Cognitive appraisals in young people with obsessive-compulsive disorder.
A number of cognitive appraisals have been identified as important in the manifestation of obsessive-compulsive disorder (OCD) in adults. There have, however, been few attempts to explore these cognitive appraisals in clinical groups of young people. This study compared young people aged between 11 and 18 years with OCD (N = 28), young people with other types of anxiety disorders (N = 28) and a non-clinical group (N = 62) on three questionnaire measures of cognitive appraisals. These were inflated responsibility (Responsibility Attitude Scale; Salkovskis et al., 2000), thought-action fusion--likelihood other (Thought-Action Fusion Scale; Shafran, Thordarson & Rachman, 1996) and perfectionism (Multidimensional Perfectionism Scale; Frost, Marten, Luhart & Rosenblate, 1990). The young people with OCD had significantly higher scores on inflated responsibility, thought-action fusion--(likelihood other), and one aspect of perfectionism, concern over mistakes, than the other groups. In addition, inflated responsibility independently predicted OCD symptom severity. The results generally support a downward extension of the cognitive appraisals held by adults with OCD to young people with the disorder. Some of the results, however, raise issues about potential developmental shifts in cognitive appraisals. The findings are discussed in relation to implications for the cognitive model of OCD and cognitive behavioural therapy for young people with OCD.


1-D cobalt(II) spin transition compound with strong interchain interaction: [Co(pyterpy)Cl(2)].X.
Cobalt(II) compounds [Co(pyterpy)Cl(2)].MeOH (1.(MeOH)) and [Co(pyterpy)Cl(2)].2H(2)O (1.(2H(2)O)) were synthesized. The compound 1.(MeOH) forms the quasi 3-D networks by making pi-pi stacking between the 1-D chains. The methanol molecules from 1.(MeOH) can be removed by heating, and substituted by absorption of water molecules. The MeOH molecules in 1.(MeOH) are removed by heating at 410 K, and they are substituted by water molecules to form 1.(2H(2)O). 1.(2H(2)O) exhibits a S = (3)/(2) (HS) left arrow over right arrow S = (1)/(2) (LS) spin transition with a thermal hysteresis. We have succeeded in constructing a guest dependent 1-D spin-crossover cobalt(II) compound.


Functional Titanium Substrates Synergetic Photothermal Therapy for Enhanced Antibacterial and Osteogenic Performance via Immunity Regulation.
Implant-associated infections (IAIs) significantly impair the integration between titanium (Ti) implants and bone tissues. Bacteria colonized on the surface of the implant can induce innate immune suppression of the host to resist clearance. Herein, an interfacial functionalization strategy is employed to introduce FeIII TA nanoparticles (NPs) and acetyl Bletilla striata polysaccharide (acBSP) on the Ti substrate to obtain the Ti-TF-acBSP system. Under near-infrared (NIR) irradiation, the hyperthermal effect induced by FeIII TA NPs directly killed bacteria. Meanwhile, macrophages are induced by acBSP to polarize into pro-inflammatory M1 phenotype, which enhanced the phagocytosis ability of macrophages and activated host innate immunity. Moreover, the asBSP instructed macrophages to secrete pro-osteogenic cytokine, which promoted osteogenic differentiation of MSCs. The results of the animal experiment in vivo confirmed that the Ti-TF-acBSP implant effectively eliminated bacterial infection under NIR irradiation, enhanced the expression of pro-inflammatory cytokine, and induced the production of bone-forming related factors. In a word, the functionalized Ti implant not only have a direct bactericidal effect but also regulate macrophage polarization as well as macrophage-mediated bactericidal and osteogenic effect. The strategy of combining photothermal therapy with immunoregulation will present a potential candidate for the development of novel antibacterial orthopedic devices.


Prolastin aerosol therapy and sputum taurine in cystic fibrosis.
Neutrophil elastase in the cystic fibrosis airways inhibits opsonophagocytosis and induces the expression of interleukin-8, a neutrophil chemoattractant. Prolastin is a therapeutic preparation of alpha-1 proteinase inhibitor (alpha1,-PI), a neutrophil elastase inhibitor. The objective of this study was to determine the effects of Prolastin aerosol therapy on airway inflammation in cystic fibrosis. The primary endpoint of this study was sputum taurine, an amino-acid present in high concentrations in neutrophils. Sputum taurine correlates with respiratory exacerbations of cystic fibrosis. Seventeen patients with cystic fibrosis were each assigned to three sequential 10-day periods including first, aerosol therapy of 5 ml saline solution bid; second, aerosol therapy of 250 mg Prolastin bid; third, no aerosol therapy. On days 8, 9 and 10 of each period, early morning sputum was collected for the quantification of alpha1-PI, neutrophil elastase activity, IL-8 and taurine. During Prolastin therapy, a 3-fold increase in sputum alpha1-PI was observed (P = 0.002). Baseline values of sputum alpha1-PI correlated with the values obtained after Prolastin aerosol (R = 0.77, P < 0.01). Sputum neutrophil elastase activity remained unchanged but taurine decreased after Prolastin therapy (during therapy P = 0.052, after therapy P = 0.026). Prolastin aerosol therapy had no adverse effect on pulmonary function. Aerosol therapy with Prolastin in patients with cystic fibrosis leads to a progressive decrease in sputum taurine. This suggests that even in the absence of sustained elastase inhibition, Prolastin aerosol therapy may have a beneficial effect on airway inflammation in patients with cystic fibrosis.


High-Yield Production of 4-Hydroxybenzoate From Glucose or Glycerol by an Engineered Pseudomonas taiwanensis VLB120.
Aromatic compounds such as 4-hydroxybenzoic acid are broadly applied in industry for a myriad of applications used in everyday life. However, their industrial production currently relies heavily on fossil resources and involves environmentally unfriendly production conditions, thus creating the need for more sustainable biotechnological alternatives. In this study, synthetic biology was applied to metabolically engineer Pseudomonas taiwanensis VLB120 to produce 4-hydroxybenzoate from glucose, xylose, or glycerol as sole carbon sources. Genes encoding a 4-hydroxybenzoate production pathway were integrated into the host genome and the flux toward the central precursor tyrosine was enhanced by overexpressing genes encoding key enzymes of the shikimate pathway. The flux toward tryptophan biosynthesis was decreased by introducing a P290S point mutation in the trpE gene, and degradation pathways for 4-hydroxybenzoate, 4-hydroxyphenylpyruvate and 3-dehydroshikimate were knocked out. The resulting production strains were tailored for the utilization of glucose and glycerol through the rational modification of central carbon metabolism. In batch cultivations with a completely mineral medium, the best strain produced 1.37 mM 4-hydroxybenzoate from xylose with a C-mol yield of 8% and 3.3 mM from glucose with a C-mol yield of 19.0%. Using glycerol as a sole carbon source, the C-mol yield increased to 29.6%. To our knowledge, this is the highest yield achieved by any species in a fully mineral medium. In all, the efficient conversion of bio-based substrates into 4-hydroxybenzoate by these deeply engineered P. taiwanensis strains brings the renewable production of aromatics one step closer.


Secretion of inducible proteinase by pathogenic Candida species.
The ability of three isolates each of seven pathogenic Candida species to grow in a liquid medium containing bovine serum albumin (BSA) as a nitrogen source was determined. All three strains of C. albicans, two strains of C. guilliermondii and one strain of C. tropicalis grew well. At any time proteinase activity was detected in the culture filtrates of only the most virulent species--C. albicans, C. tropicalis and C. parapsilosis and this observation was related to complete hydrolysis of BSA. Serologically, cross reactions were demonstrated between anti-proteinase antiserum and C. albicans and C. tropicalis culture filtrates. These results further emphasise the role of the inducible proteinase of Candida in the pathogenesis of candidosis.


Acute disseminated encephalomyelitis in North Indian children: clinical profile and follow-up.
Acute disseminated encephalomyelitis in children is not uncommon in developing countries, yet there is little systematic documentation of its clinical profile and follow-up. We studied the clinical and neuroradiologic features of acute disseminated encephalomyelitis in 52 consecutive children. Clinical details, magnetic resonance imaging (MRI) findings, and the results of other investigations were recorded, and children were followed up from 6 to 48 months. A repeat MRI was done after 3 to 4 months, and in those with persistent lesions, another MRI was done after 6 to 7 months of discharge. The mean age at presentation was 6.14 +/- 3.17 years, 73.1% were male, and 17 children had a history of antecedent infectious illness or vaccination. Most children had a meningoencephalitic presentation, with sudden-onset motor weakness in 76.9% and seizures in 36.5%. Altered sensorium and pyramidal signs were seen in 55.8% and 80.7% of children, respectively. On MRI, scattered T(2)-weighted hyperintense lesions were seen, mainly in the subcortical white matter, especially in the parietal (53.8%) and frontal (30.17%) regions. Thalamic, basal ganglia, and callosal lesions were seen in 30.76%, 17.3%, and 13.46% of cases, respectively. Variable contrast enhancement was seen in 48% of those who had contrast MRIs. The response to methylprednisolone was good, with dramatic recovery in 26.9% and marked improvement in 51.9% at discharge. On follow-up, of 44 children, residual smaller MRI lesions were seen in 30. The MRI was repeated at 6 months in children with residual lesions, and it was found that the lesions either disappeared or were significantly reduced after 6 months in 75% of cases. Four children had relapse of acute disseminated encephalomyelitis with new lesions on MRI. All of them responded to methylprednisolone. None of the clinical or neuroradiologic factors at presentation had any significant correlation with relapse. Six months after discharge, no deficits could be found in 61.3% of cases; 15.9% and 4.5% had motor and cognitive deficits, and 9% had multiple deficits. The presentation of pediatric acute disseminated encephalomyelitis in developing countries is similar to that in developed countries. In spite of an aggressive presentation, most children respond well to corticosteroids. MRI lesions disappear or are significantly reduced at 6 months in the majority of cases.


[A proposed algorithm for medicodental care of patients treated with bisphosphonates].
Bisphophonates (BP) were first named diphosphonates. They are potent inhibitors of osteoclastic activity and so reduce bone remodeling. Additionally they have anti-angiogenic properties and contribute to progressive disappearance of bony micro-vascular blood supply. Administrated orally, BP are generally used to prevent and treat osteoporosis. Injectable BP are used in patients with multiple myeloma and metastatic solid tumors. Scientific evidence dealing with a potentially devastating side effect of BP, osteochemonecrosis of the jaws, is growing rapidly. Clinical signs and symptoms include absent or delayed soft tissue healing with bony exposure following dental extraction or spontaneous gum dehiscence. Patients are usually asymptomatic but may develop pain if the bone becomes secondarily infected. At the beginning, no radiographic manifestations are seen, but in some cases a vast zone of necrotic bone can be seen on MRI, larger than what could be expected. Surgical debridements, bone curettage, local irrigation and or hyperbaric oxygen therapy have proven to be unsuccessful. Up to now, no definitive treatment strategy has been published to manage those patients leaving the dentist or the stomatologist resourceless. This article proposes recommendations for general practitioners, dentists, oral surgeons and designed for three types of patients: 1. patients to be treated with BP; 2. patients treated with BP without bisphosphonate-associated osteonecrosis; 3. patients with bisphosphonate-associated osteonecrosis. The proposed guidelines are not definitive and practitioners remain free to choose their treatment. Of upmost importance is to recognize the risks of oral complications before, during and after surgery in patients treated with BP and to inform them of such risks.


Effects of higher versus lower dexamethasone doses on pulmonary and neurodevelopmental sequelae in preterm infants at risk for chronic lung disease: a meta-analysis.
Systemic postnatal dexamethasone treatment reduces the risk of chronic lung disease in preterm infants but also may be associated with increased risk of neurodevelopmental impairment. Because it is not known whether these effects are modulated by the cumulative dexamethasone dose, we systematically reviewed the available randomized evidence on the effects of lower versus higher cumulative dexamethasone doses, in terms of death, pulmonary morbidity, and neurodevelopmental outcomes, in preterm infants. Randomized, controlled trials comparing higher- versus lower-dosage dexamethasone regimens in ventilated preterm infants were identified by searching the main electronic databases, references from relevant studies, and abstracts from the Societies for Pediatric Research (from 1990 onward). Eligibility and quality of trials were assessed, and data on study design, patient characteristics, and relevant outcomes were extracted. Six studies that enrolled a total of 209 participants were included; 2 studies contrasted cumulative dexamethasone doses in the higher ranges (>2.7 mg/kg in the higher-dosage regimen) and 4 in the lower ranges (<or=2.7 mg/kg in the higher-dosage regimen). Meta-analysis revealed no effect of dexamethasone dose on rates of death and neurodevelopmental sequelae in these 2 subgroups. Subgroup analysis of the studies contrasting dexamethasone doses in the higher ranges showed that the higher dose of dexamethasone was more effective in reducing the occurrence of chronic lung disease than was the lower dose. Interpretation of these data was hampered by the small samples of randomly assigned children, heterogeneity of study populations and designs, use of late rescue glucocorticoids, and lack of long-term neurodevelopmental data in some studies. Recommendations for optimal dexamethasone doses for preterm infants at risk for chronic lung disease cannot be based on current evidence. A well-designed, large, randomized, controlled trial is urgently needed to establish the optimal dexamethasone dosage regimen.


Antioxidant, immunomodulatory, and anti-inflammatory effects of Spirulina in disease conditions: a systematic review.
The goal of this review was to critically evaluate the research on the effects of Spirulina (SP) supplementation on pathological conditions in the general population, considering its antioxidant, immunomodulatory, and anti-inflammatory properties. On a total of 1621 records screened, eighteen studies published between 2001 and 2020 met the inclusion criteria. Despite variability in research methodology and patient conditions, the findings of these studies generally support the benefits of supplementing diet with SP in subjects with both transmittable and non-transmittable diseases. Improvements were found for all the conditions studied, with the only exception of male infertility. However, the number of clinically controlled trials examining the effects of SP on specific health conditions is still very low, and some studies show medium quality. Further research in this field is needed to confirm the possible clinical role of Spirulina supplementation in parallel with medical therapies.


Therapeutic efficacy of repeated cycles of local IL-2 injections in mice carrying slowly growing tumour grafts.
Local IL-2 administration prior to transplantation of murine sarcoma virus (MSV Harvey)-induced tumour MSVT2 provided a model of slowly growing tumours suitable for long-term investigation of the therapeutic efficacy of repeated IL-2 injection cycles. Challenge of mice with the dose of sarcoma cells, which was lethal for 20/20 untreated control recipients, revealed that 8/20 IL-2-pretreated mice were protected by the local IL-2 treatment and survival indefinitely. Nine out of twenty IL-2-pretreated mice died during the same time period as the control mice, i.e., during 36 days, and 3/20 IL-2 pretreated mice were tumour-negative until day 60, when incipient tumours arose. The three late tumours were used as a model to investigate the therapeutic efficacy of the new cycles of repeated local IL-2 administration. It was found that no resistance to IL-2 immunotherapy was induced by pretreatment of the late tumours and that the tumours were repeatedly susceptible to local IL-2 treatment. Spleen cells of the tumour-bearing mice, which were not cytotoxic for MSVT2 tumour cells in vitro, could be made cytotoxic by addition of exogenous IL-2.


Altered Vitamin D Receptor Expression in Apa-I (rs7975232) Allelic Variants-A Probable Risk Factor for Susceptibility to Hepatitis B Virus Infection and Disease Progression.
Vitamin D exerts its antiviral effect through vitamin D receptor (VDR)/retinoid X receptor-mediated host immunomodulation. Besides the downregulation of VDR expression, its polymorphism was also observed among hepatitis B virus (HBV)-positive patients. To understand the possible link between VDR polymorphism and its altered expression during HBV infection and disease progression, VDR Apa-I [rs7975232 (C>A)] single nucleotide polymorphism (SNP) was analyzed in a case-control manner. VDR Apa-I (rs7975232, C>A) polymorphism was studied using 340 HBV patients and 102 healthy controls. Genotype analysis and gene expression study was performed using restriction fragment length polymorphism and quantitative polymerase chain reaction, respectively. Statistical analysis was performed using SPSS (IBM) considering p-value <0.05 as significant for comparing the differences between the groups. Significant mean difference in VDR expression was observed between HBV-positive patients (1.6 ± 0.94) and controls (0.69 ± 0.73). Furthermore, the mean fold change of Healthy control with CC genotype (1.92 ± 0.99) was found to be marginally significant compared with mutant genotype (CA/AA) (1.08 ± 0.43/0.59 ± 0.56, p = 0.045). In HBV+ patients, the mean fold change in the CC genotype was 0.88 ± 0.38, which exhibits a significant mean difference upon comparison with other genotypes (0.52 ± 0.49, 0.113 ± 0.34; p = 0.018, p = 0.048). However, the fold change value does not differ between CA and AA genotypes. Further comparative analysis of VDR expression between the control and case also exhibits significant differences (p = 0.001) among allelic variants. Observed genotype distribution frequency exhibits a significant association with disease type. The mutant genotype was found to be significantly associated with HBV infection and disease progression, (odds ratio = 0.730, 95% confidence interval = 0.462-1.152, p = 0.06). VDR SNP rs7975232 (C>A) may affect VDR expression by controlling several other variables and suggest that deviation from wild-type genotype (CC) is associated with downregulation of expression, which in turn involved in host immunomodulation in favor of HBV infection and disease progression.


Pregnancy immunology: decidual immune cells.
Human pregnancy is a complex process. Placental development depends on the function of secretory molecules produced by placental trophoblast cells as well as by maternal uterine immune cells within the decidua. These decidual immune cells are T cells, natural killer cells, macrophages and dendritic cells. The interactions between the trophoblast cells and the maternal immune cells have an impact on the outcome of the pregnancy. Knowledge about the phenotypes and functions of the maternal immune cells in normal and pathological pregnancies including recurrent spontaneous abortions, preeclampsia and hydatidiform moles may improve our understanding of the immunobiology of the normal pregnancy as a whole and may provide approaches for improving the treatment of pathological pregnancies.


Visual impact beam plots: Analyzing research profiles and bibliometric metrics using the following-leading clustering algorithm (FLCA).
A new approach to showcasing author publications on a website involves using a visual representation instead of the conventional paper list. The creation of an impact beam plot (IBP) as a research profile for individuals is crucial, especially when incorporating collection edges that include self-cited articles through a rare cluster analysis technique not commonly found in the literature. This study presents the application of a unique method called the following-leading clustering algorithm (FLCA) to generate IBPs for 3 highly productive authors. For the 3 highly productive authors, Sung-Ho Jang from South Korea, Chia-Hung Kao from Taiwan, and Chin-Hsiao Tseng from Taiwan, all their published articles indexed in the Web of Science Core Collection were downloaded. Sung-Ho Jang published 593 articles, Chia-Hung Kao published 732 articles, and Chin-Hsiao Tseng published 160 articles. To analyze and showcase their publications, the FLCA was utilized. This algorithm helped cluster their articles and identify representative publications for each author. To assess the effectiveness and validity of the FLCA algorithm, both network charts and heatmaps with dendrograms were employed. IBPs were then created and compared for each of the 3 authors, taking into consideration their h-index, x-index, and self-citation rate. This allowed for a comprehensive visual representation of their research impact and citation patterns. The results show that these authors' h-index, x-index, and self-citation rates were (37, 44.01, 1.66%), (42, 61.47, 0.23%), and (37, 40.3, 6.62%), respectively. A higher value in these metrics indicates a more remarkable research achievement. A higher self-citation rate with a lower cluster number indicates that manuscripts are more likely to have been self-drafted. Using the FLCA algorithm, IBPs were successfully generated for each author. The FLCA algorithm allows for the easy generation of visual IBPs based on authors' publication profiles. These IBPs incorporate 3 important bibliometric metrics: h-index, x-index, and self-citations. These metrics are highly recommended for use by researchers globally, particularly with the self-citation rate, as they offer valuable insights into the scholarly impact and citation patterns of individual researchers.


Supramolecular interaction of labetalol with cucurbit[7]uril for its sensitive fluorescence detection.
This work studied the host-guest interaction between cucurbit[7]uril (CB[7]) and labetalol in acidic aqueous solution and proposed a simple competitive method for fluorescence detection of labetalol. The binding constant of labetalol-CB[7] was (1.83 ± 0.22) × 10(6) M(-1), which was greater than those of palmatine-CB[7], berberine-CB[7], and coptisine-CB[7] complexes. The fluorescence intensity of palmatine-CB[7], berberine-CB[7], and coptisine-CB[7] complexes decreased linearly with increasing concentration of labetalol ranging from 0.014 to 2.06, 0.014 to 1.15, and 0.034 to 1.23 μM, respectively. Based on the competitive interaction, the proposed detection method for labetalol showed limits of detection of 4.9 nM, 4.9 nM, and 12.0 nM, respectively, and was successfully applied for the determination of labetalol in human urine samples with good precision and recoveries from 95.4% to 102.5%. Moreover, it could be employed to monitor the time-dependent concentration of labetalol in urine from a healthy volunteer after oral medication. The superstructure-based competitive mode provided a promising fluorescence assay strategy for various potential applications.


Data collection procedures for school-based surveys among adolescents: the Youth in Europe Study.
Collection of valid and reliable surveillance data as a basis for school health promotion and education policy and practice for children and adolescence is of great importance. However, numerous methodological and practical problems arise in the planning and collection of such survey data that need to be resolved in order to ensure the validity of the data and to maximize the response rate without being prohibitively costly. This article builds on a 15-yearlong experience of such annual data collections in Iceland and describes the preparation, process, and collection of data that provide a common methodologic framework for the school-based survey, Youth in Europe, a population-based survey of 14- to 16-year-old adolescents, being collected across 18 European cities now participating in the European Cities Against Drugs (ECAD) program. We identified 11 critical steps for developing and implementing the surveys in light of the recent literature on the preparation and implementation practices in school-based data collection among adolescents. Limiting the disruption of daily operations in schools while at the same time ensuring both quality and clarity of data collection procedures in school-based surveys are of paramount importance for researchers, school personnel, and students.


Women's health inequalities in 15 Muslim-populated countries: Evidence from population and mortality statistics.
In this study, we examined health inequalities and the status of women as evidenced in the patterns of population and mortality statistics in fifteen Muslim-populated countries. Based on WHO data, female-to-male ratios were calculated to determine differential gender ratios of population and mortality, using Western gender patterns as a baseline. The socioeconomic contexts of the analysis were the percentage of women in parliaments data by OECD and the Gross National Income Per Capita PPP by the World Bank. The study results indicate that former USSR countries had fewer girls, suggesting gender selection, whilst fewer adult women in the Gulf States population indicate health inequalities. Female children's mortality was under-reported, inferring under-valuing girls. Higher female adult deaths in Egypt, Iran, and the Gulf States show greater discrimination. Women in the richest Muslim countries face more inequalities and less representation in Parliament. The implications of the study are discussed.


The KP1_4563 gene is regulated by the cAMP receptor protein and controls type 3 fimbrial function in Klebsiella pneumoniae NTUH-K2044.
Klebsiella pneumoniae (K. pneumoniae) is an opportunistic pathogen that can adhere to host cells or extracellular matrix via type 1 and type 3 fimbriae. KP1_4563 is a gene encoding a hypothetical protein in K. pneumoniae NTUH-K2044. KP1_4563 is located between the type 1 and type 3 fimbrial gene clusters and is likely associated with fimbrial function given its putative conserved domains of unknown function (DUF1471). Cyclic AMP receptor protein (CRP) regulates virulence-related gene expression and is a crucial transcriptional regulator in many bacteria. The predicted DNA recognition motif of CRP is present in the KP1_4563 promoter region. This study aimed to investigate the function of KP1_4563 in fimbriae and its transcriptional regulation mechanism by CRP. We generated Kp-Δ4563 mutant and complementation strains. We utilized phenotype and adhesion assays to evaluate the role of KP1_4563 in fimbriae. We conducted quantitative RT-PCR (qRT-PCR), LacZ fusion, electrophoretic mobility shift, and DNase I footprinting assays to study the transcriptional regulation of KP1_4563 gene by CRP. We found that KP1_4563 negatively regulates the function of type 3 fimbriae. Compared with NTUH-K2044, the absence of KP1_4563 enhanced the ability of Kp-Δ4563 to adhere to A549 cells. CRP negatively regulates KP1_4563 by directly binding to its promoter region. KP1_4563 plays an important role in type 3 fimbrial function. This novel insight will assist in the development of strategies for preventing K. pneumoniae infection.


Changes in children's stressors over the past 30 years.
Social and political changes have introduced new sources of stressors for children. Two studies of 7- to 12-year-old children (n=194; n=402) identified stressors from their perspective, compared them to items in existing instruments, analyzed changes in stressors over the past 30 years, and determined the need for a new instrument. The children named 908 stressors that were inductively sorted into 54 mutually exclusive categories of stressors. Only 24% to 50% of the categories were represented in existing instruments. Stressors that emerged in 2000 included being alone, tests, family fighting, too many things to do, and boyfriend/girlfriend issues. These findings provide insight into differences between adults' versus children's perspectives on stressors and invite speculation on the reasons why new stressors emerge over the years. The findings also support the need for a new instrument that captures the full range of stressors that children experience today.


Inflammatory and Immune Responses to Surgery and Their Clinical Impact.
The aim of this study was to describe current understanding of the local and systemic immune responses to surgery and their impact on clinical outcomes, predictive biomarkers, and potential treatment strategies. Patients undergoing major surgery are at risk of life-threatening inflammatory complications that include infection, the systemic inflammatory response syndrome (SIRS), or sepsis. Although improvements in surgical technique and peri-operative care have resulted in reduction in the rates of these complications, they remain high, especially in patients undergoing complex abdominal procedures. There are currently no drugs licensed specifically for the treatment of sepsis nor is it possible to identify those at highest risk, which would allow pre-emptive therapy that may improve outcomes. Local immune responses to surgery lead to systemic pro-inflammatory and immunosuppressive phases that are temporally related and proportionate in magnitude. Improved understanding of these mechanisms has implications for clinical study design and has led to the emergence of novel biomarkers such as Toll-like receptor expression. These can be used to stratify patient care pathways to maximize the benefit from current therapies or to select the right target at the right phase of illness for future drug development.


Laparoendoscopic single site surgery in pediatric urology: does it require specialized tools?
To describe our experience utilizing Laparoendoscopic single site (LESS) surgery in pediatric urology. Retrospective chart review was performed on LESS urologic procedures from November 2009 through March 2013. A total of 44 patients underwent 54 procedures including: nephrectomy (23), orchiopexy (14), varicocelectomy (9), orchiectomy (2), urachal cyst excision (3), and antegrade continence enema (3) (ACE). Median patient age was 6.9 years old. Estimated blood loss (EBL), ranged from less than 5cc to 47cc for a bilateral nephrectomy. Operative time varied from 56 mins for varicocelectomy to a median of 360 minutes for a bilateral nephroureterectomy. Incision length ranged between 2 and 2.5cm. In our initial experience we used a commercial port. However, as we progressed, we were able to perform the majority of our procedures via adjacent fascial punctures for instrumentation at the single incision site. One patient did require conversion to an open procedure as a result of bleeding. Three complications were noted (6.8%), with two Clavien Grade 3b complications. Two patients required additional procedures at 1-year follow-up. The use of LESS applies to many pediatric urologic procedures, ideally for ablative procedures or simple reconstructive efforts. The use of adjacent fascial puncture sites for instrumentation can obviate the need for a commercial port or multiple trocars.


Association between blood eosinophil count and small airway eosinophils in smokers with and without COPD.
Airway eosinophilic inflammation is a pathological feature in a subgroup of patients with COPD and in some smokers with a high COPD risk. Although blood eosinophil count is used to define eosinophilic COPD, the association between blood eosinophil count and airway eosinophilic inflammation remains controversial. This cross-sectional study tested this association in smokers with and without COPD while considering potential confounders, such as smoking status and comorbidities. Lung specimens were obtained from smokers with and without COPD and non-COPD never-smokers undergoing lung lobectomy. Those with any asthma history were excluded. The infiltration of eosinophils into the small airway wall was quantified on histological sections stained with major basic protein (MBP). The number of airway MBP-positive cells was greater in smokers (n=60) than in never-smokers (n=14). Smokers with and without COPD (n=30 each) exhibited significant associations between blood eosinophil count and airway MBP-positive cells (ρ=0.45 and 0.71). When smokers were divided into the high and low airway MBP groups based on their median value, blood eosinophil count was higher in the high-MBP group, with no difference in age, smoking status, comorbidities, emphysema or coronary artery calcification on computed tomography, and inhaled corticosteroid (ICS) use. The association between greater blood eosinophil count and the high-MBP group was confirmed in multivariable models adjusted for smoking status, airflow limitation and ICS use. The blood eosinophil count may reflect eosinophilic inflammation in the small airways in smokers with and without COPD.


Skeletal muscle fibre-type shifting and metabolic profile in patients with chronic obstructive pulmonary disease.
The aim of this study was to examine the nature of fibre-type redistribution in relation to fibre metabolic profile in the vastus lateralis in chronic obstructive pulmonary disease (COPD) and COPD subtypes. Fifteen COPD patients (eight with emphysema stratified by high-resolution computed tomography) and 15 healthy control subjects were studied. A combination of myofibrillar adenosine triphosphatase staining and immunohistochemistry was used to identify pure, as well as hybrid fibre types. For oxidative capacity, fibres were stained for cytochrome c oxidase and succinate dehydrogenase activities, and glycogen phosphorylase for glycolytic capacity. The proportion of type-I fibres in COPD patients was markedly lower (16% versus 42%), especially in emphysema, and the proportion of hybrid fibres was higher (29% versus 16%) compared to controls. The proportion of fibres staining positive for oxidative enzymes was lower in COPD patients, which correlated with the proportion of type-I fibres. In COPD oxidative capacity was lower within IIA fibres. The authors conclude that fibre-type transitions are involved in the fibre-type redistribution in chronic obstructive pulmonary disease. Low oxidative capacity is closely related to the proportion of type-I fibres, but an additional reduction of oxidative enzyme activity is present within IIA fibres. Fibre-type abnormalities may be aggravated in emphysema.


T cell-mediated pathology in two models of experimental colitis depends predominantly on the interleukin 12/Signal transducer and activator of transcription (Stat)-4 pathway, but is not conditional on interferon gamma expression by T cells.
The requirements for interleukin (IL)-12/signal transducer and activator of transcription (Stat)-4 signaling and induction of T cell-specific interferon (IFN)-gamma expression in the development of T helper cell (Th)1-type pathology were examined in two different models of experimental colitis. In each model, abnormal reconstitution of the T cell compartment in immunodeficient mice by adoptive cell transfer leads to a wasting syndrome and inflammation of the colon, induced by IFN-gamma and tumor necrosis factor (TNF)-alpha-producing T cells. We show here that treatment with anti-IL-12 antibodies in one of the models, or reconstitution with T cells from Stat-4-deficient (Stat-4(null)) mice in both models resulted in a milder disease in the majority of recipient animals, compared with those that were left untreated or that had been reconstituted with wt cells. Protected mice in each group also harbored lower frequencies of IFN-gamma-producing T cells than did diseased mice, suggesting that effects on wasting and colitis resulted from the attenuation of IFN-gamma expression by T cells. To test whether the development of pathogenic T cells in the two colitis models was directly dependent on T cell-specific IFN-gamma expression, IFN-gammanull donors were used for T cell reconstitution in each system. Surprisingly, large numbers of IFN-gammanull-reconstituted mice developed wasting and colitis, which in many cases was of comparable severity to that seen in animals reconstituted with wt cells. Furthermore, T cells from these animals expressed TNF-alpha, demonstrating that they had retained the ability to produce another proinflammatory cytokine. Taken together, these results demonstrate that in some forms of chronic experimental colitis the development of pathogenic T cells is influenced predominantly, though not exclusively, by IL-12 via the actions of Stat-4 proteins. Furthermore, our data suggest that in the models of colitis studied here the effects of IL-12/Stat-4 or other Th1 promoting pathways are not limited to the induction of IFN-gamma gene expression in T lymphocytes.


Animal Models: New Biosensors and Technologies?Including Those for Bone Regrowth and Wound Healing?Advance Animal Health Care While Also Providing a Fertile Testing Ground for Human Health.
Dogs have bad breath. But when Montana sheep rancher Katy Harjes noticed her collie, Hoshi, had particularly bad breath and facial swelling, she was concerned that the symptoms might be a sign of something serious. She was right; ten-year-old Hoshi had squamous cell carcinoma, a common type of oral tumor found in dogs. The cancer had not metastasized, but the damage was extensive enough that part of Hoshi's lower jaw needed to be removed. Luckily, Hoshi was a suitable candidate for a stateof-the-art bone regrowth procedure developed by Frank Verstraete, B.V.Sc, Dr.Med.Vet., M.Med.Vet., and Boaz Arzi, D.V.M., oral surgeons at the University of California (UC), Davis, School of Veterinary Medicine. Consequently, Katy and Hoshi embarked on a 15-hour road trip to California.


Rate and predictors of HIV virological failure among adults on first-line antiretroviral treatment in Dar Es Salaam, Tanzania.
Monitoring resistance to first line Antiretroviral therapy (ART) is crucial in preventing accumulation of viral mutations following the implementation of the World Health Organization "treat all" initiative. We estimated the rate and predictors of virological treatment failure among adults living with HIV/AIDS in Dar es Salaam, Tanzania. A retrospective cohort study involving adults aged 18 and above receiving first line ART in Dar as Salaam between 2016 and 2018 were recruited using multistage random sampling. Clinical and laboratory data were extracted from Care and Treatment Clinic database-2 (CTC2) followed by participant's interviews. Adjusted Cox-regression modelling was used to determine independent predictors of treatment failure. A total of 340 participants with mean age of 37 were recruited. Overall, 10.59% had virological failure and the rate of failure was 5.24 (95% CI:3.72; 7.27) per 100 person-months at risk with a median failure time of 18 months. Independent predictors of treatment failure were being a male (Adjusted hazard ratio (aHR) 2.78, 95%CI:1.16;6.63), having used treatment for less than two years (aHR, 12.48, 95%CI:3.64-22.71) and co-infection with Tuberculosis (aHR 2.1, 95%CI: 1.0;5.9). HIV virological failure occurs early during treatment in this population. Male clients, co-infected with Tuberculosis were at higher risk of ART failure within two years of treatment. Substantial stride has been made towards the achievement of the last UNAIDS 90 goal but tailored counseling and close monitoring of HIV/TB co-infected male clients following ART initiation could accelerate efforts to close the gap. Further studies on pre-treatment drug resistance mutations are called for.


Clinical development of RTS,S/AS malaria vaccine: a systematic review of clinical Phase I-III trials.
The first clinical Phase III trial evaluating a malaria vaccine was completed in December 2013 at 11 sites from seven sub-Saharan African countries. This systematic review assesses data of Phase I-III trials including malaria-naive adults and adults, children and infants from malaria endemic settings in sub-Saharan Africa. The main endpoint of this systematic review was an analysis of the consistency of efficacy and immunogenicity data from respective Phase I-III trials. In addition, safety data from a pooled analysis of RTS/AS Phase II trials and RTS,S/AS01 Phase III trial were reviewed. The RTS,S/AS01 malaria vaccine may become available on the market in the coming year. If so, further strategies should address challenges on how to optimize vaccine efficacy and implementation of RTS,S/AS01 vaccine within the framework of established malaria control measures.


Comparative development in Moenkhausia pittieri and Paracheirodon innesi (Ostariophysi: Characiformes) with comments on heterochrony and miniaturization in the Characidae.
Developmental heterochronies between the small-sized Moenkhausia pittieri and the miniature Paracheirodon innesi were determined by describing and comparing external morphological development in both species. Despite being classified as miniature, P. innesi was larger and more developed in the flexion and post-flexion larval stages than M. pittieri. Broader patterns within the Characidae suggest that body size reduction in P. innesi is tied to accelerated development and higher growth rate along with early maturation and precocious cessation of growth, which are characteristic of miniature species. Comments on adipose-fin loss in Characidae as a consequence of developmental truncation are included.


Characterizing Early Inpatient Death After Trauma.
There is a paucity of data to predict early death or futility after trauma. The objective of this study was to characterize the laboratory values, blood product administration, and hospital disposition for patients with trauma who died within 72 h of admission. All deaths within 72 h of admission over a 5-y period at a level I trauma center were reviewed. Blood transfusion within the first 4 h of arrival and patient disposition from the emergency department to the operating room (OR), surgical intensive care unit, or the neuroscience intensive care unit (NSICU) were analyzed. Kaplan-Meier curves were generated to determine time to death. A total of 622 subjects were identified; 39.5% died in the emergency department, 10.6% went directly to the OR, 13.6% were admitted to the surgical intensive care unit, and 29.7% admitted to the NSICU. Of these subjects, 201 (32.2%) patients received blood within the first 4 h. By 24 h, early blood transfusion was associated with more rapid death for patients who were admitted to the NSICU (80% versus 60% mortality, P = 0.01) but not for patients taken directly to the OR (80% versus 70% mortality, P = 0.2). Admission coagulopathy by international normalized ratio (P < 0.01), but not anemia (P = 0.64) or acidosis (P = 0.45), correlated with a shorter time to death. In contrast, laboratory values obtained at 4 h after admission did not correlate with time to death. Our data demonstrate that admission coagulation derangement and need for early blood product transfusion are the two factors most associated with early death after injury, particularly in those patients with traumatic brain injury. These data will help construct future models for futility of continued care in patients with trauma.


[Transcatheter aortic valve-in-valve implantation for the treatment of a failed surgical bioprosthesis of small size].
In the last decades, the wide use of surgical aortic bioprostheses translated into an increased rate of valve degeneration and dysfunction. However, most patients are excluded from surgical re-do due to high or prohibitive operative risk. In this clinical context, valve-in-valve transcatheter aortic valve implantation (ViV-TAVI) has been shown to be effective. Nevertheless, some relevant concerns, such as elevated residual transvalvular gradient and coronary artery obstruction, still remain. Detailed information regarding the degenerated surgical bioprosthesis and a thorough analysis of the computed tomography scan are essential for accurate pre-procedural planning and may avoid dramatic acute complications. Moreover, in difficult cases, the use of some tips and tricks may help expert operators to achieve better results.Here we report the case of a patient affected by structural degeneration of a small label size surgical aortic valve, who was excluded from surgical reintervention due to high operative risk. Therefore, we decided to perform a ViV-TAVI despite the presence of challenging features.


Interlocked circle formation by group I introns: structural requirements and mechanism.
Precursor RNA transcribed from the yeast mitochondrial gene coding for the large ribosomal RNA contains a group I intron that can excise itself in vitro. Apart from group I specific sequence elements the intron also contains a gene encoding a DNA endonuclease involved in intron dispersal. A precursor RNA derivative from which this gene has been removed self-splices efficiently, but due to activation of cryptic opening sites located in the 5' exon, the 3' part of this exon is sometimes co-excised with the intron. Upon further reaction, this enlarged intron molecules give rise to interlocked circles, comprising small circles derived from 5' exon parts and large circles of the intron. Sequence comparison between cryptic opening sites and authentic splice sites reveals in most cases homology with the 3' exon part that is capable of interacting with the Internal Guide Sequence. The role of the IGS was further substantiated by replacing the cryptic opening sites with well defined sequences of authentic splice sites: one corresponding to the 3' splice site and its mutant derivatives, the other to a fragment containing the natural 5'-3' exon junction. Precursor RNAs derived from these constructs give rise to interlocked circles, and mutation studies confirm that the 3' exon nucleotides flanking a 3' splice site are essential for their formation. The results underline the crucial role of the IGS in interlocked circle formation which behaves similarly as in the normal self-splicing reactions. It has been proposed that the two short helices formed by basepairing of the IGS with the 5' and 3' exon can co-axially stack on top of each other forming a quasi continuous RNA double helix or pseudoknot. We present a model explaining how transesterification reactions of a mutant precursor RNA in such a pseudoknot can lead to interlocked circles. The experiments support the notion that a similar structure is also operative in splicing of wild type precursor RNA.


Transcriptome analysis in maritime pine using laser capture microdissection and 454 pyrosequencing.
Maritime pine (Pinus pinaster Aiton) is one of the most advanced conifer models for genomics research. Conifer genomes are extremely large and major advances have recently been made in the characterization of transcriptomes. The combination of laser capture microdissection (LCM) and next-generation sequencing is a powerful tool with which to resolve the entire transcriptome of specific cell types and tissues. In the current work, we have developed a protocol for transcriptomic analyses of conifer tissue types using LCM and 454 pyrosequencing. Tissue sections were isolated using non-fixed flash-frozen samples processed by LCM. Complementary DNA synthesis and amplification from tiny amounts of total RNA from LCM samples was performed using an adapted protocol for C: onifer R: NA A: mplification (CRA+). The cDNA amplification yield and cDNA quality provided by CRA+ were adequate for 454 pyrosequencing. Furthermore, read length and quality results of the 454 runs were near the optimal parameters considered by Roche for transcriptome sequencing. Using the CRA+ protocol, non-specific amplifications were prevented, problems derived from poly(A:T) tails in the 454 sequencing technology were reduced, and read length and read number considerably enhanced. This technical approach will facilitate global gene expression analysis in individual tissues of conifers and may also be applied to other plant species.


[Diagnostic unit EKS-K1200].
The paper presents the block diagram of the unit, describes some assemblies and its operation as a whole, gives information on the software and technical data. The computer-based EKS-K 1200 unit integrates a pacemaker, a 12-channel electrocardiograph with a monitor, and a database.


[Heart function in chronic pressure overload caused by aortic stenosis: the role of collagen tissue].
The purpose of this study was to evaluate left ventricular (LV) structure-function interplay in aortic stenosis. LV structure was assessed from endomyocardial biopsies obtained in 27 patients with aortic stenosis. Total collagen volume fraction, orthogonal collagen fiber meshwork (cross-hatching) and endocardial fibrosis were determined by morphologic-morphometric evaluation. Control biopsy data were obtained from 6 pre-transplantation donor hearts whereas other 11 patients with normal cardiac function served as hemodynamic controls. LV biplane cineangiography and high-fidelity LV pressure measurement were carried out in all patients. Systolic function was assessed by LV biplane ejection fraction, diastolic function by time constant of relaxation, peak filling rates and passive elastic properties. Total collagen volume fraction (7.3 versus 1.6%, p < 0.01) as well as the degree of cross-hatching (1.7 versus 0.8 grade, p < 0.01) were significantly increased in patients with aortic stenosis with respect to controls. Endocardial fibrosis was present in 11/27 patients with aortic stenosis and in no patients of control group. In aortic stenosis in presence of increased total collagen volume fraction there were no changes in systolic and diastolic function, whereas in presence of changes in collagen architecture ejection fraction was depressed and passive elastic properties increased. In conclusion, in aortic stenosis, changes in collagen architecture are associated with altered systolic function and passive diastolic properties. The sole increase in total collagen volume fraction without a change in architecture leaves systolic and passive diastolic function unaltered. A prolongation of relaxation was present in aortic stenosis and appears to be mediated by muscle hypertrophy per se.


Myopic phakic STAAR collamer posterior chamber intraocular lenses for keratoconus.
To evaluate the efficacy, predictability, and safety of myopic phakic posterior chamber Implantable Collamer Lens (ICL) to correct myopia associated with keratoconus. In a prospective, noncomparative, interventional case series, outcomes in 25 consecutive keratoconus eyes of 16 patients, myopia from -3.00 to -18.00 diopters (D) and astigmatism from -0.50 to -3.00 D, were analyzed 12 months after implantation of myopic ICLs (STAAR Surgical). All eyes showed best spectacle-corrected visual acuity (BSCVA) > or =20/50, central keratometric values < or =52.00 D, and refractive cylinder < or =3.00 D. Myopic ICL implantation was performed through a corneal incision in the steepest meridian. The mean uncorrected visual acuity and BSCVA after ICL implantation were 0.17+/-0.19 and 0.12+/-0.12 logMAR, respectively. The efficacy index was 0.98. No eyes lost 2 or more lines of visual acuity, 2 eyes lost 1 line, 18 eyes did not change after surgery, and 5 eyes gained 1 or more lines. The safety index was 1.05. Best spectacle-corrected visual acuity after ICL implantation was statistically significantly better than preoperative BSCVA (P=.0021). Spherical equivalent refraction was within +/-1.00 D of the desired refraction in all cases and 84% of cases were within +/-0.50 D. Mean postoperative spherical equivalent refraction was 0.32+/-0.55 D at 12 months. Myopic ICL implantation was a safe, effective, and predictable procedure for the correction of myopia associated with keratoconus.


[Mixed gonadal dysgenesis--a rare cause of primary infertility. Report of 2 cases and review of the literature].
We report on two cases of primary male infertility attributable to mixed gonadal dysgenesis. In one case the diagnosis was made when the patient was 30 years old; in the other, the 44-year-old patient had been evaluated 4 years before for erectile dysfunction, which had been treated with a penile prosthesis without the diagnosis being made. These two cases are life following exceptional in that the diagnosis was made in adult-life following an uncomplicated male adolescence.


Dynamic Expression Patterns of Progenitor and Neuron Layer Markers in the Developing Human Dentate Gyrus and Fimbria.
The molecular mechanisms that orchestrate the development of the human dentate gyrus are not known. In this study, we characterized the formation of human dentate and fimbrial progenitors and postmitotic neurons from 9 gestational weeks (GW9) to GW25. PAX6+ progenitor cells remained proliferative until GW16 in the dentate ventricular zone. By GW11, the secondary dentate matrix had developed in the intermediate zone, surrounding the dentate anlage and streaming toward the subpial layer. This secondary matrix contained proliferating PAX6+ and/or TBR2+ progenitors. In parallel, SOX2+ and PAX6+ fimbrial cells were detected approaching the dentate anlage, representing a possible source of extra-dentate progenitors. By GW16, when the granule cell layer could be delineated, a hilar matrix containing PAX6+ and some TBR2+ progenitors had become identifiable. By GW25, when the 2 limbs of the granule cell layer had formed, the secondary dentate matrix was reduced to a pool of progenitors at the fimbrio-dentate junction. Although human dentate development recapitulates key steps previously described in rodents, differences seemed to emerge in neuron layer markers expression. Further studies are necessary to better elucidate their role in dentate formation and connectivity.


Hyperandrogenism and phenotypes of polycystic ovary syndrome are not associated with differences in obstetric outcomes.
To investigate obstetric outcomes in Danish women with different phenotypes of polycystic ovary syndrome (PCOS) and isolated hyperandrogenism (HA) and describe the risk of adverse obstetric outcomes in women with PCOS and HA compared to controls. Cohort study. Odense University Hospital, Denmark. Women with PCOS were identified prospectively starting in 1997. Singleton pregnancies in women with PCOS and HA during 2003-2011 were included (n = 199). A control group was matched to the patient cohort according to date of childbirth (n = 995). Data on clinical characteristics and obstetric outcomes were collected in patients with PCOS and HA and controls. In PCOS and HA, total and free testosterone, sex hormone binding globulin, and hemoglobin A1c were measured outside pregnancy. During pregnancy, oral glucose tolerance tests were performed in 39 patients and 123 controls according to Danish national guidelines. PCOS phenotypes were based on the Rotterdam criteria. Gestational diabetes mellitus, pregnancy-induced hypertension, preeclampsia, delivery by emergency cesarean section, preterm delivery and anthropometric measures in the newborn. The incidence of adverse obstetric outcomes and anthropometric measures among the newborns were comparable between different phenotypes of PCOS and patients with HA. In the oral glucose tolerance test, patients had a higher risk of gestational diabetes mellitus compared with controls; the odds ratio (95% confidence interval) was 3.3 (1.5-6.9) after adjustment for age, parity, and body mass index (p = 0.002). The incidence of other adverse obstetric outcomes was similar in patients and controls. Obstetric outcomes were comparable in women with different PCOS phenotypes.


Appropriate in vitro methods for genotoxicity testing of silver nanoparticles.
We investigated the genotoxic effects of 40-59 nm silver nanoparticles (Ag-NPs) by bacterial reverse mutation assay (Ames test), in vitro comet assay and micronucleus (MN) assay. In particular, we directly compared the effect of cytochalasin B (cytoB) and rat liver homogenate (S9 mix) in the formation of MN by Ag-NPs. Before testing, we confirmed that Ag-NPs were completely dispersed in the experimental medium by sonication (three times in 1 minute) and filtration (0.2 µm pore size filter), and then we measured their size in a zeta potential analyzer. After that the genotoxicity were measured and especially, S9 mix and with and without cytoB were compared one another in MN assay. Ames test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains revealed that Ag-NPs with or without S9 mix did not display a mutagenic effect. The genotoxicity of Ag-NPs was also evaluated in a mammalian cell system using Chinese hamster ovary cells. The results revealed that Ag-NPs stimulated DNA breakage and MN formation with or without S9 mix in a dose-dependent manner (from 0.01 µg/mL to 10 µg/mL). In particular, MN induction was affected by cytoB. All of our findings, with the exception of the Ames test results, indicate that Ag-NPs show genotoxic effects in mammalian cell system. In addition, present study suggests the potential error due to use of cytoB in genotoxic test of nanoparticles.


Blocking an N-terminal acetylation-dependent protein interaction inhibits an E3 ligase.
N-terminal acetylation is an abundant modification influencing protein functions. Because ∼80% of mammalian cytosolic proteins are N-terminally acetylated, this modification is potentially an untapped target for chemical control of their functions. Structural studies have revealed that, like lysine acetylation, N-terminal acetylation converts a positively charged amine into a hydrophobic handle that mediates protein interactions; hence, this modification may be a druggable target. We report the development of chemical probes targeting the N-terminal acetylation-dependent interaction between an E2 conjugating enzyme (UBE2M or UBC12) and DCN1 (DCUN1D1), a subunit of a multiprotein E3 ligase for the ubiquitin-like protein NEDD8. The inhibitors are highly selective with respect to other protein acetyl-amide-binding sites, inhibit NEDD8 ligation in vitro and in cells, and suppress anchorage-independent growth of a cell line with DCN1 amplification. Overall, our data demonstrate that N-terminal acetyl-dependent protein interactions are druggable targets and provide insights into targeting multiprotein E2-E3 ligases.


Reliability evaluation of sampling plan fixed by Council Directive 91/68/EEC for the maintenance of officially brucellosis-free flock status.
The European Union (EU) strategy with respect to sheep and goat brucellosis aims to eradicate the infection and achieve officially brucellosis-free (OBF) status in all EU holdings and territories. Council Directive 91/68/EEC of 28 January 1991 states that to maintain OBF status of ovine or caprine holdings located outside an OBF territory, only a representative number of animals need to be tested annually. However, depending on the number of animals in a holding, this testing method risks non-detection of the infection, thereby reducing the efficacy of the brucellosis control plan. The recommended sampling procedure has a low sensitivity for detecting infection in medium-sized flocks; furthermore, the risk of not detecting re-infection in OBF flocks, particularly in territories that have not yet gained OBF status, is also not acceptable. Moreover, in large-sized flocks, the Directive sampling procedure entails taking an excessive number of samples, which can be very expensive. The authors evaluated, by using statistical analyses and a simulation model based on field data, the possible consequences of the current EU strategy. It is suggested that the sampling criteria for the maintenance of OBF status in the EU should be modified and that a statistically based sampling method should be applied instead of the fixed percentage method that is currently in use.


Carboxylation of terminal alkynes with CO2 using novel silver N-heterocyclic carbene complexes.
Four novel N-heterocyclic carbene (NHC) silver complexes, , have been synthesized and characterized. The single X-ray crystal diffraction data indicate a dinuclear solid-state structure for and and a mononuclear structure for and . These complexes have been successfully used as efficient catalysts for the C-H activating carboxylation of terminal alkynes with CO2. A wide range of substrates with various functional groups afforded the corresponding aryl or alkyl substituted propiolic acids in good yields under mild conditions. Moreover, the role of bases and the reaction mechanism is thoroughly discussed.


Hemodynamic effects of n-acetylprocainamide compared with procainamide in conscious dogs.
We examined the hemodynamic actions of clinically relevant i.v. doses (20 mg/kg and 10 mg/kg) of n-acetyl procainamide (NAPA) in conscious dogs preinstrumented with a left ventricular (LV) micromanometer, LV and aortic catheters, and ultrasonic crystals for measurement of LV internal diameter shortening (% delta D). Within 30 seconds after the 20-mg/kg dose, there were significant increases in heart rate (27 +/- 7 beats/min, mean +/- SEM; n = 6), maximum dP/dt (655 +/- 206 mm Hg/sec), and % delta D (2.2 +/- 0.9%; all p less than or equal to 0.05). However, by 6 hours after the dose there were reductions compared with control in peak LV pressure (19 +/- 9 mm Hg), dP/dt (610 +/- 210 mm Hg/sec), and % delta D (2.3 +/- 0.6%; all p less than or equal to 0.05). In contrast, equimolar doses of procainamide or drug vehicle alone evoked no response, as did NAPA after pretreatment with reserpine (0.25 mg/kg/day for 2 days) or hexamethonium (10-15 mg/kg). These data suggest NAPA produces a biphasic hemodynamic response with enhancement of LV performance early and a decrease later; this response is different from that of the parent compound, procainamide. These effects are likely mediated by the adrenergic nervous system at either a ganglionic or a central level.


Social conditions and mental health during COVID-19 lockdown among people who do not identify with the man/woman binomial in Spain.
Evidence suggests that non-binary people have poorer mental and physical health outcomes, compared with people who identify within the gender binomial (man/woman). Research on the impact of the COVID-19 pandemic on mental health has been conducted worldwide in the last few months. It has however overlooked gender diversity. The aim of our study was to explore social and health-related factors associated with mental health (anxiety and depression) among people who do not identify with the man/woman binomial during COVID-19 lockdown in Spain. A cross-sectional study with online survey, aimed at the population residing in Spain during lockdown, was conducted. Data were collected between the 8th of April until the 28th of May 2020, the time period when lockdown was implemented in Spain. Mental health was measured using the Generalised Anxiety Disorder 7-item (GAD-7) scale for anxiety, and the Patient Health Questionnaire (PHQ-9) for depression. The survey included the question: Which sex do you identify with? The options "Man", "Woman", "Non-binary" and "I do not identify" were given. People who answered one of the last two options were selected for this study. Multivariate regression logistic models were constructed to evaluate the associations between sociodemographic, social and health-related factors, anxiety and depression. Out of the 7125 people who participated in the survey, 72 (1%) identified as non-binary or to not identify with another category. People who do not identify with the man/woman binomial (non-binary/I do not identify) presented high proportions of anxiety (41.7%) and depression (30.6%). Poorer mental health was associated with social-employment variables (e.g., not working before the pandemic) and health-related variables (e.g., poor or regular self-rated health). These findings suggest that social inequities, already experienced by non-binary communities before the pandemic, may deepen due to the COVID-19 pandemic.


Differentiation of retinal ganglion cells and photoreceptor precursors from mouse induced pluripotent stem cells carrying an Atoh7/Math5 lineage reporter.
The neural retina is a critical component of the visual system, which provides the majority of sensory input in humans. Various retinal degenerative diseases can result in the permanent loss of retinal neurons, especially the light-sensing photoreceptors and the centrally projecting retinal ganglion cells (RGCs). The replenishment of lost RGCs and the repair of optic nerve damage are particularly challenging, as both RGC specification and their subsequent axonal growth and projection involve complex and precise regulation. To explore the developmental potential of pluripotent stem cell-derived neural progenitors, we have established mouse iPS cells that allow cell lineage tracing of progenitors that have expressed Atoh7/Math5, a bHLH transcription factor required for RGC production. These Atoh7 lineage reporter iPS cells encode Cre to replace one copy of the endogenous Atoh7 gene and a Cre-dependent YFP reporter in the ROSA locus. In addition, they express pluripotent markers and are capable of generating teratomas in vivo. Under anterior neural induction and neurogenic conditions in vitro, the Atoh7-Cre/ROSA-YFP iPS cells differentiate into neurons that co-express various RGC markers and YFP, indicating that these neurons are derived from Atoh7-expressing progenitors. Consistent with previous in vivo cell lineage studies, the Atoh7-Cre/ROSA-YFP iPS cells also give rise to a subset of Crx-positive photoreceptor precursors. Furthermore, inhibition of Notch signaling in the iPSC cultures results in a significant increase of YFP-positive RGCs and photoreceptor precursors. Together, these results show that Atoh7-Cre/ROSA-YFP iPS cells can be used to monitor the development and survival of RGCs and photoreceptors from pluripotent stem cells.


Penicillium marneffei in bronchoalveolar lavage fluid.
Penicillium marneffei is a rare human pathogen that may infect either healthy or immunocompromised hosts. In methenamine silver-stained preparations of bronchoalveolar lavage fluid from a patient with dermatomyositis on steroid treatment, round-to-oval intracellular and extracellular microorganisms were found. The finding of occasional septate and elongated forms established the microorganism as probably P marneffei, which was confirmed on culture. Distinguishing this rare fungus from Pneumocystis carinii is important because these two diseases require different forms of treatment.


Novel avenues for treating diabetic nephropathy: new investigational drugs.
At present, treatment of diabetic kidney disease (DKD) is still mainly based on drugs acting on glycemic and blood pressure control, as there is no validated therapy able to halt the progression of renal failure. Because of the high incidence of DKD, due to the increase of diabetes mellitus in general population, new therapeutic strategies are needed. Areas covered: We analysed ongoing and already completed clinical trials, from clinicaltrials.gov and PubMed, dealing with new therapies for DKD. Expert opinion: Among the drugs currently being explored, the most promising molecules are those that interfere with glucose-dependent pathways, in particular polyol, protein kinase, hexosamine and AGEs metabolic pathways, and impaired renal vascular regulation. One of the recent goals achieved by molecular biology is the development of monoclonal antibodies able to interfere with extracellular matrix accumulation and fibrosis. Other interesting therapies are under investigation and further studies with a greater number of patients will establish a better approach for diabetic nephropathy.


Therapeutic implications of inhibition versus killing of Mycobacterium avium complex by antimicrobial agents.
Patients with the acquired immune deficiency syndrome (AIDS) with disseminated Mycobacterium avium infection have responded poorly to treatment with rifabutine (Ansamycin) and clofazimine, in spite of the good in vitro response of M. avium to these antimicrobial agents. We compared the ability of these and other antimicrobial agents to kill versus the ability to inhibit the growth of strains of the M. avium complex isolated from patients with AIDS. Killing curve experiments showed that the concentrations of rifabutine and clofazimine needed to kill two log units of M. avium are at least 32 times greater than the concentrations needed to inhibit growth. Little or no killing occurred at concentrations of these antimicrobial agents that are achievable in serum. In contrast, five of seven strains tested were killed by ciprofloxacin at concentrations that can be achieved in serum. Ciprofloxacin should be studied further for possible use in the treatment of M. avium infections.


Concise Total Synthesis of (+)-Bionectins A and C.
The concise and efficient total synthesis of (+)-bionectins A and C is described. Our approach to these natural products features a new and scalable method for erythro-β-hydroxytryptophan amino acid synthesis, an intramolecular Friedel-Crafts reaction of a silyl-tethered indole, and a new mercaptan reagent for epipolythiodiketopiperazine (ETP) synthesis that can be unravelled under very mild conditions. In evaluating the impact of C12-hydroxylation, we have identified a unique need for an intramolecular variant of our Friedel-Crafts indolylation chemistry. Several key discoveries including the first example of permanganate-mediated stereoinvertive hydroxylation of the α-stereocenters of diketopiperazines as well as the first example of a direct triketopiperazine synthesis from a parent cyclo-dipeptide are discussed. Finally, the synthesis of (+)-bionectin A and its unambiguous structural assignment through X-ray analysis provides motivation for the reevaluation of its original characterization data and assignment.


[Implication of enhanced recovery after surgery in the surgical management of hypopharyngeal squamous cell carcinoma].
Objective: To investigate the safety and efficacy of enhanced recovery after surgery (ERAS) in the clinical management of hypopharyngeal squamous cell carcinoma (HSCC). Methods: In this retrospective study, a total of 168 patients with pyriform sinus carcinoma in Qilu Hospital of Shandong University from January 2015 to January 2019 were divided into two groups, based on the different perioperative interventions that patients received, i.e. the ERAS group (n=64) and the conventional group (n=104), including 164 males and 4 females, whose ages ranged from 42 to 84 years old. The difference between two groups in the operative time, postoperative nutritional status, incidences of postoperative complications and postoperative hospitalization time were compared using the student's t test, Chi-squared test or Fisher's exact test. Results: Compared with the conventional group, patients in the ERAS group had significantly shorter operative time [(166.8±58.2) min vs. (183.3±39.9) min,t=-2.72, P=0.031], higher levels of postoperative serum albumin [(38.3±4.2) μmol/L vs. (36.6±3.3) μmol/L, t=2.73, P=0.007] and more body weight [(65.4±9.4) kg vs. (62.1±9.4) kg, t=2.22, P=0.028], lower incidences of postoperative subcutaneous infection [7.8% (5/64) vs. 20.2% (21/104), χ²=4.64, P=0.03] and severe pneumonia [4.7% (3/64) vs. 15.4% (16/104), χ²=4.52, P=0.03], and shorter postoperative hospitalization time [(16.5±3.9) d vs. (18.2±4.3) d, t=-2.65, P<0.05]. Conclusion: ERAS is effective and safe in the surgical management of HSCC, with benefits in reducing the operative stress via saving operation time, shortening the hospitalization time, ameliorating nutritional status and decreasing the incidences of complications.


Development of an inductively coupled MR coil system for imaging and spectroscopic analysis of an implantable bioartificial construct at 11.1 T.
Developing a method to noninvasively monitor tissue-engineered constructs is critical for the optimization of construct design and for assessing therapeutic efficacy. For this purpose, NMR is a powerful technique that can be used to obtain both images and spectroscopic data. But the inherent sensitivity of NMR limits the observation of a bioartificial construct with current NMR surface coil technology. In this study, we address this limitation through the development of an inductively coupled, implantable coil system, demonstrate its use at high field (11.1 T), and investigate the use of this coil system for monitoring a bioartificial construct in vitro and in vivo. The results establish that large gains in signal to noise can be obtained with this coil system over that obtainable with a surface coil. This coil system provides a means to quantitatively analyze the structure and function of implanted bioartificial organs.


Bowel ischemia: a rare complication of thiopental treatment for status epilepticus.
Refractory status epilepticus (RSE) treatment is usually performed with coma induction using an appropriate general anesthetic. Most frequent complications are represented by hypotension and infection. Other side-effects may however be encountered. We describe two patients suffering from acute bowel ischemia after thiopental (THP) treatment for RSE. A 73-year-old man with a complex-patial RSE following an acute stroke received THP (303 mg/kg over 48 h); 36 h after THP discontinuation, he presented abdominal tenderness and lactate elevation. Necrosis of the terminal ileum and colon was seen during surgical exploration; he deceased shortly thereafter. A 21 year-old woman had a cryptogenic de novo generalized-convulsive RSE resistant to 5 attempts of EEG burst-suppression. During the 6th attempt, after THP (840 mg/kg over 150 h) together with mild hypothermia, she developed an ileus with elevated serum lactate; caecum necrosis was observed during surgery. Hypernatremia, acidosis and hyperlactatemia heralded this complication in both patients. In these two patients, mechanical vascular ischemia may have resulted from drug-induced paralytic ileus. To our knowledge, this is the first report describing this potential fatal side effect in adults with RSE.


The toxicology of benzene.
Benzene is metabolized, primarily in the liver, to a series of phenolic and ring-opened products and their conjugates. The mechanism of benzene-induced aplastic anemia appears to involve the concerted action of several metabolites acting together on early stem and progenitor cells, as well as on early blast cells, such as pronormoblasts and normoblasts to inhibit maturation and amplification. Benzene metabolites also inhibit the function of microenvironmental stromal cells necessary to support the growth of differentiating and maturing marrow cells. The mechanism of benzene-induced leukemogenesis is less well understood. Benzene and its metabolites do not function well as mutagens but are highly clastogenic, producing chromosome aberrations, sister chromatid exchange, and micronuclei. Benzene has been shown to be a multi-organ carcinogen in animals. Epidemiological studies demonstrate that benzene is a human leukemogen. There is need to better define the lower end of the dose-response curve for benzene as a human leukemogen. The application of emerging methods in biologically based risk assessment employing pharmacokinetic and mechanistic data may help to clarify the uncertainties in low-dose risk assessment.


Fat graft myringoplasty--a prospective trial.
A prospective trial to determine the success rate of the fat graft myringoplasty technique is reported. A success rate of 76% overall was attained at review 1 year postoperatively with increased success for smaller perforations of the tympanic membrane.


The selective estrogen receptor modulator tamoxifen protects against subtle cognitive decline and early markers of injury 24 h after hippocampal silent infarct in male Sprague-Dawley rats.
Silent infarcts (SI) are subcortical cerebral infarcts occurring in the absence of typical ischemia symptoms and are linked to cognitive decline and dementia development. There are no approved treatments for SI. One potential treatment is tamoxifen, a selective estrogen receptor modulator. It is critical to establish whether treatments effectively target the early consequences of SI to avoid progression to complete injury. We induced SI in the dorsal hippocampal CA1 of rats and assessed whether tamoxifen is protective 24 h later against cognitive deficits and injury responses including gliosis, apoptosis, inflammation and changes in estrogen receptors (ERs). SI led to subtle cognitive impairment on the object place task, an effect ameliorated by tamoxifen administration. SI did not lead to detectable hippocampal cell loss but increased apoptosis, astrogliosis, microgliosis and inflammation. Tamoxifen protected against the effects of SI on all measures except microgliosis. SI increased ERα and decreased ERβ in the hippocampus, which were mitigated by tamoxifen. Exploratory data analyses using scatterplot matrices and principal component analysis indicated that SI rats given tamoxifen were indistinguishable from controls. Further, SI rats were significantly different from all other groups, an effect associated with low levels of ERα and increased apoptosis, gliosis, inflammation, ERβ, and time spent with the unmoved object. The results demonstrate that tamoxifen is protective against the early cellular and cognitive consequences of hippocampal SI 24 h after injury. Tamoxifen mitigates apoptosis, gliosis, and inflammation and normalization of ER levels in the CA1, leading to improved cognitive outcomes after hippocampal SI.


Identification and Characterization of Small Molecule Compounds That Modulate Trichothecene Production by Fusarium graminearum.
From the RIKEN Natural Products Depository (NPDepo) chemical library, we identified small molecules that alter trichothecene 15-acetyldeoxynivalenol (15-ADON) production by Fusarium graminearum. Among trichothecene production activators, a furanocoumarin NPD12671 showed the strongest stimulatory activity on 15-ADON production by the fungus cultured in a 24-well plate. NPD12671 significantly increased the transcription of Tri6, a transcription factor gene necessary for trichothecene biosynthesis, in both trichothecene-inducing and noninducing culture conditions. Dihydroartemisinin (DHA) was identified as the most effective inhibitor of trichothecene production in 24-well plate culture; DHA inhibited trichothecene production (>50% inhibition at 1 μM) without affecting fungal mass by suppressing Tri6 expression. To determine the effect of DHA on trichothecene pathway Tri gene expression, we generated a constitutively Tri6-overexpressing strain that produced 15-ADON in YG_60 medium in Erlenmeyer flasks, conditions under which no trichothecenes are produced by the wild-type. While 5 μM DHA failed to inhibit trichothecene biosynthesis by the overexpressor in trichothecene-inducing YS_60 culture, trichothecene production was suppressed in the YG_60 culture. Regardless of a high Tri6 transcript level in the constitutive overexpressor, the YG_60 culture showed reduced accumulation of Tri5 and Tri4 mRNA upon treatment with 5 μM DHA. Deletion mutants of FgOs2 were also generated and examined; both NPD12671 and DHA modulated trichothecene production as they did in the wild-type strain. These results are discussed in light of the mode of actions of these chemicals on trichothecene biosynthesis.


Dielectric barrier discharge and jet type plasma surface modifications of hybrid polymeric poly (ε-caprolactone)/chitosan scaffolds.
In this study, dry air plasma jet and dielectric barrier discharge Ar + O2 or Ar + N2 plasma modifications and their effects on wettability, topography, functionality and biological efficiency of the hybrid polymeric poly (ε-caprolactone)/chitosan scaffolds were reported. The samples treated with Ar + O2 dielectric barrier discharge plasma (80 sccm O2 flow rate, 3-min treatment) or with dry air plasma jet (15-cm nozzle-sample distance, 13-min treatment) had the closest wettability (49.11 ± 1.83 and 53.60 ± 0.95, respectively) to the commercial tissue culture polystyrene used for cell cultivation. Scanning electron microscopy images and X-ray photoelectron spectrometry analysis showed increase in topographical roughness and OH/NH2 functionality, respectively. Increased fluid uptake capacity for the scaffolds treated with Ar + O2 dielectric barrier discharge plasma (73.60% ± 1.78) and dry air plasma jet (72.48% ± 0.75) were also noted. Finally, initial cell attachment as well as seven-day cell viability, growth and proliferation performances were found to be significantly better for both plasma treated scaffolds than for untreated scaffolds.


Effects of supervisory style and experientially based frames of reference on organizational alcoholism programs.
Theoretical works concerning the influence of personal characteristics of work supervisors on their referral of subordinates to organizational alcoholism programs are reviewed. These works hypothesize two barriers to such referrals: supervisory style and experientially based frames of reference. The effects of these behavioral and attitudinal characteristics on supervisors' perceptions of subordinates' work readjustment are analyzed with a sample of 84 supervisors who had utilized the Federal Employee Alcoholism Policy (FEAP). Zero-order hypothesis tests illustrated that supervisory style, operationalized by the Least Preferred Co-worker Measure, was inversely related to perceived readjustment. Hence interpersonally oriented and more cognitively complex supervisors were less likely to perceive improvement. Experientially based frames of reference were operationalized by the Attitude toward the Recovering Alcoholic Scale and the Attitudes toward FEAP Scale. Only the latter was a significant influence, with high policy evaluation being positively associated with perceived readjustment. Multivariate analysis illustrated a significant main effect only for attitudes toward the alcoholism policy. These findings are discussed in terms of a need to reappraise the influence of supervisory style on referral to organizational alcoholism programs and the potential positive effects of organizational engineering on the success of such programs.


Cross-resistance testing of next-generation nucleoside and nucleotide analogues against lamivudine-resistant HBV.
Several next-generation nucleoside and nucleotide analogues are currently in clinical development for the treatment of chronic hepatitis B. However, the efficacy of newer agents against lamivudine-resistant hepatitis B virus (HBV) has not been fully explored. To investigate this in vitro, we generated novel stable cell lines expressing HBV encoding the four major patterns of lamivudine resistance mutations (rtL180M+rtM204V, rtV173L+rtL180M+rtM204V, rtM204I and rtL180M+ rtM204I). Using these cell lines, we assessed the susceptibility of all four strains of lamivudine-resistant HBV to eleven nucleoside analogues in various stages of clinical development. Our studies indicate that lamivudine-resistant HBV remain sensitive to acyclic phosphonate nucleotides (adefovir, tenofovir, and alamifovir), have reduced susceptibility to entecavir, and have high-level cross-resistance to all L-nucleosides tested including emtricitabine, telbivudine, clevudine, and torcitabine.


Using Explicit Thresholds were valuable for judging Benefits and Harms in partially contextualized GRADE Guidelines.
Guideline panels must assess the magnitude of health benefits and harms to develop sensible recommendations. However, they rarely use explicit thresholds. In this paper we report on the piloting and the use thresholds for benefits and harms. We piloted the use of thresholds in a Chilean COVID-19 living guideline. For each of the critical outcomes, we asked panelists to suggest values of the thresholds for large, moderate, small, or trivial or no effect. We collected this information through a survey and an on-line discussion. Twelve panelists decided on thresholds for three critical outcomes (mortality, need for mechanical ventilation and serious adverse events). For all outcomes, an absolute risk reduction was considered larger with more than 50 events, moderate with less than 50 events, small with less than 25 events, and trivial with less than 10 events. Having these a priori thresholds in place significantly impacted on the development of recommendations. Explicit thresholds were a valuable addition to the judgment of the certainty in the evidence, to decide the direction and strength of the recommendation and to evaluate the need for update. We believe this is a line of research worth perusing.


[Sorafenib in combination with chemotherapy as first-line therapy for FLT3-ITD positive acute myeloid leukemia].
Objective: To analyze the clinical features of acute myeloid leukemia patients with Fms-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD) mutation and the therapeutic effect of sorafenib in combination with chemotherapy as first-line therapy for these patients. Methods: Clinical features and therapeutic effect were retrospectively analyzed in 53 AML patients with FLT3-ITD mutation diagnosed in Henan Cancer Hospital from January 2013 to August 2016. The biological characteristics and clinical efficacy of chemotherapy in combination with or without Sorafeinb were analyzed. Results: FLT3-ITD mutation was identified in 53 AML patients, 22 cases (41.5%) were M(5) subtype. The median of the peripheral WBC was 61.00 (0.98-920.00) ×10(9)/L, and there were 50 (94.3%) patients with WBC>10×10(9)/L. The median of blast cell in bone marrow was 0.730 (0.234-0.966) . The total remission rate of all these 53 patients was 56.6% (30/53) . The complete remission (CR) rates in patients treated with chemotherapy in combination with sorafenib and patients with chemotherapy alone were 86.4% (19/22) and 35.5% (11/31) , respectively. The 1-year overall survival rates of the two groups were 78.3%% and 50.0% (P=0.041) , and 1-year progression free survival rates were 75.9% and 42.4% (P=0.044) , respectively. Conclusion: AML patients with FLT3-ITD mutation have the characteristics of high peripheral WBC, high blast cells in bone marrow and accompanying with M(5) subtype. Sorafeinb combined with chemotherapy can significantly improve CR rate and short term survival.


Effect of bacterial inoculation, plant genotype and developmental stage on root-associated and endophytic bacterial communities in potato (Solanum tuberosum).
Beneficial bacteria interact with plants by colonizing the rhizosphere and roots followed by further spread through the inner tissues, resulting in endophytic colonization. The major factors contributing to these interactions are not always well understood for most bacterial and plant species. It is believed that specific bacterial functions are required for plant colonization, but also from the plant side specific features are needed, such as plant genotype (cultivar) and developmental stage. Via multivariate analysis we present a quantification of the roles of these components on the composition of root-associated and endophytic bacterial communities in potato plants, by weighing the effects of bacterial inoculation, plant genotype and developmental stage. Spontaneous rifampicin resistant mutants of two bacterial endophytes, Paenibacillus sp. strain E119 and Methylobacterium mesophilicum strain SR1.6/6, were introduced into potato plants of three different cultivars (Eersteling, Robijn and Karnico). Densities of both strains in, or attached to potato plants were measured by selective plating, while the effects of bacterial inoculation, plant genotype and developmental stage on the composition of bacterial, Alphaproteobacterial and Paenibacillus species were determined by PCR-denaturing gradient gel-electrophoresis (DGGE). Multivariate analyses revealed that the composition of bacterial communities was mainly driven by cultivar type and plant developmental stage, while Alphaproteobacterial and Paenibacillus communities were mainly influenced by bacterial inoculation. These results are important for better understanding the effects of bacterial inoculations to plants and their possible effects on the indigenous bacterial communities in relation with other plant factors such as genotype and growth stage.


Generalized calcinosis cutis associated with disseminated paecilomycosis in a dog.
A 4-year-old spayed female mixed breed dog was referred to the Michigan State University, Veterinary Teaching Hospital (MSU-VTH) with vomiting, lethargy and anorexia of 2 weeks duration. Abdominal radiographs and ultrasonography showed hepatosplenomegaly. Cytological evaluation of ultrasound-guided fine needle aspirates of the liver and spleen revealed fungal organisms and pyogranulomatous inflammation; fungal culture documented Paecilomyces variotii infection. The dog received antifungal therapy and supportive care. Multiple firm plaque-like skin lesions, predominantly involving the inguinal region, developed 18 days after initial presentation and were diagnosed histopathologically as calcinosis cutis. While generalized calcinosis cutis has been reported in three dogs with blastomycosis and one dog with leptospirosis, the association with disseminated Paecilomyces spp. infection is novel.


Escaping but not the inactive X-linked protein complex coding genes may achieve X-chromosome dosage compensation and underlie X chromosome inactivation-related diseases.
X chromosome dosage compensation (XDC) refers to the process by which X-linked genes acquire expression equivalence between two sexes. Ohno proposed that XDC is achieved by two-fold upregulations of X-linked genes in both sexes and by silencing one X chromosome (X chromosome inactivation, XCI) in females. However, genes subject to two-fold upregulations as well as the underlying mechanism remain unclear. It's reported that gene dosage changes may only affect X-linked dosage-sensitive genes, such as protein complex coding genes (PCGs). Our results showed that in human PCGs are more likely to escape XCI and escaping PCGs (EsP) show two-fold higher expression than inactivated PCGs (InP) or other X-linked genes at RNA and protein levels in both sexes, which suggest that EsP may achieve upregulations and XDC. The higher expressions of EsP possibly result from the upregulations of the single active X chromosome (Xa), rather than escaping expressions from the inactive X chromosome (Xi). EsP genes have relatively high expression levels in humans and lower dN/dS ratios, suggesting that they are likely under stronger selection pressure over evolutionary time. Our study also suggests that SP1 transcription factor is significantly enriched in EsP and may be involved in the up-regulations of EsP on the active X. Finally, human EsP genes in this study are enriched in the toll-like receptor pathway, NF-kB pathway, apoptotic pathway, and abnormal mental, developmental and reproductive phenotypes. These findings suggest misregulations of EsP may be involved in autoimmune, reproductive, and neurological diseases, providing insight for the diagnosis and treatment of these diseases.


Alterations in systemic concentrations of progesterone during the early luteal phase affect RBP4 expression in the bovine uterus.
Systemic progesterone affects the timing and duration of uterine endometrial gene and protein expression and has significant effects on conceptus development. The objective of the present study was to examine how changes in progesterone concentrations during the early luteal phase affect retinol-binding protein (RBP4) mRNA and protein concentrations in the uterus. Endometrial tissue and uterine flushings were recovered on Days 7 and 13 of the oestrous cycle in heifers with high, normal and low progesterone concentrations. RBP4 mRNA and protein concentrations were higher (P<0.05) on Day 13 compared with Day 7 in heifers with high and control progesterone concentrations. However, there was no difference in RBP4 protein concentrations between Days 7 and 13 in heifers with low progesterone (P>0.05). On Day 7, although heifers with low progesterone had lower RBP4 mRNA expression compared with controls (P<0.05) there was no difference in protein concentrations between treatment groups. On Day 13, RBP4 mRNA was 2-fold higher (P<0.001) in heifers with high and control progesterone compared with their low-progesterone counterparts and RBP4 protein concentrations were over 2-fold higher (P<0.001) in heifers with high compared to low progesterone. In conclusion, progesterone modulates uterine RBP4 mRNA and protein abundance in a time- and concentration-dependent manner.


Young adult veteran perceptions of peers' drinking behavior and attitudes.
Social norms-based interventions have shown promise in reducing drinking behavior and the resulting consequences in young adults. Although most research has focused on young civilians (i.e., college students), some studies have investigated social norms-based interventions with active-duty military and veteran samples. Yet, research has not yet determined how to maximize the effectiveness of social norms-based interventions in this heavy-drinking population. As an initial step toward this goal, the current study utilized a community sample of 1,023 young adult veterans to examine (a) whether veteran perceptions of the drinking behavior of their veteran peers differ from their perceptions of civilian drinking behavior, (b) whether perceptions of specific veteran groups differ from the actual drinking behavior of veterans within those groups, (c) what levels of specificity in reference groups (same-gender civilians, same-branch veterans, same-gender veterans, or same-branch-and-gender veterans) are most strongly associated with veterans' own drinking, and (d) whether perceptions about others' attitudes toward drinking also contribute independently of perceived behavioral norms to veteran drinking. Findings indicated that participants perceived that other veterans drank more than civilians and that veteran groups drank more than veterans in the sample actually drank. Veteran-specific perceived behavioral norms were similar in their associations with drinking outcomes, whereas same-gender civilian perceived behavioral norms exhibited little or no associations with drinking. Veteran-specific perceived attitudinal norms exhibited little or no association with drinking behavior after controlling for perceived behavioral norms. These findings can be used to inform the development of social norms interventions for young adult veterans.


Clustering of cardio-metabolic risk factors in parents of adolescents with type 1 diabetes and microalbuminuria.
To evaluate the association between a clustering of cardio-metabolic risk factors in parents and the development of microalbuminuria (MA) in their offspring with childhood-onset type 1 diabetes (T1D). The study population comprised 53 parents (mean age [±SD]: 56.7±6.2 years) of 35 T1D young people with MA (MA+) and 86 parents (age: 56.1±6.3 years) of 50 matched offspring with normoalbuminuria (MA-), who underwent clinical, biochemical and cardiovascular imaging assessments. The primary study endpoint was the difference between parents from the MA+ and MA- groups in a cardio-metabolic risk score, calculated as the average value of the standardized measures (z-scores) for waist circumference, blood pressure, fasting glucose, insulin, HDL-cholesterol and triglycerides levels. Cardiovascular parameters, including carotid intima-media thickness (cIMT), flow-mediated dilatation (FMD) and pulse wave velocity (PWV), were also assessed. A DXA scan was performed to assess body composition. The cardio-metabolic risk score was significantly higher in parents of MA+ compared to parents of MA- offspring (mean [95% CI]: 1.066[0.076; 2.056] vs -0.268[-0.997; 0.460], P = .03). Parents of MA+ offspring had slightly higher values of waist circumference, lipids, insulin and blood pressure, although only diastolic blood pressure was statistically different between the 2 groups (P = .0085). FMD, cIMT, PWV (all P > .3), and DXA parameters (all P > .2) were not significantly different between the 2 groups. Parents of young offspring with childhood-onset T1D and MA showed an abnormal metabolic profile, reflected by a calculated risk score. The finding supports the role of a familial predisposition to risk of developing diabetic nephropathy.


Inhibition of growth of hair follicles by a lectin-like substance from rat skin.
A small molecular weight (5 000-10 000) substance has been isolated from rat skin by affinity chromatography on a column of acid-hydrolysed Sepharose. The substance agglutinates rabbit red blood cells, inhibits DNA synthesis in rat hair follicles, and causes the appearance of autophagic vacuoles in the epithelial cells of the lower follicle bulb.