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      • Apoptosis Induction in Human Leukemic Promyelocytic HL-60 and Monocytic U937 Cell Lines by Goniothalamin

        Petsophonsakul, Ploingarm,Pompimon, Wilart,Banjerdpongchai, Ratana Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.5

        Goniothalamin is an active compound extracted from Goniothalamus griffithii, a local plant found in northern Thailand. Goniothalamin inhibits cancer cell growth but is also toxic to normal cells. The aims of this study were to identify the cytotoxic effect of goniothalamin and the mechanism of cell death in human HL-60 and U937 cells. Cytotoxicity was determined by MTT assay and cell cycle profiles were demonstrated by staining with propidium iodide (PI) and flow cytometry. Apoptosis was confirmed by staining with annexin V-FITC/propidium iodide (PI) and flow cytometry. Reduction of mitochondrial transmembrane potential was determined by staining with dihexyloxacarbocyanine iodide and flow cytometry and expression of Smac, caspase-8 and -9 was demonstrated by Western blotting. Goniothalamin inhibited growth of HL-60 and U937 cell lines. An increase of SubG1 phase was found in their cell cycle profiles, indicating apoptosis as the mode of cell death. Apoptosis was confirmed by the flip-flop of phosphatidylserine using annexin V-FITC/PI assay in HL60 and U937 cells in a dose response manner. Furthermore, reduction of mitochondrial transmembrane potential was found in both cell types while expression of caspase-8, -9 and Smac/Diablo was increased in HL-60 cells. Taken together, our results indicate that goniothalamin-treated human leukemic cells undergo apoptosis via intrinsic and extrinsic pathways.

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        Production of polyclonal antibody against kidney antigens; a model for studying autoantibody in feline chronic kidney diseases

        Nisakorn Songaksorn,Wilaiwan Petsophonsakul,Kidsadagon Pringproa,Kannika Na Lampang,Nattawooti Sthitmatee,Nuttawan Sriphawattana,Kriangkrai Thongkorn 대한수의학회 2019 Journal of Veterinary Science Vol.20 No.6

        Chronic kidney disease is considered to be most common in geriatric domestic cats. It has been reported that the feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine prepared from the Crandell-Rees feline kidney (CRFK) cell line can induce cross-reactions of antibodies with feline kidney tissues. As an anti-cat kidney antibody was not available commercially for this study of autoantibody in cats, the purpose of this study was to produce anti-cat kidney antibody in rabbits for further study of autoantibody in cats after FVRCP vaccination. Kidney proteins from cadaveric cats were extracted and immunized into rabbits using Montanide as the adjuvant. All immunized rabbits produced high levels of anti-cat kidney antibodies, and, based on ELISAenzyme-linked immunosorbent assay testing, some began to produce antibodies as early as 2 weeks after immunization. Immunofluorescence staining of rabbit sera showed kidney-bound antibodies in glomerulus, Bowman’'s capsule, apical surface of the proximal convoluted tubule, peritubular surface, and interstitial cells. Western blot analysis of cat kidney proteins revealed molecular weights (M.W.) of 72, 55, 47, and 31 kDa, while binding to the CRFK cell proteins was observed at M.W. of 43 and 26 kDa. The antibody that recognized the 47 kDa protein was similarly detected in cats with autoantibody presence after FVRCP vaccination. The kidney-bound antibody profile at different time points and its patterns in rabbits could be used as a model for the study of autoantibody to cat kidney in feline chronic kidney diseases.

      • KCI등재

        Prevalence of autoantibodies that bind to kidney tissues in cats and association risk with antibodies to feline viral rhinotracheitis, calicivirus, and panleukopenia

        Nisakorn Songaksorn,Wilaiwan Petsophonsakul,Kidsadagon Pringproa,Kannika Na Lampang,Nattawooti Sthitmatee,Nuttawan Srifawattana,Kakanang Piyarungsri,Kriangkrai Thongkorn 대한수의학회 2021 Journal of Veterinary Science Vol.22 No.3

        Background: The feline viral rhinotracheitis, calicivirus, and panleukopenia (FVRCP) vaccine, prepared from viruses grown in the Crandell-Rees feline kidney cell line, can induce antibodies to cross-react with feline kidney tissues. Objectives: This study surveyed the prevalence of autoantibodies to feline kidney tissues and their association with the frequency of FVRCP vaccination. Methods: Serum samples and kidneys were collected from 156 live and 26 cadaveric cats. Antibodies that bind to kidney tissues and antibodies to the FVRCP antigen were determined by enzyme-linked immunosorbent assay (ELISA), and kidney-bound antibody patterns were investigated by examining immunofluorescence. Proteins recognized by antibodies were identified by Western blot analysis. Results: The prevalences of autoantibodies that bind to kidney tissues in cats were 41% and 13% by ELISA and immunofluorescence, respectively. Kidney-bound antibodies were observed at interstitial cells, apical border, and cytoplasm of proximal and distal tubules; the antibodies were bound to proteins with molecular weights of 40, 47, 38, and 20 kDa. There was no direct link between vaccination and anti-kidney antibodies, but positive antibodies to kidney tissues were significantly associated with the anti-FVRCP antibody. The odds ratio or association in finding the autoantibody in cats with the antibody to FVRCP was 2.8 times higher than that in cats without the antibody to FVRCP. Conclusions: These preliminary results demonstrate an association between anti-FVRCP and anti-cat kidney tissues. However, an increase in the risk of inducing kidney-bound antibodies by repeat vaccinations could not be shown directly. It will be interesting to expand the sample size and follow-up on whether these autoantibodies can lead to kidney function impairment.

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