Identification of stable reference genes for quantitative PCR in koalas

To better understand host and immune response to diseases, gene expression studies require identification of reference genes with stable expression for accurate normalisation. This study describes the identification and testing of reference genes with stable expression profiles in koala lymph node tissues across two genetically distinct koala populations. From the 25 most stable genes identified in transcriptome analysis, 11 genes were selected for verification using reverse transcription quantitative PCR, in addition to the commonly used ACTB and GAPDH genes. The expression data were analysed using stable genes statistical software - geNorm, BestKeeper, NormFinder, the comparative ΔCt method and RefFinder. All 13 genes showed relative stability in expression in koala lymph node tissues, however Tmem97 and Hmg20a were identified as the most stable genes across the two koala populations

Production and separation of a trehalolipid biosurfactant

This paper reports on a study of the foaming characteristics of trehalolipid biosurfactants in fermentation broths and the utilisation of foaming to enable product recovery. Trehalolipids were produced by a recently isolated marine bacterium, Rhodococcus sp. PML026, in shake flasks and bioreactors. Fermentations were conducted with hexadecane as the carbon substrate. Hexadecane is an antifoam agent which supresses foam formation during fermentation, an advantageous effect during the growth and production phases. The aim of this work was to improve trehalolipid production and identify a suitable media formulation and process conditions to ensure the hexadecane substrate was depleted by the end of the fermentation to enable foaming to occur, allowing for product separation by foam fractionation. The results show that at a threshold biosurfactant concentration and with a residual amount of hexadecane vigorous foaming of the broth commenced in the bioreactor. Foam overflowed through the bioreactor condenser exit was found to separate 23–58% of the total trehalolipids with a product enrichment of 2.3–1.4. This study demonstrates for the first time a potential inexpensive and environmentally friendly strategy to separate trehalolipids from an emulsified fermentation broth

Psychosoziale Faktoren bei koronarer Herzkrankheit : wissenschaftliche Evidenz und Empfehlungen für die klinische Praxis

Psychosocial risk factors like low socio-economic status, lack of social support and social isolation, chronic work or family stress, as well as negative emotions, e. g. depression and hostility, contribute significantly to the development and adverse outcome of coronary heart disease (CHD). Negative effects of psychosocial risk factors are conveyed via behavioural pathways including unhealthy lifestyle, e. g. food choice, smoking, sedentary life, inadequate utilisation of medical resources, and psychobiological mechanisms like disturbed autonomic and hormonal regulation: all these factors contribute to metabolic dysfunction and inflammatory and haemostatic processes, which are directly involved in the pathogenesis of CHD. Interventions to improve pychosocial factors are available and have demonstrated positive effects on risk factors and - at least in part - on CHD morbidity and mortality. The prevention of CHD should therefore include screening for psychosocial risk factors and adequate interventions. Recommendations for the screening of risk factors, behavioural change and further management of psychosocial risk factors in clinical practice are pointed out

Global transcriptomic response of bovine endometrium to blastocyst-stage embryos

The aims of this study were (i) to investigate changes in the global transcriptome of bovine endometrial explants induced by exposure to blastocysts, (ii) to investigate if male and female blastocysts elicit a differential response in the endometrial transcriptome in vitro and (iii) to determine whether bovine endometrium responds to the presence of murine embryos. In Experiment 1, endometrial explants from the same uterus were cultured for 6 h with or without 20 in vitro-produced bovine blastocysts. In Experiment 2, endometrial explants were cultured with male or female bovine blastocysts produced in vitro by IVF either using sex-sorted semen or conventional unsorted semen followed by embryo sexing based on a biopsy. In Experiment 3, endometrial explants were cultured alone or in the presence of bovine blastocysts (n = 25) or murine blastocysts (n = 25). Following culture, explants were snap frozen and stored at -80 degrees C until RNA extraction, qPCR or RNA-Seq. Culture with bovine blastocysts increased endometrial expression of 40 transcripts, all of which were interferon-tau induced. Culture with male or female bovine blastocysts increased transcript abundance of five classic interferon-stimulated genes (MX1, MX2, ISG15, OASY1, RSAD2) in explants; however, there was no difference in abundance of transcripts previously reported to be related to embryonic sex (IFNAR1, IFNAR2, CTGF, ARTN, SLC2A1, SLC2A5). Exposure to murine blastocysts did not elicit any detectable change in transcript abundance. These findings, coupled with our previous data, indicate that very local, interferon-tau-induced changes in endometrial gene expression occur in response to blastocysts; whether such changes play any role in subsequent pregnancy recognition remains to be established

Phobic anxiety in 11 nations: Part I: Dimensional constancy of the five-factor model

The Fear Survey Schedule-III (FSS-III) was administered to a total of 5491 students in Australia, East Germany, Great Britain, Greece, Guatemala, Hungary, Italy, Japan, Spain, Sweden, and Venezuela, and submitted to the multiple group method of confirmatory analysis (MGM) in order to determine the cross-national dimensional constancy of the five-factor model of self-assessed fears originally established in Dutch, British, and Canadian samples. The model comprises fears of bodily injury-illness-death, agoraphobic fears, social fears, fears of sexual and aggressive scenes, and harmless animals fears. Close correspondence between the factors was demonstrated across national samples. In each country, the corresponding scales were internally consistent, were intercorrelated at magnitudes comparable to those yielded in the original samples, and yielded (in 93% of the total number of 55 comparisons) sex differences in line with the usual finding (higher scores for females). In each country, the relatively largest sex differences were obtained on harmless animals fears. The organization of self-assessed fears is sufficiently similar across nations to warrant the use of the same weight matrix (scoring key) for the FSS-III in the different countries and to make cross-national comparisons feasible. This opens the way to further studies that attempt to predict (on an a priori basis) cross-national variations in fear levels with dimensions of national cultures. (C) 2002 Elsevier Science Ltd. All rights reserved

Evolution of a histone variant involved in compartmental regulation of NAD metabolism

NAD metabolism is essential for all forms of life. Compartmental regulation of NAD(+) consumption, especially between the nucleus and the mitochondria, is required for energy homeostasis. However, how compartmental regulation evolved remains unclear. In the present study, we investigated the evolution of the macrodomain-containing histone variant macroH2A1.1, an integral chromatin component that limits nuclear NAD(+) consumption by inhibiting poly(ADP-ribose) polymerase 1 in vertebrate cells. We found that macroH2A originated in premetazoan protists. The crystal structure of the macroH2A macrodomain from the protist Capsaspora owczarzaki allowed us to identify highly conserved principles of ligand binding and pinpoint key residue substitutions, selected for during the evolution of the vertebrate stem lineage. Metabolic characterization of the Capsaspora lifecycle suggested that the metabolic function of macroH2A was associated with nonproliferative stages. Taken together, we provide insight into the evolution of a chromatin element involved in compartmental NAD regulation, relevant for understanding its metabolism and potential therapeutic applications. MacroH2A histone variants originated before the split of fungi and animals. ADP-ribose binding is an ancestral feature of their macrodomains and is linked to the compartmental regulation of NAD metabolism. This function was selected for during the evolution of metazoans

The Short-EMBU in Australia, Spain, and Venezuela - Factorial invariance, and associations with sex roles, self-esteem, and Eysenckian personality dimensions

The short(s)-EMBU (Swedish acronym for Egna Minnen Betraffande Uppfostran [My memories of upbringing]) consists of 23 items, is based on the early 81-item EMBU, and was developed out of the necessity of having a brief measure of perceived parental rearing practices when the clinical and/or research context does not adequately permit application of time-consuming test batteries. The s-EMBU comprises three subscales: Rejection., Emotional Warmth, and (Over)Protection. The factorial and/or construct validity and reliability of the s-EMBU were examined in samples comprising a total of 1950 students from Australia, Spain, and Venezuela. The data were presented for the three national groups separately. Findings confirmed the cross-national validity of the factorial structure underlying the s-EMBU. Rejection by fathers and mothers was consistently associated with high trait-neuroticism and low self-esteem in recipients of both sexes in each nation, as was high parental emotional warmth with high femininity (humility). The findings on factorial validity are in keeping with previous ones obtained in East Germany, Greece, Guatemala, Hungary, Italy, and Sweden. The s-EMBU is again recommended for use in several different countries as. a reliable, functional equivalent to the original 81-item EMBU