Identification and Intraspecific Typing of Plague Microbe Strains with Their Potential Virulence Determination Using PCR

Developed is the method of identification and intraspecific typing of plague microbe strains along with their potential virulence determination. Intraspecific differentiation and focal attribution of the examined plague microbe strains can be determined by monolocus VNTR-PCR, and main virulence determinants (chromosomal pigmentation region and calcium-dependence plasmid genes) - by multiplex PCR

Cholinesterase in different types of the muscle tissue during the early postmortem period for diagnosis of death coming

The purpose of the research: consisted in study of postmortem regularitiesin the content of cholinesterase in different types of muscle tissue (MT) for improving accuracy of determination of the prescription of death coming PDC.Materials and methods: The activity/level of cholinesterase was determined in homogenates of the myocardial (MMH), oesophageal (OMH), diaphragm (DMH) and intercostal muscles (IMH)within the early PMP (3-13 hours after the coming of death) on 30 human corpses. MT was sampled in conditions of postmortem biopsy with use of special instruments; MT homogenates were prepared following the standard technique with subsequent determination of cholinesterase content in MT homogenates. Results and discussion: The analysis of postmortem changes in the content of cholinesterase in MT depending upon PDC revealed that after 3 hours from the moment of death coming its highest content was in muscles of the oesophagus, the least one being in MT of the intercostal muscles (respectively, (2,717.1±37.1) and (883.5±6.2) U/g, р<0.001). Levels of cholinesterase content in MT of the myocardium and diaphragm were rather close, though they differed (respectively, (1,213.8±8.8) and (1,512.8±11.5) U/g, р<0.05), and occupied an intermediate place between the corresponding values of MT of the intercostal muscles and oesophagus.A common pattern for the content of cholinesterase in different types of MT was characterized by a decrease of this content with an increase in PDC terms; besides, the dynamic lines of its changes, that we obtained, became basic ones for substantiating quantitative time dependencies and construction of relevant nomograms for forensic diagnosis of PDC by cholinesterase content in MT. Conclusions: It was proved that the content of cholinesterase in all MT homogenates, which we studied, changed regularly (and nonlinearly), but the initial and final levels of cholinesterase content differed depending upon the type of MT. Besides, the dynamics in changes of the content of cholinesterase within the time period of 3÷13 hours from the moment of death coming differed upon the type of MT too. Advantages of the technique consist in theintegrity of biochemical examination of different types of MT and simplicity in interpretation of findings. The application of the nomogram technique for assessing PDC by cholinesterase content in MT makes it possible to improve the accuracy of diagnosis for terms of the coming of death up to 60 minutes

Peculiarities of Pigmentation Expression and Structural Differences of hms Operon Genes in <i>Y. pestis</i> and <i>Y. pseudotuberculosis</i> Strains of Diverse Origin

Pigmentation was examined in 214 plague microbe strains and 68 pseudotuberculosis agent strains. Plague microbe strains of different subspecies and pseudotuberculosis microbe strains demonstrated different ability of hemin and acidic dyes sorption. Genes of hms operon in typical Y. pestis strains of five subspecies and in talassica group strains, as well as in Y. pseudotuberculosis O1 serotype strains, were sequenced and analyzed using PCR. Single nucleotide substitutions were detected in all genes of the operon in plague microbe strains as compared with the operon of pseudotuberculosis agent strains. Determined were single nucleotide substitutions promising for intraspecific differentiation of plague microbe strains

Dental status and oxidative homeostasis state in patients exposed to occupational vibration: superoxide dismutase and catalase content in oral fluid

Aim: The aim of the research involved determination of catalase and superoxide dismutase content in oral fluid of patients exposed to occupational vibration depending upon their dental status. Materials and methods: The assessment of dental status (DS) and superoxide dismutase (SOD) and catalase (CAT) content in oral fluid (OF) was performed in three groups of patients: control group (n0=129) included the persons exposed to occupational vibration and whose results of combined medical examination excluded the presence of vibration disease (VD); the second (n1=63 patients with VD stage I) and the third (n1=66 patients with VD stage II) groups consisted of the patients, who underwent treatment at the clinical department of the Research Institute of Occupational Hygiene and Occupational Diseases of Kharkiv National Medical University of the Ministry of Health of Ukraine. DS determination was carried out according to the method of K. M. Kosenko (Patent No. 57512, Ukraine) for in-patients and controls (during medical checkups) using the following indices: PMA, OHI-S, DMFT, with assessment of vacuum-pressory resistance of gingival capillaries (VPRC) (according to V. I. Kulazhenko) and community periodontal index of treatment needs (CPITN). SOD content was determined by the nonenzymatic method; CAT content was revealed spectrophotometrically. Primary data were statistically processed with the determination of accuracy by Student's test. Results: SOD content depending upon PMA intensity in VD patients ranged from 14.1±0.2 U/min to 15.7±0.5 U/min , was reliably (p2.1) and did not differ depending upon VD severity (15.7±0.5 U/min in VD stage I and 15.3±0.3 U/min in VD stage II, respectively). SOD content in OF in patients depending upon their OHI-S ranged from 13.5±0.3 U/min to 16.3±0.2 U/min and was reliably (p40 sec it was equal to 7.8±0.2 U/min, and in VPRC≤40 sec it was 8.6±0.1 U/ min) in VD stage I with decreased VPRC. Conclusions: A trend (p>0.05) towards an increase in SOD activity in VD stage I versus the controls was revealed, whereas VD stage II demonstrated a reliable (p0.05) towards an increase of SOD activity in VD stage I versus the controls was revealed, whereas VD stage II demonstrated a reliable (p<0.05) reduction of the above activity. The assessment carried out in cases requiring combined treatment with surgical or non-surgical debridement and also in patients with supraor subgingival dental calculus found out that SOD activity was reliably reduced only in cases with VD stage II. CAT activity assessment in OF in VD patients having different levels of CPITN showed that the above activity in persons requiring combined treatment (including prosthodontic treatment; CPITN≥3.1points) was markedly and reliably reduced. All the above facts determine peculiarities in oral treatment strategies for this group of patients