[en] Various approaches have been utilized in attempting to cryopreserve oocytes, beginning with slow cooling and more recently the advent of technique of vitrification. Now it seems that oocyte cryopreservation is no longer an experimental technique and it is being increasingly utilized in clinics around the world. As successful outcome in oocyte cryopreservation can be assessed by survival through the freeze-thaw process, potential for fertilization, embryo development and dynamics of meiotic spindles. This study aimed to analyse these features in context of vitrification and slow freezing. Methods: In this laboratory based study, mature MII mouse oocytes from F1(C57BL6/J X CBA) mice (n=43) were divided randomly into two groups of equal numbers and were cryopreserved by slow freezing and by vitrification. Upon re-warming these oocytes were assessed for survival and for fertilization potential. Oocytes were fixed and stained to compare the effect of both protocols on spindle reassembly and chromosome configuration 10min, 1h and 3h after warming. Unfrozen oocytes were used as controls. Results: A greater number of vitrified oocytes survived cryopreservation than slow frozen oocytes (70.3% vs. 12.5%, p=0.024). After insemination, fertilization rates were higher for vitrified oocytes as compared to slow frozen oocytes (15.86% vs. 4.6%, p=0.046). Morphology of the meiotic spindle was found to be in a disorganized configuration in slow frozen oocytes at all-time points 10 mins, 1 h and 3h), whereas in vitrified oocytes the spindles were found to be aligned at all-time points. Chromosomes were seen to be displaced from equatorial region in both groups. Conclusion: Cryopreservation of mouse oocytes was conducted with greater success using vitrification, compared to slow freezing, with survival, fertilization, and spindle assembly more favourable to a successful outcome in this model. (author)

[en] To evaluate the maternal and neonatal complications in terms of genital tract trauma to mother, perinatal mortality. Apgar score at 5 minutes and neonatal trauma in all singleton term breech cases delivered vaginally. A 100 consecutive patients with singleton breech presentation, whether booked or unbooked, were admitted and delivered vaginally in hospital during the study period. They were studied for maternal and neonatal complications. Maternal complication included any genital tract trauma during delivery while neonatal complications were perinatal mortality, low Apgar score (less than eight at 5 minutes) and birth trauma. There were a total 3977 deliveries during this study period, 145 breech presentation (incidence 3.6%). Out of those, 100 were included in the study. All were unbooked cases, 87% were multigravida while 13% were primigravida. An Apgar score of eight was recorded in 87% babies while 10% had an Apgar score of less than eight after 5 minutes. There were 3 still births and one neonatal death. Aggregated perinatal mortality rate was 40/1000 live births. Only one baby had birth trauma (Erbs paralysis) during vaginal breech delivery. Ninety seven (97%) mothers had no complications while only 3 (3%) had complications. Out of these, one had cervical tear and 2 had vaginal tears. Offering a trial of vaginal breech delivery to strictly selected and well-counselled patients remains an appropriate option without compromising perinatal and maternal outcome. It also decreases the rate of operative delivery for this malpresentation. (author)

[en] The present paper describes flow injection procedures for a-tocopherol determination in pharmaceuticals and infant milk powder based on spectrophotometric detection. These two procedures are based on the reduction of iron(III) to iron(II) and of Mo(Vi) to Mo(V) by a-tocopherol and subsequent formation of iron(II)-o-phenanthroline, and phospho molybdenum complexes which were monitored at 510 and 695 nm respectively. Calibration graphs were linear over the concentration range of 0.21-43 mu g/mL for iron(III) and 0.25-10 mu g/mL for Mo(Vi) reduction methods respectively. The limits of detection (3s) of 0.05 and 0.1 mu g/mL with sample throughput of 30 and 50/h respectively were obtained. The relative standard deviations of 1.2- 3.8% (n = 4) was achieved in the concentration range studied. The developed methods were applied to pharmaceuticals and infant milk powders and the results obtained were compared with official method and no significant difference between these methods was observed at 95% confidence level. (author)

[en] Background: Treatment of Chronic Hepatitis C is now well established with conventional interferon or pegylated interferon in combination with ribavirin. Peginterferon Alfa and Ribavirin for 6 to 12 months is currently approved initial therapy, which is expensive. Response of our patients to standard Interferon-alpha-2b and ribavirin for 24 weeks have been studied. The objective of this study was to asses Sustained Viral Response (SVR) with standard Interferon A2b and Ribavirin combination treatment in chronic Hepatitis C patients. Methods: This quasi-experimental study was conducted at Combined Military Hospital, Quetta from Jan 2006 to Jun 2007. One hundred and three patients, with 20-60 years of age suffering from chronic Hepatitis C were selected on the basis of raised ALT, positive anti-HCV antibodies, evidence of viraemia by quantitative PCR for HCV RNA and liver biopsy. All patients were started on same brand of Interferon alpha-2b, 3 MIU subcutaneously, thrice weekly and oral Ribavirin (1,000-1,200 mg/day) for 24 weeks. End treatment response (ETR) after completion of treatment and SVR six months after ETR were recorded. Results: The 103 patients, 85 males and 18 females with mean age of 21-48 years completed the treatment for 24 weeks. Mean ALT was 96.17 (SD +- 49.98). End treatment response (ETR) was 89.3% (p=0.032). Sustained Viral Response after 6 months of treatment was 86.4% (p=0.034). Conclusion: Standard Interferon and Ribavirin had excellent SVR. It is effective as well as economical treatment in Chronic Hepatitis C patients. (author)

[en] Objective: To correlate and quantitate Iymphocyte subsets with clinically diagnosed smear-negative pulmonary tuberculosis and severity of disease. Subjects and Methods: Freshly diagnosed, well-characterized smear-negative patients (n=15) of pulmonary tuberculosis were selected. Non-induced three-consecutive negative smears of sputum with simultaneous culture for AFB for 6-8 weeks, positive Mauntoux test (= or > 0 mm), blood complete picture with ESR and chest x-rays were done. Selected panel of monoclonal antibodies against specific CD markers were used. Statistical analysis done by student t-test or Mann-Whitney rank-sum test with the help of Sigma State software. Results: Hemoglobin and total lymphocyte counts were significantly reduced whereas total leukocyte counts with absolute neutrophil counts were increased. Fraction of CD4+ and CD8+ T lymphocytes with HLA-DR expression was reduced while no significant change in rest of the TB and NK lymphocytes. Conclusion: Low hemoglobin level, high neutrophil count and low total lymphocytes suggest possible direct relationship with extent of disease. The number of activated CD4+, CD8+, alpha beta and gamma delta TCR T cells have tendency to increase during Mycobacterium infection. This seems to have a potential of being a good, non-invasive prognostic indicator for patients with pulmonary tuberculosis. (author)