Bacillus anthracis strain differentiation based on SNP and VNTR loci.

Bacillus anthracis is the anthrax causative agent. For its epidemiology, it is important not only to identify the etiological agent but also to determine the patterns of its evolution and spread. Modern methods of molecular biology make it possible to detect a number of genetic markers suitable for indicating and differentiating the strains of B. anthracis, including the loci arranged as variable number tandem repeats (VNTRs) and SNPs, one nucleotide-sized differences in the DNA sequence of the loci being compared. The objective of the present study was to examine the effectiveness of SNP analysis and PCR amplif ication of VNTR loci combined with the high-resolution amplicon melting analysis for identif ication and differentiation of the anthrax agent strains. In the study, seven strains of B. anthracis obtained from soil samples and animal carcasses were investigated using vaccine strain STI-1 as a reference. For molecular genetic characterization of these bacteria, analysis of 12 SNPs and variability analysis of eight VNTR loci were carried out. To detect the differences between the strains, their PCR product melting points were measured in the presence of the EvaGreen (Sintol, Russia) intercalating dye. For SNP detection, a PCR assay with double TaqMan probes was applied. It was found that the studied virulent strains, except for B. anthracis No. 1 and 3, could not be attributed to any phylogenetic subgroup of the anthrax agents. The proposed method made it possible to differentiate four out of the seven investigated strains. Strains No. 5-7 had identical SNP and HRM prof iles and, as a result, formed a single cluster. Our investigation has conf irmed that the proposed method can be successfully used for preliminary analysis of an epizootic situation in the case of anthrax.

[Extended endoscopic endonasal posterior (transclival) approach to tumors of the clival region and ventral posterior cranial fossa. Part 1. Topographic and anatomical features of the clivus and adjacent structures]. / Éndoskopicheskii éndonazal'nyi zadnii rasshirennyi (transklival'nyi) dostup k opukholiam oblasti skata i ventral'nykh otdelov zadnei cherepnoi iamki. Chast' 1. Topografo-anatomicheskie osobennosti skata i pogranichnykh s nim obrazovanii.

OBJECTIVE: to describe the main topographic and anatomical features of the clival region and its adjacent structures for improvement and optimization of the extended endoscopic endonasal posterior (transclival) approach for resection of tumors of the clival region and ventral posterior cranial fossa. MATERIAL AND METHODS: We performed a craniometric study of 125 human skulls and a topographic anatomical study of heads of 25 cadavers, the arterial and venous bed of which was stained with colored silicone (the staining technique was developed by the authors) to visualize bed features and individual variability. Currently, we have clinical material from more than 120 surgical patients with various skull base tumors of the clival region and ventral posterior cranial fossa (chordomas, pituitary adenomas, meningiomas, cholesteatomas, etc.) who were operated on using the endoscopic transclival approach. RESULTS: We present the main anatomical landmarks and parameters of some anatomical structures that are required for performing the endoscopic endonasal posterior approach. The anatomical landmarks, such as the intradural openings of the abducens and glossopharyngeal nerves, may be used to arbitrarily divide the clival region into the superior, middle, and inferior thirds. The anatomical landmarks important for the surgeon, which are detected during a topographic anatomical study of the skull base, facilitate identification of the boundaries between the different clival portions and the C1 segments of the internal carotid arteries. The superior, middle, and inferior transclival approaches provide an access to the ventral surface of the upper, middle, and lower neurovascular complexes in the posterior cranial fossa. CONCLUSION: The endoscopic transclival approach may be used to access midline tumors of the posterior cranial fossa. The approach is an alternative to transcranial approaches in surgical treatment of clival region lesions. This approach provides results comparable (and sometimes better) to those of the transcranial and transfacial approaches.

[Cerebrovascular reactivity in patients with rheumatoid arthritis concurrent with and without hypertension].

AIM: To compare cerebrovascular reactivity (CVR) in patients with rheumatoid arthritis (RA) concurrent with essential hypertension (Group 1) and in those with RA and normal blood pressure (BP) (Group 2). SUBJECTS AND METHODS: During the study of Groups 1 (n = 37) and 2 (n = 12), the investigators estimated the prevalence of traditional cardiovascular risk factors, performed 24-hour BP monitoring, investigated CVR by transcranial Doppler (TCD) of the middle cerebral arteries (MCA) by hyperoxic and hypercapnic tests, and endothelium-dependent vasodilation (EDV) and endothelium-independent vasodilation of the brachial artery. The groups were matched for gender, age, RA activity and stage, and antirheumatic therapy volume. RESULTS: According to the results of MCA TCD, the hyperoxic test recorded impaired CVR in 34 (92%) and 10 (83%) patients in Group 1 and 2, respectively; the hypercapnic test revealed this condition in 19 (51%) and 6 (50%) patients in these groups, respectively. The hyperoxic test most commonly showed an insufficient decrease in MCA linear blood flow velocities (LBFV) in 31 (84%) and 8 (66%) patients in Groups 1 and 2, respectively; the hypercapnic test did an excessive increase in MCA LBFV in 12 (32%) and 4 (33%) patients, respectively. There was a high rate of impaired EDV in 32 (86%) and 9 (75%) patients in Groups 1 and 2, respectively. CONCLUSION: According to the results of MCA TCD, there were high and similar rates of impaired CVR in patients with RA concurrent with and without essential hypertension during the hyperoxic and hypercapnic tests.

[The determination of the type of structural stability of the human skull]. / Opredelenie tipa konstruktsionnoi ustoichivosti mozgovogo cherepa cheloveka.

A total of 819 male and female skulls (frontal sections of 250 of these) were examined by traditional craniometry and MBS-2 microscope in order to assess the variability of morphogeometric parameters of biomechanical resistance of human brain skull and designing its structure typology. The following types of construction resistance of human brain skull (craniotypes) were distinguished: 1) structure resistant; 2) configuration resistant; 3) morphologically resistant; and 4) morphologically unstable.

[The stereotopometry of the trabecular and parachordal sections of the human cerebral cranium]. / Stereotopometriia trabekuliarnogo i parakhordal'nogo otdelov mozgovogo cherepa cheloveka.

169 skulls were examined in adults and 58 in children to study variability of parameters and shape of anterior trabecular (sphenoethmoidal) and posterior parachordal (auriculo-occipital) regions of human skull, the border between which is drawn through openings of trigeminal nerve branches. Spatial coordinates of these foramina and anterior and posterior points of trabecular and parachordal regions were obtained using stereocraniobasiometer. The maps of all the regions were plotted. Age, sex, individual and bilateral variability of these structures was examined. The point, located in medial sagittal projection on the line that unites right and left round openings is of minimal variability and can be used as a central one in comparative cranium examination.

[Influenza virus penetration into the cells of a continuous dog-kidney line]. / Proniknovenie virusa grippa v kletki perevivaemoi linii pochek sobak.

The penetration of the influenza virus (WSN) into MDCK cells and its intracellular fate was studied using electron microscopy of ultrathin sections of infected cells at early steps of infection, and biochemical analysis of intracellular subviral components. In electron micrographs, the virus particles absorbed on the cells and located inside phagosomes were detected. In both the cases, the fusion of the virus with cell envelopes was distinctly seen. The fusion took place with incubation at 37 degrees C and 4 degrees C, thus suggesting a non-enzymatic mechanism. The inner virus component released into the cytosol after fusion represented virus cores, i. e. ribonucleoproteins (the final products of uncoating) surrounded by the inner protein membrane consisting of matrix (M) protein. The cores were accumulated in perinuclear cytoplasm of infected cells and were most probably intermediates in virus uncoating.