Determination of DNA damage after exposure to inhalation anesthetics in human peripheral lymphocytes and sperm cells in vitro by comet assay.

In this study, genotoxic activities of four halogenated anesthetics (halothane, isoflurane, sevoflurane and desflurane) were investigated in human peripheral blood lymphocytes (PBLs) and sperm cells in vitro by alkaline comet assay. For this purpose, sperm or lymphocyte suspension was exposed to different concentrations (0.1 mM, 1 mM, 10 mM and 100 mM) of anesthetic agents and 1% dimethyl sulfoxide (DMSO) or phosphate-buffered saline (PBS) as controls. The DNA strand breaks as well as alkali-labile sites were measured as percentage tail intensity with comet assay. The results of this study demonstrate that all analyzed drugs were capable of inducing DNA damage on PBLs in a dose-dependent manner in vitro. However, the results in sperm cells were slightly different since we did not observe any genotoxic effect for desflurane in any of the exposure doses, and the genotoxic effect of halothane was not dose dependent. This experimental study points out to the presence of DNA damage after exposure to halogenated anesthetics in both PBLs and sperm cells, although this effect seems to be higher in PBLs.

Appendicoumbilical fistula: a rare reason for neonatal umbilical mass.

The normal umbilicus is a simple structure, but the intrauterine development of the umbilicus is highly complex. Neonatal umbilical mass anomalies usually represent failure of obliteration of the vitelline duct or the allantois which results in persistence of remnants, which can lead to a wide variety of disorders. In this paper, we present a case of an appendicoumbilical fistula in a neonate along with the differential diagnosis and management options. Embryologic explanation of the etiology was discussed with the possible association with different forms of malpositioning and rotation of the gut.

Determination of DNA damage by alkaline halo and comet assay in patients under sevoflurane anesthesia.

In this study, we report data on the possible genotoxic effect of inhalation anesthetic sevoflurane (SVF) by comparing two techniques, comet and alkaline halo assay, in peripheral blood lymphocytes (PBL) of patients before, during, and after anesthesia and in controls. DNA single strand breaks were detected in PBL of malignant breast cancer diagnosed patients (stage II-III), who were undergoing mastectomy. Blood samples were taken before the induction of anesthesia, at 120 min of SVF anesthesia, and on the postoperative fifth day. The nuclear spreading factor (NSF) for each cell was assessed by alkaline halo assay, and the total comet score (TCS) was evaluated by comet assay. A statistically significant increase (P < 0.0001) was observed in the mean NSF at 120 min of anesthesia (38.24 +/- 14.14) as compared with samples before anesthesia (12.33 +/- 6.14), and the mean NSF was significantly decreased after the postoperative fifth day (17.89 +/- 9.44). Similar results were obtained by the comet assay with significant increase (P < 0.0001) in DNA damage at 120 min of anesthesia (79.66 +/- 15.28) as compared with samples before anesthesia (36.30 +/- 11.39). The DNA damage was almost with the preoperative damage rates after the fifth day of anesthesia (43.40 +/- 12.19). In conclusion, the study points out a reversible genotoxic effect of SVF and the similar DNA damage levels obtained by comet and alkaline halo assay indicate that although halo assay has a completely different principle, it can conveniently be utilized for the assessment of DNA single strand breakage in individual mammalian cells with its experimental advantages.

Oxidative DNA damage and total antioxidant status in rats during experimental gram-negative sepsis.

Sepsis and septic shock remains as leading cause of death in adult intensive care units. It is widely accepted that gram-negative bacteria and their endotoxins cause sepsis and septic shock, predominantly. Enhanced generation of reactive oxygen species may be responsible for tissue injury in septic shock and endotoxemia. The aim of this study was to assess oxidative DNA damage and the total antioxidant status (TAS) in peripheral lymphocytes of rats during different intraperitoneal gram-negative sepsis stages. Adult male Sprague-Dawley rats were divided randomly into four groups. Control group was intraperitoneally inoculated with 2 mL of pyrogene-free saline (Group I, n = 6), and the other rats received an intraperitoneal inoculum with 2 mL of saline containing 2 x 10(8) CFU of Escherichia coli. The animals were killed at time zero (Group I, n = 6), at 6th (Group II, n = 7), 12th (Group III, n = 7), and 24th (Group IV, n = 7) hour after the E. coli inoculation. Oxidative DNA damage in peripheral lymphocytes of rats was evaluated by modified comet assay (single-cell gel electrophoresis). Formamidopyrimidine DNA glycosylase (Fpg) and Endonuclease III (Endo III) were used to detect oxidized purines and pyrimidines, respectively. Total antioxidant quantification was carried out using ABTS+ (2,2'-Azino-di-[3 ethylbenzthiazoline sulphonate]) radical formation kinetics (Randox kit) in serum samples. Significant elevations of basal levels of strand breaks (SB) in Group IV were observed as compared with Group I, II, and III. There was a significant increase in Fpg sites in Group III as compared with Group I and II. However, there was no significant difference in terms of Endo III sites in any of the groups. Although the TAS was decreased with the stages of sepsis, this moderate decrease was significant in only Group IV as compared with Group I. There was no statistically significant correlation between DNA damage and TAS for any of the groups.

The role of antioxidant supplementation in occupational exposure to waste anaesthetic gases.

OBJECTIVES: Although the genotoxicity related to waste anaesthetic gases is controversial, a consistent number of observations have provided evidence for an increased level of DNA strand breaks. The goal of the research was to investigate this hypothesis and estimate the genoprotective role of antioxidant supplementation in technical anaesthesiology staff working in operating theatres. METHODS: Heparinized venous blood samples were collected from 17 exposed technical anaesthesiology staff (mean age 34.3 +/- 3.5 years) and non-exposed control group (mean age 32.2 +/- 3.4 years) and examined in the alkaline comet assay for DNA strand breakage. Vitamin E (300 mg/day) plus vitamin C (500 mg/day) were supplemented to the technical anaesthesiology staff for 12 weeks and blood samples were retaken and evaluated by comet assay. RESULTS: The DNA breakage observed in the lymphocytes of the technical anaesthesiology staff was 21.5 +/- 5.0, as calculated by total comet score (TCS). This score was significantly higher (P<0.001) than in the controls (8.6 +/- 4.7) before antioxidant treatment. Supplementation of vitamins E plus C significantly (P<0.01) reduced the mean TCS as 14.2 +/- 6.1. CONCLUSION: The results of our study indicate that occupational exposure to anaesthetic gases induces oxidative DNA damage. Supplementation of the diet for 12 weeks with vitamin C and vitamin E resulted in a significant decrease in the DNA damage.