Experimental visceral leishmaniasis in golden hamsters

As a result of a long passage of L. donovani isolate on golden hamsters (21 passages were observed), in transplanting the agent from animals with a distinct clinical picture there was formed a highly virulent strain "G" of L. donovani for this species of animals. The weight arrest and then body mass losses were the most early signs of the disease. Parasites were regularly accumulated in spleen and liver and to a less extent in bone marrow. The main manifestations of visceral leishmaniasis in hamsters are cachexia, lienal syndrome, polyglandular deficiency on the background of hypoplasia of lymphoid tissue and defects of the system of monocytic phagocytes. Such symptom-complex can be a result of neuroendocrine deficiency during visceral leishmaniasis. Pathohistological picture of experimental visceral leishmaniasis is similar to that of man, so L. donovani infection in hamsters can serve as a model for studies of different medical and biological aspects of leishmaniasis.     

Mapping of the transferrin gene in laboratory rats, mice and man by direct in situ hybridization

Mapping of the gene coding for transferrin was carried out in metaphase chromosomes from bone marrow of laboratory mice and rats as well as from PHA-stimulated human lymphocytes using direct in situ hybridization technique. Plasmid pRTf-17 carrying the insert of rat transferrin cDNA was nick-translated with [125I]dCTP and used as a specific hybridization probe. The total number of silver grains and their distribution along differentially stained chromosomes were determined in 464 metaphase plates (114, 263 and 87 from rat, mouse and man, respectively). The data obtained enable us to assign transferrin gene to chromosome 3 in human and chromosome 9 in mouse. For the first time, the rat transferrin gene was localized on chromosome 7. The most probable sites of transferrin gene localization are 7q31-34, 9F1-3 and 3q21 in rat, mouse and human chromosomes, respectively.     

Peptide Aβ(16-25) forms nanofilms in the process of its aggregation

A method for the synthesis and high purification of fragments of Aβ(1-42) peptide has been elaborated. We have synthesized the amyloidogenic fragment Aβ(16-25) predicted by us and studied the process of its aggregation by electron microscopy and X-ray analysis. Electron microscopy images show that the peptide forms a film, which is not characteristic of amyloid fibrils. At the same time, according to the X-ray diffraction data, its preparations display the presence of two main reflections (4.6-4.8 and 8-12 Å) characteristic of cross-β structure of amyloid fibrils. Thus, the fragment Aβ(16-25) that we predicted is a promising object not only for studying the process of polymerization of the peptides/proteins, but also for using it as a nanomaterial to study a number of biological processes.     

Ectopic cervical thymi and no thymic involution until midlife in naked mole rats

Immunosenescence is a hallmark of aging and manifests as increased susceptibility to infection, autoimmunity, and cancer in the elderly. One component of immunosenescence is thymic involution, age‐associated shrinkage of the thymus, observed in all vertebrates studied to date. The naked mole rat (Heterocephalus glaber) has become an attractive animal model in aging research due to its extreme longevity and resistance to disease. Here, we show that naked mole rats display no thymic involution up to 11 years of age. Furthermore, we found large ectopic cervical thymi in addition to the canonical thoracic thymus, both being identical in their cell composition. The developmental landscape in naked mole rat thymi revealed overt differences from the murine T‐cell compartment, most notably a decrease of CD4⁺/CD8⁺ double‐positive cells and lower abundance of cytotoxic effector T cells. Our observations suggest that naked mole rats display a delayed immunosenescence. Therapeutic interventions aimed at reversing thymic aging remain limited, underscoring the importance of understanding the cellular and molecular mechanisms behind a sustained immune function in the naked mole rat.     

Comparative characteristics of distribution and spatial heterogeneity of ant and mammal populations in northeastern Altai (a cartographic analysis)

Cluster analysis and plotting based on censuses of ant nests and existence energy of mammals were used to classify their communities and species by similarity of distribution. As a result, eight maps were drawn up, with characteristics of the areas populated by the animal groups analyzed. Changes of population density and species abundance with altitude zones have been traced. The force and similarity of relationship between the heterogeneity of communities and environmental factors are estimated. Similarities and differences in the distribution and ecological habitat requirements of the animal groups under study are determined.     

Cells with unusual mitochondria in the thyroid gland of an old rat]

Thyroid glands of 3 year old rats have been studied by electron microscopy. The cells with changed mitochondria were found next to the typical thyrocytes in the follicle walls. The large mitochondria with dense matrix and tubular cristae often occupy the major part of the cytoplasm in these cells. On this basis, the author considers the changed cells to be the Askanazy cells, or oncocytes. Mitochondria-rich cells were found also among calcitocytes. The origin of oncocytes from usual thyrocytes and calcitocytes is under discussion.     

Establishing Primary Adult Fibroblast Cultures From Rodents

The importance of using primary cells, rather than cancer cell lines, for biological studies is becoming widely recognized. Primary cells are preferred in studies of cell cycle control, apoptosis, and DNA repair, as cancer cells carry mutations in genes involved in these processes. Primary cells cannot be cultured indefinitely due to the onset of replicative senescence or aneuploidization. Hence, new cultures need to be established regularly. The procedure for isolation of rodent embryonic fibroblasts is well established, but isolating adult fibroblast cultures often presents a challenge. Adult rodent fibroblasts isolated from mouse models of human disease may be a preferred control when comparing them to fibroblasts from human patients. Furthermore, adult fibroblasts are the only available material when working with wild rodents where pregnant females cannot be easily obtained. Here we provide a protocol for isolation and culture of adult fibroblasts from rodent skin and lungs. We used this procedure successfully to isolate fibroblasts from over twenty rodent species from laboratory mice and rats to wild rodents such as beaver, porcupine, and squirrel.Download video file.^{(71M, mov)}     

Analysis of DNA Double-strand Break (DSB) Repair in Mammalian Cells

DNA double-strand breaks are the most dangerous DNA lesions that may lead to massive loss of genetic information and cell death. Cells repair DSBs using two major pathways: nonhomologous end joining (NHEJ) and homologous recombination (HR). Perturbations of NHEJ and HR are often associated with premature aging and tumorigenesis, hence it is important to have a quantitative way of measuring each DSB repair pathway. Our laboratory has developed fluorescent reporter constructs that allow sensitive and quantitative measurement of NHEJ and HR. The constructs are based on an engineered GFP gene containing recognition sites for a rare-cutting I-SceI endonuclease for induction of DSBs. The starting constructs are GFP negative as the GFP gene is inactivated by an additional exon, or by mutations. Successful repair of the I-SceI-induced breaks by NHEJ or HR restores the functional GFP gene. The number of GFP positive cells counted by flow cytometry provides quantitative measure of NHEJ or HR efficiency.Download video file.^{(82M, mov)}