X-Message-Number: 32427 Date: Thu, 25 Feb 2010 19:23:02 -0800 (PST) From: Subject: genistein suppresses cryopreservation-induced damage [Genistein is an effective cryopreservation solution additive. However, based on cost, it is hard to beat filtered green tea.] Hum Reprod. 2009 Sep;24(9):2061-70. Epub 2009 Jun 12. Cryopreservation-induced human sperm DNA damage is predominantly mediated by oxidative stress rather than apoptosis. Thomson LK, Fleming SD, Aitken RJ, De Iuliis GN, Zieschang JA, Clark AM. Fertility First, PO Box 807, Hurstville, NSW 2220, Australia. BACKGROUND: Whereas studies have revealed that the cryopreservation of human semen increases sperm DNA fragmentation, the mechanisms involved in this type of cryo-injury are largely unknown. Elucidation of these mechanisms may provide insight into preventing such injury. METHODS: We obtained 60 semen samples from 60 men and conducted experiments to determine the cause of cryopreservation-induced DNA fragmentation using 8-oxo-7,8-dihydro-2'deoxyguanosine (8OHdG) as a biomarker of oxidative stress, percentage caspase positive cells as an indicator of apoptosis, the potential antioxidant genistein and the caspase inhibitor Z-VAD(OMe)-FMK. RESULTS: Cryopreservation led to a significant increase in percentage DNA fragmentation, percentage 8OHdG and percentage caspase positive cells (P < 0.001). Percentage DNA fragmentation was positively correlated with percentage 8OHdG before (r = 0.756, P < 0.001) and after cryopreservation (r = 0.528, P = 0.017). The addition of 50 and 100 microM genistein to the cryoprotectant had a significant protective effect on sperm DNA (P < 0.001) although the caspase inhibitor demonstrated no difference to the control. CONCLUSIONS: Human sperm DNA fragmentation is associated with an increase in oxidative stress during cryopreservation, rather than the activation of caspases and apoptosis. The estrogenic compound genistein may be useful in reducing this effect but larger trials are needed to confirm this. PMID: 19525298 J Med Food. 2009 Apr;12(2):429-34. Cell viability of normal human skin fibroblast and fibroblasts derived from granulation tissue: effects of nutraceuticals. Borawska MH, Czechowska SK, Markiewicz R, Hayirli A, Olszewska E, Sahin K. Department of Bromatology, Medical University of Bialystok, Bialystok, Poland. The effects of lycopene, genistein, and epigallocatechin-3-gallate (EGCG) on cell viability were tested in vitro using a normal human skin fibroblast (NHSF) cell line (CRL-1474) and granulation tissue fibroblasts (GTFs) obtained from a patient with middle ear cholesteatoma. Cell cultures were added with lycopene (1, 5, and 10 microM), genistein (1, 5, 10, 25, and 50 microM), and EGCG (1, 5, 10, 25, and 50 microM) and their respective control cultures were established by adding 5 mL/L tetrahydrofuran (THF), 5 mL/L dimethyl sulfoxide (DMSO), and 5 mL/L DMSO. A colorimetric assay was employed for determining cell viability using thiazolyl blue tetrazolium bromide. Cell viability was expressed as a percentage of the control. Data were analyzed using two-way analysis of variance separately for each compound. Lycopene addition decreased viability of NHSFs and GTFs compared with THF addition (64.1%, 60.5%, and 100%, respectively, P < .0001). Genistein addition also increased viability of both NHSFs and GTFs compared with DMSO addition (P < .02). Increasing EGCG concentration tended to cause a linear increase in viability of NHSFs but did not alter viability of GTFs (P < .10). Our data suggest that genistein and EGCG but not lycopene could help maintaining or improving skin health through enhancing viability of skin fibroblasts. PMID: 19459748 Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=32427