DNA damage in homocystinuria: 8-oxo-, 8-dihydro-2'-deoxyguanosine levels in cystathionine-ß-synthase deficient patients and the in vitro protective effect of N-acetyl­L­cysteine

Introduction: Homocysteine (Hcy) tissue accumulation occurs in a metabolic disease characterized biochemically by cystathionine ß-synthase (CBS) deficiency and clinically by mental retardation, vascular problems, and skeletal abnormalities. Previous studies indicate the occurrence of DNA damage secondary to hyperhomocysteinemia and it was observed that DNA damage occurs in leukocytes from CBS-deficient patients. This study aimed to investigate whether an oxidative mechanism could be involved in DNA damage previously found and investigated the in vitro effect of N-acety-L-cysteine (NAC) on DNA damage caused by high Hcy levels. Methods: We evaluated a biomarker of oxidative DNA damage in the urine of CBS­deficient patients, as well as the in vitro effect of NAC on DNA damage caused by high levels of Hcy. Moreover, a biomarker of lipid oxidative damage was also measured in urine of CBS deficient patients. Results: There was an increase in parameters of DNA (8-oxo-7,8-dihydro-2'- deoxyguanosine) and lipid (15-F2t-isoprostanes levels) oxidative damage in CBS-deficient patients when compared to controls. In addition, a significant positive correlation was found between 15-F2t-isoprostanes levels and total Hcy concentrations. Besides, an in vitro protective effect of NAC at concentrations of 1 and 5 mM was observed on DNA damage caused by Hcy 50 µM and 200 µM. Additionally, we showed a decrease in sulfhydryl content in plasma from CBS-deficient patients when compared to controls. Discussion: These results demonstrated that DNA damage occurs by an oxidative mechanism in CBS deficiency together with lipid oxidative damage, highlighting the NAC beneficial action upon DNA oxidative process, contributing with a new treatment perspective of the patients affected by classic homocystinuria.

Milk protein quantity and quality in low-birth-weight infants. III. Effects on sulfur amino acids in plasma and urine.

Well, appropriate-for-gestational age, low-birth-weight infants weighing 2,100 gm or less were divided into three gestational age groups and assigned randomly within each age group to one of five feeding regimens: pooled human milk; formula 1 (F1) = 1.5gm/dl protein, 60 parts bovine whey proteins: 40 parts bovine caseins; F2 = 3.0 gm/dl, 60:40; F3 = 1.5 gm/dl, 18:82; F4=3.0 gm/dl, 18:82. Plasma and urine concentrations of methionine and of cystathionine were higher in the infants fed F1 to F4 than in the infants fed BM. The plasma cystine concentrations of infants fed F2 (which had a cystine content at least twice that of any of the other formulas) were significantly higher than those of infants fed BM. Plasma taurine concentrations of infants fed F1 or F4, which were virtually devoid of taurine, decreased steadily during the course of study becoming lower than those of infants fed BM. Urine taurine concentrations of infants fed F1, F3, or F4 (but not F2 which had more taurine than F1, F3, or F4) were lower than those of infants fed BM. These results provide further evidence for the limited capacity of the preterm human infant to convert methionine to cystine, owing to delayed maturation of cytathionase, and suggest a limited capacity to convert cystine to taurine. The latter suggestion is consistent with low human hepatic cysteinesulfinic acid decarboxylase activity 0.26 (fetal) and 0.32 (adult) nmoles/mg protein/hour vs 468 in rat liver.