Efficiency of recombinant thymosin ß4 in spontaneous mouse model of chronic dermatitis.

The therapeutic efficiency of recombinant thymosin ß4 (rTß4) synthesized by us was studied in vivo on spontaneous CBRB mouse model that is adequate to human chronic dermatitis. Three applications of the drug during a week significantly alleviated symptoms of the disease in female mice, and in complex with subsequent antibacterial and antifungal therapy led to a pronounced and lasting (2 months) therapeutic effect. The results attest to a possibility of using rTß4 in combination with the known treatment protocols for chronic inflammatory diseases of the skin.

[Rational approach to administration of combination therapy in arterial hypertension: modern recommendations and personal experience].

Modern treatment of arterial hypertension (AH) in based on concept of necessity of arterial pressure (AP) lowering to target levels for prevention of cardiovascular (CV) diseases (D) and reduction of cardiovascular mortality. AP <140/90 and <140/80-85 mm Hg are target levels for general population and patients with diabetes, respectively. Most patients should be initially prescribed combination therapy as in ambulatory practice mainly patients from high and very high risk groups are observed. Prescribing combination therapy one should take into consideration not only category of risk of CVD development but also AP level, i.e. degree of AH. It is not expedient to always start combination therapy with low doses of preparation because administration of such therapy in patients with 2-3 degree AH inevitably leads to necessity of further elevation of doses and lengthening of time to achievement of target AP. It should be mentioned that achievement of target AP is possible with continuation of therapy with higher dose of same combination without addition of third drug. Rational pharmacotherapy of AH implies concentration of efforts on consideration of not only AP but also of factors of risk of development of CV complications (C) especially on detection of symptomless target organs damage and clinical complications for assessment of total of CVC development because of recent update of data on prognostic significance of symptomless damage of target organs including heart, blood vessels, kidney, eyes, and brain.

[1,10-phenantroline europium complexes: their inclusion in liposomes and cytotoxicity].

For a series of 1,10-phenantroline tris-beta-diketonate europium complexes (EuC), cytotoxic activity on the HBL-100 human breast carcinoma cells was determined. Liposomal preparation of the most active EuC, V12, was also tested for cytotoxicity. Testing of this preparation in vivo on starting lethal murine model of T cell leukemic lymphoma ASF-LL showed that the inclusion of V12 in liposomes did not increase its antitumour activity in a local mode of administration.

[Criteria for the effectiveness of interleukin-2 immunotherapy in a spontaneous murine mammary tumor model].

A novel approach is suggested to identify more homogenous subgroups involved in the follow-up of growth of spontaneous mammary tumors in mice (116, history-based analysis). That depends on subclinical period (preneoplastic and non-invasive stages of tumor growth) as well as rate of growth after clinical manifestation. An analysis of tumor growth rate versus survival of experimental and control animals after primary diagnosis and clinical manifestation of tumor showed that following a single peritumoral 2.5 x 10(6) IU IL-2 treatment tumor growth slowed down (n = 29; p < or = 0.05) while survival tended to improve. Originally fast-growing tumors without significant subclinical stage continued to grow but slowly. Females with such tumors survived longer than untreated controls without showing, however, any improvement on that parameter.

[Antitumor activity of the plant remedy peptide extract PE-PM in a new mouse T-lymphoma/eukemia model].

A new mouse ASF-LL model of adult T-lymphoma/leukemia (ATLL) in humans was characterized by cytological, histopathological, and flow cytometry analyses. Encouraging similarities of morphological, pathological, and clinical signs were found. These included characteristic flower appearance of leukemic cells, lymphadenopathy and hepatosplenomegaly, multiple growths in the skin, urogenital tissues, lungs and pituitary gland, CD4+CD25+ phenotype of the majority of tumor cells that were selectin-L positive, a rapid clinical course, and poor response to standard chemotherapy. Plant peptides obtained from the traditional Russian herbal medicine have gradually gained considerable attention as a new source of anticancer drugs. We have tested antitumor activity of a peptide extract PE-PM obtained from a mixture of Chelidonium majus L., Inula helenium L., Equisetum arvense L. and Inonotus obliquus in new mouse T-lymphoma/leukemia model ASF-LL. Distinct antitumor activity of two local injections of the peptide extract PE-PM was detected by tumor growth inhibition and survival improvement of 33% of recipients bearing intraperitoneal form of ASF-LL.

Evaluation of antitumor activity of peptide extracts from medicinal plants on the model of transplanted breast cancer in CBRB-Rb(8.17)1Iem mice.

We studied antitumor effects of peptide extracts from plants on slowly growing mammary adenocarcinoma in CBRB-Rb(8.17)1Iem mice used as a model of breast cancer in humans. The antitumor effect of a single injection of the test peptides was evaluated by the delay of the appearance and growth of palpable breast cancer in mice over 4 weeks. Peptides from Hypericum perforatum and a mixture of Chelidonium majus L., Inula helenium L., Equisetum arvense L., and Inonotus obliquus exhibited maximum activity. Peptide extracts from Frangula alnuc Mill. and Laurus nobilis L. were less active. No antitumor effect of Camelia sinesis Kuntze was detected.

Polymer-bound 6' sialyl-N-acetyllactosamine protects mice infected by influenza virus.

To develop a mouse model for testing receptor attachment inhibitors of human influenza viruses, the human clinical virus isolate in MDCK cells A/NIB/23/89M (H1N1) was adapted to mice by serial passaging through mouse lungs. The adaptation enhanced the viral pathogenicity for mice, but preserved the virus receptor binding phenotype, preferential binding to 2-6-linked sialic acid receptors and low affinity for 2-3-linked receptors. Sequencing of the HA gene of the mouse-adapted virus A/NIB/23/89-MA revealed a loss of the glycosylation sites in positions 94 and 163 of HA1 and substitutions 275Asp-->Gly in HA1 and 145Asn-->Asp in HA2. The four mouse strains tested differed significantly in their sensitivity to A/NIB/23/89-MA with the sensitivity increasing in the order of BALB/cJCitMoise, C57BL/6LacSto, CBA/CaLacSto and A/SnJCitMoise strains. Testing of protective efficacy of the polyacrylamide conjugate bearing Neu5Acalpha2-6Galbeta1-4GlcNAc trisaccharide under conditions of lethal or sublethal virus infection demonstrated a strong protective effect of this preparation. In particular, aerosol treatment of mice with the polymeric attachment inhibitor on 24-110 h after infection completely prevented mortality in sensitive animals and lessened disease symptoms in more resistant mouse strains.

Galectins as markers of aggressiveness of mouse mammary carcinoma: towards a lectin target therapy of human breast cancer.

Galectins, beta-galactoside binding proteins, expressed selectively in human breast carcinoma are attractive targets to employ lectin-aimed therapeutics. We examined beta-galactoside binding potency of neoplastic cells using fluorescein-labelled synthetic glycoconjugates as probes for flow cytometry. As a result, surface beta-galactoside binding proteins/galectins were discovered on mouse mammary carcinoma cells in vitro and in vivo unlike non-malignant cells from the several tissues; and asialo-GM1 ganglioside carbohydrate part--containing probe was the most specific one. However, in liver and lung metastatic cells galectins seem to be expressed within cytoplasm and/or nuclei. Galectin expression correlated directly with aggressive tumour potential in the A/Sn transplantable model similar to findings in several human breast carcinoma cell lines. However, galectin expression was reduced during tumour progression in more aggressive forms of spontaneous BLRB mammary carcinomas like it was shown for human breast carcinoma specimens. Analysis of the histopathological data led, however, to the conclusion that galectin expression hardly might be a suitable marker of aggressiveness of heterogeneous mammary carcinomas as the observed level of galectin expression is influenced by the amount of the stroma in a tumour sample and/or probably, galectin expression inversely correlates with tumour aggressiveness during the initial and advanced steps of mammary tumour progression. We conclude that surface beta-galactoside binding proteins/galectins that are selectively expressed during mouse mammary carcinoma progression, similarly to human breast carcinomas, seem to be proper targets for asialo-GM1-vectored cytotoxics and our mouse model system might be a relevant instrument to further test novel modes of anti-breast cancer therapy.

Antitumour activity of cytotoxic liposomes equipped with selectin ligand SiaLe(X), in a mouse mammary adenocarcinoma model.

The overexpression of lectins by malignant cells compared with normal ones can be used for the targeting of drug-loaded liposomes to tumours with the help of specific carbohydrate ligands (vectors). Recently we have shown that liposomes bearing specific lipid-anchored glycoconjugates on a polymeric matrix bind in vitro to human malignant cells more effectively and, being loaded with a lipophilic prodrug of merphalan, reveal higher cytotoxic activity compared with unvectored liposomes. In this study, carbohydrate-equipped cytotoxic liposomes were tested in vivo in a mouse breast cancer model, BLRB-Rb (8.17)1Iem strain with a high incidence of spontaneous mammary adenocarcinoma (SMA). Firstly, a cell line of the SMA was established which was then used to determine the specificity of the tumour cell lectins. After screening of the lectin specificity of a number of fluorescent carbohydrate probes, SiaLe(X) was shown to be the ligand with the most affinity, and a lipophilic vector bearing this saccharide was synthesised. Then different liposomal formulations of the synthetic merphalan lipid derivative and SiaLe(X) vector were prepared and applied in the treatment of mice with grafted adenocarcinomas. The results of the tumorigenesis data show that the therapeutic efficacy of merphalan increases sharply after its insertion as a lipophilic prodrug into the membrane of SiaLe(X)-vectored liposomes.

Immunostimulatory activity of Milife, a novel immunomodulator of fungus origin.

Milife is a novel immunomodulator derived from the fungus Fusarium Sambucium. In this study we examined immunomodulatory properties of Milife in 10 months-old BLRB mice. Milife was given to mice orally in a daily dose of 1 mg per mouse, for 2 to 6 days. Groups of mice were sacrificed on days 2, 4, and 6 of treatment, and 3 weeks after completion of a 6 days treatment with Milife, and lymphoid organs were obtained for analysis. Milife administration led to rapid and significant increase in total leukocyte and lymphocyte numbers in peripheral blood that persisted for at least 3 weeks after a 6 days treatment. Cellularity of lymph nodes, bone marrow and thymus increased significantly at days 4 and 6 of treatment, but returned to pretreatment levels after Milife discontinuation. Though total splenocyte numbers did not change dramatically, there occurred delayed increase in CD4+ cells in the spleen 3 weeks following treatment. Preferential accumulation of CD4+ cells was also consistently found in peripheral blood, with the peak being observed at day 6 of treatment. As a result, CD4/CD8 ratio in blood and spleen was significantly higher in treated than in untreated mice. Splenocytes from treated mice proliferated more vigorously in response to Con A. When added in vitro, Milife also mildly costimulated Con A-induced proliferation of splenocytes from intact animals. In conclusion, we have found that Milife can stimulate leuko- and lymphopoesis in BLRB mice, in particular, accumulation of CD4+ T cells in peripheral lymphoid organs. We conclude that Milife may represent an immunomodulator with the potential to correct T cell dysfunction in patients with immunodeficiency.

The genetic activity of N6-hydroxyadenine and 2-amino-N6-hydroxyadenine in Escherichia coli, Salmonella typhimurium and Saccharomyces cerevisiae.

The genetic activity of 2-amino-N6-hydroxyadenine or 2-amino-N-hydroxylaminopurine (AHA) and N6-hydroxyadenine or 6-N-hydroxylaminopurine (HAP) was studied in S. typhimurium, E. coli and Saccharomyces cerevisiae strains. AHA was a more potent mutagen for bacteria and a less potent mutagen for yeast than HAP. The mutagenic activity of analogs was not influenced by excision, mutagenic or double-strand DNA repair mutations. On the other hand, the uvrBdel mutation has a drastic effect on the mutagenicity and toxicity of both analogs in the Salmonella strains studied. HAP was a very potent mutagen in yeast with a low capability of inducing mitotic recombination contrary to common mutagens, possessed unique intergenic specificity and was able to induce mutations in diploids at rather high frequency.

[The role of the serum interleukin-2 inhibitor in the rejection of allogeneic hematopoietic tissue in sublethally irradiated mice]. / Rol' syvorotochnogo ingibitora interleikina-2 v protsesse ottorzheniiaallogennoi krovetvornoi tkani v organizme subletal'no obluchennykh myshei.

The influence of ionizing radiation (5 Gy) on the interleukin-2 inhibitor in mouse serum has been investigated. It has been shown that the concentration of IL-2 inhibitor decreases on days 3-6 and increases considerably on days 10-15 after irradiation. A correlation has been found between the number of T-helpers in spleens of exposed allogenic chimeras and low IL-2 inhibitor content of serum. An attempt has been made to use the increased IL-2 inhibitor level for improving the acceptance of allogenic cells in the sublethally exposed mice.

[Effect of administration of allogeneic lymphocytes on the survival of sublethally irradiated mice]. / Vliianie vvedeniia allogennykh limfotsitov na vyzhivanie subletal'no obluchennykh myshei.

The administration of allogenic (CBA----C57B1/6) and semi-allogenic (CBA----F1) lymphocytes to sublethally exposed recipient mice either stimulates or inactivates endogenous colony-formation depending on the dose of lymphocytes administered. The stimulation of endogenous colony-formation correlates with the increased survival rate after radiation doses that decrease the survival rate of the control recipients.