Increased levels of comet-detected spermatozoa DNA damage following in vivo isotopic- or X-irradiation of spermatogonia.

To investigate whether DNA damage arising in spermatogenic germ cells can be detected in resultant sperm, we have irradiated murine testis and collected spermatozoa from the vas deferens 45 days later. These cells were derived from spermatogonia present at the time of irradiation. Two forms of irradiation were used, external X-rays (4Gy) and internal auger electrons from contamination of the male mouse with the isotope Indium-114m (1.85MBq), which was localised in the testis. Both forms of irradiation produced a profound fall in vas deferens sperm count and testis weight, Indium-114m being more effective. Using the neutral Comet assay for double strand break detection, significant increases in sperm comet tail length and moment were observed. The levels of damage were similar for both treatments. Care had to be taken during the assay to distinguish between sperm and somatic cells as the proportion of the latter increased after irradiation. We conclude that the comet assay can detect DNA damage in spermatozoa after the in vivo exposure of male germ cells to a known testicular genotoxic agent. The assay may be useful for the assessment of sperm DNA damage (double stranded) associated with male infertility and post-fertilization developmental abnormalities in the offspring.

Testicular and sperm DNA damage after treatment with fludarabine for chronic lymphocytic leukaemia.

This study investigated whether chemotherapy using fludarabine (FLU) caused testicular damage and if cytotoxicity could be detected as sperm DNA damage in the single cell Comet assay. A patient with chronic lymphocytic leukaemia requesting preservation of fertility was treated with seven monthly cycles of fludarabine (45.8 mg total dose per cycle). Testicular assessments, serum follicle stimulating hormone (FSH), luteinizing hormone (LH), and testosterone measurements, semen analysis and sperm Comet assays were carried out at presentation (pre-FLU therapy), after 1 and 7 months of FLU treatment, and finally at 11 months after completion of chemotherapy. We found that testicular damage occurred within a month, as indicated by reduced testicular volume, oligozoospermia, elevated FSH and LH, and lower testosterone concentrations. Spermatozoa with a large range of DNA damage were detected in the samples from both the control and treated men. DNA damage in the spermatozoa was marked by 7 months of FLU treatment. The high levels of sperm DNA damage seen during and possibly persisting after treatment suggests that caution should be exercised if the ejaculates from these men are used for in-vitro fertility treatment. Further experiments are needed to assess the biological significance of these DNA changes; it may, however, be prudent at present to be cautious when counselling these patients.

Popliteal artery entrapment syndrome: arteriographic findings and thrombolytic therapy.

PURPOSE: To review the arteriographic appearance of popliteal artery entrapment syndrome (PAES) and functional popliteal artery entrapment, and determine the role of thrombolysis in the treatment of these disorders. MATERIALS AND METHODS: Retrospective review of hospital records from 1991 to 1998. RESULTS: Seven patients with PAES and one with functional entrapment were identified. The popliteal artery was occluded in two limbs and compressed in 13. Active plantar flexion was necessary to demonstrate impingement in nine limbs. Medial deviation of the popliteal artery was evident in six of 14 patent popliteal arteries, and lateral deviation was observed in one limb. "Classic" abrupt medial angulation of the popliteal artery was observed in one limb. Both limbs were involved in all six patients who underwent bilateral popliteal exploration. Thrombolytic therapy was performed in three limbs. In two instances, it permitted a less extensive surgical procedure than would otherwise have been required. CONCLUSIONS: There is considerable variability in the arteriographic appearance of PAES, which is arteriographically indistinguishable from functional entrapment. It is frequently bilateral. Thrombolytic therapy does not obviate surgery but may permit a less extensive procedure to be performed.

The effect of allopurinol and catalase on cardiovascular hemodynamics during hemorrhagic shock.

The primary objective of this study was to determine the effect that the xanthine oxidase inhibitor allopurinol (ALLO) and the hydrogen peroxide scavenger catalase (CAT) have on the cardiovascular compensatory ability of the dog to respond to severe hemorrhagic hypotension. Twenty-four mongrel dogs were anesthetized with sodium pentabarbitol and surgically prepared to monitor 1) average arterial blood pressure (AAP), 2) central venous pressure (CVP), 3) heart rate (HR), 4) cardiac index (CI = CO/kg), and hindlimb skeletal muscle blood flow (MBF). Total body vascular conductance (TBC) and skeletal muscle vascular conductance (MVC) were calculated by dividing the CI or MBF by the difference between the AAP and CVP. Eight animals were placed into each of the following three groups, bled over a 1-hr period of time to an AAP of 50 mm Hg and monitored for an additional 2 hr. Group I controls received an intravenous volume of lactated Ringer's equivalent to that volume given to groups II and III. Group II was pretreated 24 hr prior to hemorrhage with 100 mg/kg ALLO orally and received a bolus injection of 25 mg/kg 15 min prior to hemorrhage plus an intravenous infusion of 5 mg/kg/hr over the 3-hr study. Group III was given the same ALLO treatment as group II plus an additional 5-mg/kg/hr intravenous infusion of CAT throughout the duration of the 3-hr study. The results show that the intense compensatory increase in total body vascular tone which occurs during severe hypovolemia is significantly reduced at the 60-, 120-, and 180-min periods in the ALLO/CAT group; however, when ALLO alone was used this effect lasted only through the 120-min period. A similar, but statistically less convincing, picture was seen in the skeletal muscle vascular bed. Thus, the ALLO/CAT group seemed to inhibit some free radical mechanisms better than the ALLO group during and immediately following hemorrhage. Allopurinol alone lost its effectiveness before the 3 hr, which suggests that a free radical mechanism may play an early role in the pathophysiologic shock sequence. As shock continues, however, other factors seem to override the free radical mechanism. One possible explanation for this early tissue protective action of allopurinol and catalase is the inhibition of the oxygen free-radical-induced microvascular swelling and plugging.