Regulation of proliferation and viability of tumor cells in vitro by alloferon-1 and allostatin-1

Alloferon-1 (AF) and allostatin-1 (AS) cytotoxic and growth modulating activities have been compared. AF is cationic oligopeptide isolated from the hemolymph of experimentally infected blow fly Calliphora vicina. AS is AF synthetic analog that differs from the parent molecule in two amino acids substituted. It has been shown that both AF and AS have no direct cytotoxic activity in concentrations ranging from 1 x 10(-1) to 10 microg/ml, however, the peptides demonstrated significant effect on tumor cells proliferation in vitro. Both peptides displayed growth modulating activity in mass cell cultures and boosted growth inhibiting activity of doxorubicin in the course of P388D1 cells cloning, although AS potentated doxorubicin cytostatic activity to a greater extent. Similarly, AS boosted anti-clonogenic activity of cyclophosphamide applied in a subthreshold concentration. Experiments with peptide-fluorescein complex have demonstrated that AF and AS belong to the group of cell-penetrating peptides. Moreover, the experiments displayed AF ability to bind with chromosomes.     

Specific and integrated roles of Lmx1a, Lmx1b and Phox2a in ventral midbrain development

The severe disorders associated with a loss or dysfunction of midbrain dopamine neurons (DNs) have intensified research aimed at deciphering developmental programs controlling midbrain development. The homeodomain proteins Lmx1a and Lmx1b are important for the specification of DNs during embryogenesis, but it is unclear to what degree they may mediate redundant or specific functions. Here, we provide evidence showing that DN progenitors in the ventral midbrain can be subdivided into molecularly distinct medial and lateral domains, and these subgroups show different sensitivity to the loss of Lmx1a and Lmx1b. Lmx1a is specifically required for converting non-neuronal floor-plate cells into neuronal DN progenitors, a process that involves the establishment of Notch signaling in ventral midline cells. On the other hand, lateral DN progenitors that do not appear to originate from the floor plate are selectively ablated in Lmx1b mutants. In addition, we also reveal an unanticipated role for Lmx1b in regulating Phox2a expression and the sequential specification of ocular motor neurons (OMNs) and red nucleus neurons (RNNs) from progenitors located lateral to DNs in the midbrain. Our data therefore establish that Lmx1b influences the differentiation of multiple neuronal subtypes in the ventral midbrain, whereas Lmx1a appears to be exclusively devoted to the differentiation of the DN lineage.     

Modifying effect of the repeated experience of agonistic confrontations on effect of naltrexone in male mice

In mice with different experience of agonistic confrontations: victories or social defeats during 3 and 10 days (T3 and T10 winners and T3 and T10 losers, resp.), T10 winners displayed a lesser aggression and a more hostile behaviour than T3 winners. Naltrexone dose-dependently decreased attacks in the T3 winners and did not affect aggressive grooming, diggings, autogrooming, and exploratory activity. Naltrexone was ineffective in T10 winners. The naltrexone effects were similar in T3 and T10 losers and its high and low doses contrarily affected different parameters of submissive behaviour. The repeated experience of agonistic confrontations seems to modify the naltrexone effects depending on a neurochemical background, differing in winners and losers.     

Effect of ectopic superexperssion of the anti-apoptotic gene bcl-2 on the level and character of karyotypic instability in the CHLL V-79 RJK Chinese hamster cell line

Karyotypic destabilization in cells of Chinese hamster fibroblasts CHL V-79 RJK with ectopically overexpressed antiapoptotical human bcl-2 gene from pSFFV-bcl-2 vector has been analysed. Analysis of G-banded metaphase chromosomes from 4 clones with different levels of bcl-2 expression revealed an increased level of chromosomal instability in bcl-2-transfected cells. Besides, an increased percentage of aneu- and polyploid cells and high level of cells with different chromosomal aberrations was observed. The degree of karyotypic instability positively correlated with the level of bcl-2 expression in bcl-2-transfected cells. Cells of a clone with the highest bcl-2 expression at the 13th passage of cultivation displayed an almost 100% polyploidization and the presence of specific aberrations and a tricentric marker chromosome. Selection of cells with non-random specific chromosome changes was observed in pSFFV-bcl-2-transfected CHL V-79 RJK cells in the process of their long-term cultivation. By contrast, cells of the parental cell line, as well as the control pSFFV-neo transfectants, displayed a stable karyotype throughout the long period of cultivation. It is important that the presence of morphological markers of gene amplifications--DOO, DM, MH--was observed in bcl-2-transfected cells. These findings suggest that the overexpression of antiapoptotic human bcl-2 gene may result in destabilization of the karyotype structure in cells of Chinese hamster fibroblasts CHL V-79 RJK. The character and level of destabilization correlate with the level of ectopic overexpression of this gene.     

Glutamate induces formation of free radicals in rat brain synaptosomes

The influence of glutamate and agonists of its ionotropic receptors on free radical formation in rat brain synaptosomes was investigated using the fluorescent dye DCFDA. Glutamate at concentrations of 100 μM and 1 mM increased the production of reactive oxygen species. This phenomenon was eliminated by removing calcium from the incubation medium. Addition of NMDA (100 μM) or kainate (100 μM) to a suspension of synaptosomes also led to free radical formation. The influence of glutamate receptor agonists was blocked by the specific antagonists MK-801 and NBQX. Thus, activation of NMDA and AMPA/kainate receptors can lead to oxidative stress in neuronal presynaptic endings.     

Genetic Regulatory Mechanisms of Evolution and Embryogenesis in a Distorting Mirror of Carcinogenesis

The article is an attempt to provide an overview of the relationships between the regulatory genetic mechanisms of three fundamental processes of biology, i.e., development, evolution, and cancer. The problems of the evolutionary inevitability of the onset of cancer, the evolutionarily developed systems for protecting the body against tumors, and the general systems of regulation used in evolution, development, and cancer are considered.     

Cytogenetic peculiarities of the cervical epithelium in inflammatory diseases

The functional status of epithelial cells in inflammatory diseases of the cervix has been studied with the use of cytogenetic methods of detection of chromosome nucleolar organizers. The highest level of proliferation and rRNA synthesis was detected in cylindrical epithelial cells using indexes of compact and transitional nucleolonemic types of nuclear organizer regions; an elevated level of proliferation and rRNA synthesis was found in the squamous epithelial cells of the intermediate layer, and a low level of proliferation and rRNA synthesis was found in the squamous epithelial cells of the superficial layer.     

The NADPH oxidase activity of pea seedling roots in rhizobial infection depending on abiotic and biotic factors

The changes in NADPH activity was studied in the roots of 3–4-day-old etiolated pea (cultivar Aksaiskii usatyi) seedlings depending on plant inoculation with Rhizobium leguminosarum bv. viceae (strain CIAM 1026), adverse environmental factors (low temperature and high dose of a mineral nitrogen fertilizer), chemical substances (sodium nitroprusside and methyl viologen, or paraquat), and a biotic factor—the bacterium Escherichia coli (strain XL-1Blue). It was demonstrated that all exogenous factors increased the activity of microsomal NADPH oxidase. Rhizobial infection removed the activation caused by exogenous factors only in the case of high nitrogen content in the medium, thereby displaying an antagonistic effect. A synergistic action on the enzyme activity was observed in the variants with combined action of rhizobia + paraquat and rhizobia + E. coli. An increased NADPH oxidase activity coincided with a growth inhibition of pea seedling roots. The results are discussed from the standpoint of the roles of NADPH oxidase and reactive oxygen species in the legume-rhizobium symbiosis.     

Cytomorphological characteristics of dendritic cells

Five morphological degrees of maturity of dendritic cells induced from monocytes of peripheral blood in oncological patients have been revealed using originally prepared and stained cytological preparations. Cytological examination of dendritic cells can be used for their identification in production of dendritic cell antitumor vaccines.     

Kanamycin concentrations in liver tissue and bile in acute experimental cholecystocholangiohepatitis

The levels of kanamycin in the bile and liver tissue were studied on 10 healthy dogs and 10 dogs with experimental acute abacterial cholecystocholangiohepatitis. It was shown that bile excretion of kanamycin in the dogs increased 6-7 times with development of acute inflammation in the biliary system.