Search

Although vitrification is a useful technique for preservation of bovine oocytes, the yield of blastocysts derived from the vitrified oocytes is still low. We have recently reported a new type of cryoinjury, multiple aster formation, by which pronuclear migration and development of vitrified–warmed and in vitro-fertilized bovine oocytes are impaired. The aim of the present study was to investigate the effect of glutathione (GSH) content of vitrified bovine oocytes on multiple aster formation and subsequent in vitro development. Treatment of bovine cumulus–oocyte complexes with β-mercaptoethanol (βME) and l-cysteine (Cys) during in vitro maturation resulted in 2.5-fold higher GSH content not only in fresh control but also in vitrified–warmed oocytes. The percentage of normally fertilized zygotes exhibiting sperm aster(s) was >95% in all four groups (with or without βME/Cys × fresh control or vitrified). The frequency of multiple aster formation in vitrified oocytes (three-fold higher than that in fresh control oocytes) was not affected by the increased level of intracellular GSH with βME/Cys. Consequently, the migration and development of pronuclei as well as the yield of blastocysts from vitrified–warmed oocytes (17 versus 41%) were not improved. In addition, there was no effect of increased GSH level on the yield of blastocysts in fresh control groups. [ABSTRACT FROM PUBLISHER]

The aim of the work was to evaluate the in vitro developmental competence of in vitro-matured buffalo oocytes after Cryotop vitrification (CTV) and in vitro fertilization (IVF). To optimize parameters, two cryoprotectant (CP) concentrations and two warming-dilution procedures were applied. Oocytes were vitrified in 16.5% ethylene glycol (EG), 16.5% dimethylsulphoxide (DMSO) and 0.5 sucrose in Groups A and C, and in higher CP concentrations (20% EG, 20% DMSO and 0.5 sucrose) in Groups B and D. Warming was performed in 1.25 sucrose for 1 min, then in 0.62, 0.42 and 0.31 sucrose, 30 s each (Groups A and B), or in 0.25 sucrose for 1 min and in 0.15 sucrose for 5 min (Groups C and D). After warming, the oocytes were fertilized and cultured in vitro. Survival rate post-warming was lower in Group D (83.6%) than in Groups A and B (92.4 and 92.8%, respectively), while intermediate values were found in Group C (85.7%). Survival rates at 24 h decreased in Groups C and D (52.0% and 50%, respectively) and remained high in Groups A and B (84.0% and 85.6%, respectively), thus indicating that the dilution of CP after warming is critical for buffalo oocyte cryopreservation. Similar differences were also observed in cleavage rates (42.7%, 55.3%, 28.4% and 36.3% for Groups A, B, C and D, respectively) whereas no differences in blastocyst rates were found among groups (6.4%, 7.8%, 5.9% and 6.9% for Groups A, B, C and D, respectively). Blastocyst production after IVF of vitrified oocytes proves the feasibility of CTV in buffalo species. [ABSTRACT FROM AUTHOR]

The purpose of this study was to analyze the relationship between the reproducibility of holmium laser enucleation of the prostate (HoLEP) and prostate size over the learning curve. We compared the outcome among three institutions in three subgroups on the basis of the weight of tissue retrieved. There were no significant differences in operating time, efficiency of the procedure, decrease in hemoglobin level and postoperative urinary incontinence among three institutions, only in patients with prostates 20 g–<40 g retrieved. Our data indicate that HoLEP is more reproducible in patients with a moderate-sized prostate over the learning curve. [ABSTRACT FROM AUTHOR]

We have studied the prevalence of the subgenus F adenoviruses and the molecular characteristics of adenovirus type 41 in Hiroshima Prefecture, Japan, as a limited area during the period of 1997–2004. Subgenus F adenoviruses were detected in 30 (3.4%) of 892 fecal specimens by enzyme immunoassay (EIA), and 80.0% (24 of 30) of positive patients were <36 months old. One (3.3%) and 29 (96.7%) of the 30 EIA-positive specimens were adenoviruses type 40 (Ad40) and 41 (Ad41), respectively. The genomes of Ad41 strains amplified by PCR were divided into two genomic type clusters (GTC1 and GTC2) based on the hexon gene as described by Li et al. (J Clin Microbiol 42: 4032–4039, 2004.). Twenty-one (95.5%) of 22 Ad41 strains detected between 2000 and 2004 belonged to GTC1, whereas all seven strains detected between 1997 and 1999 belonged to GTC2. These genomic typings were the same for the hexon and fiber genes except for one strain. This strain contained a hexon gene belonging to GTC1 and a fiber gene belonging to GTC2 and was considered to be a recombinant between adenoviruses of these types. [ABSTRACT FROM AUTHOR]

Background: Human T-cell lymphotropic virus type I (HTLV-I) is linked to adult T-cell leukemia/lymphoma (ATL) and HTLV-I-associated myelopathy (HAM; also known as tropical spastic paraparesis [TSP]), a chronic neurodegenerative disorder. Worldwide, several million HTLV-I carriers are at risk for disease, with an estimated lifetime cumulative risk of 1%-5%. However, the determinants of disease progression are relatively unknown. We studied human leukocyte antigens (HLA class II) that have been implicated in the pathogenesis of HTLV-I-related diseases.Methods: We analyzed HLA class II alleles among asymptomatic HTLV-I carriers (n = 45), patients with ATL (n = 49) or HAM/TSP (n = 54), and HTLV-I seronegative control subjects (n = 51). All participants were of African descent and were enrolled in epidemiologic studies conducted at the University of the West Indies, Kingston, Jamaica. We used standard microlymphocytotoxicity assays for HLA antigen serotyping and polymerase chain reaction-based methods to examine HLA class II DRB1 and DQB1 alleles.Results: Two antigens determined by serotyping, DR15 and DQ1, occurred at significantly increased frequency among HTLV-I carriers compared with seronegative control subjects (42% versus 22% for DR15 [odds ratio [OR] = 2.7; 95% confidence interval [CI] = 1.0-7.2] and 78% versus 53% for DQ1 [OR = 3.1; 95% CI = 1.2-8.5]). Asymptomatic carriers were shown to have an HLA class II allele distribution similar to that of patients with ATL, and the frequencies of the alleles DRB1*1501, DRB1*1101, and DQB1*0602 were significantly greater in patients with ATL and asymptomatic carriers than in patients with HAM/TSP. In addition, haplotypes DRB1*1101-DQB1*0301 and DRB1*1501-DQB1*0602 were significantly increased among patients with ATL compared with patients with HAM/TSP.Conclusions: These data suggest that host genetic background is an important factor in determining whether HTLV-I carriers develop either ATL or HAM/TSP. [ABSTRACT FROM AUTHOR]

A high concentration of serum supplements has been contained in basic vitrification solutions to protect plasma membranes. The objective of this study was to examine if vitrification of immature porcine oocytes could be achieved successfully in a chemical-defined solution containing 0.6 mg mL[sup -1] hydroxypropyl cellulose (HPC). Cumulus-oocyte complexes (COC) were aspirated from follicles 3 to 6 mm in diameter in abattoir-derived porcine ovaries. The COC or denuded oocytes were vitrified according to a commercial protocol of Cryotop (Kitazato BioPharma Co. Ltd., Shizuoka, Japan) with original solutions or modified ones that had serum supplement replaced with HPC. After vitrification and warming, viabilities of oocytes and cumulus cells were evaluated under a fluorescent microscope after staining with fluorescein diacetate and propidium iodide (Table 1). Statistical analyses of results from 4 replicated trials were performed by ANOVA with a Bonferroni/Dunn post hoc test (significance, P < 0.05). Although viabilities of vitrified/warmed oocytes in all groups were significantly lower (82.7-89.1%) than those of fresh controls (99.5%), there were no significant differences among vitrified groups. When COC were vitrified, viability of cumulus cells (54.3%) in HPC group was not different from that of nonvitrified controls (72.5%) but higher than that in the original solution group (48.1%). In conclusion, these results demonstrate that HPC rather than serum supplement could be a suitable chemically defined supplement for vitrification of immature porcine COC. [ABSTRACT FROM AUTHOR]