Sequence and expression of mRNAs encoding the alpha 1 and alpha 2 subunits of a DHP-sensitive calcium channel

Complementary DNAs were isolated and used to deduce the primary structures of the alpha 1 and alpha 2 subunits of the dihydropyridine-sensitive, voltage-dependent calcium channel from rabbit skeletal muscle. The alpha 1 subunit, which contains putative binding sites for calcium antagonists, is a hydrophobic protein with a sequence that is consistent with multiple transmembrane domains and shows structural and sequence homology with other voltage-dependent ion channels. In contrast, the alpha 2 subunit is a hydrophilic protein without homology to other known protein sequences. Nucleic acid hybridization studies suggest that the alpha 1 and alpha 2 subunit mRNAs are expressed differentially in a tissue-specific manner and that there is a family of genes encoding additional calcium channel subtypes.     

Perforin gene defects in familial hemophagocytic lymphohistiocytosis

Familial hemophagocytic lymphohistiocytosis (FHL) is a rare, rapidly fatal, autosomal recessive immune disorder characterized by uncontrolled activation of T cells and macrophages and overproduction of inflammatory cytokines. Linkage analyses indicate that FHL is genetically heterogeneous and linked to 9q21.3-22, 10q21-22, or another as yet undefined locus. Sequencing of the coding regions of the perforin gene of eight unrelated 10q21-22-linked FHL patients revealed homozygous nonsense mutations in four patients and missense mutations in the other four patients. Cultured lymphocytes from patients had defective cytotoxic activity, and immunostaining revealed little or no perforin in the granules. Thus, defects in perforin are responsible for 10q21-22-linked FHL. Perforin-based effector systems are, therefore, involved not only in the lysis of abnormal cells but also in the down-regulation of cellular immune activation.     

Determination of five macrolide antibiotic residues in eggs using liquid chromatography/electrospray ionization tandem mass spectrometry

A method using liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) for the determination of trace levels of five macrolide antibiotics (spiramycin, tilmicosin, oleandomycin, erythromycin, and tylosin) in eggs is presented. Data acquisition under MS/MS was achieved by applying multiple reaction monitoring (MRM) of two or three fragment ion transitions to provide a high degree of sensitivity and specificity for both quantification and confirmation. Matrix-matched standard calibration curves were used to achieve the best accuracy of the method. A fully nested experimental design was used to study the measurement uncertainty arising from intermediate precision and trueness or proportional bias. The overall recoveries, that is, those determined by the nested experiments, of spiramycin, tilmicosin, oleandomycin, erythromycin, and tylosin at fortified levels of 60, 100, 200, and 300 microg/kg were 96.8, 98.2, 98.3, 98.8, and 95.4%, respectively. The LC/ESI-MS/MS method detection limits (S/N > or = 3:1) of five macrolides were <1.0 microg/kg.     

Evaluation of bread wheat (Triticum aestivum L.) genotypes for yield and related traits under drought stress conditions

ABSTRACT

Drought is a major factor threatening crop production worldwide. Developing wheat varieties that are adapted to drought prone environments is a sustainable strategy to improve wheat production and productivity. The aim of this study was to evaluate and select bread wheat genotypes for yield and yield components, and for stability under drought stress and non-stress conditions. One hundred and twenty genotypes were evaluated at five test sites in the 2018/19 cropping season using a 10 x 12 alpha lattice design with two replicates. The level of drought stress was imposed using different sowing dates (early planting representing non-stressed, while late planting as drought stressed conditions) following the onset of the main rain at each site. Grain yield and yield components were recorded, and drought indices were calculated for each genotype. Among the drought tolerance indices, GMP, MP, HM, STI and YI were found to be the most suitable for predicting drought tolerance because they had significant and positive correlations with yield under drought stress and non-stress conditions. Rank sum analysis identified the most drought tolerant genotypes as ‘YS-34', ‘YS-85' and ‘YS-82’. The selected wheat genotypes are useful genetic resources for future drought tolerance breeding programmes in Ethiopia or similar agro-ecologies.     

Microbial Indicators of Faecal Contamination in Water: A Current Perspective

It is well documented that faecal contamination of drinking water has caused numerous disease outbreaks. Because the risks of disease outbreaks correlate with the incidence of faecal contamination, faecal bacteria are used as indicators of faecal contamination and hence, the possible presence of disease-causing organisms. However, different microbiological faecal indicators are used in different countries and jurisdictions. Therefore, it is important to understand the potentials and limitations of these indicator organisms before realistically implementing guidelines and regulations to safeguard our water resources. This review considers the history of indicator organisms, the evolution of the analytical methodologies (biochemical and molecular) and addresses the advantages and limitations of current faecal indicator microorganisms.     

Multiple statistical approaches of community fingerprint data reveal bacterial populations associated with general disease suppression arising from the application of different organic field management strategies

Multiple statistical analyses of terminal restriction fragment length polymorphism (T-RFLP) data were used to screen and identify bacterial populations involved in general disease suppression in an organically managed soil. Prior to sampling three different management strategies (i.e. mixed hay (H), tilled fallowing and open-field vegetables production) were used during the transition from conventional to organic farming, with and without compost amendment. The H transition strategy consistently led to the lowest damping-off disease incidence on two different crops in separate greenhouse and field experiments. Bacterial population structure in bulk soil and the rhizosphere of both crops was characterized using T-RFLP analyses of amplified 16S rDNA sequences. First, principal component analysis (PCA) revealed changes in the relative abundance of bacterial terminal restriction fragments (TRF) in response to transition strategy and/or compost amendment in eight different experimental contexts. In each context, a different subset of TRF substantially contributed to the variation along the first two principal components. However, terminal restriction fragment M148 contributed significantly to the observed variation in 6 out of the 8 experiments, and moderately in the remaining 2 experiments. As a second approach, nonparametric analyses of variance revealed that the relative abundance of TRF differed among treatments. While the responsive subsets identified varied somewhat by experimental context, M137, M139 and M141 were more abundant in samples from the H transition strategy in multiple experimental contexts. Subsequent correlation analyses revealed that TRF associated with disease suppressive treatments (i.e. H with and without compost) were frequently negatively correlated with damping-off disease incidence. As a group, these TRF were disproportionately associated with lower disease levels further indicating their role in disease suppression. Interestingly, in silico analysis of the bacterial 16S rDNA sequence database revealed that the TRF identified in this study (e.g. M137, M139, M141, and M148) might correspond to well-characterized genera of bacterial biological control agents.     

Detection of nodavirus in barramundi, Lates calcarifer (Bloch), using recombinant coat protein-based ELISA and RT–PCR

The coat protein encoded by the nodavirus RNA2 gene originally isolated from greasy grouper, Epinephelus tauvina , was cloned, expressed as a recombinant polyhistidine-tailed fusion protein and characterized by immunoblot analysis. The purified recombinant protein was used to develop an indirect enzyme-linked immunosorbent assay (ELISA) to detect body exudate and plasma antibodies against the coat protein in both experimentally infected and commercial barramundi. In addition, the nucleotide sequence was employed to develop a RT–PCR detection assay based on the T4 region. The results showed that the virus could be detected as early as 3 days post-infection by RT–PCR while antibodies against the recombinant coat protein were detectable on day 6 post-infection. Among 112 commercial barramundi samples collected from October 1999 to April 2000, 9% showed positive ELISA results which were further verified by Western blot.     

Schisandrin B protects against solar irradiation-induced oxidative stress in rat skin tissue

Schisandrin B (Sch B) and schisandrin C (Sch C), but not schisandrin A and dimethyl diphenyl bicarboxylate, protected rat skin tissue against solar irradiation-induced oxidative injury, as evidenced by a reversal of solar irradiation-induced changes in cellular reduced glutathione and α-tocopherol levels, as well as antioxidant enzyme activities and malondialdehyde production. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production in rat skin microsomes. Taken together, Sch B or Sch C, by virtue of its pro-oxidant action and the subsequent eliciting of a glutathione antioxidant response, may prevent photo-aging of skin.     

Survival of various ERIC-genotypes of Shiga toxin-producing Escherichia coli in well water

Recently, there has been a surge of interest in understanding the survival of Shiga toxin-producing Escherichia coli (STEC) in aquatic environments. Fifteen strains of STEC were monitored, individually, in untreated well water samples incubated at 10 and 22^{^}C for 56 days. The strains were selected from three serogroups (O26, Ol11 and O157) and represented five distinct ERIC (enterobacterial repetitive intergenic concensus)-genotypes. The microcosms were prepared in triplicate and inoculated at an initial cell density of about 7.0 log CFU/ml well water. At 10^{^}C, the cell density of five STEC strains fell below the detection limit of 0.8 log CFU/ml by day 56. Of the ten persisting strains, four showed superior survival with cell densities decreasing to an average of about 5 log CFU/ml while the remaining six strains showed moderate levels of survival, decreasing to an average cell density of about 3 log CFU/ml. At 22^{^}C, strain H32 (genotype I) and H15 (genotype B) persisted at 1.1 log CFU/ml and 2.2 log CFU/ml in 56 days, respectively. The other 13 STEC strains dropped below the detection limit between weeks 3 to 8. The 15 strains demonstrated highly variable levels of survival with no correlation between ERIC-genotypes or serogroups and the strains' ability to persist in the well water samples. Although strain H32 (O157:H7) persisted significantly longer than strain H22 (O157:H7) in natural well water at both 10 and 22^{^}C, both strains survived equally well in sterile well water, indicating that individual STEC strains vary in their ability to compete with background microbial populations.