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Biomedical subjects

V F Dima

Publications and source records attributed to V F Dima.

At least 37 records · Page 2Linked to original sources

New approach to the adoptive immunotherapy of Walker-256 carcinosarcoma with activated macrophages combined with photodynamic therapy.

Experiments were performed on five batches of Wistar inbred rats with Walker-256 carcinoma receiving sole (PDT, MAK) or combined therapy (PDT + MAK-A; PDT + MAK-B); the control batch (HBSS) consisted of animals with untreated tumors. The results were as follows: a) the sole treatment (PDT, MAK) gave survival rates between 37.7 and 47.5%, b) the "combined" therapy in five doses increased significantly (70.8%) the survival rate of tumor bearing rate as well as the rate of complete regression (82.1%). The cell-mediated immunity test and histopathological as well as the electron microscopy observations were in full agreement with the results above. Summing up, these results demonstrate that "combined" photodynamic therapy with intra- and peritumoral MAK infusion stimulated cell-mediated antitumoral activity, increased survival rates and reduces incidence of Walker-256 carcinoma in rat model.

Animals↗

Development of an irradiated vaccine that protects against enterotoxigenic Escherichia coli diarrhoea.

In the pathogenesis of diarrhoea in man bacteria adhesion to enterocytes is mediated by specific CFA/I or CFA/II antigens. A perorally administered vaccine was prepared from E. coli H10407 (078:H11) by irradiation with electrons with high energy (EHE). Two hours after cimetidine administration rats were immunized per os with 5 irradiated vaccine doses at 4-day intervals. Seven days after the last immunization animals were infected by inoculating 1 x 10(9) germs in the ligated intestinal loop. Reduction of the intestinal secretion by over 50% 18 hours after inoculation was considered an efficient protection marker. The obtained results have proved a significant reduction of the intestinal secretion in immunized animals infected with serotypes 078:H11(63 +/- 4%) and 078:H12(59 +/- 5%) as compared to non-immunized animals. Experimental induction of the intestinal protection against Escherichia coli enterotoxigenic (ETEC) strains points to the possibility of using this type of irradiated vaccine in the prophylaxis of diarrhoea in man.

Animals↗

Photodynamic therapy and some clinical applications in oncology.

Photodynamic therapy (PDT), a new treatment modality for localized cancers involving the selective interaction of visible light with photosensitizers, such as hematoporphyrin derivatives (HpD) or dihematoporphyrin ether/ester (DHE) (Photofrin II). Photodynamic therapy of malignant tumors includes biological, photochemical and photophysical processes. These processes involve: (i) absorption of photosensitizing agent; (ii) selective retention of photosensitizer in tumors and (iii) irradiation of sensitized tumor by laser irradiation. This paper provides a review of photosensitizers, photochemistry, subcellular targets, side effects and lasers involved in photodynamic therapy. In addition, gradual increase in knowledge related to in vivo and in vitro mechanisms of action of PDT, as well as some clinical applications of photodynamic therapy are presented.

Humans↗

In vitro effects of pulsed near-ultraviolet laser exposure on human larynx carcinoma cells.

Effects of pulsed near-ultraviolet laser beam on structural characteristics and macromolecular synthesis of carcinoma HEp2 cells were investigated. Laser irradiation damage induced in these eukaryotic cells could be characterized by two development stages: a) a reversible stage with minor morphological damages (1.5 kJ/m2) and 2) an irreversible one, at higher fluences, characterized by cellular membrane damage, necrobiosis and cells detachment from the substrate (4.5 kJ/m2). A. Studies performed referring to macromolecular syntheses of low laser fluences (1.5 kJ/m2)--irradiated HEp2 cells showed the following aspects: a) syntheses inhibiton phase in the first cycles of cellular replication and b) syntheses stimulation phases in the following cycle with total repair of laser-induced molecular lesions. B. At high laser fluences (3-4.5 kJ/m2), metabolic lesions repair was partially or totally blocked after prolonged culturing at 37 degrees C. Ths paper suggests some mechanisms of laser action on macromolecular synthesis and correlates them with morphological changes induced by laser exposure of carcinoma cells.

Carcinoma↗

DNA synthesis in monkey kidney cellular culture, hematoporphyrin sensitized and He--Ne laser irradiated.

He--Ne laser irradiation for 1-45 J/cm2 has four actions upon monkey kidney cell cultures: a) a stimulation of nuclear DNA synthesis after 20 J/cm2; b) a diminution of replication after higher dose (45 J/cm2); c) inhibition of nuclear DNA synthesis and d) a marked cytotoxic effect. At lower radiation fluences (1-5 J/cm2) the influence on the biosynthesis of DNA is not significant.

Animals↗

Characterization of colonization factor antigen CFA/I from an enterotoxigenic Escherichia coli strain (0128:H12).

CFA/I antigen was isolated and purified from E. coli, mutant 279 B-1-14, serotype 0128:H12, and had the following biochemical and biological features: a) amino-acid content was similar to that of purified antigen prepared from strain H10407; b) latex particles sensitization with purified CFA/I antigen produced bovine and human erythrocytes group A/II hemagglutination in carbohydrates presence; c) purified anti-CFA/I specific antibodies agglutinated CFA/I-positive enterotoxigenic E. coli strains; d) 3H-leucine-labelled CFA/I antigen adhered to rabbits intestinal mucosa at significant values; e) intestinal mucosa pretreating with purified CFA/I antigen, followed by 3H-leucine labelled enterotoxigenic bacteria infection, had a least 3 local effects: 1) intestinal mucosa protection against parental enterotoxigenic bacteria; 2) inhibition of CFA/I-positive bacteria adherence to intestinal mucosa; 3) release of approximately 96% intraluminally inoculated bacteria.

Amino Acids↗

Studies of the effects of associated photodynamic therapy and drugs on macromolecular synthesis of tumoral cells grown in vitro.

HeLa S3 tumoral cells were used as an experimental model for studying the association of photodynamic therapy (PDT) and antitumoral agents. Tumoral monolayer cultures were incubated 18 hours at 37 degrees C with Photofrin II, trypsinized and suspended in Eagle medium supplemented with 10% FCS and then treated with antitumoral agents 90 minutes before He-Ne laser exposure. The tumoral cells were exposed to antitumoral agents in the following concentrations (equivalent to ED70): adriamycin (0.0297 micrograms); mitomycin C (0.0199 micrograms); 5-FU (0.4937 micrograms) and vinblastine (0.0109 micrograms) per 10(5) cells. Macromolecular syntheses (DNA, RNA and proteins) were investigated by use of radioactive precursors: 3H-thymidine, 3H-uridine and 3H-leucine, as expressed in percent referring to Photofrin II-pretreated controls; they were exposed to He-Ne laser but not treated with antitumoral agents. All experiments were followed for 72 hours incubation at 37 degrees C. The conclusions of the results of PDT associated with antitumoral agents sustain the following aspects: a) the antitumoral agents activity (adriamycin, mitomycin C, 5-FU, vinblastine) was more noticeable when applied 90 minutes before He-Ne laser irradiation; b) inhibition of radioactive precursors uptake in DNA, RNA and proteins was accompanied by suppression of in vitro tumoral cells development and c) PDT association with antitumoral agents could manifest at least three positive effects upon animals; 1) PDT potentiating effects with antitumoral agents; 2) suppressing effects on tumoral macromolecular synthesis; 3) antitumoral agents cytotoxic elimination (due to the low doses used).

Antineoplastic Combined Chemotherapy Protocols↗

Laser effects on cellular adhesion and influence on the interrelation between HeLa S3 and human diploid cells.

The interactions between HeLa S3 tumoral cells and human fibroblasts after nitrogen-laser irradiation (337.1 nm) have been studied by using an in vitro cell invasion model. For the quantitative and morphological evaluation of nitrogen-laser radiation action upon tumoral adhesion to the fibroblast monostrate, we used: a) 3H-thymidine labelling of HeLa S3 tumoral cells; b) morphological modifications studies by phase contrast and scanning electron microscopy. The results emphasized the following aspects: 1. In non-irradiated cell cultures we noticed three interaction stages: adhesion, tumoral spreading and displacement with fibroblasts destruction; on the other side, we found a reduced adhesion to non-irradiated human fibroblasts of laser irradiated tumoral cells. 2. Significant percent increasing of non-irradiated tumoral cells adhesion to fibroblast monostrate, irradiated with various laser fluences (e.g. 0.2 kJ/m2--48.1%; 0.8 kJ/m2--63.8% and for 1.6 kJ/m2--79.5%). This phenomenon evidenced the close interrelation between irradiation fluences and tumoral adhesion rates. 3. The importance of numerical ratio between tumoral cells and fibroblasts in tumoral adhesion and invasion processes (e.g. ratio 1:10 tumoral adhesion reached 8.1%; in 1:5--25.9%; in 1:1--59.4% and for 2:1--83.9%). 4. Marked cytotoxic effects for both cell types after exposure to high and very high laser fluences (1.6--6.4 kJ/m2). Our results emphasize near UV-laser irradiation effects upon some of tumoral adhesion and invasion mechanisms and demonstrate the interrelations between cell populations manifesting a different vital potential.

Cell Adhesion↗

Reaction and response of newborn guinea pigs to experimental Salmonella typhi infection.

Newborn guinea pigs, orally infected with Salmonella typhi were examined at various intervals of time in order to determine bacterial distribution in tissues and to establish possible correlation with the clinical aspects manifested. Histopathological examination evidenced typical lesions in jejunum, ileum, caecum and especially in regional lymphatic tissues. Spleen, liver and mesenteric lymph nodes presented granulomatous lesions similar to those observed in in human typhoid fever. After oral administration, the animals reacted with anorexia, febrile reactions, bacteremia, diarrhoea, positive stool cultures, dehydration, lethargy and antibodies too were produced. Our results indicate that typhoid infection may be induced in newborn guinea pigs; the model may be used for an assessment of attenuated live typhoid vaccine control.

Animals↗

Activation of guinea pig peritoneal macrophages by ribosomal extract of Salmonella typhi strain. Electron microscopy studies.

Normal macrophages and Salmonella typhi ribosomal antigen-activated (immune) ones were studied by electron microscopy. Examination of fine sections from samples incubated between 30 and 120 minutes at 37 degrees C, emphasized phagocytic ability of activated macrophages (29-69% activated macrophages phagocytized 3.7-5.6 bacteria/cell, in contrast to 23-43% with 2.6-4.4 bacteria/phagocytized by non-immune cells). The ultrastructural studies of activated macrophages, performed by electron microscopy, showed the following aspects: a) macrophages with pseudopodiform prolongations, presenting the tendency of bacterial sequestration: b) phagosomes with ingested virulent germs; c). bacteria presenting various degrees of wall and cytoplasm alterations; d). mitochondria with multiple cristae; e). cells with well developed Golgi apparatus and the presence of lysosomes in great numbers; f). existence of a space between the phagosome membrane and the bacterial wall. Experimental results demonstrated: I. an increased phagocytic activity of Salmonella typhi ribosomal antigen-activated macrophages and II. activation expressed as ultrastructural modifications at the level of immune macrophages and also of the bacteria phagocytized by them.

Animals↗

Immunoprotection of guinea pigs against experimental acute pyelonephritis after the administration of Escherichia coli purified fimbriae.

Experimental results of the present study evidenced the following aspects: a) the antigen, prepared from type I Escherichia coli purified fimbriae (H. 2946 strain), induced immunity at urinary tract level; b) the immunoprotection induced by oral vaccination with multiple doses of fimbriated antigen produced a significant decrease of acute pyelonephritis in newborn guinea pigs and at the same time, a local protection of the urinary tract; c) the immunoprophylaxis by vaccine prepared from fimbriae represents a preferential solution for urinary tract infections prevention in general and especially in children; d) the frequency distribution differences between "protected" and "non-protected" animals were evaluated by chi-square--test with YATES correction and proved to be statistically significant at probability levels.

Acute Disease↗