THE EFFECT OF HIGH INTENSITY WHITE NOISE ON THE ULTRASTRUCTURE OF AXO-DENDRITIC SYNAPSES IN COLLICULUS INFERIOR OF ADULT MALE CATS. QUANTITATIVE ELECTRON MICROSCOPIC STUDY.

Environmental noise is a serious problem for the society and medicine. Chronic loud noise is known to produce numerous adverse effects on different levels of the organism. In addition to behavioral changes, the involvement of different regions of the brain was documented. The analysis of structural modifications provoked by noise in brain give the possibility to suggest that one of the effects of noise may be the alterations in neurotransmission. Therefore, of special interest should be the elucidation of the effects of chronic noise on the fine structure of synapses of brain areas participating in the processing of auditory information. In the present study, using transmission electron microscope. We elucidate the effects of high intensity chronic white noise on the morphology of axo-dendritic synapses, and size and diameter of synaptic vesicles in auditory region, inferior colliculus of adult male cats. Experimental animals were exposed to 100 dB (5-20 kHz) white noise for one hour per day, for 10 consecutive days. On 11th day, after special procedures, the area of interest was examined under electron microscope. In ~ 15% large synaptic terminals the clustering of synaptic vesicles, as well as swelling, partial vacuolization or degeneration of presynaptic mitochondria were detected. Morphometric analysis of docked (with presynaptic membrane) and undocked synaptic vesicle size revealed that docked vesicles are smaller than undocked vesicles. It was observed in both control and experimental animals, however, in experimental animals, such difference was more significant. Such results suggest that due to continuous transmission, the majority of vesicles are unable to replenish their cargo via transporters. Evaluation of synaptic vesicles size undertaken in the current electron microscopic study has advanced the understanding of the pathophysiology of white noise exposure on auditory brain processing regions, in addition to our understanding of fractional neurotransmitter release at the nerve terminal and on overall brain function.

LINE-1 METHYLATION IN BLOOD AND TISSUES OF PATIENTS WITH BREAST CANCER.

Methylation is an epigenetic alteration proved to be involved in many disease processes including cancer. This change affects mainly gene promoters and repetitive sequences in genome. Long Interspersed Nuclear Element-1 (LINE-1) is a family of retrotransposons - repetitive elements that modify gene activity and can themselves be targeted by epigenetic mechanisms. LINE-1 methylation level is a surrogate marker for global methylation. In many conditions this parameter is found to be altered not only in affected cell groups, but also throughout other tissues. The aim of our study was to compare LINE-1 methylation pattern in DNA extracted from blood of the patients with benign and malignant breast tissue. In addition, we investigated correlation of LINE-1 methylation in blood and tissues of same patients and relationship of all variables with histopathologic and phenotypic characteristics of tumors. Patients with biopsy-proved ductal invasive carcinoma of breast and no preoperative chemo/radiotherapy were chosen for the study group. Another pool of patients with various benign breast lesions represented controls. Blood samples from both group members were collected preoperatively. Tumor tissue sections were processed for pathology report and part of remaining tissue was used for methylation study. LINE-1 methylation level was quantified using ELISA-based assay. It was analyzed in combination with histologic and phenotypic tumor parameters and compared between different tissues and different study groups. LINE-1 was found to be significantly hypomethylated in breast cancer tissue compared to blood. Blood samples of patients with malignant tumors showed slightly lower methylation level, than samples obtained from control group members. Lymphovascular invasion was the only aggressiveness-determining factor that was found to be at least weakly correlated with LINE-1 hypomethylation in blood. We can conclude, that global hypomethylation measured by LINE-1 methylation level is significant in tumor tissue. But there is no significant difference between LINE-1 methylation levels in blood of patients with benign and malignant breast tumors; therefor LINE-1 hypomethylation in blood cannot be used as a marker for early tumor detection. Neither is it valid for determination of tumor behavior.

UNUSUAL MANIFESTATION OF NEUROBORELIOSIS (CASE REPORT).

The paper reported the verified case of neuroboreliosis with unusual clinical presentation of Parkinsonism. Study aimed at establishing the significance of a precise differential diagnosis with substantial analysis of the symptoms of several diseases to avoid the false diagnosis and to conduct the opportune and adequate therapeutic management. We described the case of the diagnosed neuroboreliosis with clinical expression of Multiple Sclerosis (MS) and Parkinsonism. A 44 years old man was diagnosed as MS according to the McDonald's Criteria, who within two years developed typical clinical signs of Parkinsonism. Patient investigated neurologically, Brain contrast MRI (1.5 Tesla) was performed; Cerebrospinal fluid was researched for oligoclonal bands. Blood IgM and IgG were researched against Chlamidia pneumonie, Micoplasma pneumonie, Borrelia Burgdorferi, Herpes simplex 1/2, Cytomegalovirus by ELISA method. Clinically the patient expressed amimic face, oligobradikinesia, extrapiramidal rigidity in all limbs, resting tremor in upper limb fingers, horizontal nystagmus. Brain MRI showed multiple gadolinium enhanced demyelization lesions in periventricular and sub-cortical white matter. CSF oligoclonal bands were positive without dysfunction of blood-brain barrier. Blood IgM, IgG detected to be negative against Chlamidia pneumonie, Micoplasma pneumonie, cytomegalovirus, Herpes simplex ½, while the blood IgG was strongly positive against Borrelia burgdorferi, confirmed by followed Western blot test. Patient was stabilized by puls-therapy with 1 gr/intravenous Solumedrol (5 days) along with Rocephin treatment (2 gr /iv) for 21 days followed by long term treatment with Antiparkin (Carbidopa 250 mg, Levodopa 25 mg). MS and even Parkinsonism in suspicious cases should thoroughly be investigated for differentiation from chronic Neuroboreliosis.

BACTERIOLOGICAL EXAMINATION OF THE ABDOMINAL EFFUSION IN BACTERIAL PERITONITIS.

This study provides a contemporary epidemiology of aspirates taken during surgery from the abdominal cavity among patients with bacterial peritonitis to identify the isolates and study their sensitivity to antibiotics. Our bacteriology investigations included isolation of poor cultures, and detection of microbes was conducted using a rapid identification system (API20E, API Staph, API Strep, API Ana, BioMerieux). Rapid tests for detection of oxidase and catalase activity were also used. Susceptibility of microorganisms to antibiotics was defined by the disc-diffusion method using standard discs (EUCAST guidelines 2015) according to Clinical Laboratory Standard Institute (CLSI) protocols (ATB strips: ATB G, ATB Staph, ATBANA, ATBPse, ATBStrep. BioMerieux). The recovery rate from the clinical samples was good, likely because our protocol immediately inoculated study material into the thioglycollate broth which is an appropriate medium both for aerobic and anaerobic bacteria. Among the 36 patients with monomicrobial growth by bacteriological investigation, Gram-negative bacteria prevailed; Escherichia coli was recovered in 14 patients and Enterobacter cloacae in 9 patients. Among the Gram-positive bacteria, D-group Streptococci were prevalent, Enterococcus faecalis was found in six patients, Staphylococcus aureus in three patients, Candida albicans in two patients. In one patient, we observed dual colonization of two Gram-negative anaerobes Bacteroides fragilis and Fusobacterium spp. Polymicrobial growth was evident in three cases in the following combinations: Candida albicans and Escherichia coli, Enterobacter cloacae and Candida albicans, Escherichia coli and Bacteroides fragilis. Antibiotic susceptibility testing indicated that 12% of Gram-negative bacteria were resistant to quinolones and 19% to third-generation cephalosporins. No evidence of methicillin-resistant Staphylococcus aureus was found in Gram-positive specimens. The timely identification of microbes and administration of appropriate therapy based on antibiotic sensitivity profiles is important to optimizing clinical outcomes in bacterial peritonitis.

CLOSTRIDIAL PARAPROCTITIS WITH GAS GANGRENE OF FRONT-LATERAL ABDOMINAL WALLS AND NECROTIC FASCIO-MYOSITIS (CASE REPORT).

Anaerobic clostridial infection is the most severe form of paraproctitis. The incubation period is very short, from 3 to 6 hours, sometimes lasting for 1-2 days. Clostridial infection spreads rapidly and induces gas gangrene, causes destruction of cells and other intermediate substances, and impedes blood circulation. This paper presents a case study of an extremely severe form of anaerobic infection with spontaneous gas gangrene, cellulitis, fasciomyositic necrosis, severe intoxication and septic shock on the abdominal front and lateral surfaces. This patient presented as infected with Clostridium septicum, a rare and highly toxic Gram-positive, spore-forming, obligate anaerobic bacillus that progresses and migrates rapidly, affecting all soft tissues (muscle, fascia), and produces four toxins which cause gas gangrene, intravascular hemolysis, tissue necrosis, and septic shock. The mortality rate is typically 80%. In this case study, a positive clinical outcome was achieved by aggressive identification of the microbe, appropriate and immediate therapy, and vigorous surgical intervention. Specifically, immediate surgery was conducted to ensure a wide excision of damaged tissues, necrectomy, curettage, wide drainage, readjustment, oxygenation through drainages, further additional surgical corrections through CT control with wide bandages in the operating area. Further, the diagnostic workup was thorough, identifying the microbe through a properly constructed diagnostic algorithm, ultrasound and CT studies, infectious agent assessments, and bacteriological monitoring carried out on the 1st-2nd-5th-7th-12th-15th-21st-25th days. Rational antibiotic therapy with permanent susceptibility testing informed the selection of an appropriate agent. Finally, markers for the evaluation of severity (Apache scale) were assessed, as they were for stage of infection (prokalcitonin), inflammation (CRP) and other indicators.

Headaches and cerebral blood flow disturbances in patients with chronic nasal pathology.

Study aimed at research of cerebral blood flow and blood NO contents in patients with chronic non-surgical nasal pathology with accompanying headaches. Eighty five patients with non-surgical chronic nasal pathology with accompanying headaches were investigated. Patients were divided in two age categories: 1st group of 58 patients (18 to 55 years), and 2nd group of 27 patients (>55 years). Control consisted of 30 age-matched healthy individuals. CBF was studied by means of Transcranial Doppler Imaging in patients and control. Free blood NO and NO - complexes were measured by Electron Paramagnet Resonance (EPR) method. Statistics performed by SPSS-11.0. In both age categories of patients the mean blood velocity in middle cerebral artery, anterior cerebral artery, basilar artery of patients was significantly increased against control (p<0.05). Pulsation index (PI) found to be decreased compared to control. The blood free NO signals were non-significantly increased against control. The intensive EPR signals of HbNO and FeSNO were revealed in patients and no signals were detected in control. No differences were revealed between clinical groups regarding the blood EPR signals of NO and NO-complexes (p<0.5). Negative correlation was found between the PI and intensity of headaches (r=-0.37; p<0.01). Multivariate linear regression analysis found the significance of longevity of chronic nasal pathology, Pulastion Index and blood hemoglobin contents for Intensity of headaches (p<0.05). It is concluded that CBF disturbances in chronic nasal pathology may result in intensive headaches.

Epidemiology of nodular goiter in South Georgia and selection of some diagnostic criteria from the view point of prevention.

Mountainous regions of Georgia have been long known as a center of goiter endemia. Study of thyroid gland diseases in this contingent of population represents a key target. The object of the conducted examinations was study of epidemiology of goiter in the South Georgia. According to the results of our study high index of spreading endemic goiter in the population of the South Georgia is conditioned by iodine-deficiency. In addition, the factor providing spreading of thyroid gland pathology in the country is improper feeding. At the same time we aimed to study in these populations such diagnostic indices of various pathologies of thyroid gland as carcinoma - embryonic antigen CEA, which has been less studied. There is scanty information on these markers during thyroid gland pathology and in populations of Georgia it is absent at all. Due to the fact that the mentioned marker makes possible to conduct monitoring of the disease, we consider the study of its infomativity to be important during various forms of nodular goiter. For comparison of the results study of oncomarker REA was conducted in group of healthy individuals and non-nodular goiter. On the base of obtained data it can be supposed that study of oncomarker CEA is informative for diagnosis of various forms of nodular goiter as additional criteria, especially in case of obscure or not conducted morphological diagnosis.

Probable role of immunological tolerance to ischemia injury in brain.

To address the issues of immunological tolerance to ischemia injury in the brain we have researched ischemic stroke patients with and without prodromal transitional ischemic attacks (TIAs) for several blood acute phase reactants involved in inflammatory reactions in respect to initial infarct size, clinical course of disease and functional outcome at 1 month. The study involved 54 ischemic stroke patients aged 45 to 70 years, 46 female and 38 male admitted within 24 hours of symptoms onset in neurological clinic of Georgian State Medical University during 2000-2006. Exclusion criteria comprised severe somatic pathology, liver and renal dysfunctions. Control subjects were aged-matched 15 healthy volunteers, who did not reveal any significant signs of cerebrovascular disease according to the anamnesis, clinical and instrumental investigations. Etiology of stroke was classified according to TOAST criteria. Patients were divided into three groups: the first group - 22 patients with first-ever stroke, the second -17 patients with prodromal TIAs from one to three months before stroke and the third - 15 patients with prodromal TIAs within 4 weeks before stroke. Initial neurological impairment assessed immediately after admission by NIHSS score. All three groups selected with the same initial severity of stroke with mean NIHSS score = 12+/-3.5; for evaluation of clinical course of disease patients were assessed by NIHSS on 7th day of stroke. In 48 hours from stroke onset the blood levels of (IL-1beta, IL-6, TNF-alpha, IL-10) were significantly elevated against control (p<0.05). At this time, no statistical differences were detected between groups regarding the initial blood levels of IL-1beta and TNF-alpha, while the level of IL-6 was significantly lower in the third group (p<0.05). Blood contents of IL-10 and TGF-beta1 found to be non-significantly elevated in the third group against two other groups, while blood TGF-beta1 was significantly increased compared to control. Significant positive correlation was found between IL-6 blood contents and clinical course of disease (r=+0.32, p<0.05). Multivariate logistic regression found the significance of initial blood IL-6 contents for probability of stroke functional outcome at 1 month. It can be supposed that relatively mild blood inflammatory response in third group can be related to occurring of immunological tolerance.

Prognostic significance of p27(Kip 1) expression in renal cell carcinoma.

The importance of cyclin-dependent kinase inhibitor genes (CDKIG) in benign and malignant urological diseases is a subject of intense ongoing investigation. The goal of the current study was to analyze the expression of p27(Kip 1) CDKIG in benign and malignant renal cells and assess their possible association with different clinical parameters. Expression of p27(Kip 1) CDKIG was evaluated and compared in 24 normal human kidneys and in 46 renal cell carcinoma (RCC) samples. Intensity of the gene expression was compared between the groups and possible association was analyzed with the cancer clinical parameters. The gene was significantly higher expressed in normal than in RCC tissue samples (p=0.0045). Intensity of the marker expression in RCC was negatively correlated with tumor size (Rho=-0.438, p=0.0051) and associated with stage and grade (p=0.0488 and <0.0001, respectively). The patients with symptomatic disease had significantly less marker expression than incidentally discovered tumors (p=0.0301). The marker expression was significantly higher in oncocytomas as compared with conventional RCCs (p=0.0378) The baseline p27(Kip 1) expression level in these patients was significantly lower than in non-recurrent tumors (p=0.04). Disease-related related death was observed in 4 cases. The baseline p27(Kip 1) expression level in these patients was significantly lower than in alive patients (p=0.0106). The Log-Rank analysis showed that loss of p27(Kip 1) expression were the risk-factors of oncological patient death (p=0.005). Expression of p27 is significantly decreased in RCC as compared with normal kidney tissue. Intensity of the gene expression is associated with clinical parameters: tumour size, stage, grade and disease presentation. Loss of p27 expression is a risk-factor for disease progression.

Down-regulation of p27(Kip 1) cyclin-dependent kinase inhibitor in prostate cancer: distinct expression in various prostate cells associating with tumor stage and grades.

The goal of the study was to analyze the expression of p27(Kip 1) cyclin-dependent kinase inhibitor protein (CDKI) in different cells of benign, malignant and hormonally treated prostate cancer tissue and assess their possible association with different clinical parameters. Expression of p27(Kip 1) CDKI was evaluated and compared in: 32 BPH, 20 prostate cancer (PCa) and 6 hormonally treated prostate cancer (HTPCa) tissues. Intensity of the expression was compared between the groups and association was soughed with the cancer clinical parameters. Total expression of p27(Kip 1) was significantly higher in BPH as compared with PCa (p=0.0002) and HTPCa (p=0.0324). The difference between PCa and HTPCa was not significant. p27(Kip 1) was higher expressed by epithelial, ductal and vascular prostatic cells of BPH as compared with PCa (p=0.0001, 0.0101 and 0.0224, respectively). The stromal expression of the marker was not different between the groups. Epithelial marker expression was significantly increased in HTPCa as compared with PCa (p=0.0460). In the PCa group, the intensity of the protein expression was negatively associated with the tumor stage, Gleason scores 1, 2, and the Gleason sum (p=0.0453, 0.0202, 0.0074 and 0.0098, respectively). This difference was found in epithelial, vascular and ductal prostatic cells. Down-regulation of p27(Kip 1) CDKI in PCa is detected in epithelial, vascular and ductal, but not the stromal cells. The intensity of the expression in these cells is associated with tumor stage and grades. The hormonotherapy is causing up-regulation of p27(Kip 1) expression in prostate adenocarcinoma cells.

[Significance of homocysteines in the development of acute myocardial infarction].

The aim was the study of the connection of the changes in serum homocysteine concentration with acute myocardium infarction, in order to establish whether the increase in homocysteine concentration is one of the risk-factors in the development of the cardiac insufficiency. The patients with acute myocardial infarction were studied. The following tests were performed: determination of the homocysteine concentration in serum by ELISA; determination of the main characteristics of hemostasis by automatic coagulometer (AMELUNG); determination of lipid spectrum by biochemical analyzer (Johnson & Johnson Vitros DT). The results obtained allowed us to suggest that the changes in the serum homocysteine concentration in patients with myocardial infarction occur in accordance with the changes in the protective mechanisms of the organism, and it may be used for the diagnostics of weakening and strengthening of the processes of arteriosclerosis and thrombosis.