[Analysis of indocyanine green plasma disappearance rate in clinical practice]. / Vybor metodiki opredeleniya skorosti plazmennoi eliminatsii indotsianina zelenogo v klinicheskoi praktike.

OBJECTIVE: To justify the optimal method for determining indocyanine green plasma disappearance rate (PDRICG). MATERIAL AND METHODS: We analyzed PDRICG in intensive care units. Indocyanine green was administered intravenously at a dose of 0.25 mg/kg. PDRICG was analyzed simultaneously by using of three methods: 1) PDD (PiCCO2 LiMON device), 2) SBS with analysis of plasma samples on precise spectrophotometer, 3) SBS with analysis of plasma samples on simple experimental photometer. RESULTS: PDD method was used for 346 PDRICG tests in 256 patients. Of these, 14.3% of measurements were erroneous. Paired tests using PDD and SBS methods were performed in 299 cases. SBS method resulted erroneous data in 0.6% of cases. Certain correlation (r=0.79, p<0.001) was found between the reference method (SBS with spectrophotometry) and the PDD method. Bland-Altman plot for these two methods showed that proportional bias of mean difference was caused by extremely high PDRICG of the PDD method (for example, more than 30%/min). Comparison of two SBS variants (spectrophotometer and experimental photometer) revealed good correlation (r=0.91, p<0.001). CONCLUSION: SBS method for measuring PDRICG ensures accurate results under mechanical interferences in patients with impaired capillary blood flow. This eliminates the need for redo measurement. Duplication of the PDD and SBS methods is recommended when repeating the test is not possible (organ donors).

CHARACTERISTICS OF A172 AND T98G CELL LINES.

During continuous cultivation cell lines can lose a number of innate characteristics or acquire new ones. In this work we compared growth and phenotypic characteristics of human glioblastoma À172 and Ò98G lines from cell culture collection of Research Institute of Influenza of Ministry of Health of Russian Federation (St. Petersburg). The activity of genes encoding intracellular proteins that determine cell lines belonging to mesenchymal type, as well as several growth factor genes and extracellular matrix genes were estimated. Cell lines A172 and T98G varied in morphology and surface markers expression. A172 cells were characterized by higher expression of mesenchymal markers CD90, CD105, fibroblast activation protein, and tenascin C. Both cell lines showed high level of a2 smooth muscle actin expression. The obtained data indicating high activity of genes encoding major inductors of angiogenesis (VEGF, FGF2 (b), TGFb1) and thrombospondin-1 in foregoing cell lines are in agreement with published data. Reduction of fetal serum in culture medium from 10 to 5 % in both cell lines resulted in the increase of proportion of cells with surface antigens CD73 and CD105. Both A172 and T98G cell lines sustain the main features of glioblastomas and therefore can serve as research objects in investigation of this kind of neoplasms.

[Oncolytic potential of recombinant influenza A virus vectors on a model of malignant glioma in vivo].

Malignant glioma is the most frequently occurring primary brain tumor. Despite significant progress in the diagnostics and treatment of neoplastic diseases the prognosis for patients with III-IV grade gliomas, remains extremely unfavorable. Rapidly developing area in oncology is the employment of therapeutic viruses with natural or genetically engineered oncolytic activity. In the present study we demonstrated the oncolytic potential of a recombinant influenza A virus vector with impaired interferon antagonism function of NS1 protein in treatment of malignant glioma. Recombinant influenza A virus (HA-DS-GFP) expressing green fluorescent protein from the NS1 open reading frame was used as a model vector. HA-DS-GFP virus has shown infectivity towards glioma cells both in vitro, and in vivo (experimental glioma model in rats). Intratumoral inoculation of HA-DS-GFP resulted in a substantial inhibition or complete regression of tumor growth. Our data demonstrate that recombinant influenza vectors have promising potential in therapy of malignant gliomas.

Characteristics of mesenchymal stromal cells isolated from patients with breast cancer.

Mesenchymal stromal cells were isolated from the adipose tissue obtained during surgery for breast cancer and cultured under conditions of normal or low oxygen concentrations. In patients that had received a course of radiation and polychemotherapy prior to surgery, the proliferative potential of mesenchymal stromal cells was irreversibly disturbed. In patients receiving no therapy prior to surgery, the morphological, growth, phenotypic, and differentiation characteristics of mesenchymal stromal cells did not differ from the corresponding parameters of mesenchymal stromal cells from healthy donors. Culturing under hypoxic conditions increased adipogenic differentiation potencies of mesenchymal stromal cells from donors and patients.

[Influence of mesenchymal stromal cells on B-cell line growth and immunoglobulin synthesis].

A number of publications contain contradictory data about influence of mesenchymal stromal cells (MSC) on B-lymphocyte growth, differentiation and production of immunoglobulins (Ig). The aim of the study was to investigate the influence of MSC derived from adipose tissue of healthy donors and cancer patients on the proliferation and Ig synthesis of lymphoblastoid cell line Namalva and myeloma cell line U266. Co-cultivation of Namalva cells with MSC stimulated their proliferation, decreased the doubling time and the minimal effective seeding dose and therefore made cloning of these lymphoblastoid cells possible. The presence of MSC supported the survival and proliferation of Namalva cells cultivated in growth factor deficient medium. MSC also stimulated proliferation of U266 myeloma cells. Both MSC derived from adipose tissue from the healthy donors and from patients with breast cancer effectively stimulated B-cell lines proliferation. Presence of MSC in mixed cultures had no influence on the production of IgM or IgE by Namalva or U266 cells respectively. Co-cultivation of Namalva or U266 with MSC resulted in the formation of close intercellular contacts between cells of both types.

[Main milestones of development and application of ductal cytometry at the Russian Federal Research Center for Radiology and Surgical Technology, St. Petersburg].

Ductal cytometry provides data on cellular DNA and RNA levels and overall profile of specific proteins identifiable by monoclonal antibodies. Results of its long-term use in clinical and oncological research are presented. Application of dosage ranging 0.28-1.1 mGy/sec was followed by stable 1.8-2-fold increase in the myelokariocyte profile cbering DNA synthesis. Bone marrow proliferation did not increase until relatively low dosage was used. A study of combined effects of prolonged gamma irradiation and lead and cadmium ions on rat's hemopoiesis pointed to radiation as the sole causative factor when cadmium chloride was used. Hemopoietic characteristics came back to normal when a combination of lead acetate and ionizing radiation was used, as a result of the oppositely directed action of the two factors. Standard monoclonal antibodies should not be employed for evaluating immunological vigor of patients with malignant gliomas due to the presence of a specific pathological link in their immune system.

[The combained effects of the long-term gamma-irradiation and heavy metall ions on the haematopoietic system of rats].

The combiened effects of different dose rates (0.625 microGy/s - 1.1 mGy/s) of gamma-irradiation and of cuprum and of cadmium ions on the haematopoietic system of rats were studied. It was found that only low dose rates (0.625-10 microGy/s, summary doses 0.5-2.0 Gy) of gamma-irradiation yields in the increasing proliferative activity of bone marrow. The number of myelocariocytos in S-phase was increased at 1.5-1.8 times. In case of the treatment with both cadmium chloride and radiation the changes in proliferative activity of bone marrow are completely due to the radiation factor. Combination of cuprum acetate and ionizing radiation induce opposite effects providing formal normalization of the haematopoietic characteristic of bone marrow up to 3, 6 and 12 months after the end of the radiation and the chemical exposure of the animal.