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Lymphocytes are routinely used in human biomonitoring to assess the potential toxic and cytoprotective effects of diet on both DNA damage and repair and, by implication, health. Logistically, samples may require to be cryopreserved and stored. How this affects cells used in human biomonitoring is often not considered. In this study we have evaluated the influence of cryopreservation on endogenous and induced DNA strand breakage, altered bases (oxidized purines, oxidized pyrimidines and misincorporated uracil), antioxidant capacity and DNA repair capability in human peripheral blood lymphocytes. Neither isolation nor freezing increased DNA strand breakage above endogenous levels found in freshly isolated human lymphocytes. Oxidized bases (both pyrimidines and purines) and misincorporated uracil, were similar for fresh and frozen lymphocytes. Fresh and frozen lymphocytes responded almost identically to hydrogen peroxide. Quercetin-mediated cytoprotection against hydrogen peroxide-induced strand breakage was maintained in cryopreserved lymphocytes after short-term (24 h) and longer term (2 months) storage compared with freshly isolated and treated cells. Hydrogen peroxide-induced DNA strand breakage was repaired in fresh lymphocytes. Cryopreserved lymphocytes were unable to repair oxidant-induced DNA strand breaks. Frozen human lymphocytes can therefore be successfully used for most aspects of DNA damage biomonitoring, but not for repair. [ABSTRACT FROM PUBLISHER]

DNA damage in lymphocytes, as measured by alkaline single cell gel electrophoresis (pH 12.7), is greatly increased by the concurrent lysis of whole blood in both freshly isolated samples and in PHA-stimulated cultures over a period of 7 days. Further, there is a marked progressive increase in DNA damage with time in PHA-stimulated lymphocytes cultured in whole blood even when the lymphocytes are separated before analysis; no such increase is seen in lymphocytes cultured alone. This indicates that there are components in whole blood that can cause DNA damage in lymphocytes, with granulocytes and lysis of red blood cells likely candidates. The DNA damage is greatly reduced in granulocyte-depleted whole blood cultures, but even in these significant increases are seen at later sampling times. Consequently, careful sample preparation is of paramount importance if the Comet assay is to be successfully used to assess DNA damage in human peripheral blood lymphocytes. Further, the progressive increase in DNA damage in whole blood cultures may influence other methods using lymphocytes for population biomonitoring and may be significant for in vitro genotoxicity testing. [ABSTRACT FROM PUBLISHER]

Presents information on a study which determined the absorption and DNA protective effects of flavonoid glycosides from an onion meal. Methodology of the study; Results and discussion on the study.

Provides information on a study which examined the effects of folate deficiency on DNA stability and DNA methylation in rat colonocytes in vivo. Materials and methods used in the study; Results and discussion.

Poor folate status may be important in the aetiology of several epithelial cell malignancies including cancer of the uterine cervix. Folic acid is essential in the synthesis of purine nucleotides and the pyrimidine nucleoside thymidine and it is probable that imbalances in these DNA precursors negatively effect DNA stability and may ultimately lead to malignant transformation. The development of a modified 'comet assay' using the bacterial DNA repair enzyme uracil DNA glycosylase, to detect misincorporated uracil in human DNA is reported here. The effect of perturbing folic acid and deoxyuridine levels on uracil misincorporation in normal human lymphocytes and cultured human tumour cells was investigated using this assay. HeLa cells and peripheral human lymphocytes incubated as agarose-embedded nucleoids, with 1 unit of uracil DNA glycosylase per microg of DNA, contained low levels of uracil in their DNA. Both HeLa cells and stimulated human lymphocytes cultured in folate-deficient medium were growth arrested. Incubating human lymphocytes in folate-deficient medium significantly increased the level of uracil detected compared with control cells. HeLa cells showed an increase in non-specific DNA damage (strand breaks). Deoxyuridine (100 microM) significantly increased the level of uracil detected in the DNA of both folate-deficient and control HeLa cells. It appears that this modified comet assay specifically detects misincorporated uracil in single human cells. It should, therefore, prove valuable in determining the role of folic acid status in DNA instability and cancer. [ABSTRACT FROM PUBLISHER]

We set out to study the value of cardiotocography (CTG) performed before induction of labour for prolonged pregnancy in predicting obstetric intervention. This was a prospective observational study, based in a district general hospital. We studied 100 consecutive patients who underwent induction of labour for prolonged pregnancy. Cardiotocographs (CTGs) were performed before induction of labour on the tenth day after the estimated date of confinement (290 days). The CTGs were then reviewed without knowledge of the outcome of the induction of labour. Obstetric outcomes were then compared. The main outcome measures were the intrapartum presence or absence of meconium stained liquor and the necessity for obstetric intervention. Ninety per cent of CTGs were normal. There was no difference found between the two groups for operative delivery or the presence of meconium liquor. Caesarean section was more likely in the group with an abnormal CTG before induction of labour, but the possibility of this being due to chance is high in this study. There was one case of undiagnosed growth restriction in the abnormal CTG group. These results may be due to a true lack of difference in obstetric intervention between women with normal or abnormal CTGs prior to induction of labour or more probably an inability to detect a difference in our small study. These baseline data allow the calculations necessary for a more substantive trial. [ABSTRACT FROM AUTHOR]