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 (p<0.05) lower in patients with VD versus the controls (17.8±0.2 U/min and 14.2±0.2 U/min respectively, when PMA>2.1) 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 (p<0.05) lower in patients with OHI-S≥1.7 U. A comparative analysis showed that the activity of the enzymatic protection of the periodontal membrane could be also determined by the state of hard tissues, in particular by such DS index as DFTM. The activity of SOD in VD stage II was found to be reliably (p<0.05) reduced in patients with DFTM index exceeding 15 pts. A somewhat different pattern of SOD activity was found in OF in patients with VD stage I: SOD activity in OF was similar in all DFTM indices and it became reduced depending upon an increase of DFTM index. SOD content depending upon VPRC index in patients with VD ranged from 10.7±0.5 U/min to 16.8±0.3 U/min and was reliably (p<0.05) lower in cases with VPRC index ≤40 sec. CAT content depending upon PMA intensity in VD patients ranged from 4.6±0.4 U/min to 11.3±0.3 U/min and was reliably (p<0.05) higher in patients with VD stage I versus the controls and differed according to the severity. CAT content in OF in patients depending upon OHI-S ranged from 5.2±0.2 U/min to 10.1±0.3 U/min, was reliably (p<0.05) lower in cases with OHI-S ≥1.7 U, did not differ from the indices observed in the control group and was also found to be reliably lower in patients with VD stage II versus those with VD stage I (7.3±0.3 U/min and 8.6±0.2 U/min, respectively, when OHI-S ranged within 0.7÷1.6 U). CAT content in OF depending upon VPSC index in patients with VD ranged from 5.8±0.2 U/min to 8.6±0.6 U/min and was reliably (p<0.05) lower in cases with VPSC index ≤40 sec. Thus, CAT activity in OF in patients was reliably (p<0.05) reduced (in VPRC>40 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 (p<0.05) reduction of the above activity. At the same time, an unsatisfactory state of oral hygiene was shown to promote inhibition of the enzymatic protection of their periodontal membrane in patients with VD stage I. A trend (p>0.05) 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.

Dental status and oxidative homeostasis state in patients exposed to occupational vibration: reduced glutathione content in oral fluid.

AIM: The aim of the research involved determination of reduced glutathione 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 reduced glutathione (RG) content in oral fluid (OF) was carried out in three groups of patients: control group (n=129) included the persons exposed to occupational vibration whose results of comprehensive medical examination excluded the presence of vibration disease (VD); the second group (n=63 patients with the VD stage I) and the third group (n=66 patients with VD stage II), 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). RG content (activity) in OF was determined according to Garishvili. Primary data were statistically processed with the determination of accuracy by Student's test. RESULTS: Assessment of metabolic indices, which characterize the state of oxidative homeostasis enzyme chain, showed that RG content in OF depending upon VD severity reliably (p<0.05) reduced. RG content depending upon PMA intensity in patients with VD ranged from 23.5±0.8 mg/cm3 to 28.6±0.3 U/min and was reliably (p<0.05) lower in patients with VD stage I versus controls (23.2±1.0 U/min and 26.7±0.3 U/min respectively, when PMA>2.1) and also reliably lower in patients with VD stage II versus patients with VD stage I (29.9±0.9 U/min and 26.2±0.4 U/min respectively, when PMA>1.0). RG content depending upon OHI-S values in patients with VD ranged from 33.4±1.2 U/min to 24.1±1.1 U/min and was reliably (d<0.05) lower in patients with OHI-S values equal to 1.7 U and higher versus controls (24.1±1.1 U/min and 27.1±0.2 U/min, respectively) and also reliably lower in patients with VD stage II versus patients with VD stage I (27.3±0.4 U/ min and 33.4±1.2 U/min, respectively, with OHI-S≤0.6 U). A comparative analysis showed that the activity of the enzymatic protection of the periodontal membrane could be also determined by the state of hard tissues, in particular by such DS index as DFMT. The activity of RG in VD stage I was shown to be reliably (p<0.05) reduced in patients with DFMT index exceeding 15 pts (in DFMT≤10 pts RG activity was 32.2±0.4 U/min, whereas in DFMT exceeding 15 pts it was equal to 26.7±0.6 U/min). Somewhat different pattern of RG activity in OF was found in patients with VD stage II: RG activity in OF was reduced in all DFMT values in these patients and its reduction was shown to be dependent on an increase in DMFT index. In DMFT≤10 the patients with VD stage II were found to have a reliable (p<0.05) reduction in RG activity in OF versus the control group (30.1±0.3 U/ min and 23.8±0.5 U/min, respectively) and this activity was shown to be inhibited in DFMT increase (23.8±0.5 U/min with DFMT≤10 and 19.3±0.9 U/min with DFMT≥20 pts, respectively). RG content depending upon VPRC in VD patients ranged from 29.2±0.1 U/min to 29.2±0.1 U/min and was reliably (p<0.05) lower in patients with their VPRC values ≤40 sec. Assessment of RG activity in OF of VD patients with different levels of CPITN showed that RG content in persons requiring comprehensive treatment (also including prosthetic treatment; CPITN≥3.1 pts) was reliably reduced (versus corresponding groups of patients but with low CPITN values) both in VD stages I and II (24.1±1.0 U/min and 19.3±0.9 U/min, respectively). CONCLUSIONS: Increases in the rate and expression of periodontal lesions (by PMA index) depending upon the presence and severity of VD with a proper decrease in the level of RG content in OF were registered. An activation of the enzymatic chain of antioxidant protection in patients with VD under low HI values and a simultaneous inhibition of enzymatic activity under high HI values were found out. VD stage I revealed an increased RG activity versus controls (p<0.05), while VD stage II demonstrated a reliable (p<0.05) reduction of the above activity. Moreover, an unsatisfactory state of the oral cavity hygiene contributed to enzymatic protection of their periodontium in patients with VD (irrespective of its stage). Regularities were revealed, which supported the benefit of pathogenetic relationships between the state of the periodontal microcirculatory bed and enzymatic activity of OF in patients with VD. Both patients with VD and persons, who are exposed to occupational vibration, need for diagnosis of activity of enzymes in OF since, as our analysis of findings has shown, DS indices are interdependent with activity of the enzymatic chain of the antioxidant protection of OF.

Informativeness of structural and biochemical changes in the muscle tissue of myocardium in the early postmortem period.

Postmortem structural and biochemical changes in the muscle tissue (MT) of myocardium from the positions of forensic examinations (FE) of the prescription of death coming (PDC) were not studied systematically, this fact determining the purpose of the present research. AIM: The aim of the research consisted in study of structural and biochemical changes in the tissue of myocardium during the early postmortem period (PMP). MATERIALS AND METHODS: The muscle tissue of myocardium within the early PMP (3-13 hours) after the coming of death was studied on 30 human corpses. Six BCM in myocardium muscle homogenates (MMH) were determined: BCM1 - the content of glycogen, BCM2 - the content of acid phosphatase, BCM3 - the content of lactate, BCM4 - the content of lactate dehydrogenase (LDH), BCM5 - the content of lipofuscin, BCM6 - the content of cholinesterase. MT was taken with use of special instruments, MT homogenates were prepared following the standard technique. Cytological studies of MT preparations of myocardium as well as their photographic recording were made on an Axiostar microscope (Zeiss, FRG). The optic density (OD) of nuclei and cytoplasm of cardiomyocytes (CMC) in conventional units of OD was measured using VideoTest program (Russia). RESULTS: It was found out that changes in MT of myocardium during the early PMP were characterized by the morphological, biochemical and biophysical regularities that we revealed; their most demonstrative features were as follows: - a gradual and constant reduction of the relative OD of CMC nuclei (YM-7) and cytoplasm (YM-8) during 3-13 hours from the moment of death, the rate and stage of these dynamics depending nonlinearly upon PDC; we substantiated and received quantitative regularities (polynomials) for the above biophysical indicators. CONCLUSIONS: A comparative morphological study of the ultrastructure of CMC at the early PMP depending upon PDC was performed; - the early PMP is characterized by proper biochemical changes in MT, the most demonstrative of them are as follows: a reduction in the content of glycogen (YM-1)and a dynamic increase in the content of lipofuscin(YM- 5).For all six BCM, representative absolute and relative values of their content in MMH depending upon PDC were obtained; - paired correlative values between biochemical and biophysical markers of the state of MT of myocardium were examined in their systemic relationships and proper SCC were determined by six time intervals of the early PMP, thereby making it possible to substantiate those of them that were criterially significant for increasing the accuracy of diagnosis of PDC.

The content of lactate in muscle tissue of different types in the early post-mortal period.

It's known from the basics of clinical biochemistry and pathophysiology, lactate is the end product of anaerobic glycolysis. During exercise lactate leaves the muscles and is converted by the liver into pyruvate or metabolized by brain tissue and heart muscle. The level of lactate in muscle tissue from the standpoint of forensic examination to determine the age of death (PDC) during the early postmortem period (PMP) has not been previously studied. AIM: The aim of the study was to study the postmortem patterns of lactate content in muscle tissue (MT) of various types to increase the accuracy of determining the age of death. MATERIALS AND METHODS: Determination of lactate content was performed in homogenates of myocardial muscles, esophagus, diaphragm and intercostal muscles in early PMP (3-13 hours after death) in 30 human corpses. MT collection was performed in sectional biopsy using special tools, preparation of MT homogenates - according to standard methods with subsequent determination of lactate content in MT homogenates by enzymatic photometric method. RESULTS: Analysis of postmortem changes in lactate content in MT, depending on the time periods of the prescription of death coming (PDC)revealed that after 3 hours. since the onset of death, the highest content was in the intercostal muscles, the lowest - in the MT of the esophagus (respectively - (6,847 ± 0,042) mmol/g and (3,266 ± 0,031) mmol/g, p<0,001. in MT of different types was characterized by fluctuations with increasing terms of PDC, in addition, our time series became the basis for substantiating the quantitative time dependences and construction of appropriate nomograms for forensic diagnosis of PDC by lactate content in MT. CONCLUSIONS: t is proved that the lactate content naturally (and nonlinearly) changed in all studied homogenates of MT, but the initial and final level of lactate content differs depending on the type of MT. In addition, the dynamics of changes in lactate content in the time period 3÷13 hours. from the moment of death, depending on the type of MT also varies. The quantitative analytical and graphical dependences of the change in the lactate content in MT in the early PMP revealed in the study allowed to substantiate the corresponding nomograms.

The nomogram technique for determining the prescription of death coming by the content of acid phosphatase in the muscle tissue.

Criminologists and medical examiners in practice are interested in the existence of reliable, stable criteria that would allow to unambiguously interpret certain post-mortem phenomena observed in the body, and which would allow to determine the age of death. AIM: The aim of the study was to study the postmortem patterns of acid phosphatase in muscle tissue (MT) of various types to improve the accuracy of determining the age of death. MATERIALS AND METHODS: Determination of acid phosphatase content is performed in homogenates of myocardial, esophageal, diaphragm and intercostal muscles in the early postmortem period (PMP) (3-13 hours after death) in 30 human corpses. MT sampling was performed in the conditions of sectional biopsy with the use of special tools, preparation of MT homogenates - according to the standard method with subsequent determination of MT phosphatase content in MT homogenates by kinetic method. RESULTS: Analysis of postmortem changes in the content of acid phosphatase in MT depending on the time periods of the determining the age of death revealed that after 3 hours from the moment of death its content was the highest in a myocardium, the smallest - in MT of intercostal muscles (accordingly - 3,475±0,057 units/g and 2,662±0,028 units/g, p <0,001). The general pattern of acid phosphatase content in MT of different types was characterized by an increase in the content with increasing the age of death terms. In addition, the time series of changes in the acid phosphatase content obtained by us became the basis for substantiating the quantitative time dependences and constructing appropriate nomograms for forensic diagnosis of determining the age of death by the acid phosphatase content in MT. CONCLUSIONS: It is proved that the content of acid phosphatase naturally (and nonlinearly) changed in all studied homogenates of MT, but the initial and final level of acid phosphatase, depending on the type of MT differs. In addition, the dynamics of changes in the content of acid phosphatase in the time period 3÷13 hours. from the moment of death, depending on the type of MT, also varies. The quantitative analytical and graphical dependences of the change in the content of acid phosphatase in MT in the early PMP revealed in the study allowed to substantiate the corresponding nomograms.

Lipofuscin content in the muscle tissue during the early postmortem period: improvement of forensic diagnosis of the prescription of death coming.

Forensic medicine is naturally supported by fundamental sciences, its integration with them contributing to its improvement on the whole, particularly in diagnosis of the prescription of death coming. Scientific achievements and foreign specialists in forensic medicine during recent years have made it possible to significantly deepen our sound knowledge of postmortem phenomena. AIM: The aim of the research consisted in study of postmortem regularities in the content of lipofuscin in different types of the muscle tissue (MT) for improving accuracy of determination of prescription of death coming. MATERIALS AND METHODS: The content of lipofuscin was determined in homogenates of the myocardial, oesophageal, diaphragm and intercostal muscles within the early postmortem period on 30 human corpses. MT was sampled in conditions of postmortem biopsy with use of special instruments; homogenates were prepared following the standard technique with subsequent determination of lipofuscin content in MT homogenates according. Presentation of revealed regularities in changes of lipofuscin content in each type of MT homogenates is provided by building dynamic lines with polynomials of different (2-5) stages and accuracy of reproduction R2>0.95. RESULTS: By results of biochemical examination of lipofuscin in different types of the MT within different terms of the early postmortem period (PMP) it was proved that its content regularly changed in all types of the above tissue. The analytical and graphical dependences of the change in the content of lipofuscin in muscle tissue within the early PMP made it possible to substantiate relevant nomograms. Limitations for using the nomogram technique are as follows: prescription of death coming more than 13 hours, unknown conditions of the stay of a corpse after the coming of death. Advantages of the technique consist in the integrity of biochemical examination of different types of MT and simplicity in interpretation of findings. CONCLUSIONS: The application of the nomogram technique for assessing PDC by lipofuscin content in MT makes it possible to improve the accuracy of diagnosis for terms of the coming of death up to 60 minutes.