Monkey Renal Cells Research Articles

Tetroxanes as New Agents against Leishmania amazonensis.

Leishmaniasis is a neglected disease, caused by a parasite of Leishmania genus and widespread in the tropical and subtropical areas of the world. Currents drugs are limited due to their toxicity and parasite resistance. Therefore, the discovery of new treatment, more effective and less toxic, is urgent. In this study, we report the synthesis of six gem-dihydroperoxides (2a-2f), with yields ranging from 10 % to 90 %, utilizing a new methodology. The dihydroperoxides were converted into ten tetroxanes (3a-3j), among which six (3b, 3c, 3d, 3g, 3h and 3j) showed activity against intracellular amastigotes of Leishmania amazonensis. The cytotoxicity of all compounds was also evaluated against canine macrophages (DH82), human hepatoma (HepG2) and monkey renal cells (BGM). Most compounds were more active and less toxic than potassium antimonyl tartrate trihydrate, used as positive control. Amongst all tetroxanes, 3b (IC50 =0.64 μm) was the most active, being more selective than positive control in relation to DH82, HepG2 and BGM cells. In summary, the results revealed a hit compound for the development of new drugs to treat leishmaniasis.

Evaluation of acute oral toxicity, embryotoxicity and cytotoxicity of the polar fraction of Parkinsonia aculeata aerial parts extract.

Ethnopharmacobotanical information reports that Parkinsonia aculeata infusion is used to control diabetes-related complications and dyslipidemia. However, few studies are reported on the safe use of this species. The aim of this study is to evaluate the acute toxicity, embryotoxicity and cytotoxicity of a polar fraction obtained from hydroethanolic extract of P. aculeata (PfrHEPA). For the acute toxicity test, we considered the Up and Down method which the guidelines are described by the Organization for Economic Cooperation and Development (OECD N°425). The animals were treated with PfrHEPA (2000mg/kg) or with distilled water (10ml/kg) by gavage and observed from Day 1 to14. For embryotoxicity assay, zebrafish embryos were exposed to PfrHEPA (100mg/L) and toxicity parameters were observed during four consecutive days. The cytotoxicity of PfrHEPA (5, 10, 25, 50, 75 and 100μg/ml, respectively) was performed on normal cell lines (mesenchymal stem cells, African green monkey renal cells and mouse pre-adipocytes 3T3-L1 using the MTT salt reduction assay. In the acute toxicity test, no mortality was observed in mice treated with PfrHEPA (2000mg/kg), as well as behavioral changes, histopathological abnormalities and hematological and biochemical variables. In the embryotoxicity test, no abnormal changes related to the toxicological parameters were observed in the period of 96h. Regarding the cytotoxicity assay, PfrHEPA showed no cytotoxic effect on the normal cell lines tested, with an IC50 value>100μg/ml. These results suggest the safe use of P. aculeata, however, more trials are needed for PfrHEPA to be presented as new safe therapeutic proposal for the control of metabolic disorders.

The Antitumoral Activity and the Cytotoxicity on Renal Cells of Ethanolic Extracts from the Leaves of Four Varieties of Olea europaea L. Grown in Morocco

The antitumoral activity and the cytotoxicity on renal cells of ethanolic extracts prepared from the leaves of four varieties of Olea europaea L. grown in Morocco such as Picholine marocaine, Menara, Picholine longuedoc and Leuccino was studied and compared. The antitumoral activity on Human Rhabdmyosarcom cells (RD) (line CCL-136) of all samples was stronger than their cytotoxicity toward African Monkey renal cells (Vero) (line CCL-81). The P. marocaine extract showed the best therapeutic index (32.6) and a higher antitumoral (IC50 = 5.5 μg/mL) and cytotoxicity (IC50 = 75.6 μg/mL) activity than those of the other varieties. In addition, this extract is characterize by a higher content of active compounds (oleanolic acid, maslinic acid, ursolic acid, tyrosol, vanillin, caffeic acid) comparing to the other ones. The P. marocaine leaves extract is a promising agent combating tumors.

Hypoxic stress-enhanced expression and release of adrenomedullin (AM) and up-regulated AM receptors, while glucose starvation reduced AM expression and release and down-regulated AM receptors in monkey renal cells

The proliferative peptide adrenomedullin (AM) has a wide distribution in a variety of tissues and cells. The mechanism how the AM gene is regulated in cells is not yet known. The renal cortex, renal vascular smooth muscles, glomeruli and tubular epithelial cells are very sensitive to hypoxia. Renal hypoxia produces acute renal tubular necrosis and markedly induces AM expression in damaged cells. However, little information is available regarding the possible pathophysiological production and release of renal tubular AM. Regulation of membrane-bound AM receptors in renal cells has not yet been systematically studied. To elucidate the potential pathological role of human AM we examined the production and release of AM, as well as the characteristics of surface membrane AM receptors in cultured monkey renal tubular epithelial cells (RC) exposed to hypoxia, induced with endothelin-1, and subjected to glucose deprivation. Exposure of RC to hypoxia (1 % O(2), 5 % CO(2) in N(2)), and to phorbol 12-myristate 13-acetate (PMA) increased production and secretion of AM and increased specific [(125)I]AM binding on RC. Metabolic stress (1 % glucose in the cultivation medium) and preincubation of RC with rival peptide endothelin-1 significantly reduced immunoreactive-AM in a conditioned medium and whole cell surface membrane AM binding on RC. Altogether, our data suggest that the AM is involved in the adaptation of renal tubular cells to hypoxia. Increased expression of AM mRNA and regulation of AM receptors in metabolic stress may function as an important autocrine/paracrine regulator(s) of renal tubular epithelial cells.

Development of IgHV family specific nucleic acid vaccine against lymphoma by construction of fusion gene of immunoglobulin heavy chain variable region and cytokine

To construct the gene co-expression vector of immunoglobulin heavy chain variable region (IgHV) gene and GM-CSF or IL-2 as IgHV family specific nucleic acid vaccine to lymphoma, and study the immune response of the immunized mice against lymphoma. Six clones with significantly different length in gene fragments were selected from a gene bank constructed from normal fetal umbilical cord blood. These gene fragments in the clones were sequenced. The sequences were translated into IgHV peptides. T cell epitopes in the IgHV were predicted by bioinformatics. Meanwhile 6 clones of IgHV1 constructed before were analyzed. The most typical clone of IgHV1 and IgHV3 containing most of the T cell epitopes were selected. The IgHV gene fragments whose complementary determining region 3 (CDR3) was cut out and composed mainly of framework region were cloned into eukaryotic expression vector. The gene fragments of framework region of IgHV linking with gene of GM-CSF or IL-2 were cloned into pcDNA3.0 to form fusion genes of IgHV (FR)-GM-CSF/IL-2. Then they were transected into COS cells, African green monkey renal cells, by Lipofectin and their transient expression product was detected by ELISA. Twenty male Balb/c mice were randomly divided into 5 groups of 4 mice to be injected with different vectors at the weeks 0, 2, and 4: with IgHV3 (FR)/pcDNA3.0, with IgHV3 (FR)-IL-2/pcDNA 3.0, with blank vector pcDNA3.0 as control, with IgHV1 (FR)/pcDNA3.0, and with IgHV1 (FR)-IL-2/pcDNA3.0. At the weeks 0, 2, 4, 6, and 8 serum was collected from the mice to undergo indirect immunofluorescence examination to detect the antibodies against Namalwa cells, lymphoma cells from Africa green monkey, and normal lymphocytes. ELISA was used to examine the serum interferon (IFN)-gamma. About 30 of T cell epitopes existed in each IgHV peptides predicted by bioinformatics. Among the bioinformatics prediction, about 90% of the T cell epitopes were in the framework region (FR) of IgHV, and the first 10% with higher prediction score were in FR. The CDR3 of two typical IgHV sequences were cut down and the remaining sequences, mainly composed of FR, were used to construct the IgHV (FR)/pcDNA3.0 expression vectors. The 3' end of IgHV1 (FR) and IgHV3 (FR) were linked to GM-CSF or IL-2 gene successfully. The expression of GM-CSF in the group transfected with IgHV (FR)-GM-CSF/pcDNA3.0 were 200 times higher than in the group transfected with control pcDNA3.0. The expression of IL-2 in the group transfected with IgHV (FR)-IL-2/pcDNA3.0 were 60 times higher than in the group transfected with control pcDNA3.0. The antibody against IgHV1 family lymphoma cell line Namalwa could be detected since the second week in the mice immunized with IgHV1 (FR)-IL-2/pcDNA3.0. The antibody could be detected since the fourth week in the mice immunized with IgHV1 (FR). The antibody against lymphocyte line of IgHV3 family could be detected since the second week in the mice immunized with IgHV3 (FR)-IL-2/pcDNA3.0. The antibody could be detected since the fourth week in the mice immunized with IgHV3 (FR)/pcDNA3.0. The contents of serum IFN-gamma the mice immunized with IgHV1 (FR)-IL-2/pcDNA3.0 and with IgHV3 (FR)-IL-2/pcDNA3.0 was significantly higher than those of the mice immunized with IgHV (FR)/pcDNA3.0 or pcDNA3.0 (all P < 0.01). The gene fragments of IgHV (FR) can be used to construct IgHV family specific nucleic acid vaccine that induces anti-lymphoma immune response in mice by muscle injection. The expressing vectors of IgHV (FR)-GM-CSF/IL-2 induces stronger immunoreaction.

Mammalian Cells in Space Weightlessness

Human clearly perceive and respond to microgravity as an individual unit and manifest such abnormalities as space sickness, and decrease in Calcium in the bones. It is not fully understand, however, whether each of the cells perceives and responds to microgravity. This is a problem of a major scope in gravitational and space biology. I discussed here methods of studying ceIIular response to microgravity cellular response in the space, and others. The cell culture can be applied to elucidate the mechanism of gravity response in cells. FMPT was conducted in the space shuttle Endeavor launched on September 12, 1992, and I and my colleagues had a study concerned the effects of microgravity on the ultrastructure and function of mammalian cells in culture. The study was conducted with the following three major objectives: The first objective was to study the change under microgravity in the cytoskeletons using Monkey's renal cells JTC-12. The second objective was to study energy consumption by cultured cells under microgravity and production of urokinase of the ceIIs. The third objective was to establish the basic techniques of cell culture in the space. Dr. Mouri, payload scientist, conducted the experiments during the flight as scheduled. Cells cultured in culture chambers ACCl and ACC2 were used for the objective 1 and phase contrast microscopic photographs taken in the space laboratory. After trypsinization, the cells were cultured again and fixed with glutaraldehyde after a prescribed period of time. Culture chambers ACC2 and ACC4 were for the objective 2 and 3 and the used cells were brought back to the earth alive. Analysis of photos taken by Dr. Mouri is in progress. Cultured cells were in good conditions under microgravity, and satisfactory results for establishing cell culture techniques, were obtained. Cultured human and animal cells have been used in more than 20 space experiments since Discoverer 17 Iaunched in 1960 and the cellular response to microgravity may differ depending on the cell types. It is possible to conduct experiments on earth using short term microgravity by a drop tower or simulated microgravity using clinostat. Hypergravity using a centrifuge is also useful for studying the cell response to gravity. Since opportunities for space laboratory experiments are Iimited, these equipments are believed to be most useful for sophistication of space experiments.

Susceptibility to viruses of monkey renal cells before and after freezing: Chagnon, A., Cofture, B., and U. Pavilanis, Ann. Inst. Pasteur, 143: 455, 1967

Susceptibility to viruses of monkey renal cells before and after freezing: Chagnon, A., Cofture, B., and U. Pavilanis, Ann. Inst. Pasteur, 143: 455, 1967

A NEUROTROPIC VARIANT OF MEASLES VIRUS IN SUCKLING MICE.

Strain Sugiyama, adapted to FL cells after 6 passages in primary monkey renal cells, acquired the ability to propagate serially in the brain of suckling mice somewhere between the 49th and 76th FL passage. The virus produced a lethal spastic paralysis in about half of mice at the early passages and in almost 100% from the 9th passage on. Mice were nearly 100% susceptible within 5 days after birth and almost 100% resistant at the age of 7 days or over. Extraneural routes of inoculation were ineffective for infection. Histologic changes were found only in the cerebrum, not in the cerebellum or spinal cord, nor in other viscera organs, and consisted of degeneration of nerve cells with or without formation of inclusions characteristic of measles, multinuclear giant cell formation by fusion of nerve cells, and proliferation of glia cells. Fluorescent antibody study revealed remarkable specificity of nerve cell involvement. Specific measles antigens were demonstrated predominantly in nerve cells, only rarely in glia cells, but not in other types of cell.