Rhesus Macaques: VNTR Polymorphism of the FCGRT Gene.

Variable number tandem repeat (VNTR) polymorphisms of the human neonatal IgG Fc receptor α-chain gene (FCGRT) are known to influence the expression levels of FCGRT and IgG in serum. Monkeys are considered to be a relevant biological model for studying the effects of immunobiological drugs. The study determined the functional VNTR polymorphisms of the FCGRT gene in 109 male rhesus macaques from the nursery of the Kurchatov Complex of Medical Primatology. PCR amplification of samples was carried out followed by electrophoretic separation of DNA fragments in a 2% agarose gel. Individual DNA amplification products were sequenced (according to Sanger system) in forward and reverse directions to confirm the specificity. The genotyping showed that the VNTR polymorphism of the FCGRT gene in the studied population of rhesus macaques is presented by 9 variants. The frequency of the VNTR5 allele associated with lower IgG levels was 14.2%, and the most common one was the VNTR7 allele (25.2%). We also identified alleles that have not been previously reported: VNTR3, VNTR4, VNTR6, VNTR8, and VNTR9. The study allows to consider rhesus macaques as a potential model for studying the immunological response depending on the genetic VNTR variant of FCGRT.

[Indicators of spontaneous behavior of rhesus monkeys with short-term course alcohol self-administration under free choice]. / Pokazateli spontannogo povedeniya makak-rezus pri subkhronicheskom potreblenii alkogolya v modeli so svobodnym vyborom.

OBJECTIVE: To analyze spontaneous behavior in 36 male rhesus macaques during formation of alcoholic motivation under free choice. MATERIAL AND METHODS: The model composed the initiation stage with the provision of an aqueous ethanol solution of increasing concentration from 1 to 4% with a sweetener and restricted access to the tap water supply and the subsequent stage of the formation of alcoholic motivation with a free choice between 4% ethanol solution without sweetener and water. The behavior was recorded by the «One-Zero¼ method with ethogram compiled in accordance with the behavioral peculiarities of rhesus monkeys when housed individually. Three subgroups of high, medium and low-level ethanol consumption were distinguished. RESULTS: In the subgroup of high ethanol consumption median consumption was 1.70 g/kg/day at initiation (p<0.05, compared to other subgroups) and 1.79 g/kg/day (p<0.05) at free choice stage. Animals of high consumption subgroup had significantly higher frequency being at the cage bottom and in a sitting posture. We observed significant changes in a number of indicators of spontaneous behavior depending on the level of ethanol consumption, which included displacement behavior, stereotypic behavior and posture of animals. In the high consumption subgroup, there was a significant increase in the frequency of stereotypic behavior, atypical behavior, being on four legs, as well as a decrease in the frequency of being at the back side of the cage and of displacement behavior. At the same time, the inhibitory effect of ethanol on the hypothalamic-pituitary-adrenal system was revealed. CONCLUSION: In the model of free choice alcohol self-administration rhesus monkeys demonstrate a significant change in a number of indicators of spontaneous behavior depending on the level of ethanol consumption, which includes displacement, stereotypic behaviors and animal posture.

Genotyping of Macaque Population on the Brain-Derived Neurotrophic Factor Gene Polymorphism by Mismatch Amplification Mutation Assay (MAMA)-PCR.

Human rs6265 (196G>A) polymorphism in the BDNF gene is associated with many clinically significant phenotypic manifestations. Rhesus monkey (Macaca mulatta) has a functionally significant rs309950446 ( 136G>A) polymorphism. To determine this polymorphism in macaques, we used mismatch amplification mutation assay (MAMA)-PCR method with non-complementary nucleotide to the template chain at the 3rd position from the 3'-end of the allele-specific primers (mismatch primers), which allowed the best discrimination of the alleles. Genotyping of male rhesus monkeys (n=178) and cynomolgus monkeys (Macaca fascicularis) (n=90) was carried out. The A/A, G/G, and G/A genotypes were found in 16, 34, and 50% rhesus macaques, respectively. In the cynomolgus macaques, the mutant polymorphic allele was not detected. The study results allow considering rhesus macaques as a potential biological model for assessment of the gen-environment interaction of the BDNF gene polymorphism.

[Changes in telomere length in leukocytes of male rhesus macaques of different ages.]

Telomeres are specialized terminal sections of chromosomes that ensure the stability of the latter. DNA duplication during cell division is associated with telomere shortening due to the phenomenon of terminal underreplication. As cells divide, shortening of telomere length is considered to be one of the most important causes of cell aging. Estimation of telomere length still remains the subject of scientific research in gerontology and it is not used in clinical practice. Most often, rodents are used as a model object for studying the aging process, but the neuroendocrine mechanisms that influence, among other things, the regulation of the aging process differ in rodents and humans. The model objects closest in phylogenetic relation to humans are monkeys. In particular, Rhesus macaques is one of the representatives of the Old World most often used in biomedical research. However, data on age-related changes in telomere length in monkeys are extremely scarce. We studied the absolute average length of telomeres in DNA from blood leukocytes of 29 clinically healthy male rhesus monkeys aged from 4 to 24 years using quantitative PCR-method. The data obtained did not correspond to the normal distribution and the correlation analysis showed the absence of a significant dependence of telomere length on the age of the animals (rs=0,27; p>0,05). Thus, our study does not confirm the dependence of changes in the average length of telomeres of blood leukocytes with age.

Formation of Alcohol Motivation in Monkeys (Macaca mulatta) Kept in Isolation.

The effects of environmental factors (flavors, different ethanol concentration, alcoholic deprivation, and food reinforcement) on the formation of alcohol motivation was studied in rhesus macaques (Macaca mulatta, n=6). Motivation for alcohol intake was induced in two stages: initiation (sessions 1-160) and formation of motivation (sessions 161-516). Monkeys preferred multifruit flavor and 4% ethanol solution, while ethanol deprivation did not stimulate alcohol consumption. The pronounced individual differences in the pattern of alcohol motivation were revealed: the intake of 4% ethanol solution ranged from 0.21±0.03 to 0.43±0.06 g/kg without food reinforcement and increased from 0.78±0.03 to 1.22±0.09 g/kg with food reinforcement. The results suggest that the proposed method is valid and can be used as an experimental model of alcohol dependence in non-human primates.

[Spectrum, clinical features, diagnosis of opportunistic and comorbid pathology in HIV-infected patients admitted to infection hospital of Moscow].

AIM: To analyse structure, clinical features, diagnosis of opportunistic and concomitant diseases in patients with HIV infection admitted to infection hospital of Moscow. MATERIAL AND METHODS: A total of 4155 patients with HIV infection (1518 of them with AIDS) most of them (89%) at the age of 20-39 years were treated in Moscow AIDS hospital in 2006-2007. The examination included standard blood and urine tests, device diagnosis, immunological, bacteriological and molecular investigations of biological materials for detection of opportunistic infections. Cell-mediated immunity was also studied. HIV infection resulted in a lethal outcome in 255 (6.1%) inpatients. RESULTS: Leading causes of hospitalization of patients at early stages of HIV infection were bacterial bronchitis or pneumonia, hepatic pathology (chronic viral hepatitides, alcohol-associated diseases), sepsis. One-third of the inpatients were at AIDS stage characterized by tuberculosis (66.3%), visceral candidosis (12%), manifest cytomegalovirus infection (10.1%), cerebral toxoplasmosis (9.2%), pneumocystic pneumonia (5.5%). The number of HIV-infected persons with atypical mycobacteriosis, lymphoproliferative diseases, brain tumors increased. Chronic hepatitis C prevailed among liver damage cause in HIV infection, it also often caused hospitalization and death of patients. 60.3% patients having HIV infection who died without AIDS stage had hepatic cirrhosis. Tuberculosis was a leading cause of severe pulmonary pathology, most frequent opportunistic disease, main cause of death in patients with HIV infection. One-third of patients had generalized tuberculosis. Tuberculosis was diagnosed in more than 40% HIV-infected patients with pulmonary lesion, in 65% AIDS patients, 36% dead AIDS patients. CONCLUSION: To render effective anti-HIV treatment, infection hospital must be equipped with facilities providing device tests, molecular diagnosis, modern etiotropic and pathogenetic medication.

Oxidative stress in patients with beta-thalassemia major.

The aim of this work is to study the level of oxidative stress in blood of beta-thalassemia major patients with transfusional iron overload and chelation therapy as a central pathological process. Beta-thalassemia major results in an increase in the concentration of lipid peroxidation products in blood plasma of more than 100% and in the intensity of chemiluminescence - about 20% in comparison to healthy controls. The activity of the antioxidant enzyme superoxide dismutase in the blood of beta-thalassemia major patients is decreased by more than 30% and the total antioxidant activity is diminished by about 70% compared to controls. Experimental data confirm the progression of oxidative stress in patients with beta-thalassemia major: activation of free radical processes and lipid peroxidation, decreased antioxidant capacity. Strong oxidative damage and essential alternations define these parameters as sensitive markers of oxidative stress in patients with beta-thalassemia major. The combination of effective iron-chelatory agents with natural or synthetic antioxidants can be extremely helpful in clinical practice in the regulation of the antioxidant status of patients with beta-thalassemia major.

[Differential diagnosis of lymphadenopathy in HIV/AIDS].

AIM: To determine the role of histological diagnosis of lymphadenopathy (LAP) associated with clinico-laboratory picture in patients with HIV infection/AIDS. MATERIAL AND METHODS: Target biopsy of the peripheral lymph node was made in 80 HIV-infected patients from 2002 to 2005. Histological diagnosis was made in all the patients with light microscopy, in some patients at immunohistological examination. RESULTS: Most of the patients had peripheral blood CD4 lymphocytes under 200 cell/mcl. Viral load was hundred thousands copies in mcl. Tuberculosis was diagnosed in 33 (41%) patients, lymphomas--in 23(29%), lymphogranulomatosis--in 5 (6%), reactive lymphadenopathy--in 15 (19%), germinogenic tumors--in 3 (4%), sarcoidosis--in 1 (1%). Histologically, LAP was represented by follicular hyperplasia (n = 9), involution (n = 2), bacterial lymphadenitis with necrosis (n = 4). CONCLUSION: Biopsy of peripheral lymph nodes is an early, safe, reliable and cost-effective method of LAP diagnosis in patients with AIDS.