We evaluated normal blood glucose levels in two species of monkeys, namely rhesus monkeys (Macaca mulatta) and cynomolgus macaques (Macaca fascicularis) kept in the Breeding Center of the Institute of Medical Primatology. The animals were either under moderate stress due to catching or after recovery of defensive and orienting reactions by habituation to the blood sampling. Based on these data, seasonal, gender- and age-dependent variations in glucose levels were found. Glucose levels in rhesus monkeys accustomed to blood sampling procedures were shown to be substantially lower compared with those in non-adapted gender- and age-matched animals.
Three injections of doxorubicin to rhesus macaques cause severe intoxication, characterized by anemia, cachexia, and degeneration of the viscera. The life span of monkeys injected with the drug and receiving after 24 h mesenchymal stem cell transplantation varied from 96 to 120 days in comparison with 50-74 days in animals receiving stem cells before doxorubicin. Controls received doxorubicin and saline; the lifespan of one monkey was 24 days, of the other - 1 year and 8 months. The increase in activity of proinflammatory cytokine IL-6 was paralleled by an increase in the level of C-reactive protein.
Changes in telomere length and mutations in p53 and RB genes were analyzed in 6 children with pre-B acute lymphoblastic leukemia. Telomere length was reduced in 4 patients. One patient had mutation in p53 and in one patient loss of heterozygosity of the BR gene was found; in both samples telomere length was reduced.Key Words: acute lymphoblastic leukemia; telomeres; antioncogens Acute lymphoblastic leukemia (ALL), a clonal malignant disease of the blood [1], is the most frequent form of leukemia in children. ALL is accompanied by molecular changes in DNA of blast cells [3,4,11], in particular, inactivation of p53 and RB antioncogens.Telomeres forming the_ ends of eukaryotic chromosomes protect them from degradation, fusion, and recombination. Telomere length in normal ceils decreases with each cell division. At the same time, in immortalized cells with unlimited division potential, telomere length remains unchanged. It is likely that maintenance of telomere length is essential for infinite cell proliferation.The aim of the present study was to analyze changes in telomere length and activity of p53 and RB genes and possible associations between these parameters in children with pre-B ALL.
MATERIALS AND METHODSBlood and bone marrow samples were obtained from 6 children aged 1.5-11 years (patients of the Hematology Department, Sochi Children's Hospital) after Institute of Medical Primatology, Russian Academy of Medical Sciences, Sochi diagnosis of pre-B ALL. Lymphocytes and bone marrow cells were separated by density gradient centrifugation (Lymphocyte Separation Medium, Flow). All samples were analyzed by panning with monoclonal antibodies to differentiation antigens. DNA from bone marrow cells was isolated as described previously [7] and its purity was controlled spectrophotometrically by the ratio of optical densities at 260 and 280 nm.Telomere length was measured as described elsewhere [8]. Cell DNA (5 pg) was digested by Alu I and Hinf I endonucleases (Boehringer Mannheim) and restriction fragments were separated by gel electrophoresis in 0.7% agarose. After standard depurination, denaturation, and neutralization, the DNA fragments were transferred to nylon membrane and fixed. The telomere-specific probe [TrAGGG],~ was synthesized by PCR using [GGGTrA]~ and [TAACCC]I~ primers. The probe was labeled with uridine-digoxygenin (Boehringer Mannheim) during synthesis. Hybridization was performed in 50% formamide and 50% hybridization buffer. The spots were visualized using a Dig luminescent detection kit (Boehringer Mannheim) in accordance with manufacturer's protocol.Mutations at codons 3-9 of p53 antioncogene was analyzed by denaturing gradient gel electrophoresis (DGGE). Primers tbr PCR and DGGE conditions were 0007-4888/99/0004-0398522,00 ~1999 Kluwer Academic/Plenum Publishers
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