Heterotopic autotransplantation of vitrified mouse ovary

Purpose  

The aim of this study was to investigate the survival and development of premature follicles and oocytes from a vitrified-transplanted ovary in a murine experimental model.     

Effect of fragment removal on blastocyst formation and quality of human embryos

Blastomere fragmentation is one of the most significant defects in cleaving embryos. Scientists believed that removing the fragments was a possible way to reduce their unwanted effects. This hypothesis has been tested in some studies in which the development of human fragmented embryos was followed in vivo after all fragments were removed, but little is known about the potential for in-vitro development of such embryos, which is the subject of the present study. For this purpose, 4-6 cell surplus human embryos were scored according to the degree and pattern of fragmentation into four grades, allocated into two groups of control and fragmentation removal (experimental) and cultured sequentially. At the end of day 6 of culture, in the experimental group especially in grade IV blastocyst rate, size and number of blastomeres in each blastocyst were all improved compared with those of the control group (42.3 versus 20.0%; 19,205.7 +/- 1060.3 versus 15,825.9 +/- 448.7 microm(2) and 100.14 +/- 13.48 versus 63.75 +/- 19.79 respectively, P < 0.05). In the grade IV embryos, apoptotic index was also significantly reduced after embryo fragmentation removal (3.40 +/- 0.88 versus 22.99 +/- 4.45, P < 0.05). In conclusion, fragmentation removal had a positive effect on human fragmented embryos and produced the best quality blastocysts.     

Achieving high survival rate following cryopreservation after isolation of prepubertal mouse spermatogonial cells

Purpose

Isolating spermatogonia cells with high purity and viability and achieving better survival rate following cryopreservation

Methods

Isolating the cells by Magnetic Activating Cell Sorting (MACS) method using anti CD49f (α6 integrin) antibody and Dynabeads and freezing in DMSO-based freezing mediums containing three different FBS concentrations of 50%, 60% and 70%.

Results

The mean (±SD) purity of the isolated cells was 92.52?±?3.57 (range 92.43–98.25). The cells frozen in group I, II and III had mean 39.60?±?1.48 (range 37.98–41.62), 89.05?±?3.83 (range 80.83–90.33) and 90.52?±?1.71 (range 89.07–92.52) viability, respectively.

Conclusion

Higher viable cell counts and purity can be attained by the use of α6 integrin and magnetic beads. After the thawing of spermatogonial cells, optimum viability was achieved in freezing media containing 60% FBS.     

Emergency Medicine Residents and Statistics: What is the Confidence?

The objective of this study was to assess whether residents have the essential tools and a sense of competency when evaluating published studies, especially the statistics. Questionnaires were mailed to emergency medicine (EM) residency programs in the United States querying residents' demographics and training in statistics as well as their impressions and use of statistics in the current literature; a five-question statistical quiz was also included. Possible responses of—almost always, more than ½ time, ½ time, less than ½ time, almost never—were tallied individually as well as compared in groups of polarized answers: over 1/2 time (almost always + more than ½ time) vs. under ½ time (less than ½ time + almost never). There were 495 questionnaires returned from 42 centers. No significant difference was found when comparing quiz performance with participants' self-reported statistical knowledge. There were considerable differences in the polarized answers (Over vs. Under), whether statistics: were used appropriately (40% vs. 15%, respectively); were used to enhance weak data (54% vs. 13%, respectively); enhanced their understanding of information (38% vs. 24%, respectively); simplified complex data (26% vs. 41%, respectively); were understood by them (23% vs. 38%, respectively); confused them (37% vs. 24%, respectively); were skipped (52% vs. 23%, respectively). Participants felt there should be more statistical training (49% vs. 22%, Over vs. Under, respectively). There was no difference in respondents who did or did not read the statistics (39% vs. 34%, Over vs. Under, respectively). Many EM residents surveyed do not trust, read, or understand statistics presented in current journal articles. Residency programs may want to consider enhanced training in statistics.     

Effect of laser-assisted hatching and necrotic blastomere removal on the development of vitrified–warmed four-cell mouse embryos

PURPOSE: To investigate the effect of laser-assisted hatching and necrotic blastomere removal on the development of vitrified-warmed mouse embryos. METHODS: The vitrified-warmed four-cell stage mouse embryos were divided into five groups; vitrified intact with no laser-assisted hatching, vitrified intact with laser-assisted hatching, vitrified damaged with neither laser assisted hatching nor necrotic blastomere removal, vitrified damaged with laser-assisted hatching, and vitrified damaged with necrotic blastomere removal. Thereafter blastocyst formation, blastomere and apoptotic cell number within all groups were statistically compared. RESULTS: The rate of blastocyst formation showed a significant improvement in the group vitrified intact with laser-assisted hatching. However, neither laser-assisted hatching nor necrotic blastomere removal can improve a delayed vitrified-warmed damaged embryos in term of blastocyst formation and total cell number. Nevertheless, apoptotic cell number was significantly reduced after application of both techniques. CONCLUSIONS: Laser-assisted hatching can improve the development of vitrified-warmed intact four-cell stage mouse embryos, whereas necrotic blastomere removal has no significant effect on the development of vitrified-warmed four-cell stage damaged embryos.     

Effect of <Emphasis Type="Italic">Papaver rhoeas</Emphasis> extract on in vitro maturation and developmental competence of immature mouse oocytes

Purpose  

This experiment examined the effect of Papaver rhoeas L. extract on in vitro maturation, in vitro fertilization (IVF) and subsequent developmental competence of mouse oocytes.     

Efficacy of a human embryo transfer medium: a prospective, randomized clinical trial study

Purpose: The aim of this prospective, randomized trial was to evaluate the efficacy of Embryo-Glue^® as a human embryo transfer medium in IVF/ICSI cycles. Method: A total of 815 nonselected patients undergoing IVF/ICSI treatment between September 2003 and February 2004 were randomly allocated into the test (417 patients) and the control (398 patients) groups. In both groups, embryos were cultured in G-1™ver 3, supplemented with 10% recombinant human albumin. On the day of embryo transfer (day 3), the best or good quality embryos were selected for intrauterine transfer. In the test group, the selected embryos were treated with EmbryoGlue^® prior to the transfer, whereas in the control group they were transferred without any treatment. Results: The patients’ characteristics such as age and the number of ART cycles and also the number of patients in each indication of infertility and the number of embryos selected for transfer were all similar between the two groups. In the test group, the clinical pregnancy rate in the tubal factors and the implantation rate in the tubal factors and recurrent implantation failures increased significantly compared with those in the control group. In the test group, life birth and the triplet delivery rates increased significantly compared with those in the control group. Conclusion: EmbryoGlue^® is a useful embryo transfer medium, and at least in some infertile patients it can improve clinical implantation and ongoing pregnancy rates.     

Colorimetric detection of multidrug-resistant or extensively drug-resistant tuberculosis by use of malachite green indicator dye

The malachite green microtube (MGMT) susceptibility assay was performed directly on sputum specimens (n = 80) and indirectly on Mycobacterium tuberculosis clinical isolates (n = 60). The technique is based on the malachite green dye, which changes color in response to M. tuberculosis growth. The MGMT assay is simple and rapid and does not require expensive instruments.     

Light and electron microscopic study of epithelial cells from the human oviduct and uterus subcultured on extracellular matrix gel

OBJECTIVE: To investigate the structure of epithelial cells from the human oviduct and uterus on extracellular matrix (ECM) gel in the first passage. STUDY DESIGN: Human oviducts and endometrial tissues were obtained from patients undergoing total hysterectomy; the epithelial cells, having been isolated by enzyme digestion, were cultured on polystyrene plastic surfaces. The epithelial nature of the cells was confirmed by immunocytochemistry, and their morphology was examined by microscopy. Cells of an epithelial nature were then trypsinized and cultured on an ECM gel-coated filter insert for 5 days. The cells, in parallel with the tissues, were subsequently prepared for transmission electron microscopy. RESULTS: Plastic-cultured cells had no sign of differentiation and appeared as elongated spindle cells in sections. These cells looked columnar and highly polarized after being cultured on ECM gel surfaces. They were similar to epithelial cells from the corresponding tissue fragment. Cultured on ECM gel, the ciliated epithelial cells of human oviducts appeared ultrastructurally similar to glandular cells from the human uterus. Cilia did not form under culture conditions. CONCLUSIONS: It seems that human uterine and oviduct epithelial cells can acquire polarized morphology and differentiated states on ECM gel after having lost it on plastic surfaces and that ECM gel by itself is not enough to induce cilia formation in culture.