Intoxication with some pesticides induce release of cytochrome C and DNA fragmentation in human cell line Huh 7

USMF „N. Testemiţanu” Catedra Biochimie şi Biochimie Clinică; Universitatea din Bucureşti, Facultatea de Biologie, Catedra BiochimieLindane and deltametrine are insecticides with very large utilization in agriculture and public health wich are dangerous contaminants for human organism. We studied the capacity of lindane and deltametrin to induce release of cytochrome C and DNA fragmentation in human cell line Huh 7, as a posibele mecanism of toxiticity. Our results sugest that intoxication with lindane and deltametrin induce biochemical modifications related to apoptosis. Lindanul şi deltametrinul sunt insecticide pe larg utilizate în agricultură şi sănătate publică, periculoase ca contaminanţi pentru organismul uman. Noi am studiat capacitatea acestor insecticide de a provoca eliberarea citocromului c şi fragmentatrea ADN în culturi de hepatocite umane transformate Huh7, ca un posibil mecanism al toxicităţii lor urmat de declansarea prcesului apoptotic. Rezultatele obţinute, sugerează că intoxicaţia cu aceste pesticide induce modificările biochimice caracteristice apoptozei

Characterization of Sucrose Thin Films for Biomedical Applications

Sucrose is a natural osmolyte accumulated in the cells of organisms as they adapt to environmental stress. In vitro sucrose increases protein stability and forces partially unfolded structures to refold. Thin films of sucrose (C12H22O11) were deposited on thin cut glass substrates by the thermal evaporation technique (P∼10−5 torr). Characteristics of thin films were put into evidence by Fourier Transform Infrared Spectroscopy (FTIR), X-ray Photoelectron Spectroscopy (XPS), scanning electron microscopy (SEM), and differential thermal analysis and thermal gravimetric analysis (TG/DTA). The experimental results confirm a uniform deposition of an adherent layer. In this paper we present a part of the characteristics of sucrose thin films deposited on glass in medium vacuum conditions, as a part of a culture medium for osteoblast cells. Osteoblast cells were used to determine proliferation, viability, and cytotoxicity interactions with sucrose powder and sucrose thin films. The osteoblast cells have been provided from the American Type Culture Collection (ATCC) Centre. The outcome of this study demonstrated the effectiveness of sucrose thin films as a possible nontoxic agent for biomedical applications

PP1γ2 and PPP1R11 Are Parts of a Multimeric Complex in Developing Testicular Germ Cells in which their Steady State Levels Are Reciprocally Related

Mice lacking the protein phosphatase 1 gamma isoforms, PP1γ1 and PP1γ2, are male-sterile due to defective germ cell morphogenesis and apoptosis. However, this deficiency causes no obvious abnormality in other tissues. A biochemical approach was employed to learn how expression versus deficiency of PP1γ2, the predominant PP1 isoform in male germ cells, affects spermatogenesis. Methods used in this study include column chromatography, western blot and northern blot analyses, GST pull-down assays, immunoprecipitation, non-denaturing gel electrophoresis, phosphatase enzyme assays, protein sequencing, and immunohistochemistry. We report for the first time that in wild-type testis, PP1γ2 forms an inactive complex with actin, protein phosphatase 1 regulatory subunit 7 (PPP1R7), and protein phosphatase 1 regulatory subunit 11 (PPP1R11), the latter, a potent PP1 inhibitor. Interestingly, PPP1R11 protein, but not its mRNA level, falls significantly in PP1γ-null testis where mature sperm are virtually absent. Conversely, both mature sperm numbers and the PPP1R11 level increase substantially in PP1γ-null testis expressing transgenic PP1γ2. PPP1R11 also appears to be ubiquitinated in PP1γ-null testis. The levels of PP1γ2 and PPP1R11 were increased in phenotypically normal PP1α-null testis. However, in PP1α-null spleen, where PP1γ2 normally is not expressed, PPP1R11 levels remained unchanged. Our data clearly show a direct reciprocal relationship between the levels of the protein phosphatase isoform PP1γ2 and its regulator PPP1R11, and suggest that complex formation between these polypeptides in testis may prevent proteolysis of PPP1R11 and thus, germ cell apoptosis

Analysis of ionizing radiation induced effects in radioresistant tumours and bystander normal cells

International audienceChondrosarcoma is a chemo and radioresistant tumour for which the main treatment remains surgery. Hadron therapy has shown better specificity and lower toxicity for the surrounding normal tissue when compared to conventional photon therapy. This study focuses on analysing cellular mechanisms involved in stress and response to DNA damage induced by ionizing radiation in radioresistant tumour cells and bystander normal cells. Two different chondrosarcoma cell lines (SW1353 and L835) were irradiated with X-ray and low energy protons at doses in the range 0.1-2 Gy. To study the bystander phenomena we used a medium transfer protocol. The bystander supernatant containing signals emitted by irradiated chondrosarcoma cells, was transferred to non-irradiated normal chondrocyte (T/C-28a2) and endothelial (EA.hy926) cells. In order to investigate the oxidative stress induced by irradiation we used flow cytometry to quantify the mitochondrial and cellular reactive oxygen species at 3h and 24h following exposure to X-ray. Similar, for the bystander cells, we investigated the oxidative stress induced by the irradiated cells at 3 and 24h after media transfer. Chondrosarcoma cells present an increase in mitochondrial stress 3h after irradiation in contrast with the bystander cells that present modifications 24h after media transfer. To understand the mechanisms behind the cellular response in bystander cells we used a proteomic assay for the chondrocyte cells receiving medium from the irradiated chondrosarcoma SW1353 cells at the dose of 0.1 Gy. The proteomic assay of the bystander chondrocytes receiving media from either X-ray and proton irradiated cells highlighted a series of modified proteins involved in several cellular mechanisms such as oxidative stress responses, cellular motility, cell death, cancer pathways and exosomes pathways. Our results showed the potential for these proteins to be good biomarker-candidates involved in cellular response in bystander cells

Analysis of ionizing radiation induced effects in radioresistant tumours and bystander normal cells

International audienceChondrosarcoma is a chemo and radioresistant tumour for which the main treatment remains surgery. Hadron therapy has shown better specificity and lower toxicity for the surrounding normal tissue when compared to conventional photon therapy. This study focuses on analysing cellular mechanisms involved in stress and response to DNA damage induced by ionizing radiation in radioresistant tumour cells and bystander normal cells. Two different chondrosarcoma cell lines (SW1353 and L835) were irradiated with X-ray and low energy protons at doses in the range 0.1-2 Gy. To study the bystander phenomena we used a medium transfer protocol. The bystander supernatant containing signals emitted by irradiated chondrosarcoma cells, was transferred to non-irradiated normal chondrocyte (T/C-28a2) and endothelial (EA.hy926) cells. In order to investigate the oxidative stress induced by irradiation we used flow cytometry to quantify the mitochondrial and cellular reactive oxygen species at 3h and 24h following exposure to X-ray. Similar, for the bystander cells, we investigated the oxidative stress induced by the irradiated cells at 3 and 24h after media transfer. Chondrosarcoma cells present an increase in mitochondrial stress 3h after irradiation in contrast with the bystander cells that present modifications 24h after media transfer. To understand the mechanisms behind the cellular response in bystander cells we used a proteomic assay for the chondrocyte cells receiving medium from the irradiated chondrosarcoma SW1353 cells at the dose of 0.1 Gy. The proteomic assay of the bystander chondrocytes receiving media from either X-ray and proton irradiated cells highlighted a series of modified proteins involved in several cellular mechanisms such as oxidative stress responses, cellular motility, cell death, cancer pathways and exosomes pathways. Our results showed the potential for these proteins to be good biomarker-candidates involved in cellular response in bystander cells

International Congress on Animal Husbandry “ New

Original scientific paper, presented at

BASED ON 16S ARN MITOCHONDRIAL GENES**

Abstract: The vertebrate mitochondrial genome has been an important model system for studying molecular evolution, organismal phylogeny, and genome structure. Phylogenetic relatioships were inferred from analysis of 570 base pairs (bp) of mithocondrial DNA (mtDNA), representing a conserved region of 16S rRNA. We sequenced 13 cyprinids species and one putative outgroup (Misgurnus fossilis) from Romania. Based upon nucleotide sequence comparisons of cyprinid mitochondrial 16SRNA genes, we established the phylogenetic relationships between analyzed species. The phylogenetic trees obtained by two different methods (neighbor-joining and maximum parsimony) have the same topology and show that most species examined have supported the traditional division of the Cyprinidae into two subfamilies: Cyprininae and Leuciscinae