Identification of Bacillus anthracis by multiprobe microarray hybridization

We have developed a rapid assay based on microarray analysis of amplified genetic markers for reliable identification of Bacillus anthracis and its discrimination from other closely related bacterial species of the Bacillus cereus group. By combining polymerase chain reaction (PCR) amplification of six B. anthracis-specific genes (plasmid-associated genes encoding virulence factors (cyaA, pagA, lef, and capA, capB, capC) and one chromosomal marker BA-5449) with analysis of amplicons by microarray hybridization, we were able to unambiguously identify and discriminate B. anthracis among other closely related species. Bacillus identification relied on hybridization with multiple individual microarray oligonucleotide probes (oligoprobes) specific to each target B. anthracis gene. Evaluation of the assay was conducted using several B. anthracis strains (with or without pXO1 and pXO2 plasmids) as well as over 50 other species phylogenetically related to B. anthracis, including B. cereus, B. thuringiensis, B. mycoides, and B. subtilis. The developed microarray analysis of amplified genetic markers protocol provides an efficient method for (i) unambiguous identification and discrimination of B. anthracis from other Bacillus species and (ii) distinguishing between plasmid-containing and plasmid-free Bacillus anthracis strains.     

An Automated System for Leukofiltration of Blood Components

Biomedical Engineering - An automated system for filtering blood transfusion media through special filters, monitoring and logging the filtration parameters, and processing filtration protocols is...     

Phenotypic and genetic analysis of the cerebellar mutant tmgc26, a new ENU‐induced ROR‐alpha allele

ROR‐alpha is an orphan nuclear receptor, inactivation of which cell‐autonomously blocks differentiation of cerebellar Purkinje cells with a secondary loss of granule neurons. As part of our ENU mutagenesis screen we isolated the recessive tmgc26 mouse mutant, characterized by early‐onset progressive ataxia, cerebellar degeneration and juvenile lethality. Detailed analysis of the tmgc26?/? cerebella revealed Purkinje cell and granule cell abnormalities, and defects in molecular layer interneurons and radial glia. Chimera studies suggested a cell‐autonomous effect of the tmgc26 mutation in Purkinje cells and molecular layer interneurons, and a non‐cell‐autonomous effect in granule cells. The mutation was mapped to a 13‐Mb interval on chromosome 9, a region that contains the ROR‐alpha gene. Sequencing of genomic DNA revealed a T‐to‐A transition in exon 5 of the ROR‐alpha gene, resulting in a nonsense mutation C257X and severe truncation of the ROR‐alpha protein. Together, our data identify new roles for ROR‐alpha in molecular layer interneurons and radial glia development and suggest tmgc26 as a novel ROR‐alpha allele that may be used to further delineate the molecular mechanisms of ROR‐alpha action.     

Intracavity Thermometry in Medicine

Biomedical Engineering - We present here an analytical review of technical means used for intracavity thermometry of biological objects. Radiometers can be effective both in medicine and veterinary...     

Development and malformations of the cerebellum in mice

The cerebellum is the primary motor coordination center of the CNS and is also involved in cognitive processing and sensory discrimination. Multiple cerebellar malformations have been described in humans, however, their developmental and genetic etiologies currently remain largely unknown. In contrast, there is extensive literature describing cerebellar malformations in the mouse. During the past decade, analysis of both spontaneous and gene-targeted neurological mutant mice has provided significant insight into the molecular and cellular mechanisms that regulate cerebellar development. Cerebellar development occurs in several distinct but interconnected steps. These include the establishment of the cerebellar territory along anterior-posterior and dorsal-ventral axes of the embryo, initial specification of the cerebellar cell types, their subsequent proliferation, differentiation and migration, and, finally, the interconnection of the cerebellar circuitry. Our understanding of the basis of these developmental processes is certain to provide insight into the nature of human cerebellar malformations.     

Molecular follow-up of preneoplastic lesions in bronchial epithelium of former Chernobyl clean-up workers

Ionizing radiation is a potent lung carcinogen, but the precise molecular damage associated with it is still unknown. In this study we investigated cancer-related molecular abnormalities including K-ras (codon 12) mutation, p16(INK4A) promoter hypermethylation and microsatellite alterations at seven chromosomal regions in successive biopsies obtained from former Chernobyl cleanup workers in comparison with smokers and nonsmokers who have never had radiation exposure. Our results indicate that prolonged persistence of inhaled radioactive particles is associated with appearance of allelic loss at 3p12, 3p14.2 (FHIT), 3p21, 3p22-24 (hMLH1) and 9p21 (p16INK4A) in bronchial epithelium of former Chernobyl clean-up workers. The prevalence of 3p14.2 allelic loss was associated with decreased expression of the FHIT mRNA in their bronchial epithelium in comparison with control group of smokers. During several years of our monitoring samples of epithelium were collected from the same area of bronchial tree. In epithelium exposed to carcinogens (tobacco smoke and/or radioactivity) the total number of molecular abnormalities was significantly higher in dysplasia and in morphologically normal foci progressed later to dysplasia than in these samples which never showed evidence of such progression. Our findings indicate that extensive cancer-related molecular abnormalities sequentially occur in radiation damaged bronchial epithelium of former Chernobyl clean-up workers.     

Specified mechanisms of action of preimmunisation with diphtherial anatoxin on myocardial pathology after critical care

Chronic experiments for 5 weeks made in 41 mongrel adult dogs have shown that prevaccination with diphtheria antitoxin has a negative effect on myocardial and hemodynamic restoration after resustitation. The dogs have arrhythmia, survival of the animals is worse. However, such prevaccination improved survival of the animals due to reduction of reperfusion trauma in animals after hemorrhagic shock in spite of conduction disorders and myocardial ischemia.