CONT). Then, tissue samples were analyzed by biochemical, histological and stereological methods.Results: The mean number of the neurons in the arcuate nucleus of the INH group was significantly less; but, that is significantly more in the STI group compared to the other groups. The neuronal density of ventromedial nucleus in the STI group was higher; while the density in the INH group was lower than the other groups. In the dorsomedial nucleus, neuron density of the INH group was lower than the other groups. In terms of the myenteric plexus volumes, that of the INH group was lowest. The myelinated axon number in the INH group was significantly highest. The myelin sheath thickness and axon area of the INH group was significantly lower than the other groups.Discussion: The results of the study show that the vagal inhibition is more effective than the vagal stimulation on the weight loss in the obesity.


Maier-Saupe-type theory of ferroelectric nanoparticles in nematic liquid crystals.
Several experiments have reported that ferroelectric nanoparticles have drastic effects on nematic liquid crystals-increasing the isotropic-nematic transition temperature by about 5 K, and greatly increasing the sensitivity to applied electric fields. In a recent paper [Lopatina and Selinger, Phys. Rev. Lett. 102, 197802 (2009)], we modeled these effects through a Landau theory, based on coupled orientational order parameters for the liquid crystal and the nanoparticles. This model has one important limitation: Like all Landau theories, it involves an expansion of the free energy in powers of the order parameters, and hence it overestimates the order parameters that occur in the low-temperature phase. For that reason, we now develop a new Maier-Saupe-type model, which explicitly shows the low-temperature saturation of the order parameters. This model reduces to the Landau theory in the limit of high temperature or weak coupling, but shows different behavior in the opposite limit. We compare these calculations with experimental results on ferroelectric nanoparticles in liquid crystals.


Assessment of extradural degenerative disease with Gd-DTPA-enhanced MR imaging: correlation with surgical and pathologic findings.
To test whether gadolinium-DTPA-enhanced MR would increase the conspicuity of extradural degenerative disease in the previously unoperated patient, we prospectively studied a group of 30 patients with symptoms suggestive of disk disease. Surgical findings and pathologic correlations were used as an objective measure of accuracy. Gadolinium-DTPA increased the confidence of diagnosis at one of eight operated cervical levels (six patients) and changed the diagnosis from extradural degenerative disease to tumor in one patient. The mechanism of enhancement of the epidural space and peridiskal region appears to be related to accumulation of contrast material within the epidural venous plexus, as well as to epidural fibrosis associated with disk disruption and herniation. While the immediate clinical utility of gadolinium-DTPA for morphologic analysis seems limited to difficult cervical spine cases, the presence of enhancement as a marker of epidural fibrosis and attempted healing may prove to be of great importance in studying the natural history and basic pathophysiology of degenerative disk disease.


A structure-specific nucleic acid-binding domain conserved among DNA repair proteins.
SMARCAL1, a DNA remodeling protein fundamental to genome integrity during replication, is the only gene associated with the developmental disorder Schimke immuno-osseous dysplasia (SIOD). SMARCAL1-deficient cells show collapsed replication forks, S-phase cell cycle arrest, increased chromosomal breaks, hypersensitivity to genotoxic agents, and chromosomal instability. The SMARCAL1 catalytic domain (SMARCAL1(CD)) is composed of an SNF2-type double-stranded DNA motor ATPase fused to a HARP domain of unknown function. The mechanisms by which SMARCAL1 and other DNA translocases repair replication forks are poorly understood, in part because of a lack of structural information on the domains outside of the common ATPase motor. In the present work, we determined the crystal structure of the SMARCAL1 HARP domain and examined its conformation and assembly in solution by small angle X-ray scattering. We report that this domain is conserved with the DNA mismatch and damage recognition domains of MutS/MSH and NER helicase XPB, respectively, as well as with the putative DNA specificity motif of the T4 phage fork regression protein UvsW. Loss of UvsW fork regression activity by deletion of this domain was rescued by its replacement with HARP, establishing the importance of this domain in UvsW and demonstrating a functional complementarity between these structurally homologous domains. Mutation of predicted DNA-binding residues in HARP dramatically reduced fork binding and regression activities of SMARCAL1(CD). Thus, this work has uncovered a conserved substrate recognition domain in DNA repair enzymes that couples ATP-hydrolysis to remodeling of a variety of DNA structures, and provides insight into this domain's role in replication fork stability and genome integrity.


Relationship of body fat distribution with cardiovascular risk factors in healthy Chinese.
The relationships between six body girths (shoulder, midarm, waist, hip, thigh, and calf) and cardiovascular risk factors (systolic and diastolic blood pressures and glucose, triglyceride, lipoprotein cholesterol, and apolipoprotein levels) were examined in 407 healthy Chinese urban workers in Taipei, Taiwan who were between 40 to 59 years old. Canonical correlation analysis revealed significant associations of upper body adiposity (shoulder, midarm, and waist girths) with cardiovascular risk factors in all subgroups assessed: men, premenopausal women, and postmenopausal women. Waist girth and hip girth were consistent and important variables, and weighted in the opposite direction. Waist-hip ratio (WHR) was the best descriptor of centralized adiposity. Centralized fat distribution was positively associated with blood pressure and glucose, triglyceride, and apolipoprotein (apo) B levels, and negatively associated with high-density-lipoprotein (HDL) cholesterol and apo A-I levels in this population. Body fat distribution had an effect independent of body mass index and accounted for some of the differences in triglyceride, HDL cholesterol, apo A-I, and apo B concentrations among men, premenopausal women, and postmenopausal women. Our findings in a Chinese population are similar to data from other studies in Western populations, and are consistent with the hypothesis that centralized adiposity is related to cardiovascular risk factors independent of general obesity.


Typologies in GPs' referral practice.
GPs' individual decisions to refer and the various ways of working when they refer are important determinants of secondary care use. The objective of this study was to explore and describe potential characteristics of GPs' referral practice by investigating their opinions about referring and their self-reported experiences of what they do when they refer. Observational cross-sectional study using data from 128 Norwegian GPs who filled in a questionnaire with statements on how they regarded the referral process, and who were invited to collect data when they actually referred to hospital during one month. Only elective referrals were recorded. The 57 participants (44,5 %) recorded data from 691 referrals. The variables were included in a principal component analysis. A multiple linear regression analysis was conducted to identify typologies with GP's age, gender, specialty in family medicine and location as independent variables. Eight principal components describe the different ways GPs think and work when they refer. Two typologies summarize these components: confidence characterizing specialists in family medicine, mainly female, who reported a more patient-centred practice making priority decisions when they refer, who confer easily with hospital consultants and who complete the referrals during the consultation; uncertainty characterizing young, mainly male non-specialists in family medicine, experiencing patients' pressure to be referred, heavy workload, having reluctance to cooperate with the patient and reporting sparse contact with hospital colleagues. Training specialists in family medicine in patient-centred method, easy conference with hospital consultant and cooperation with patients while making the referral may foster both self-reflections on own competences and increased levels of confidence.


Comparative study of efficacy of l-5-hydroxytryptophan and fluoxetine in patients presenting with first depressive episode.
Role of l-5-hydroxytryptophan (l-5-HTP) in depression is relatively less studied but the literature has shown its robust role in depression. The present randomized double blind study was undertaken to assess the role of l-5-HTP as an antidepressant and to compare its antidepressant efficacy with fluoxetine in first depressive episode patients of Indian population. A total of 70 patients of first depressive episode, all of whom were diagnosed with ICD-10 criteria, were recruited but only 60 patients completed the study and were randomly divided into two groups, receiving l-5-HTP and fluoxetine, respectively, for a period of 8weeks. All patients were administered Hamilton Rating Scale for Depression (HAM-D) to assess severity of depression at baseline, 2weeks, 4weeks and 8weeks. The efficacy of treatment was assessed by comparing HAM-D scores obtained at these examinations with the baseline examination; final evaluation of both efficacy and tolerance was assessed using the Clinical Global Impression (CGI) scale at the end of study. Both treatment groups showed significant and nearly equal reduction in HAM-D scores beginning at week two and continuing through week eight. Twenty-two patients (73.33%) in the l-5-HTP group and 24 patients (80%) in the fluoxetine group showed positive response at the end of the study. l-5-HTP has definitely got antidepressant effect in patients of depression. Antidepressant effect was seen within 2weeks of treatment and was apparent in all degrees of depression. The therapeutic efficacy of l-5-HTP was considered as equal to that of fluoxetine.


Prevalence of intimate partner violence across medical and surgical health care settings: a systematic review.
Intimate partner violence (IPV) is a serious health problem and a leading cause of nonfatal injury in North American females. Prevalence of IPV has ranged from less than 20% to more than 50% across primary care, emergency medicine, and family medicine. We conducted a systematic review and meta-analysis of the literature to examine best estimates of IPV prevalence as opportunities for targeted interventions in health care specialties. We included 37 articles in this study. Based on our pooled data, best estimates of the lifetime prevalence of any type of IPV were 38% in family medicine and 40% in emergency medicine.


Table olives from Portugal: phenolic compounds, antioxidant potential, and antimicrobial activity.
The phenolic compounds composition, antioxidant potential, and antimicrobial activity of different table olives from Portugal, namely, natural black olives "Galega", black ripe olive "Negrinha de Freixo", Protected Designation of Origin (PDO) "Azeitona de Conserva Negrinha de Freixo" olives, and "Azeitona de Conserva de Elvas e Campo Maior" Designation of Origin (DO) olives, were investigated. The analysis of phenolic compounds was performed by reversed-phase HPLC/DAD, and seven compounds were identified and quantified: hydroxytyrosol, tyrosol, 5-O-caffeoilquinic acid, verbascoside, luteolin 7-O-glucoside, rutin, and luteolin. The antioxidant activity was assessed by the reducing power assay, the scavenging effect on DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals, and the beta-carotene linoleate model system. The antioxidant activity was correlated with the amount of phenolics found in each sample. The antimicrobial activity was screened using Gram-positive (Bacillus cereus, Bacillus subtilis, Staphylococcus aureus) and Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae) and fungi (Candida albicans, Cryptococcus neoformans). PDO and DO table olives revealed a wide range of antimicrobial activity. C. albicans was resistant to all the analyzed extracts.


Pregnant women living with HIV (WLH) supported at clinics by peer WLH: a cluster randomized controlled trial.
Throughout Africa, Peer Mentors who are women living with HIV (WLH) are supporting pregnant WLH at antenatal and primary healthcare clinics (McColl in BMJ 344:e1590, 2012). We evaluate a program using this intervention strategy at 1.5 months post-birth. In a cluster randomized controlled trial in KwaZulu-Natal, South Africa, eight clinics were randomized for their WLH to receive either: standard care (SC), based on national guidelines to prevent mother-to-child transmission (4 clinics; n = 656 WLH); or an enhanced intervention (EI; 4 clinics; n = 544 WLH). The EI consisted of four antenatal and four postnatal small group sessions led by Peer Mentors, in addition to SC. WLH were recruited during pregnancy and 70 % were reassessed at 1.5 months post-birth. EI's effect was ascertained on 16 measures of maternal and infant well-being using random effects regressions to control for clinic clustering. A binomial test for correlated outcomes evaluated EI's overall effectiveness. Among EI WLH reassessed, 87 % attended at least one intervention session (mean 4.1, SD 2.0). Significant overall benefits were found in EI compared to SC using the binomial test. However, it is important to note that EI WLH were significantly less likely to adhere to ARV during pregnancy compared to SC. Secondarily, compared to SC, EI WLH were more likely to ask partners to test for HIV, better protected their infants from HIV transmission, and were less likely to have depressed mood and stunted infants. Adherence to clinic intervention groups was low, yet, there were benefits for maternal and infant health at 1.5 months post-birth.


Anti-hyperliposis effect of maitake fruit body (Grifola frondosa). I.
Experimental rat models (5-week-old Sprague-Dawley rats) with hyperlipemia were prepared by feeding high-cholesterol feed containing sodium cholate and casein as a protein source. Dried maitake (Grifola frondosa) powder was mixed with the basic high-cholesterol feed and the serum lipids were periodically measured. Values of cholesterol, triglyceride and phospholipid in serum of rats in the maitake-feed group were suppressed by 0.3-0.8 times those in animals fed the basic feed, the latter values being close to those in rats given normal feed. The value of high density lipoprotein (HDL)-cholesterol in serum which is generally reduced by the ingestion of high-cholesterol feed remained the level it was at the beginning of the experiment. Weights of extirpated liver and epididymal fat-pads were significantly less (0.6-0.7 times) than those in the basic feed group, indicating that maitake inhibits lipid accumulation in the body. Liver lipids were also measured and the values were found to be decreased by maitake administration as true of serum lipid, suggesting maitake has an anti-liver lipid activity. Measurement of the amount of total cholesterol and bile acid in feces showed, the ratio of cholesterol-excretion had increased 1.8 times and bile acid-excretion 3 fold by maitake treatment. From these results, it is believed that maitake helps to improve the lipid metabolism as it inhibits both liver lipid and serum lipid which are increased by the ingestion of high-fat feed.


Prediction of celiac disease at endoscopy.
Celiac disease is increasingly recognized worldwide, but guidelines on how to detect the condition and diagnose patients are unclear. In this study the prevalence and predictors of celiac disease were prospectively determined in a cross-sectional sample of Lebanese patients undergoing esophagogastroduodenoscopy (EGD). Consecutive consenting patients (n = 999) undergoing EGD answered a questionnaire and had blood taken for serologic testing. Endoscopic markers for celiac disease were documented and duodenal biopsies were obtained. The diagnosis of celiac disease was based on abnormal duodenal histology and positive serology. Risk factors were used to classify patients to either high or low risk for celiac disease. Independent predictors of celiac disease were derived via multivariate logistic regression. Villous atrophy (Marsh 3) and celiac disease were present in 1.8 % and 1.5 % of patients, respectively. Most were missed on clinical and endoscopic grounds. The sensitivity of tissue transglutaminase (tTG) testing for the diagnosis of villous atrophy and celiac disease was 72.2 % and 86.7 %, respectively. The positive predictive value of the deamidated gliadin peptide (DGP) test was 34.2 % and that of a strongly positive tTG was 80 %. While the strongest predictor of celiac disease was a positive tTG (odds ratio [OR] 131.7, 95 % confidence interval [CI] 29.0 - 598.6), endoscopic features of villous atrophy (OR 64.8, 95 %CI 10.7 - 391.3), history of eczema (OR 4.6, 95 %CI 0.8 - 28.8), anemia (OR 6.7, 95 %CI 1.2 - 38.4), and being Shiite (OR 5.4, 95 %CI 1.1 - 26.6) significantly predicted celiac disease. A strategy of biopsying the duodenum based on independent predictors had a sensitivity of 93 % - 100 % for the diagnosis of celiac disease, with an acceptable (22 % - 26 %) rate of performing unnecessary biopsies. A strategy that excluded pre-EGD serology produced a sensitivity of 93 % - 94 % and an unnecessary biopsy rate of 52 %. An approach based solely on standard clinical suspicion and endoscopic findings is associated with a significant miss rate for celiac disease. A strategy to biopsy based on the derived celiac disease prediction models using easily obtained information prior to or during endoscopy, maximized the diagnosis while minimizing unnecessary biopsies.


[Work days lost due to health problems in industry].
This cross-sectional study estimated the prevalence of work days lost due to health problems and associated factors among industrial workers. The study population was a simple random cluster sample of 3,403 workers from 16 to 65 years of age in the city of Salvador, Bahia State, Brazil. Data were collected with individual home interviews. Among industrial workers, one-year prevalence of work days lost to health problems was 12.5%, of which 5.5% were directly work-related and 4.1% aggravated by work. There were no statistically significant differences when compared to other worker categories. Self-perceived workplace hazards, history of work-related injury, and poor self-rated health were associated with work days lost due to work-related injuries/diseases. The findings showed that work days lost are common among both industrial and non-industrial workers, thereby affecting productivity and requiring prevention programs.


Electron transfers in a TiO2-containing MOR zeolite: synthesis of the nanoassemblies and application using a probe chromophore molecule.
New assemblies constituted by a microporous matrix of mordenite (MOR) zeolite on which TiO2 nanoclusters are deposited were synthesized using ionic oxalate complexes and TiCl3 titanium precursors. The samples were used to investigate the transfer of electrons produced by spontaneous or photo-induced ionization of a guest molecule (t-stilbene, t-St) occluded in the porous volume towards the conduction band of a conductive material placed nearby, in the pores or at least close to their entrance. The reaction mechanisms were compared in these Ti-rich solids and in a Ti-free mordenite sample. The characterization by XRD, N2 physisorption, TEM, XPS and DRIFT spectroscopy of the supramolecular TiO2/MOR systems before t-St adsorption showed the preservation of the crystalline structure after Ti addition and thermal activation treatments. They also revealed that titanium is mainly located at the external surface of the zeolite grains, in the form of highly dispersed and/or aggregated anatase. After incorporation of the guest molecule in the new assemblies, diffuse reflectance UV-visible and EPR spectroscopies indicate that the electron transfer processes are similar with and without TiO2 but strongly stabilized t-St˙(+) radicals are detected in the TiO2-MOR samples whereas such species were never detected earlier in TiO2-free mordenite using these techniques. The stabilization process is found to be more efficient in the sample prepared with TiCl3 as the precursor than with titanium oxalates. It is proposed that the proximity of TiO2 with the formed t-St˙(+) radicals provokes the stabilization of the radical through capture of the ejected electron by the semi-conductor and that confinement effects can also play a role.


LGBTQ Youth in Unstable Housing and Foster Care.
: media-1vid110.1542/5984244876001PEDS-VA_2017-4211Video Abstract BACKGROUND AND OBJECTIVES: Lesbian, gay, bisexual, transgender, and questioning (LGBTQ) youth are suggested to be overrepresented in unstable housing and foster care. In the current study, we assess whether LGBTQ youth are overrepresented in unstable housing and foster care and examine disparities in school functioning, substance use, and mental health for LGBTQ youth versus heterosexual youth in unstable housing and foster care. A total of 895 218 students (10-18 years old) completed the cross-sectional California Healthy Kids Survey from 2013 to 2015. Surveys were administered in 2641 middle and high schools throughout California. Primary outcome measures included school functioning (eg, school climate, absenteeism), substance use, and mental health. More youth living in foster care (30.4%) and unstable housing (25.3%) self-identified as LGBTQ than youth in a nationally representative sample (11.2%). Compared with heterosexual youth and youth in stable housing, LGBTQ youth in unstable housing reported poorer school functioning (Bs = -0.10 to 0.40), higher substance use (Bs = 0.26-0.28), and poorer mental health (odds ratios = 0.73-0.80). LGBTQ youth in foster care reported more fights in school (B = 0.16), victimization (B = 0.10), and mental health problems (odds ratios = 0.82-0.73) compared with LGBTQ youth in stable housing and heterosexual youth in foster care. Disparities for LGBTQ youth are exacerbated when they live in foster care or unstable housing. This points to a need for protections for LGBTQ youth in care and care that is affirming of their sexual orientation and gender identity.


A UGT2B17-positive donor is a risk factor for higher transplant-related mortality and lower survival after bone marrow transplantation.
We recently identified a human minor histocompatibility (H) antigen, encoded by UDP glycosyltransferase 2 family, polypeptide B17 (UGT2B17), whose immunogenicity results from differential expression in donor and recipient cells as a consequence of a homozygous deletion of the UGT2B17 gene. UGT2B17 is highly expressed in the liver and colon, which are major targets for graft-versus-host disease (GVHD). To assess the significance of homozygous UGT2B17 gene deletion in allogeneic haematopoietic stem cell transplantation (HSCT), we analysed DNA from 435 stem cell transplant recipients with a haematological malignancy and their human leucocyte antigen-identical unrelated bone marrow donors using sequence-specific primer polymerase chain reaction. Homozygous deletion of the UGT2B17 gene was observed in 85% of normal donors and in 82% of patients. The analysis showed no significant association between UGT2B17 mismatch in the GVHD direction and the incidence of acute GVHD, chronic GVHD, relapse, or survival. However, the use of a UGT2B17-positive donor was an independent risk factor for higher transplant-related mortality and lower survival after transplantation. UGT2B17 is a metabolic enzyme for hormones, drugs, and potentially toxic exogenous compounds and is expressed in subsets of haematopoietic cells. Thus, the enzyme function of UGT2B17 in donor cells may affect the outcome of allogeneic HSCT.


Mother-offspring and nest-mate resemblance but no heritability in early-life telomere length in white-throated dippers.
Telomeres are protective DNA-protein complexes located at the ends of eukaryotic chromosomes, whose length has been shown to predict life-history parameters in various species. Although this suggests that telomere length is subject to natural selection, its evolutionary dynamics crucially depends on its heritability. Using pedigree data for a population of white-throated dippers (Cinclus cinclus), we test whether and how variation in early-life relative telomere length (RTL, measured as the number of telomeric repeats relative to a control gene using qPCR) is transmitted across generations. We disentangle the relative effects of genes and environment and test for sex-specific patterns of inheritance. There was strong and significant resemblance among offspring sharing the same nest and offspring of the same cohort. Furthermore, although offspring resemble their mother, and there is some indication for an effect of inbreeding, additive genetic variance and heritability are close to zero. We find no evidence for a role of either maternal imprinting or Z-linked inheritance in generating these patterns, suggesting they are due to non-genetic maternal and common environment effects instead. We conclude that in this wild bird population, environmental factors are the main drivers of variation in early-life RTL, which will severely bias estimates of heritability when not modelled explicitly.


Appraisal of Meconium at Delivery: A look at intervention strategies.
A critical appraisal of the scientific literature on managing mesconium in labor identified 15 studies which were used to evaluate intervention strategies. Only four were randomized trials: two on the use of amnioinfusion in labor, one on the technique of bulb versus DeLee catheter suction of the newborn, and one on the need for endotracheal intubation and suction in meconium-stained neonates. Current practice is dictated by the most favorable tradeoff between benefit and risk because of limited scientific evidence.


Aberrant repair initiated by the adenine-DNA glycosylase does not play a role in UV-induced mutagenesis in Escherichia coli.
DNA repair is essential to counteract damage to DNA induced by endo- and exogenous factors, to maintain genome stability. However, challenges to the faithful discrimination between damaged and non-damaged DNA strands do exist, such as mismatched pairs between two regular bases resulting from spontaneous deamination of 5-methylcytosine or DNA polymerase errors during replication. To counteract these mutagenic threats to genome stability, cells evolved the mismatch-specific DNA glycosylases that can recognize and remove regular DNA bases in the mismatched DNA duplexes. The Escherichia coli adenine-DNA glycosylase (MutY/MicA) protects cells against oxidative stress-induced mutagenesis by removing adenine which is mispaired with 7,8-dihydro-8-oxoguanine (8oxoG) in the base excision repair pathway. However, MutY does not discriminate between template and newly synthesized DNA strands. Therefore the ability to remove A from 8oxoG•A mispair, which is generated via misincorporation of an 8-oxo-2'-deoxyguanosine-5'-triphosphate precursor during DNA replication and in which A is the template base, can induce A•T→C•G transversions. Furthermore, it has been demonstrated that human MUTYH, homologous to the bacterial MutY, might be involved in the aberrant processing of ultraviolet (UV) induced DNA damage. Here, we investigated the role of MutY in UV-induced mutagenesis in E. coli. MutY was probed on DNA duplexes containing cyclobutane pyrimidine dimers (CPD) and pyrimidine (6-4) pyrimidone photoproduct (6-4PP). UV irradiation of E. coli induces Save Our Souls (SOS) response characterized by increased production of DNA repair enzymes and mutagenesis. To study the role of MutY in vivo, the mutation frequencies to rifampicin-resistant (RifR) after UV irradiation of wild type and mutant E. coli strains were measured. We demonstrated that MutY does not excise Adenine when it is paired with CPD and 6-4PP adducts in duplex DNA. At the same time, MutY excises Adenine in A•G and A•8oxoG mispairs. Interestingly, E. coli mutY strains, which have elevated spontaneous mutation rate, exhibited low mutational induction after UV exposure as compared to MutY-proficient strains. However, sequence analysis of RifR mutants revealed that the frequencies of C→T transitions dramatically increased after UV irradiation in both MutY-proficient and -deficient E. coli strains. These findings indicate that the bacterial MutY is not involved in the aberrant DNA repair of UV-induced DNA damage.


Composition of the adult digestive tract bacterial microbiome based on seven mouth surfaces, tonsils, throat and stool samples.
To understand the relationship between our bacterial microbiome and health, it is essential to define the microbiome in the absence of disease. The digestive tract includes diverse habitats and hosts the human body's greatest bacterial density. We describe the bacterial community composition of ten digestive tract sites from more than 200 normal adults enrolled in the Human Microbiome Project, and metagenomically determined metabolic potentials of four representative sites. The microbiota of these diverse habitats formed four groups based on similar community compositions: buccal mucosa, keratinized gingiva, hard palate; saliva, tongue, tonsils, throat; sub- and supra-gingival plaques; and stool. Phyla initially identified from environmental samples were detected throughout this population, primarily TM7, SR1, and Synergistetes. Genera with pathogenic members were well-represented among this disease-free cohort. Tooth-associated communities were distinct, but not entirely dissimilar, from other oral surfaces. The Porphyromonadaceae, Veillonellaceae and Lachnospiraceae families were common to all sites, but the distributions of their genera varied significantly. Most metabolic processes were distributed widely throughout the digestive tract microbiota, with variations in metagenomic abundance between body habitats. These included shifts in sugar transporter types between the supragingival plaque, other oral surfaces, and stool; hydrogen and hydrogen sulfide production were also differentially distributed. The microbiomes of ten digestive tract sites separated into four types based on composition. A core set of metabolic pathways was present across these diverse digestive tract habitats. These data provide a critical baseline for future studies investigating local and systemic diseases affecting human health.


Vitamin D3 and its nuclear receptor increase the expression and activity of the human proton-coupled folate transporter.
Folates are essential for nucleic acid synthesis and are particularly required in rapidly proliferating tissues, such as intestinal epithelium and hemopoietic cells. Availability of dietary folates is determined by their absorption across the intestinal epithelium, mediated by the proton-coupled folate transporter (PCFT) at the apical enterocyte membranes. Whereas transport properties of PCFT are well characterized, regulation of PCFT gene expression remains less elucidated. We have studied the mechanisms that regulate PCFT promoter activity and expression in intestine-derived cells. PCFT mRNA levels are increased in Caco-2 cells treated with 1,25-dihydroxyvitamin D(3) (vitamin D(3)) in a dose-dependent fashion, and the duodenal rat Pcft mRNA expression is induced by vitamin D(3) ex vivo. The PCFT promoter region is transactivated by the vitamin D receptor (VDR) and its heterodimeric partner retinoid X receptor-alpha (RXRalpha) in the presence of vitamin D(3). In silico analyses predicted a VDR response element (VDRE) in the PCFT promoter region -1694/-1680. DNA binding assays showed direct and specific binding of the VDR:RXRalpha heterodimer to the PCFT(-1694/-1680), and chromatin immunoprecipitations verified that this interaction occurs within living cells. Mutational promoter analyses confirmed that the PCFT(-1694/-1680) motif mediates a transcriptional response to vitamin D(3). In functional support of this regulatory mechanism, treatment with vitamin D(3) significantly increased the uptake of [(3)H]folic acid into Caco-2 cells at pH 5.5. In conclusion, vitamin D(3) and VDR increase intestinal PCFT expression, resulting in enhanced cellular folate uptake. Pharmacological treatment of patients with vitamin D(3) may have the added therapeutic benefit of enhancing the intestinal absorption of folates.


Relationship of ketamine's antidepressant and psychotomimetic effects in unipolar depression.
Ketamine and other NMDA (N-methyl-D-aspartate) antagonists produce fast-acting antidepressant-like effects, although the underlying mechanism is unclear. Furthermore, high affinity NMDA antagonists such as ketamine are associated with psychotomimetic effects. To date the link between the antidepressant and psychotomimetic effects of ketamine has not been explored. We examined the relationship between the antidepressant and psychotomimetic effects of a single ketamine infusion in subjects diagnosed with major depressive disorder. In a double-blind, cross-over, placebo-controlled, two weeks clinical trial we studied the effects of ketamine (0.54 mg/kg within 30 min) in a group of 27 hospitalized depressive patients. Higher intensity of psychotomimetic symptoms, measured using BPRS, during ketamine administration correlated with alleviation in mood ratings during the following week with maximum on day seven. Ketamine was superior to placebo in all visits (day 1, 4, and 7) assessed by MADRS with effect size (Cohen´s d) of 0.62, 0.57, and 0.44 respectively. There was no significant correlation between ketamine and nor-ketamine plasma levels and MADRS score change at any study time point. The substantial relationship between ketamine's antidepressant and psychotomimetic effects was found. This relationship could be mediated by the initial steps of ketamine's action, trough NMDA receptors, shared by both ketamine's clinical effects.


Effect of Non-invasive Spinal Stimulation on Self-sustained Firing Motoneuron Model: In-Silico Study Using Human Body Model.
Transcutaneous spinal electrical stimulation (tSCS) is a non-invasive neuromodulation approach using a low intensity direct current. Recent developments in the technique have opened the possibility that tSCS can help restore motor function after spinal cord injury (SCI). However, the exact mechanism of action tSCS has on the spinal circuits is still unknown. Due to the complexity of experimental synthesis in a human model to delineate the mechanisms, models that link the stimulation paradigm and circuit behaviors are advantageous. Thus, this study aims to simulate the underlying changes in motor circuit firing rates in response to external stimuli induced by tSCS. Serial stimulations combining a high-fidelity finite element model with the human torso and spinal cord with a lumped motor neuron model is constructed. The parameters for both components of the model were derived from previous studies. We focused our analysis on a lumped motor neuron model that describes sustained firing behavior of the motor neuron driven primarily by persistent inward current (PIC), a signature behavior of the motor neuron after SCI. Modulation of the PIC behaviors was achieved by stimulating voltage-dependent calcium and sodium channels in the dendrite using a tSCS-induced electric field (E-field) expressed at different a spatial locations of the motor neuron in the gray matter. The PIC behaviors of spinal motor neurons in the left ventral horn were suppressed, while for the most part invariant in the right ventral horn. These initial simulations will provide a steppingstone for future examinations that incorporate additional neuronal models of inhibitory and excitatory interneurons to access the circuit-level effect of spinal stimulation.


Signaling effectors underlying pathologic growth and remodeling of the heart.
Cardiovascular disease is the number one cause of mortality in the Western world. The heart responds to many cardiopathological conditions with hypertrophic growth by enlarging individual myocytes to augment cardiac pump function and decrease ventricular wall tension. Initially, such cardiac hypertrophic growth is often compensatory, but as time progresses these changes become maladaptive. Cardiac hypertrophy is the strongest predictor for the development of heart failure, arrhythmia, and sudden death. Here we discuss therapeutic avenues emerging from molecular and genetic studies of cardiovascular disease in animal models. The majority of these are based on intracellular signaling pathways considered central to pathologic cardiac remodeling and hypertrophy, which then leads to heart failure. We focus our discussion on selected therapeutic targets that have more recently emerged and have a tangible translational potential given the available pharmacologic agents that could be readily evaluated in human clinical trials.


The von Hippel-Lindau protein sensitizes renal carcinoma cells to apoptotic stimuli through stabilization of BIM(EL).
von Hippel-Lindau (VHL) disease is caused by germ-line mutations in the VHL tumor suppressor gene and is the most common cause of inherited renal cell carcinoma (RCC). Mutations in the VHL gene also occur in a large majority of sporadic cases of clear-cell RCC, which have high intrinsic resistance to chemotherapy and radiotherapy. Here we show that VHL-deficient RCC cells express lower levels of the proapoptotic Bcl-2 family protein BIM(EL) and are more resistant to etoposide and UV radiation-induced death compared to the same cells stably expressing the wild-type VHL protein (pVHL). Reintroducing pVHL into VHL-null cells increased the half-life of BIM(EL) protein without affecting its mRNA expression, and overexpressing pVHL inhibited BIM(EL) polyubiquitination. Suppressing pVHL expression with RNA interference resulted in a decrease in BIM(EL) protein and a corresponding decrease in the sensitivity of RCC cells to apoptotic stimuli. Directly inhibiting BIM(EL) expression in pVHL-expressing RCC cells caused a similar decrease in cell death. These results demonstrate that pVHL acts to promote BIM(EL) protein stability in RCC cells, and that destabilization of BIM(EL) in the absence of pVHL contributes to the increased resistance of VHL-null RCC cells to certain apoptotic stimuli.


Understanding how communities respond to COVID-19: experiences from the Orthodox Jewish communities of Antwerp city.
The importance of community involvement in the response against disease outbreaks has been well established. However, we lack insights into local communities' experiences in coping with the current COVID-19 pandemic. This study explored both the impact of, and response to, COVID-19 within the Orthodox Jewish communities of Antwerp (Belgium) during the first lockdown period (March 2020 - May 2020). We conducted an explorative qualitative study using a participatory approach. First, we performed a community mapping to identify relevant stakeholders. Through the active involvement of a community advisory board and based on qualitative interviews with key-informants and community members, we elicited lived experiences, attitudes, and perceptions towards COVID-19. Interviews were conducted both face-to-face and using online web conferencing technology. Data were analyzed inductively according to the principles of thematic analysis. Government-issued outbreak control measures presented context-specific challenges to the Orthodox Jewish communities in Antwerp. They related mainly to the remote organization of religious life, and practicing physical distancing in socially and culturally strongly connected communities. Existing community resources were rapidly mobilized to adapt to the outbreak and to self-organize response initiatives within communities. The active involvement of community and religious leaders in risk communication proved to be of great importance to facilitate the coverage and uptake of pandemic control measures while protecting essential community values and traditions. Creating bottom-up and community-adapted communication strategies, including addressing language barriers and involving Rabbis in the dissemination of prevention messages, fostered a feeling of trust in government's response measures. However, unmet information and prevention needs were also identified, such as the need for inclusive communication by public authorities and the need to mitigate the negative effects of stigmatization. The experiences of Orthodox Jewish communities in Antwerp demonstrate a valuable example of a feasible community-centered approach to health emergencies. Increasing the engagement of communities in local decision-making and governance structures remains a key strategy to respond to unmet information and prevention needs.


Behavioral Ecology of Captive Species: Using Bibliographic Information to Assess Pet Suitability of Mammal Species.
Which mammal species are suitable to be kept as pet? For answering this question many factors have to be considered. Animals have many adaptations to their natural environment in which they have evolved that may cause adaptation problems and/or risks in captivity. Problems may be visible in behavior, welfare, health, and/or human-animal interaction, resulting, for example, in stereotypies, disease, and fear. A framework is developed in which bibliographic information of mammal species from the wild and captive environment is collected and assessed by three teams of animal scientists. Oneliners from literature about behavioral ecology, health, and welfare and human-animal relationship of 90 mammal species are collected by team 1 in a database and strength of behavioral needs and risks is assessed by team 2. Based on summaries of those strengths the suitability of the mammal species is assessed by team 3. Involvement of stakeholders for supplying bibliographic information and assessments was propagated. Combining the individual and subjective assessments of the scientists using statistical methods makes the final assessment of a rank order of suitability as pet of those species less biased and more objective. The framework is dynamic and produces an initial rank ordered list of the pet suitability of 90 mammal species, methods to add new mammal species to the list or remove animals from the list and a method to incorporate stakeholder assessments. A model is developed that allows for provisional classification of pet suitability. Periodical update of the pet suitability framework is expected to produce an updated list with increased reliability and accuracy. Furthermore, the framework could be further developed to assess the pet suitability of additional species of other animal groups, e.g., birds, reptiles, and amphibians.


Growth Hormone Deficiency: Health and Longevity.
The important role of GH in the control of mammalian longevity was first deduced from extended longevity of mice with genetic GH deficiency (GHD) or GH resistance. Mice with isolated GHD (IGHD) due to GHRH or GHRH receptor mutations, combined deficiency of GH, prolactin, and TSH, or global deletion of GH receptors live longer than do their normal siblings. They also exhibit multiple features of delayed and/or slower aging, accompanied by extension of healthspan. The unexpected, remarkable longevity benefit of severe endocrine defects in these animals presumably represents evolutionarily conserved trade-offs among aging, growth, maturation, fecundity, and the underlying anabolic processes. Importantly, the negative association of GH signaling with longevity extends to other mammalian species, apparently including humans. Data obtained in humans with IGHD type 1B, owing to a mutation of the GHRH receptor gene, in the Itabaianinha County, Brazil, provide a unique opportunity to study the impact of severe reduction in GH signaling on age-related characteristics, health, and functionality. Individuals with IGHD are characterized by proportional short stature, doll facies, high-pitched voices, and central obesity. They have delayed puberty but are fertile and generally healthy. Moreover, these IGHD individuals are partially protected from cancer and some of the common effects of aging and can attain extreme longevity, 103 years of age in one case. We think that low, but detectable, residual GH secretion combined with life-long reduction of circulating IGF-1 and with some tissue levels of IGF-1 and/or IGF-2 preserved may account for the normal longevity and apparent extension of healthspan in these individuals.


Clinical utility of the Vesical Imaging-Reporting and Data System for muscle-invasive bladder cancer between radiologists and urologists based on multiparametric MRI including 3D FSE T2-weighted acquisitions.
To investigate the clinical utility of the Vesical Imaging-Reporting and Data System (VI-RADS) by comparing its diagnostic performance for muscle-invasive bladder cancer (MIBC) between radiologists and urologists based on multiparametric MRI, including three-dimensional (3D) fast spin-echo (FSE) T2-weighted acquisitions. This study included 66 treatment-naïve patients (60 men, 6 women; mean age 74.0 years) with pathologically proven bladder cancer who underwent multiparametric MRI, including 3D FSE T2-weighted imaging, before transurethral bladder tumour resection between January 2010 and November 2018. The MRI scans were categorised according to the five-point VI-RADS score by four independent readers (two board-certified radiologists and board-certified urologists each), blinded to the histopathological findings. The VI-RADS scores were compared with the postoperative histopathological diagnosis. Interobserver agreement was assessed using weighted kappa coefficients. ROC analysis and generalised estimating equations were used to evaluate the diagnostic performance. Forty-nine (74.2%) and 17 (25.8%) tumours were confirmed to be non-MIBC and MIBC, respectively, based on pathological examination. The interobserver agreement was good-to-excellent between all pairs of readers (range, 0.73-0.91). The urologists' sensitivity/specificity values for DCE-MRI VI-RADS scores were significantly lower than those of radiologists. No significant differences were observed for the overall VI-RADS score. The AUC for the overall VI-RADS score was 0.94, 0.92, 0.89, and 0.87 for radiologists 1 and 2 and urologists 1 and 2, respectively. The VI-RADS score, based on multiparametric MRI including 3D FSE T2-weighted acquisitions, can be useful for radiologists and urologists to determine the bladder cancer muscle invasion status preoperatively. • VI-RADS (using multiparametric MRI including 3D FSE T2-weighted acquisitions) achieves good to excellent interobserver agreement and has similar diagnostic performance for detecting muscle invasion by both radiologists and urologists. • The diagnostic performance of the overall VI-RADS score is high for both radiologists and urologists, particularly due to the dominant effect of diffusion-weighted imaging on the overall VI-RADS score. • The sensitivity and specificity values of the T2WI VI-RADS scores for four readers in our study (using 3D FSE T2-weighted acquisitions) were similar (with slightly higher specificity values) to previously published results (using 2D FSE T2-weighted acquisitions).


Elevated right atrial pressure does not reduce collateral blood flow to ischemic myocardium.
Right atrial pressure (RAP) may become substantially elevated during heart failure and has been reported to reduce collateral flow to the ischemic myocardium of isolated hearts. The effect of elevated RAP on blood flow to collateral-dependent and normal myocardium of in situ hearts was studied in 20 open-chest anesthetized dogs with acute occlusion of the left anterior descending coronary artery. Regional myocardial blood flow was measured with radioactive microspheres while RAP was elevated by restricting right ventricular (RV) outflow with constant aortic pressure. Increasing RAP from 3.8 +/- 0.5 to 21.5 +/- 0.8 and then to 34.3 +/- 0.9 mmHg did not reduce blood flow to any transmural region of LV normal or collateral-dependent myocardium. Further elevation of RAP to 49.3 +/- 1.1 mmHg reduced subepicardial but not subendocardial collateral flow. Blood flow to normal RV increased. Retrograde flow and peripheral coronary pressure increased as RAP was elevated. Previously injected 11-micron microspheres were present in the retrograde flow when RAP was elevated; thus retrograde capillary flow contributed to the retrograde flow. The results explain discrepancies among previous reports, and they are consistent with a waterfall phenomenon in the coronary collateral circulation.


Planar tetranuclear lanthanide clusters with the Dy4 analogue displaying slow magnetic relaxation.
Two isostructural tetranuclear lanthanide clusters of general formula [Ln(III)(4)(μ(3)-OH)(2)(o-van)(4)(O(2)CC(CH(3))(3))(4)(NO(3))(2)]·CH(2)Cl(2)·1.5H(2)O (Ln = Gd (1) and Dy (2)) (o-van = 3-methoxysalicylaldehydato anion) are reported. The metallic cores of both complexes display a planar 'butterfly' arrangement. Magnetic studies show that both are weakly coupled, with 2 displaying probable SMM behaviour.


Role of Computed Tomography in Planning the Appropriate X-Ray Gantry for Quantitative Aortography of Post-transcatheter Aortic Valve Implantation Regurgitation.
The clinical robustness of contrast-videodensitometric (VD) assessment of aortic regurgitation (AR) after transcatheter aortic valve implantation (TAVI) has been demonstrated. Correct acquisition of aortic root angiography for VD assessment, however, is hampered by the opacified descending aorta and by individual anatomic peculiarities. The aim of this study was to use preprocedural multi-slice computed tomography (MSCT) to optimize the angiographic projection in order to improve the feasibility of VD assessment.Methods and Results:In 92 consecutive patients, post-TAVI AR (i.e., left ventricular outflow tract [LVOT] AR) was assessed on aortic root angiograms using VD software. The patients were divided into 2 groups: The first group of 54 patients was investigated prior to the introduction of the standardized acquisition protocol; the second group of 38 consecutive patients after implementation of the standardized acquisition protocol, involving MSCT planning of the optimal angiographic projection. Optimal projection planning has dramatically improved the feasibility of VD assessment from 57.4% prior to the standardized acquisition protocol, to 100% after the protocol was implemented. In 69 analyzable aortograms (69/92; 75%), LVOT-AR ranged from 3% to 28% with a median of 12%. Inter-observer agreement was high (mean difference±SD, 1±2%), and the 2 observers' measurements were highly correlated (r=0.94, P<0.0001). Introduction of computed tomography-guided angiographic image acquisition has significantly improved the analyzability of the angiographic VD assessment of post-TAVI AR.


Combined Metabolic Activators Accelerates Recovery in Mild-to-Moderate COVID-19.
COVID-19 is associated with mitochondrial dysfunction and metabolic abnormalities, including the deficiencies in nicotinamide adenine dinucleotide (NAD+ ) and glutathione metabolism. Here it is investigated if administration of a mixture of combined metabolic activators (CMAs) consisting of glutathione and NAD+ precursors can restore metabolic function and thus aid the recovery of COVID-19 patients. CMAs include l-serine, N-acetyl-l-cysteine, nicotinamide riboside, and l-carnitine tartrate, salt form of l-carnitine. Placebo-controlled, open-label phase 2 study and double-blinded phase 3 clinical trials are conducted to investigate the time of symptom-free recovery on ambulatory patients using CMAs. The results of both studies show that the time to complete recovery is significantly shorter in the CMA group (6.6 vs 9.3 d) in phase 2 and (5.7 vs 9.2 d) in phase 3 trials compared to placebo group. A comprehensive analysis of the plasma metabolome and proteome reveals major metabolic changes. Plasma levels of proteins and metabolites associated with inflammation and antioxidant metabolism are significantly improved in patients treated with CMAs as compared to placebo. The results show that treating patients infected with COVID-19 with CMAs lead to a more rapid symptom-free recovery, suggesting a role for such a therapeutic regime in the treatment of infections leading to respiratory problems.


Lipid-laden macrophages in induced sputum are a marker of oropharyngeal reflux and possible gastric aspiration.
The diagnostic properties of a "lipid index" of macrophages in induced sputum as a noninvasive marker of aspiration of acidic gastric contents were evaluated. In a cross-sectional study, 33 subjects (17 with symptoms suggestive of gastrooesophageal reflux) with normal chest radiographs and no symptoms of aspiration or sinus disease, underwent dual-channel 24-h ambulatory oesophageal pH recording and sputum induction. Oropharyngeal reflux, defined as at least one episode of a fall in pH to <4 at the upper oesophageal electrode, was considered indicative of aspiration of acidic gastric contents ("gold standard"). An index for the presence of intracellular lipid in sputum macrophages, detected by oil red O stain, was obtained. The sensitivity, specificity and predictive values of this "lipid index" were calculated. The "lipid index" could be calculated in 29 of 33 samples with high interobserver repeatability (intraclass correlation coefficient 0.96). Twenty subjects showed oropharyngeal reflux and nine did not. The median "lipid index" in subjects with oropharyngeal reflux (24.5) was significantly greater than that in those without reflux (1.0) (p<0.001). A "lipid index" of 7.0 had a sensitivity of 90%, a specificity of 89%, a positive predictive value of 95% and a negative predictive value of 80%. A "lipid index" of 7.0 in the macrophages of induced sputum is a good marker of oropharyngeal reflux.


Evidence Synthesis in Harm Assessment of Medicines Using the Example of Rosiglitazone and Myocardial Infarction.
The current system of harm assessment of medicines has been criticized for relying on intuitive expert judgment. There is a call for more quantitative approaches and transparency in decision-making. Illustrated with the case of cardiovascular safety concerns for rosiglitazone, we aimed to explore a structured procedure for the collection, quality assessment, and statistical modeling of safety data from observational and randomized studies. We distinguished five stages in the synthesis process. In Stage I, the general research question, population and outcome, and general inclusion and exclusion criteria are defined and a systematic search is performed. Stage II focusses on the identification of sub-questions examined in the included studies and the classification of the studies into the different categories of sub-questions. In Stage III, the quality of the identified studies is assessed. Coding and data extraction are performed in Stage IV. Finally, meta-analyses on the study results per sub-question are performed in Stage V. A Pubmed search identified 30 randomized and 14 observational studies meeting our search criteria. From these studies, we identified 4 higher level sub-questions and 4 lower level sub-questions. We were able to categorize 29 individual treatment comparisons into one or more of the sub-question categories, and selected study duration as an important covariate. We extracted covariate, outcome, and sample size information at the treatment arm level of the studies. We extracted absolute numbers of myocardial infarctions from the randomized study, and adjusted risk estimates with 95% confidence intervals from the observational studies. Overall, few events were observed in the randomized studies that were frequently of relatively short duration. The large observational studies provided more information since these were often of longer duration. A Bayesian random effects meta-analysis on these data showed no significant increase in risk of rosiglitazone for any of the sub-questions. The proposed procedure can be of additional value for drug safety assessment because it provides a stepwise approach that guides the decision-making in increasing process transparency. The procedure allows for the inclusion of results from both randomized an observational studies, which is especially relevant for this type of research.


Evidence for a Golgi-to-endosome protein sorting pathway in Plasmodium falciparum.
During the asexual intraerythrocytic stage, the malaria parasite Plasmodium falciparum must traffic newly-synthesized proteins to a broad array of destinations within and beyond the parasite's plasma membrane. In this study, we have localized two well-conserved protein components of eukaryotic endosomes, the retromer complex and the small GTPase Rab7, to define a previously-undescribed endosomal compartment in P. falciparum. Retromer and Rab7 co-localized to a small number of punctate structures within parasites. These structures, which we refer to as endosomes, lie in close proximity to the Golgi apparatus and, like the Golgi apparatus, are inherited by daughter merozoites. However, the endosome is clearly distinct from the Golgi apparatus as neither retromer nor Rab7 redistributed to the endoplasmic reticulum upon brefeldin A treatment. Nascent rhoptries (specialized secretory organelles required for invasion) developed adjacent to endosomes, an observation that suggests a role for the endosome in rhoptry biogenesis. A P. falciparum homolog of the sortilin family of protein sorting receptors (PfSortilin) was localized to the Golgi apparatus. Together, these results elaborate a putative Golgi-to-endosome protein sorting pathway in asexual blood stage parasites and suggest that one role of retromer is to mediate the retrograde transport of PfSortilin from the endosome to the Golgi apparatus.


Effect of an invasive ant and its chemical control on a threatened endemic Seychelles millipede.
The impact of invasive species on island faunas can be of major local consequence, while their control is an important part of island ecosystem restoration. Among these invasive species are ants, of which some have a disruptive impact on indigenous arthropod populations. Here, we study the impact of the invasive African big-headed ant, Pheidole megacephala, on a small Seychelles island, Cousine, and assess the impact of this ant, and its chemical control, using the commercially available hydramethylnon-based bait, Siege, on the endemic keystone Seychelles giant millipede species, Sechelleptus seychellarum. We found no significant correlations in landscape-scale spatial overlap and abundance between the ant and the millipede. Furthermore, the ant did not attack healthy millipedes, but fed only on dying and dead individuals. The chemical defences of the millipede protected it from ant predation. Ingestion of the bait at standard concentration had no obvious impact on the millipede. The most significant threat to the Seychelles giant millipede in terms of P. megacephala invasion is from possible catastrophic shifts in ecosystem function through ant hemipteran mutualisms which can lead to tree mortality, resulting in alteration of the millipede's habitat.


Linking ecosystem function and hydrologic regime to inform restoration of a forested peatland.
Drainage is a globally common disturbance in forested peatlands that impacts peat soils, forest communities, and associated ecosystem functions, calling for informed hydrologic restoration strategies. The Great Dismal Swamp (GDS), located in Virginia and North Carolina, U.S.A., has been altered since colonial times, particularly by extensive ditch networks installed to lower water levels and facilitate timber harvests. Consequently, peat decomposition rates have accelerated, and red maple has become the dominant tree species, reducing the historical mosaic of bald cypress, Atlantic white-cedar, and pocosin stands. Recent repair and installation of water control structures aim to control drainage and, in doing so, enhance forest community composition and preserve peat depths. To help inform these actions, we established five transects of 15 plots each (75 plots total) along a hydrologic gradient where we measured continuous water levels and ecosystem attributes, including peat depths, microtopography, and forest composition and structure. We found significant differences among transects, with wetter sites having thicker peat, lower red maple importance, greater tree density, and higher overall stand richness. Plot-level analyses comported with these trends, clearly grouping plots by transects (via nonmetric multidimensional scaling) and resulting in significant correlations between specific hydrologic metrics and ecosystem attributes. Our findings highlight hydrologic controls on soil carbon storage, forest structure, and maple dominance, with implications for large-scale hydrologic restoration at GDS and in other degraded forested peatlands more broadly.


A United States perspective on the challenges in nursing education.
Nursing education in the US today is at crossroads. The profession has made great strides in gaining public respect and recognition for the work that nurses do. Nurses' voices are being heard in important political debates about revolutionary changes in the problematic US health care system. Advanced practice nurses are becoming valued providers of primary care to US citizens. It is innovative educational programs and educators that have provided the foundation to help nursing use its voice and to propel the profession forward. However, nurse educators are finding that they face major challenges in keeping nursing on track to be in the forefront of health care in the future. Some of these challenges include confronting nursing and faculty shortages, eliminating inconsistent and confusing educational choices, taking responsibility for mandates to stay on the cutting edge of quality initiatives, providing excellent clinical experiences for students and being willing to step out of old comfort zones to engage in designing imaginative and innovative ways to educate nurses in the future. Nurse educators must be successful in turning these challenges into opportunities if nursing is to command a key role in an evolving US health care system.


Mental representation of the body in action in Parkinson's disease.
Mixed findings characterize studies in Parkinson's disease (PD): some studies indicate a relationship between physical impairments and the ability to mentally represent the body, while others suggest spared abilities for this cognitive function. To clarify the matter, in the present study we explored the mental representations of the body in action in the same PD patients, taking also into account lateralization of symptoms and visual imagery skills. 10 PD patients with left- (lPD), 10 with right (rPD) lateralized symptoms (lPD), and 20 matched healthy controls have been recruited for the study. All patients were screened for neuropsychological impairments. To explore a more implicit component we used the hand laterality task (HLT), while the mental motor chronometry (MMC) was used to explore a more explicit one. Two control tasks, with objects instead of body parts, were administered to control for visual imagery skills. In the HLT, we detected the effects of biomechanical constraints effects in both controls and PD patients. In the latter group, importantly, this was true independently from lateralization of symptoms. In the MMC, we found the expected positive correlation between executed and imagined movements for both hands in controls only, while all PD patients, again independently form lateralization, only showed this effect for the left hand. In terms of visual imagery, only rPD patients differed from controls when asked to implicitly rotate letters, and in terms of accuracy only. However, this difference is explained by executive functions measured through the neuropsychological assessment rather than by a "pure" visual imagery impairment. In summary, our findings suggest that two different aspects of the mental representations of the body in action, one more implicit and the other more explicit, can be differently affected by PD. These impairments are unlikely explained by a basic visual imagery deficit. When present, impairments concern a higher dimension, related to motor functions and awareness, and not driven by sensory impairments, as shown by the independence of effects from physical laterality of symptoms.


Effects of Kinesio taping and Mcconnell taping on balance and walking speed of hemiplegia patients.
[Purpose] The aim of this study was to evaluate the overlap effect of the PNF following the application of Kinesio taping and the McConnell taping, and also the impact of the taping application method on the balance and walking speed of the patients with stroke. [Subjects and Methods] Thirty-six patients who were diagnosed with hemiplegia due to stroke were selected as subjects of this study. They were randomly and evenly divided into experiment group 1 (Kinesio taping group), experiment group 2 (McConnell taping group), and the control group; each group had 12 patients. [Results] The Berg balance scale (BBS) was used to evaluate balance, and the ability in this study. A 10 m walking test (10MWT) was performed to measure the walking speed. Experiment group 1 showed a statistically significant improvement in balance and walking speed compared to experiment group 2, and the control group in week 4 and week 8. [Conclusion] Application of Kinesio taping had a more beneficial effect on the balance and walking speed than joint-fixation taping of the patients with stroke.


Alcoholic Fractions F5 and F6 from Withania somnifera Leaves Show a Potent Antileishmanial and Immunomodulatory Activities to Control Experimental Visceral Leishmaniasis.
Visceral leishmaniasis (VL) causes fatal life-threatening disease, if left untreated. The current drugs have various limitations; hence, natural products from medicinal plants are being focused in search of new drugs to treat leishmaniasis. The aim of the present study was to evaluate the antileishmanial and immunomodulatory activities of F5 and F6 alcoholic fractions from Withania somnifera leaves and purified withaferin-A in Leishmania donovani-infected peritoneal macrophages and BALB/c mice. We observed that F5 (15 µg/mL), F6 (10 µg/mL), and withaferin-A (1.5 µM) reduce amastigote count in peritoneal macrophages and induce reactive oxygen species and significant decrease in IL-10 mRNA expression compared to control upon treatment. Subsequently, in vivo study mice were treated with F5 (25 and 50 mg/kg b.wt.), F6 (25 and 50 mg/kg b.wt.) orally, and withaferin-A (2 mg/kg b.wt.) intraperitoneally for 10 consecutive days and a drastic reduction in parasite burden in both spleen and liver were observed. The treatment resulted in the reduction in IL-10, IL-4, and TGF-β mRNA expression and a significant increase in IFN-γ/IL-10 expression ratio in the treated group compared to control. The humoral response of these alcoholic fractions and withaferin-A shows increased IgG2a levels when compared with IgG1 in treated mice. Taken together, our result concludes that withanolides in alcoholic fractions demonstrate a potent antileishmanial and immunomodulatory activities in experimental VL.


Effect of fish oils containing different amounts of EPA, DHA, and antioxidants on plasma and brain fatty acids and brain nitric oxide synthase activity in rats.
The interest in n-3 polyunsaturated fatty acids (PUFAs) has expanded significantly in the last few years, due to their many positive effects described. Consequently, the interest in fish oil supplementation has also increased, and many different types of fish oil supplements can be found on the market. Also, it is well known that these types of fatty acids are very easily oxidized, and that stability among supplements varies greatly. In this pilot study we investigated the effects of two different types of natural fish oils containing different amounts of the n-3 PUFAs eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and antioxidants on plasma and brain fatty acids, blood lipids, vitamin E, and in vivo lipid peroxidation, as well as brain nitric oxide synthase (NOS) activity, an enzyme which has been shown to be important for memory and learning ability. Sprague-Dawley rats were divided into four groups and fed regular rat chow pellets enriched with 5% (w/w) of butter (control group), a natural fish oil (17.4% EPA and 11.7% DHA, referred to as EPA-rich), and a natural fish oil rich in DHA (7.7% EPA and 28.0% DHA, referred to as DHA-rich). Both of the fish oils were stabilized by a commercial antioxidant protection system (Pufanox) at production. The fourth group received the same DHA-rich oil, but without Pufanox stabilization (referred to as unstable). As an index of stability of the oils, their peroxide values were repeatedly measured during 9 weeks. The dietary treatments continued until sacrifice, after 10 days. Stability of the oils varied greatly. It took the two stabilized oils 9 weeks to reach the same peroxide value as the unstable oil reached after only a few days. Both the stabilized EPA- and DHA-rich diets lowered the triacylglycerols and total cholesterol compared to control (-45%, P < 0.05 and -54%, P < 0.001; -31%, P < 0.05 and -25%, P < 0.01) and so did the unstable oil, but less efficiently. Only the unstable oil increased in vivo lipid peroxidation significantly compared to control (+40%, P < 0.001). Most of the fatty acids in the plasma phospholipids were significantly affected by both the EPA- and DHA-rich diets compared to control, reflecting their specific fatty acid pattern. The unstable oil diet resulted in smaller changes, especially in n-3 PUFAs. In the brain phospholipids the changes were less pronounced, and only the diet enriched with the stabilized DHA-rich oil resulted in a significantly greater incorporation of DHA (+13%, P < 0.01), as well as total n-3 PUFAs (+13%, P < 0.01) compared to control. Only the stabilized DHA-rich oil increased the brain NOS activity (+33%, P < 0.01). Both the EPA- and DHA-rich diets affected the blood lipids in a similarly positive manner, and they both had a large impact on plasma phospholipid fatty acids. It was only the unstable oil that increased in vivo lipid peroxidation. However, the intake of DHA was more important than that of EPA for brain phospholipid DHA enrichment and brain NOS activity, and the stability of the fish oil was also important for these effects.


Urban minority youth swimming (in)ability in the United States and associated demographic characteristics: toward a drowning prevention plan.
To examine swimming ability and variables associated with swimming for US inner-city, minority children. Empirical research on minority children's swimming ability is non-existent, and drowning rates for this population are high. Cross-sectional survey research. Descriptive statistics were produced. Multiple regression was applied using significant demographic variables by swimming ability. Six US cities were chosen (Chicago, Illinois; Houston, Texas; Memphis, Tennessee; Miami, Florida; Oakland, California; Philadelphia, Pennsylvania). Young Men's Christian Association (YMCA) facilities were used to solicit subjects. A large sample (n = 1680) was gathered, which targeted poor, minority children. Parents of children aged 4-11 years and adolescents (12-17 years) completed surveys that research team members or trained YMCA staff supervised during non-swimming YMCA programmes. African-American respondents reported a 57.5% "at risk" (unable to swim or uncomfortable in deep end of pool) swimming ability. Hispanic/Latino children confirmed a 56.2% "at risk" level as compared with 30.9% for white subjects. Age, sex, child's lunch programme, parental education and race variables were all significantly (p<0.05) related to swimming ability. Regression analysis revealed that all demographic variables fell into a significant model (p<0.001) as predictor variables. Poor minority children, specifically African-American and Hispanic/Latino, are at a significant disadvantage concerning swimming ability. Female subjects were notably more "at risk" regarding their swimming ability than male subjects. Age, race and socioeconomic factors (lunch programme and parental education) were significantly associated with children who have low swimming ability.


Ring-tailed lemurs: a species re-imagined.
For over 50 years, ring-tailed lemurs have been studied continuously in the wild. As one of the most long-studied primate species, the length and breadth of their study is comparable to research on Japanese macaques, baboons and chimpanzees. They are also one of the most broadly observed of all primates, with comprehensive research conducted on their behaviour, biology, ecology, genetics, palaeobiology and life history. However, over the last decade, a new generation of lemur scholars, working in conjunction with researchers who have spent decades studying this species, have greatly enhanced our knowledge of ring-tailed lemurs. In addition, research on this species has expanded beyond traditional gallery forest habitats to now include high altitude, spiny thicket, rocky outcrop and anthropogenically disturbed coastal forest populations. The focus of this special volume is to 're-imagine' the 'flagship species of Madagascar', bringing together three generations of lemur scholars.


Abnormal carbohydrate composition of the major penetrating membrane protein of En(a-) human erythrocytes.
The major penetrating membrane glycoprotein (band 3) was isolated from En(a-) and normal human erythrocytes. The two proteins differed only in carbohydrate composition. Band 3 from En(a-) erythrocytes contained greater amounts of galactose and N-acetyl-glucosamine. The loss of the sialoglycoprotein sialotetrasaccharides in the En(a-) cell is not compensated by the appearance of these units in band 3 of En(a-) erythrocytes.


Analysis of multiple protein detection methods in human osteoporotic bone extracellular matrix: From literature to practice.
The punctual analysis of bone Extracellular Matrix (ECM) proteins represents a pivotal point for medical research in bone diseases like osteoporosis. Studies in this field, historically done to appreciate bone biology, were mainly conducted on animal samples and, up to today, only a few studies on protein detection in human bone are present. The challenges in bone ECM protein extraction and quantitation protocols are related to both the separation of proteins from the mineral content (i.e. hydroxyapatite) and the difficulty of avoiding protein denaturation during the extraction processes. The aim of the present work was to define appropriate protocol(s) for bone ECM protein extraction that could be applied to investigate both normal and pathological conditions. We compared and optimised some of the most used protocols present in the literature, modifying the protein precipitation method, the buffer used for resuspension and/or the volume of reagent used. Bradford and BCA assays and Western Blotting were used to evaluate the variations in the total protein recovery and the amount of selected proteins (Type I Collagen, TGF-β, IGF-1, Decorin, Osteopontin, Bone Sialoprotein-2 and Osteocalcin). Collectively, we were capable to draw-up two single-extract protocols with optimal recovery and ideal protein content, that can be used for a detailed analysis of ECM proteins in pathological bone samples. Time-consuming multi-extract procedures, optimised in their precipitation methods, are however crucial for a precise detection of specific proteins, like osteocalcin. As the matter of fact, also the demineralization processes, commonly suggested and performed in several protocols, could hinder an accurate protein detection, thus inherently affecting the study of a pathological bone ECM. This study represents a starting point for the definition of appropriate strategies in the study of bone extracellular matrix proteins involved in the onset and maintenance of bone diseases, as well as a tool for the development of customized scaffolds capable to modulate a proper feedback loop in bone remodelling, altered in case of diseases like osteoporosis.


Developmental morphology of cover crop species exhibit contrasting behaviour to changes in soil bulk density, revealed by X-ray computed tomography.
Plant roots growing through soil typically encounter considerable structural heterogeneity, and local variations in soil dry bulk density. The way the in situ architecture of root systems of different species respond to such heterogeneity is poorly understood due to challenges in visualising roots growing in soil. The objective of this study was to visualise and quantify the impact of abrupt changes in soil bulk density on the roots of three cover crop species with contrasting inherent root morphologies, viz. tillage radish (Raphanus sativus), vetch (Vicia sativa) and black oat (Avena strigosa). The species were grown in soil columns containing a two-layer compaction treatment featuring a 1.2 g cm-3 (uncompacted) zone overlaying a 1.4 g cm-3 (compacted) zone. Three-dimensional visualisations of the root architecture were generated via X-ray computed tomography, and an automated root-segmentation imaging algorithm. Three classes of behaviour were manifest as a result of roots encountering the compacted interface, directly related to the species. For radish, there was switch from a single tap-root to multiple perpendicular roots which penetrated the compacted zone, whilst for vetch primary roots were diverted more horizontally with limited lateral growth at less acute angles. Black oat roots penetrated the compacted zone with no apparent deviation. Smaller root volume, surface area and lateral growth were consistently observed in the compacted zone in comparison to the uncompacted zone across all species. The rapid transition in soil bulk density had a large effect on root morphology that differed greatly between species, with major implications for how these cover crops will modify and interact with soil structure.


Suprascapular nerve block for the treatment of frozen shoulder in primary care: a randomized trial.
Frozen shoulder is a common problem in general practice, but its treatment is difficult since none of the currently used therapies are proven to be effective. To assess the effectiveness of suprascapular nerve block to relieve pain and improve range of movement, and its suitability for use in primary care. This small study by a single practitioner aims to justify a larger multicentred trial. A randomized trial of 30 patients to compare a single suprascapular nerve block with a course of intra-articular injections. Patients' pain levels and ranges of movement were assessed over a 12-week period. Suprascapular nerve block produced a faster and more complete resolution of pain and restoration of range of movement than a series of intra-articular injections. These differences were confirmed by statistical analysis using the Mann-Whitney U-test (P < 0.01 for pain levels and P < 0.05 for range of abduction and external rotation.) This study suggests that suprascapular nerve block is a safe and effective treatment for frozen shoulder in primary care, and justifies a larger multicentred trial using independent blinded assessment. Such a study should include a third group treated by suprascapular nerve block without steroid; a more comprehensive assessment of patient debility.


Minimal clinically important rehabilitation effects in patients with osteoarthritis of the lower extremities.
To estimate minimal clinically important differences (MCID) of effects measured by the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) in patients with osteoarthritis (OA) of the lower extremities undergoing a comprehensive inpatient rehabilitation intervention. A prospective cohort study assessed patients' health by the WOMAC at baseline (entry into the clinic) and at the 3 month followup, and by a transition questionnaire asking about the change of "health in general related to the OA joint" during that time period. The WOMAC section score differences between the "equal" group and the "slightly better" and "slightly worse" groups resulted in the MCID for improvement and for worsening. In total 192 patients were followed up. The MCID for improvement ranged from 0.80 to 1.01 points on the continuous WOMAC numerical rating scale from 0 to 10, reflecting changes of 17 to 22% of baseline scores. The MCID for worsening conditions ranged from 0.29 (6%) to 1.03 points (22%). In the transition reply subjectively unchanged patients reported a "pessimistic bias" of 0.35 to 0.51 points, except for the stiffness section. Both MCID and pessimistic bias showed regression to the mean and baseline dependency. The assessment of MCID using the transition method is a heuristic and valid strategy to detect particular rehabilitation effects in patients with OA of the lower extremities with the use of the WOMAC, and it is worth implementing. The size of the MCID and of the systematic bias is comparable to that assessed by other methods and in other therapeutic settings.


Aluminum exposure and toxicity in neonates: a practical guide to halt aluminum overload in the prenatal and perinatal periods.
During the last years, human newborns have been overexposed to biologically reactive aluminum, with possible relevant consequences on their future health and on their susceptibility to a variety of diseases. Children, newborns and particularly preterm neonates are at an increased risk of aluminum toxicity because of their relative immaturity. Based on recent original publications and classical data of the literatures, we reviewed the aluminum content in mother's food during the intrauterine life as well as in breast milk and infant formula during lactation. We also determined the possible role of aluminum in parenteral nutrition solutions, in adjuvants of vaccines and in pharmaceutical products. A special focus is placed on the relationship between aluminum overexposure and the insurgence of bone diseases. Practical points of management and prevention are suggested. Aluminum sources that infants may receive during the first 6 months of life are presented. In the context of prevention of possible adverse effects of aluminum overload in fetal tissues during development, simple suggestions to pregnant women are described. Finally, practical points of management and prevention are suggested. Pediatricians and neonatologists must be more concerned about aluminum content in all products our newborns are exposed to, starting from monitoring aluminum concentrations in milk- and soy-based formulas in which, on the basis of recent studies, there is still too much aluminum.


Liquiritigenin Reduces Blood Glucose Level and Bone Adverse Effects in Hyperglycemic Adult Zebrafish.
Diabetes mellitus is a metabolic disease characterized by chronic hyperglycemia that induces other pathologies including diabetic retinopathy and bone disease. Adult Danio rerio (zebrafish) represents a powerful model to study both glucose and bone metabolism. Then, the aim of this study was to evaluate the effects of liquiritigenin (LTG) on blood glucose level and diabetes complications in hyperglycemic adult zebrafish. LTG is a flavonoid extracted from Glycyrrhiza glabra roots which possess important antioxidant, anti-inflammatory, and anti-diabetic properties. During four weeks of glucose treatment, LTG significantly prevented the onset of the hyperglycemia in adult zebrafish. Moreover, hyperglycemic fish showed increased advanced glycation end-products (AGEs) and parathormone levels whereas LTG completely prevented both of these metabolic alterations. Large bone-loss areas were found in the scales of glucose-treated fish whereas only small resorption lacunae were detected after glucose/LTG treatment. Biochemical and histological tartrate resistant acid phosphatase (TRAP) assays performed on explanted scales confirmed that LTG prevented the increase of osteoclastic activity in hyperglycemic fish. The osteoblastic alkaline phosphatase (ALP) activity was clearly lost in scales of glucose-treated fish whereas the co-treatment with LTG completely prevented such alteration. Gene expression analysis showed that LTG prevents the alteration in crucial bone regulatory genes. Our study confirmed that LTG is a very promising natural therapeutic approach for blood glucose lowering and to contrast the development of bone complications correlated to chronic hyperglycemia.


Inactivation of Hippo pathway characterizes a poor-prognosis subtype of esophageal cancer.
Identification of molecular subtypes that reflect different prognoses and treatment responses, especially immune checkpoint inhibitors (ICIs) in esophageal squamous cell carcinoma (ESCC), is essential for treatment decisions. We performed targeted sequencing in 201 patients with ESCC to discover genetic subtypes and validate our findings via multiple data sets. We identified 3 driver genes (FCGBP, GRIN2B, and FRY), and recurrent truncating mutations in FRY impaired its tumor-suppressive function and promoted tumor proliferation. A 3-gene mutation signature (FAT1, FAT3, and FRY) recognized a molecular subtype named "FAT/FRY" with frequent Hippo pathway-related mutations. In multiple ESCC cohorts, the patients with the FAT/FRY subtype had poorer prognosis than did patients in the WT group. Transcriptome analysis indicated that the FAT/FRY subtype was characterized by inactivation of the Hippo pathway, hypoxia, chemoresistance, higher infiltration of CD8+ T cells and activated DCs, and a transcriptome similar to that of cancer responders. Furthermore, the 3-gene signature predicted better survival for patients treated with ICIs, partially explained by its positive correlation with the tumor mutation burden and neoantigen burden. The 3-gene signature is a biomarker to recognize the FAT/FRY molecular subtype, evaluate prognosis, and select potential beneficiaries of ICIs in ESCC.


Home care in Italy: a system on the move, in the opposite direction to what we expect.
For a long time, the international literature has described the Italian public system of home care for frail elderly people as underfunded and mostly cash-oriented; a system, thus, relying almost entirely on informal care provided by the family and, more recently, by migrant workers. Abroad and in Italy, most experts have long shared the expectation that, if and when, public expenditure devoted to home care was to increase, the outcome would be an expansion in the provision of services in kind. This study analyses how the provision of home care actually has changed in the last decade. The analysis reveals that indeed public expenditure has risen. However, while the system of home-care provision also has changed, it has done so in the opposite direction to that expected. In fact, most of the additional resources have funded an increase in the number of users of the companion payment [Indennità di Accompagnamento (IA)], a cash benefit of €487 per month. In this respect, the Italian welfare system is more cash-oriented than it was 10 years ago. This article discusses the reasons behind the increased uptake of the IA, namely: an increase in the needs and demands of older people; the traits of the Italian welfare system; and the peculiar features of the companion payment itself. The article then looks at why services in kind rose to a lesser degree, pinpoints the main reason as being based on the politics of social care at national level, and finally focuses on the challenges that the Italian home-care system has to face within the changed policy environment with respect to quality of care, carers' conditions and support for users with high-level needs.


Effects of the human immunodeficiency virus type 1 Tat protein on the expression of inflammatory cytokines.
Increased levels of inflammatory cytokines, including tumor necrosis factor (TNF), interleukin-1 (IL-1), and IL-6, have been detected in specimens from human immunodeficiency virus type 1 (HIV-1)-infected individuals. Here we demonstrate that HIV-1 activates the expression of TNF but not of IL-1 and IL-6 in acutely and chronically infected T cells. The increase in TNF gene expression is due to activation of the TNF promoter by the viral gene product Tat. Transactivation of TNF gene expression requires the product of the first exon of the tat gene and is cell type independent. T cells chronically infected with pol-defective HIV-1 provirus constitutively express both Tat and TNF at levels significantly higher (fivefold) than those seen in control cells, and treatment with phorbol myristate acetate greatly enhances Tat expression and TNF production. As TNF can increase the production of IL-1 and IL-6 and these inflammatory cytokines all enhance HIV-1 gene expression and affect the immune, vascular, and central nervous systems, the activation of TNF by Tat may be part of a complex pathway in which HIV-1 uses viral products and host factors to increase its own expression and infectivity and to induce disease.


The nuclear receptor REV-ERBα represses the transcription of growth/differentiation factor 10 and 15 genes in rat endometrium stromal cells.
Cellular oscillators in the uterus play critical roles in the gestation processes of mammals through entraining of the clock proteins to numerous downstream genes, including growth/differentiation factor (Gdf)10 and Gdf15. The expression of Gdf10 and Gdf15 is significantly increased in the uterus during decidualization, but the mechanism underlying the regulation of Gdf gene expression in the uterus is poorly understood. Here, we focused on the function of the cellular oscillators in the expression of Gdf family by using uterine endometrial stromal cells (UESCs) isolated from pregnant Per2-dLuc transgenic rats. A significant decline of Per2-dLuc bioluminescence activity was induced in in vitro decidualized UESCs, and concomitantly the expression of canonical clock genes was downregulated. Conversely, the expression of Gdf10 and Gdf15 of the Gdf was upregulated. In UESCs transfected with Bmal1-specific siRNA, in which Rev-erbα expression was downregulated, Gdf10 and Gdf15 were upregulated. However, Gdf5, Gdf7, and Gdf11 were not significantly affected by Bmal1 silencing. The expression of Gdf10 and Gdf15 was enhanced after treatment with a REV-ERBα antagonist in the presence or absence of progesterone. Chromatin immunoprecipitation-PCR analysis revealed the inhibitory effect of REV-ERBα on the expression of Gdf10 and Gdf15 in UESCs by recognizing their gene promoters. Collectively, our findings indicate that the attenuation of REV-ERBα leads to an upregulation of Gdf10 and Gdf15 in decidual cells, in which cellular oscillators are impaired. Our results provide novel evidence regarding the functions of cellular oscillators regulating the expression of downstream genes during the differentiation of UESCs.


A novel school-based intervention to improve nutrition knowledge in children: cluster randomised controlled trial.
Improving nutrition knowledge among children may help them to make healthier food choices. The aim of this study was to assess the effectiveness and acceptability of a novel educational intervention to increase nutrition knowledge among primary school children. We developed a card game 'Top Grub' and a 'healthy eating' curriculum for use in primary schools. Thirty-eight state primary schools comprising 2519 children in years 5 and 6 (aged 9-11 years) were recruited in a pragmatic cluster randomised controlled trial. The main outcome measures were change in nutrition knowledge scores, attitudes to healthy eating and acceptability of the intervention by children and teachers. Twelve intervention and 13 control schools (comprising 1133 children) completed the trial. The main reason for non-completion was time pressure of the school curriculum. Mean total nutrition knowledge score increased by 1.1 in intervention (baseline to follow-up: 28.3 to 29.2) and 0.3 in control schools (27.3 to 27.6). Total nutrition knowledge score at follow-up, adjusted for baseline score, deprivation, and school size, was higher in intervention than in control schools (mean difference = 1.1; 95% CI: 0.05 to 2.16; p = 0.042). At follow-up, more children in the intervention schools said they 'are currently eating a healthy diet' (39.6%) or 'would try to eat a healthy diet' (35.7%) than in control schools (34.4% and 31.7% respectively; chi-square test p < 0.001). Most children (75.5%) enjoyed playing the game and teachers considered it a useful resource. The 'Top Grub' card game facilitated the enjoyable delivery of nutrition education in a sample of UK primary school age children. Further studies should determine whether improvements in nutrition knowledge are sustained and lead to changes in dietary behaviour.