LOSS OF CAS3 AND INCREASE OF BAX EXPRESSION ASSOCIATED WITH PROGRESSION OF CERVICAL INTRAEPITHELIAL NEOPLASIA.

Apoptosis plays one of the major roles in the progression of human cancers including cervical carcinoma. The aim of our study was to analyse the expression of Cas3, Bax and their correlation with the proliferation index and ER expression status during the progression of cervical intraepithelial neoplasia (CIN). Study included altogether 140 specimens, divided into two major groups, such as: cervical lesions without co-infections and with co-infections. Standard immunohistochemistry was used to detect antigens: Ki67, Cas3, Bax and ER. The study results showed that the expression of Cas3 is significantly decreased whilst the expression of Bax is significantly increased during the progression of CIN in both groups with and without co-infections. The expression of Bax negatively correlates with the expression of Cas3 (r=-42.4, p<0.05) and ER (r=-33.4, p<0.05) and positively correlates with the expression of proliferation marker Ki67 (r=56.3, p<0.05). The results indicate that the deregulated apoptosis measured as increased expression of Bax and Cas3 loss, as well as the increase in proliferation index measured as Ki67 expression is significantly related to the progression of CIN into cervical carcinoma. Therefore, the measuring of mentioned protein expression could be used as the markers of the CIN progression.

STEM CELL INDEX IN THE PROGRESSION OF CERVICAL INTRAEPITHELIAL NEOPLASIA.

Stem cells represent the small subpopulation of healthy and cancerous tissues, which are characterised with increased proliferation and self-renewal properties. From the many different markers of stem cells, we have investigated the stem cell index during the progression of cervical intraepithelial neoplasia (CIN), based on the immunohistochemical expression of CD44 in total of 140 tissue samples from uterine cervix. In addition, we have performed the profound correlation analysis of CD44 with different epithelial-mesenchymal, proliferation, apoptosis and hormonal markers at both protein and mRNA level. The results of our study indicated that, stem cell index based on the CD44 detection is significantly increased with the progression of cervical intraepithelial neoplasia. In addition, CD44 expression significantly correlates with the epithelial-mesenchymal transition, proliferation-apoptotic features and ER status in both protein and mRNA level. Two, groups of cervical intraepithelial lesions as well as carcinoma can be identified based on the expression of CD44, Ki67, Cas3 and ER. To the best of our knowledge we are first to demonstrate such findings in CIN and cervical carcinoma and identified characteristics could be used for the early assessment of CIN patient prognosis and for the relevant clinical management.

EVALUATION OF THE RISK OF CERVICAL INTRAEPITHELIAL NEOPLASIA PROGRESSION BASED ON CELL PROLIFERATION INDEX, EPITHELIAL-MESENCHYMAL TRANSITION AND CO-INFECTIONS.

Human papilloma virus (HPV) infection, especially with high risk types, represent the major etiological factor for the development of cervical precancerous and cancerous lesions. However, other factors including cell proliferation index, epithelial-mesenchymal transition and the presence of co-infections might also influence the progression of cervical intraepithelial neoplasia (CIN). The aim of our study was to analyse, the expression of cell proliferation markers and epithelial-mesenchymal transition markers during the progression of cervical intraepithelial neoplasia, in cases with and without co-infections. Standard immunohistochemistry was used to detect, Ki67, cyclin D1, phosphohiston-H3, p63, E-cadherin, ß-catenin and vimentin. The results of our study indicated that the expression of Ki67, phosphohiston-H3 and p63 is significantly increased during the progression of CIN disease, whilst the expression of E-cadherin and ß-catenin are progressively lost. The expression of mesenchymal marker vimentin is also increased in CINIII and in invasive carcinoma. Proliferation index based on Ki67 labelling is significantly higher in cases with co-infections and the expression on E-cadherin is significantly lower in cases with co-infections compared to cases without co-infections. In conclusion, the measurement of proliferation index, based on Ki67 labelling, as well as mitotic index based on phosphohiston-H3 detection can reliably indicate high and low risk groups of the progression of CIN. Similarly, higher p63 expression, loss of E-cadherin and ß-catenin and higher vimentin expression can indicate the progression risk of CIN. The presence of co-infections is associated with the increased expression of proliferation marker Ki67 and the loss of E-cadherin and therefore it can be considered as an additional marker of CIN progression.

IMPACT OF SOCIOECONOMIC STATUS ON CARDIOVASCULAR RISK IN GEORGIAN POPULATION.

Cardiovascular diseases (CVD) continue to be the leading cause of death and disability worldwide, representing 30% of all death. In Georgia 36% of all death caused by ischemic heart disease and 23% caused by stroke. There is a limited data on association between socioeconomic status (SES) and CVD risk in Georgia. Our study aims at assessing association between SES and CVD risk factors in a cohort of adult Georgians. A cross-sectional study was conducted in the region of Western Georgia between June 2007 - June 2008. 1196 individuals aged 40-70 years were enrolled in the study. To assess SES we collected information about education, income, ownership of a private transport, and employment. CVD risk was assessed by the WHO/ISH risk classification scale. In addition, anthropometric measurements were conducted in each study participant. Low education level (adjusted odds ratio (aOR) 2.85; 95% confidence interval (CI), 1.57-5.17), obesity (aOR 2.13, 95% CI 1.16 -3.92) and abdominal obesity (aOR 2.21, 95% CI 1.26 -3.87) were statically significantly associated with more than 10% 10 year risk of a fatal or non-fatal cardiovascular event. Lower education level (not finished high school -aOR 2.21 95% CI 1.44 - 3.38, undergraduate - aOR 1.52, 95% CI 1.12 - 2,05), income 120 USD or more (aOR 0.73, 95% CI 0.54 -1.00), overweight (aOR 1.68, 95% CI 1.07 - 2.63), obesity (aOR 1.58, 95% CI 1.04 - 2.41), and abdominal obesity (aOR 1.43, 95% CI 1.01 - 2.03) were found to be statistically significant predictors of CVD in our study population. We found that education level, income, body mass index and waist to hip ratio are CVD risk factors. Public health authorities, as well as clinicians should consider this finding in primary and secondary prevention and organize multidisciplinary teams to address those risk factors.

USE OF COMPLEMENTARY AND ALTERNATIVE MEDICINE IN GEORGIA.

In Georgia, like in most countries globally, people commonly resort to complementary and alternative medicine (CAM). However, not much is known about CAM practices there. The aim of the study was to document common practices of CAM in Georgia and related patient attitudes. We collected data from peoples who commonly use CAM at 20 service provision centers in Georgia using cluster sampling from 300 patients. We admininstered a cross-sectional survey and conducted descriptive statistical analyses. People in Georgia use CAM either for prevention to improve general health (33%) or to treat chronic conditions (36%), spending about 25 Euros per month out of pocket. Most (77%) get their knowledge about CAM from family or friends , less than half (44%) from books or media, and 11% from medical providers. A close person's advice or experience was the most common rationale given for CAM use (54%). In our sample, 17% either don't trust or are unsatisfied with conventional medicine, 29% found CAM treatment "very effective" and 61% "quite/partially" effective; only 5% not effective. Conventional treatment was stopped in half of the cases. 35% of respondents informed their physicians of their CAM use, while about half did not. Public mistrust towards conventional medicine, CAM user high satisfaction, relatively low cost of such services in Georgia - are the factors letting us to suggest that CAM use will further increase. Frequent self taking decisions made by patients to stop physician prescribed treatment, not informing physicians on CAM use, as well as other factors put patients health at risk. Further research and capacity building in practice, education and other related aspects are needed to establish evidence-based regulation and standards for CAM in Georgia that ensure informed decision making and patient safety.

COMPLEMENTARY AND ALTERNATIVE MEDICINE AND MEDICAL EDUCATION IN GEORGIA: STATE OF THE ART AND FURTHER PERSPECTIVES.

Aim - complementary and alternative medicine (CAM) is popular in Georgia, but providers' training not well understood or regulated. The aim of this mixed-methods study was to inform the process of implementing quality CAM curricula in Georgia and strategies for the inclusion of effective CAM curricula into medical education in Georgia. We analyzed existing medical curricula. We conducted a contextual analysis based of qualitative data collected from relevant medical education experts, qualified physicians, CAM practitioners and other stakeholders; and administered a quantitative MD students' survey to MD students. CAM components are currently not represented in medical curricula in Georgia. Physicians largely lack adequate knowledge of CAM and its practice. All stakeholders supported that it would be beneficial to develop CAM educatory courses, both to future practitioners (medical students, initially as an elective subject) and practicing physicians (through CME). We recommend development/integration of an elective subject and/or a curricular component as a first step of CAM integration into the medical curricula in Georgia for MD students and CME courses for physicians. Interdisciplinary and international collaborations may help achieve best outcome, and safe practice of CAM in Georgia, forming a base for physician - CAM practitioner collaborations for quality care for patients in Georgia.

HIV/AIDS prevalence in the Southern Caucasus.

According to statistics, Armenia, Azerbaijan and Georgia, are the countries with low prevalence of HIV (<0.3%). Yet the dramatic almost nine fold increases in registered HIV cases in the region between 2000 and 2008 (from 574 to 5323) remain a cause for serious concern. This report summarizes and compares the latest data on the epidemiology of HIV/AIDS in Southern Caucasus: Armenia, Azerbaijan and Georgia. It also provides a comparison with HIV/AIDS situation in Eastern Europe. The work is mainly based on the review of UNAIDS 2010 country progress reports and end-year reports of HIV/AIDS Surveillance in Europe. Tables and graphs have been created to summarize the obtained information and to improve the analysis. Trends in HIV/AIDS spread in Southern Caucasus are similar to trends in Eastern Europe: the number of HIV and AIDS cases continues to increase. The vast majority of people living with HIV/AIDS were aged 25-49-years at the time of diagnosis. The official information confirms that the epidemic in Southern Caucasus remains located among males (with 83.8% in Azerbaijan, 75% in Georgia, and 73% in Armenia) and most-at-risk populations. In Georgia and Azerbaijan, as in Eastern Europe, the predominant mode of transmission is through injecting drug use (58.8% and 64.2%, respectively), while in Armenia over the last several years the transmission has shifted from injecting drug use to heterosexual spread (50.2%). The second reported mode of transmission in Georgia and Azerbaijan is unprotected heterosexual activity (34.9% and 23.4%, respectively). Medical transmission, mother to child transmission, and transmission among men who have sex with men are rare, typically comprising less than 3% of total registered transmission in each country of the region. As UNAIDS contends the prevalence values are underestimated, and the registered numbers of HIV/AIDS cases in the region do not reflect the actual spread of the infection. There is also a substantial discrepancy between the cumulative number of HIV/AIDS cases reported in the national case reporting systems and the HIV prevalence reported in surveys (high prevalence observed in IDUs in Armenia and Azerbaijan, and in MSM in Georgia). In the context of such underreporting, population-based and community-based HIV surveys have far better public health utility in estimating burden of HIV/AIDS than the case reporting system.

[Comparative characteristics of membrane forms of aldehyde dehydrogenase]. / Sravnitelnaia kharakteristika membrannykh form aldegiddeidrogenazy.

It was demonstrated from the results of kinetic analysis that acetaldehyde oxidation reaction, which is catalyzed by membrane isoenzyme forms of mitochondrial and microsomal aldehyde dehydrogenases, has the ordered Bi-Bi mechanism. An attempt was made to compare the active site structures of these two isozyme forms using cyclopropylethyl-containing benzoic acid amides which are the reversible inhibitors of the studied enzyme activity. The amides competitively inhibited each enzyme activity relative to acetaldehyde. The inhibition constant values were calculated. As to NAD+ partially competitive type of inhibition was observed for aldehyde dehydrogenase of the microsomes. At low amide concentrations the mitochondrial aldehyde dehydrogenase was inhibited uncompetitively. With increased concentration of the compounds over 75 microM the character of inhibition by 2.4-dichlorine- and 2.4-dichlorine-5-methylbenzoic acid amides changed to mixed one; and when using the amides of 2.5-dichlorine-4-methylbenzoic acid it changes for pseudoinhibition. The difference of kinetic behaviour of the studied compounds with aldehyde dehydrogenase membranes forms suggests the conformation similarity of the sites for binding acetaldehyde for the both enzymes and distinction of the sites for coenzyme binding.

[Regeneration of the liver by using modified polymer compounds for the purpose of hemostasis]. / Regeneratsiia pecheni pri primenenii s tsel'iu gemostaza modifitsirovannykh polimernykh kompozitsii.

Results of experimental investigations of using polymer compositions KL-3, including modifications with ionol ("Adgenol") and levamisol in 18 rabbits with injuries of the liver are presented. It has been established that 3 and 6 months after using the mentioned compositions viable healing of liver wounds are obtained. Different degrees of intensity of inflammatory reactions and of development of the connective tissue were noted, most pronounced after using "Adgenol". Adding Levamizol to the polymer composition facilitated the recovery of the initial hepatic cells.

[Kinetic characteristics of enzymes of ethanol metabolism in rats]. / Kineticheskie kharakteristiki fermentov obmena étanola u krys]

Experimental animals were separated as for narcotic sleep length into short-sleeping and long-sleeping rat groups and as for alcohol motivation to rats preferring ethanol or water or an intermediate group as well. Alcohol- and aldehyde dehydrogenase reaction maximum speed as well as Michaelis-Menten constants for these enzymes were measured for each of these groups. Alcohol dehydrogenase affinity to cofactor in the reverse reaction was an order of magnitude higher as compared with the affinity to substrate in all the animal groups. Apparent Michaelis-Menten constant values were 1.5-2 times higher in short-sleeping and ethanol preferring rats in the direct reaction. There were no such differences for the reverse reaction. Alcohol dehydrogenase reaction maximum speed for short-sleeping group is close to that for animals preferring ethanol or water. For different forms of aldehyde dehydrogenase an increase of the apparent maximum speeds was distributed as follows: water preferring group < intermediate group < ethanol preferring group. Aldehyde dehydrogenase-1 Michaelis-Menten apparent constants for acetaldehyde were changed in the same way. Thus, data obtained suggest that ethanol metabolism speed along the path alcohol/aldehyde dehydrogenases are close in ethanol-preferring and short-sleeping rats.

Biocompatibility of a prolonged-action antialcohol preparation.

Acid and alkaline phosphatase activities in liver lysosomes and liver mitochondria, respectively, as well as in connective tissue capsule were studied for an antialcohol preparation (polyurethane carrier and disulfiram at 150, 300 and 625 mg/kg body weight) implanted subcutaneously. Increased acid phosphatase activity was observed in connective tissue capsule on days 14, 30 and 90 after implantation and in liver lysosome fraction on days 14 and 30 with preparation containing disulfiram at 300 and 625 mg/kg body weight. Alkaline phosphatase activity increased both in connective tissue capsule and in liver mitochondria up to day 30 only when sample with maximal disulfiram dose was implanted. Acid phosphatase activity is an adequate indicator for biocompatibility of prolonged-action medicine preparations. The data obtained show that the suggested medical form containing disulfiram 150 mg/kg body weight is biocompatible. Interaction between acid and alkaline phosphatase activity changes and biocompatibility and destruction of polymer implants is discussed.

[Immobilization of trypsin on polyurethane carriers]. / Immobilizatsiia tripsina na poliuretanovykh nositeliakh.

A method of covalent immobilization of trypsin on polyurethanes containing residues of 2.4-toluene or hexamethylene diisocyanates in a side chain is developed. It is shown that the amount of the immobilized enzyme is independent of isocyanate nature, but its proteolytic activity is higher on carriers containing 2.4-toluene diisocyanate residues. This is accounted for by higher mobility of enzyme molecules due to a decrease in the number of chemical bonds between trypsin and the polymer matrix. The immobilized enzyme is stable for a long time and does not lose proteolytic activity being cyclically used for many times.

[Activity of enzymes of ethanol metabolism as a test in evaluating the effectiveness of sensitizing alcohol deterrents]. / Aktivnost' fermentov obmena étanola kak test dlia opredeleniia éffektivnosti antialkogol'nykh preparatov sensibiliziruiushchego deistviia.

A procedure is developed for determination of the efficiency of the antialcoholic drugs' action on the system of ethanol oxidation in vitro: alcohol dehydrogenase by the direct and reverse reactions and aldehyde dehydrogenase of subcellular fractions of the rat liver. This procedure is also used to determine the antialcoholic activity of a number of new compounds and to compare them with disulphiram (antabus, teturam), the known antialcoholic drug.

[Changes in the supramolecular organization of polyurethaneurea-acylsemicarbasides in the process of their biodestruction]. / Izmenenie nadmolekuliarnoi organizatsii poliuretanomochevino-atsulsemikarbazidov v protsesse ikh biodestruktsii.

Using the scanning electron microscope changes in supramolecular organization of polyurethane, (polyurethanecarbamideacylsemicarbasides), having in the main polymer chain foci of tripeptide were studied. They were subjected to proteolytic enzyme action and implanted into animal organisms. It was found that erosive processes going on the polymer plate are essentially similar either under action of proteolytic enzymes or inside a living organism.

[Biodegradable co-polymers of amino acids and synthetic polymers]. / Biodestruktiruemye sopolimery aminokislot i sinteticheskikh polimerov.

The methods of synthesis and properties of copolymers of natural amino acids and of oligopeptides with synthetic polymers are considered as well as their liability to biodestruction processes both under the effect of proteolytic enzymes in vitro and under conditions of the implantation to animals. Such copolymers may be successfully used in biology and medicine as a material for manufacturing temporal endoprostheses, carriers of medicinal preparations, immobilization of biologically active compounds as well as in studies of the mechanism of their action.

[Method for the quantitative study of the growth of fibroblastic cells in tissue culture]. / Metod kolichestvennogo issledovaniia rosta fibroblasticheskikh kletok v kul'ture tkani.

The method is elaborated for a quantitative evaluation of hypodermic fibroblast element growth in tissue culture of albino rats. The method is based on determination of the growth activity and intensity for each of the explant growth zones (compact, necwork-like zones and zone of migrating elements). The method permits a rapid and an exact evaluation of the influence of different substances added to the culture medium.

[Enzymic segregation of segmented polyurethanes containing sugar residues in the composition of the main chain]. / Fermentativnoe rasshcheplenie segmentirovannykh poliuretanov, soderzhashchikh v sostave glavnoi tsepi ostatki sakharov.

It is shown that segmented polyurethanes with D-lactose and D-maltose residues in the main polymer chain are subjected to a specific effect of enzymes: beta-galactosidase and alpha-amylase, respectively. This leads to a decrease in values of characteristic viscosity and physicomechanical indices of polyurethanes after their incubation with the mentioned enzymes. The compounds obtained are subjected to acid hydrolysis, which confirms sugar residues in the main polymer chain.

Structural peculiarities of block copolyurethanes with peptide links as rigid block extenders.

X-ray crystal analysis was performed on block copolyurethanes with peptide links as rigid block extenders. The increase in peptide link length was shown to improve the microphase separation of flexible and rigid segments. The influence of the isocyanate nature on phase separation conditions in the block copolyurethanes studied was established. The correlation between mechanical and temperature characteristics and peculiarities of the microphase structure was found. The latter seems especially important because of the dependence existing between the biodegradation tendency of the polyurethanes and their supermolecular organization.