Local and systemic effects of low-dose transtympanic methotrexate: in vivo animal study.

OBJECTIVE: To evaluate the safety of low-dose transtympanic methotrexate in a rat model. DESIGN: Experimental animal study. SETTING: Tertiary training and research hospital. METHODS: Twenty-four rats were randomly divided into three study groups. Diluted methotrexate solution was administered transtympanically to fill the middle-ear cavity, twice a week in group one and three times a week in group two. Ringer lactate solution was administered transtympanically three times a week in the control group. MAIN OUTCOME MEASURES: Local and systemic effects of low-dose transtympanic methotrexate. RESULTS: In the methotrexate groups, middle-ear mucosal oedema was present in all animals. Auditory brainstem response thresholds indicated no inner-ear dysfunction in any group. Liver function and serum haemoglobin levels showed no statistically significant difference in any group. However, liver biopsies from groups one and two showed mild portal hyperaemia. CONCLUSION: These findings are encouraging, and support further investigation of the topical application of methotrexate in autoimmune hearing diseases, as an alternative or adjunct to transtympanic steroids.

Single-dose GnRH agonist administration in the luteal phase of GnRH antagonist cycles: a prospective randomized study.

This study was designed to evaluate the effect of luteal-phase administration of single-dose gonadotrophin-releasing hormone (GnRH) agonist on pregnancy, implantation and live birth rates in patients who received GnRH antagonist for pituitary suppression. The study population consisted of 164 patients who underwent intracytoplasmic sperm injection (ICSI) after ovulation induction by gonadotrophins and GnRH antagonist for the prevention of a premature LH surge. For luteal-phase support, all the cases received intravaginal 600 mg micronized progesterone. In this prospective study, patients were randomly assigned to two groups. In one group, patients received an additional single dose of GnRH agonist (0.5 mg leuprolide acetate) subcutaneously on day 6 after ICSI, whereas the patients in the other group did not. Although the number of embryos transferred and the grade of the embryos were similar in the two groups, the patients in the luteal-phase agonist group had significantly higher rates of implantation and clinical pregnancy rates ( P < 0.05). When the two groups were compared, there were also statistically significant differences in multiple pregnancy and live birth rates ( P < 0.05). Administration of single-dose GnRH agonist as a luteal-phase support in ovarian stimulation-GnRH antagonist cycles in addition to standard luteal support seems to be effective in all cycle outcome parameters.

Ischemia-modified albumin and flap viability.

Monitoring flap viability in the postoperative period is important for the prediction of partial or total flap failure. Many alternative methods such as laser Doppler flowmetry, photoplethysmography, infrared spectroscopy, fluorescein injection, evaluation of flap color, turgor, temperature and capillary reflow have been used for this purpose. Although these methods are valuable in the determination of flap condition, they are not sufficient prognostic markers for flaps. In this study, we investigated the relationship between serum ischemia-modified albumin (IMA) values and flap viability. In 20 Wistar rats, caudal-based 9 x 3 cm dorsal flaps were prepared. IMA values were measured before the flap elevation as well as 6 h and 1 week after the surgical procedure. Seven days after the operation, the viable flap areas were calculated. The relationship between changes in the IMA level during the study and viable flap area measurements was investigated. There was no statistically significant correlation between serum IMA level alterations during the preoperative to postoperative 6th hour period and necrotic flap area measurements. This lack of correlation was also present between the change in IMA levels during the preoperative to postoperative 7th day period and necrotic flap area measurements. In conclusion, serum IMA levels do not predict rat skin flap viability.

Prevalence of hepatitis E virus antibodies in patients with chronic hepatitis B and chronic hepatitis C.

OBJECTIVES: To investigate the prevalence of hepatitis E virus (HEV) among patients with chronic hepatitis B and chronic hepatitis C, serum samples were collected between January and December 2004 from patients with chronic hepatitis B and chronic hepatitis C. METHODS: There were 190 adult patients with chronic hepatitis B virus (HBV) and 174 with chronic hepatitis C virus (HCV) infection in the study group. As the control group, a cohort of 178 age- and sex-matched individuals without known liver disease was selected. RESULTS: Anti-HEV IgG antibodies were positive in 26/190 (13.7%) of chronic HBV and 94/174 (54%) of chronic HCV patients. In the control group anti-HEV positivity was 15.7% (28/178). There was no difference in the percentage of chronic HBV patients and control group who were positive for anti-HEV antibody, but the presence of HEV infection was significantly higher in chronic HCV patients. CONCLUSIONS: Our findings suggest that HEV and HCV might share a common route of transmission in our region. We recommend that preventive measures against HEV should be undertaken in chronic HCV patients as superinfection with HEV can cause a more severe pattern of disease in chronic hepatitis patients.

Fertility of ejaculated and testicular megalohead spermatozoa with intracytoplasmic sperm injection.

In this study the fertility and outcome of intracytoplasmic sperm injection (ICSI) using megalohead spermatozoa from the ejaculates and testicles was evaluated. Seventeen males with megalohead and pinhead sperm forms in their ejaculate were studied in 22 cycles. A high number of sperm heads without tails and abundant round spermatid forms were commonly observed. Round-headed spermatozoa were seldom accompanied by these severely abnormal spermatozoa. The majority of megalohead spermatozoa were observed to have multiple tails, were predominant in the sample, and were used for ICSI. Ejaculated megalohead spermatozoa were used for ICSI in 15 cycles, while testicular spermatozoa were used in seven cycles where there were no vital spermatozoa or spermatozoa of low vitality in the ejaculate. The same abnormal morphology was observed in the testicles as in the ejaculated spermatozoa in the same males. Mean (+/- SD) low motility 4.7 +/- 5.6% and sperm count (3.8 +/- 4.19 x 10(6)) were common findings in these severely teratozoospermic patients. A low fertilization rate (43.2%) was achieved by using megalohead sperm forms (group I, n = 17) in comparison with the control group (60.2%) which had zero normal sperm morphology according to strict criteria (group II, n = 30) (P <0.01). Furthermore, a low pregnancy rate (9.1%) was obtained in the megalohead sperm group in comparison with the control group (40%) (P <0.05). Low fertilization and pregnancy rates may be due to a high incidence of chromosomal abnormalities from severely defective spermatozoa in the ejaculate. Couples should be counselled and warned about possible low fertilization and pregnancy rates with ICSI when only pinhead and megalohead forms with a high number of sperm heads without tails are present in the ejaculate.

Multiple pregnancies obtained by testicular spermatid injection in combination with intracytoplasmic sperm injection.

Recent studies have shown that the injection of spermatid cells into the human oocyte can result in normal fertilization, embryo development and even delivery of live, healthy offspring. In our study, 23 azoospermic cases with severe spermatogenetic defects in their testicular biopsy are presented. The serum follicle stimulating hormone (FSH) concentrations and histopathological results of these males have been documented and compared in terms of fertilization and embryo development. The mean FSH value of the azoospermic males was 15.8 +/- 2.3 mIU/l, ranging from 1.6 to 39 mIU/l. Elongated spermatids were used in three cases only, as these more mature forms were mostly present in the testicular sample. In the remaining 20 cases, only round spermatids were found for use in intracytoplasmic sperm injection (ICSI). The fertilization rate with two pronuclei was 31.3%. The fertilization rate was found to be as high as 71% in three patients in the elongating and elongated spermatids group and as low as 25.6% in the round spermatid group. A few immature, non-motile spermatozoa were seen in only two cases from the elongated spermatid group. However, in the remaining cases, no spermatozoa were observed. The number of pronuclear (PN) arrest was quite high when only round spermatids were used (36.1%). Total fertilization failure was observed in two cases from the round spermatid group with Sertoli cell only and germ cell aplasia. A total of three pregnancies was achieved in 23 cases (13.0%), two from the elongated spermatid group and one from the round spermatid group. One biochemical pregnancy with a round spermatid resulted in an early spontaneous abortion and surprisingly, the remaining pregnancies were achieved with elongated spermatids resulting in multiple pregnancies. One twin and one triplet pregnancy were established following four embryo transfers in each patient. The twin pregnancy resulted in a live birth with two healthy babies; unfortunately, the triplet pregnancy ended in an abortion at 11 weeks. The use of testicular spermatids in the treatment of non-obstructive azoospermia may give hope by offering a novel treatment model. In cases with very severe spermatogenetic defect, even multiple pregnancies can be achieved with elongated spermatid cells by yielding a high implantation rate. However, the efficiency of round spermatids in achieving fertilization and pregnancy was disappointing.