Changes in lipid metabolism during Walker 256 tumor growth and the therapeutic effect of hyperthermia.

The dynamics of lipoprotein content during Walker 256 tumor growth in rats was studied. Moderate changes in HDL and LDL were paralleled by significant changes in VLDL level. A 2-fold increase of VLDL in comparison with the intact control was recorded on day 10 of tumor growth. Exposure to total hyperthermia additionally stimulated VLDL synthesis and this parameter increased by 4 times and more in rats with tumors in comparison with controls. This effect of hyperthermia correlated with significant subsequent decrease of rat mortality caused by the lethal effect of the tumor.

[Features of the immune proteasome expression in ascite Zajdela hepatoma after implantation into Brattleboro rats with the hereditary defect of arginine-vasopressin synthesis].

The expression of the total proteasome pool, immune proteasome subunits LMP2 and LMP7, TAP1 and TAP2 transporters, as well as RT1A molecule of MHC class I was investigated in the ascite Zajdela hepatoma at the 10th day after implantation into Brattleboro rats with the hereditary defect of hypothalamic arginine-vasopressin synthesis (AVP) and into WAG rats with normal AVP expression. In Zajdela hepatoma cells implanted into Brattleboro rats the 3-fold increase of the total proteasome pool and LMP2 level and 8-fold increase of the LMP7 level was detected by Western blotting as compared to those in WAG rats. Differences in the LMP2 and LMP7 expression suggest variations in their functions, namely the important role of LMP7 in anti-tumor immunity. The growth of Zajdela hepatoma in WAG rats was accompanied by the decreased level of total proteasome pool as well as immune proteasome expression as compared to those in Brattleboro rats during the regression of tumor. The analysis of TAP1 and TAP2 revealed the pronounced expression of these peptide transporters in Zajdela hepatoma cells implanted into Brattleboro and WAG rats. The expression level of RT1A molecule of MHC class I was increased 3 times in Zajdela hepatoma cells implanted into Brattleboro rats as compared to WAG rats. Moreover, flow cytometric analysis of CD4- and CD8-lymphocytes number in the spleen of Brattleboro and WAG rats was performed at the 10th day after implantation of Zajdela hepatoma. The increased number of CD4- and CD8-lymphocytes was observed in the spleen of Brattleboro as compared to WAG. The increased subpopulations of cytotoxic T-lymphocytes and T-helpers might promote the tumor regression in Brattleboro rats. The reduced populations of CD4- and CD8-lymphocytes in the spleen of WAG rats were accompanied by the splenomegaly and tumor progression. The data obtained suggest that AVP deficiency in Brattleboro rats leads to the increase of the immune proteasome and MHC class I expression in Zajdela hepatoma cells, resulting in tumor immunogenicity and its elimination by the adaptive immunity.

[Concordance between vasopressin gene expression and growth of Walker 256 carcinosarcoma in rats].

The growth features of Walker 256 carcinosarcoma in rats of different genotypes were investigated. The experiments has been carried out on rats of the inbred Brattleboro and WAG lines, as well as on their hybrids segregated during congenic translocation of the normal vasopressin gene to the genotype of the Brattleboro rats. Brattleboro rats do not express the vasopressin gene. It has been found that there are only two types of tumor growth dynamics. In rats of the inbred Brattleboro line and in homozygotes di/di, that were segregated by backcrossings of heterozygous offsprings from the original crossbreeding between (WAG x Brattleboro) F1 x Brattleboro and the individuals with parental Brattleboro genotype, having grown to some extent the tumor regresses and disappears. In hybrid heterozygous siblings of di/+ genotype tumor grows linearly with time and always leads to fatal outcome. It has been found that, in the congenic procedure, the tumor regression trait is stably maintained and persistently inherited in lineage concordantly with the di/di genotype and, in rats with at least one allele of a normally expressed vasopressin gene, continuous and lethal tumor growth is always observed.

[Expression of the RT1A molecule of the class I major histocompatibility complex in a Walker 256 tumor after transplantation into Brattleboro rats with a genetic defect of arginine-vasopressin synthesis].

The dynamics of expression of the RT1A antigen of the class I major histocompatibility complex (MHC) in a Walker 256 tumor after its transplantation into Brattleboro rats with a genetic defect of Arginine-Vasopressin synthesis in the hypothalamus was studied. Expression of the RT1A antigen was detected by means of Western-blotting and flow cytometry in the tumor cells on the 14th-17th days after transplantation. In addition, a simultaneous increase in the portion of cells that express the RT1A antigen and in the level of its expression per cell was observed. It is presupposed that at a deficiency of Arginine-Vasopressin, a renewal of expression of the class I MHC antigens, which results in an increase of immunogenicity of this tumor and regression, occurs in the Walker 256 tumor in the Brattleboro rats.

[Osmoregulating effect of vasopressin with prolonged action on Brattleboro rats with hypothalamic diabetes insipidus].

The particularities of urine osmotic concentration depending on hormonal background of vasopressin were studied in rats. It was found that WAG and Brattleboro lines of rats characterized respectively by normal level and absence of endogenous vasopressin, possess interline correlation of urine osmolality (p = 0.86) in various conditions between the extreme hydrating and dehydratation. Concentrating level of WAG rats varies from 747 +/- 94 to 2936 +/- 128 mOsm/kg, but that of Brattleboro rats changes more within the 160 +/- 9 being twice lower as isotonicity to 1305 +/- 142 mOcm/kg. Urine concentrating goes up to 1391 +/- 76 mOcm/kg in Brattleboro rats already on the day of the action of exogenous vasopressin secreted from ALZET minipump, however, in spite of constant work of this minipump during 4 hrs a week, further increasing of urine osmolality was not observed in Brattleboro rats.

[Hormonal dependency of Walker 256 carcinosarcoma growing in rats].

The particularities of Walker 256 carcinosarcoma growing were studied in rats depending on hormonal background. Rats of lines NISAG and Brattleboro were characterized by the increased and lowered level of endogenous corticosteroids, respectively, in contrast with rats WAG different dynamics of growing tumors. If rats NISAG are not distinguished from rats WAG on the growing tumors, then the tumor grows weakly in the rats of line Brattleboro, and it takes place during the first fortnight only but then begins a regression up to disappearance. Velocity of tumor increasing is slowed in rats of WAG line under adrenalectomy but this effect disappears after introduction of exogenous cortisol. Cortisol introduction does not correct atypical dynamics of tumors in rats Brattleboro.

[Effect of vasopressin gene expression on the growth of Walker 256 carcinosarcoma in rats].

The growth pattern of the Walker 256 solid tumor has been studied in rats with different doses of the mutant vasopressin gene. In contrast to the vasopressin gene of normal WA rats, that of mutant Brattleboro rats has a deletion in the coding region that blocks expression at the translation level. The mutation is inherited as a recessive character and is expressed in homozygous Brattleboro rats as diabetes insipidus with an increased water consumption because of the absence of vasopressin in the blood. (WAG x Brattleboro) F1 hybrids have the same normal phenotype as WAG rats, including a low water consumption. Walker 256 carcinosarcoma, which is not strain-specific, intensely grows only in WAG and (WAG x Brattleboro) F1 rats. In these groups, the growth of the tumor leads to the animal death within approximately 30 days after the inoculation of tumor cells. In Brattleboro rats, the carcinosarcoma grows less intensely: the tumor node somewhat increases only within the first two weeks, after which the tumor began to decrease and eventually disappears altogether. Both characters exhibit a 100% concordance at the individual level.

[Dynamics of renal medullary vinculin under changing of osmoregulating function].

Reduction of vinculin occurs in the renal medulla under the long-lasting dehydration. The protein content measured in inner medulla of rats of the WAG line under hydration was 92.1 +/- 6.3 in relative units, but it was only 77.6 +/- 2.3 after a 3-day water deprivation. The vinculin content in the inner medullar layer from mutant rats of Brattleboro line incapable of synthesizing vasopressin is essentially higher: it is 188.9 +/- 8.5 in hydrated conditions and drops to 148.4 +/- 7.3 under a 3-day dehydration. The same high level of vinculin is in outer medulla from rats of Brattleboro line: 222.1 +/- 11.8 in hydrated animals and 174.9 +/- 11 after a 3-day dehydration. No differences were revealed in response of vinculin to alternative osmoregulating stimulation in cortex in both rat lines.

Walker 256 tumor growth in rats with hereditary defect of vasopressin synthesis.

Stable deceleration of Walker 256 tumor growth was detected in Brattleboro rats with vasopressin synthesis defect in comparison with normal WAG rats. In contrast to continuous tumor growth typical of rats, the growth of this tumor in Brattleboro rats was negligible and was observed during the first 15-18 days after transplantation, after which the tumor regressed and disappeared. The effect was age-dependent and was more pronounced in old animals. Repeated injection of Walker 256 cells does not lead to tumor development, which attested to direct involvement of the immune system in the detected phenomenon.

[Reaction of alpha-actinin from renal inner medulla on prolonged dehydration]. / Izmenenie soderzhaniia alpha-aktina v mozgovom veshchestve pochki u krysy pri degidratatsii.

A western-blot analysis using monoclonal antibodies revealed that a reduction of alpha-actinin occurred in the renal inner medulla under the long-lasting dehydration. The ratio between protein content measured in rats of WAG line being hydrated or after 3-days water deprivation consisted of 52.7+/-6.2 against 23.9+/-3.3 as evaluated in relative units. The alpha-actinin level changes similarly in mutant rats of Brattleboro line not capable of synthesizing vasopressin. It was 57.5+/-4.6 in hydrated animals, and statistically lower in rats being under 3-day water deprivation--26.4+/-5.7 in relative units of protein.

[Interaction of the diabetes insipidus locus alleles with the renal 120 kDa protein-encoding gene in rat development]. / Vzaimodeistvie allelei lokusa diabetes insipidus i gena pochechnogo belka 120 kDa v ontogeneze krysy.

Age-dependant dynamics of the kidney inner medullary 120-kDa protein content in vasopressin-deficient Brattleboro rats with di/di mutant genotype was studied in comparison with WAG rats with genotype and normal vasopressin expression. Age-dependant dynamics of vasopressin content in neurohypophysis of WAG rats was also examined. It was shown that 10-day-old WAG rats were unable to elevate the synthesis of the 120 kDa protein in respond to long-term dehydration, while this tendency was clearly observed in the 15-day-old rats and later in the development. In WAG rats the onset of this specific feature was time correlated with the development of the respond to hydration by the elevation of vasopressin synthesis and release from neurohypophysis into blood. In the di/di rats dehydration had no effect on the kidney 120-kDa protein synthesis in all ages examined. These results point to the interaction between the di alleles and the 120-kDa protein-encoding gene in the course of development.

[Phenotypic expression of the mutant gene diabetes insipidus in rats and criteria of genotyping by phenotype]. / Fenotipicheskoe proiavlenie mutantnogo gena diabetes insipidus u krys i kriterii genotipirovaniia po fenotipy.

The autosomal semidominant mutant gene di (diabetes insipidus) is manifested in homozygotes in the form of diabetes insipidus with water consumption from 25 to 100% of body weight per day. The heterozygotes di/+ drink water at a rate higher than 5% but lower than 25%. The level of water consumption in rats with +/+ genotype does not exceed 5% of body weight per day. Segregation analysis of F1 animals yielded by various crosses showed that genotyping of di/di homozygotes is absolutely reliable at 30% and higher level of the water consumption per day.

[Functional features of actinin and tropomyosin distribution in rat kidney]. / Funktsional'nye osobennosti raspredeleniia aktinina i tropomiozina v pochke u krysy.

The intrarenal distribution of actinin and tropomyosin was studied by western blot analysis in various functional conditions. It was found that actinin content is always higher in cortex than in medulla. The highest tropomyosin content was revealed in outer medulla, but it is more than twice higher in rats of mutant Brattleboro line versus hydrated normal WAG rats. This ratio rises up to 46.12 +/- 2.14 versus 13.83 +/- 0.66 (in relative units) for rats being under dehydration that maximally activated vasopressin secretion in normal WAG rats.

Immune system in vasopressin-deficient rats during ontogeny.

Morphofunctional immune disorders were revealed in vasopressin-deficient Brattleboro rats with diabetes insipidus during ontogeny. We observed a permanent decrease in the number of blood lymphocytes, increase in neutrophil count, reduced activity of macrophages, early involution of the thymus and spleen, and suppression of antibody production. These changes reflect impaired general resistance of these animals.

[Specificity of gene di (diabetes insipidus) expression in homologous inbred rat strains]. / Osobennosti ékspressii gena di (diabetes insipidus) v gomologichnykh inbrednykh liniiakh krys.

The diabetes insipidus mutation is displayed in homozygotes in the form of diabetes insipidus with water consumption from 30 to 100% of body weight per day. We developed two inbred sublines of the di/di Brattleboro rats as well as the recombinant inbred subline by integrating genes of August rats into the di/di mutant genome. Changes in the genetic background proved to have no effect on the quantitative parameters of the diabetes insipidus. The intensity of the secondary immune response and the content of tropomyosin in the medulla of the rat kidney can serve as additional marker traits of the di/di genotype.