Efficiency of Autologous Egg Cryopreservation: Eight Years’ Experiences and Clinical Outcomes

Oocyte cryopreservation is one of the state-of-art technologies in human reproductive medicine, which brings opportunities for women to preserve their fertility. In the present study, we analyzed the efficiency and outcomes of 8 years’ autologous egg cryopreservation: Frozen oocytes were warmed from 120 cycles and oocyte survival, fertilization, blastocyst development, clinical pregnancy, embryo implantation, live birth rates and birth weights were collected based on the patients’ ages of  37 years old. The details of oocyte cryopreservation and the efficiency were further analyzed based on different patient categories. During the study period, 849 oocytes from 120 cycles were warmed. Oocyte survival, fertilization, and blastocyst development were not affected by women’s ages at the time of cryopreservation. However, number of patients without blastocyst formation was significantly (P 37 years old (31.2%) than that in patients 37 years old (28.6%) after fresh embryo transfer. Some patients did not have blastocysts mainly due to low fertilization by poor sperm or small number of oocytes warmed. These results indicate that the efficiency of oocyte cryopreservation, evaluated by live birth and embryo implantation rates is affected by women’s age, number of oocytes warmed and sperm quality

A Method for Small Number of Human Sperm Cryopreservation

Recently, some sperm vitrification devices were developed to simplify the procedures to freeze small number of human sperm. In the present study, we used these devices to further examine some factors that affect sperm motility after fast freezing. Experiments were designed to examine the effects of 1) direct immersion of the devices to liquid nitrogen and indirect immersion of the devices to liquid nitrogen in which the devices were sealed in cryogenic storage vials; 2) different freezing volumes (1–5 μl); 3) different equilibration times (1–5 min); and 4) different ratio of freezing solution (0,1-5,1) on post thawing sperm motility. It was found that fast sperm freezing in the sealed vials had high post thawing sperm motility (91.3–93.7% of recovered sperm motility rates) while direct immersion of the devices to liquid nitrogen had 0% post thawing sperm motility. No differences in the recovered sperm motility rates were observed between different freezing solution volumes (87.4–90.5%), different equilibration times (89.5–94.0%), and different freezing solution ratios (90.8–94.6%). However, only 6.8% of recovered sperm motility rate was obtained if sperm were frozen in the medium without sperm freezing solution. These results indicate that human sperm can be rapidly frozen after the devices are sealed in the vials with different equilibration time in the medium containing sperm freezing solution. High post thawing sperm motility can be recovered with this method so that ~90% of sperm are usable after freezing

RNAi-directed downregulation of OsBADH2 results in aroma (2-acetyl-1-pyrroline) production in rice (Oryza sativa L.)

<p>Abstract</p> <p>Background</p> <p>Aromatic rice is popular worldwide because of its characteristic fragrance. Genetic studies and physical fine mapping reveal that a candidate gene (<it>fgr</it>/<it>OsBADH2</it>) homologous to <it>betaine aldehyde dehydrogenase </it>is responsible for aroma metabolism in fragrant rice varieties, but the direct evidence demonstrating the functions of <it>OsBADH2 </it>is lacking. To elucidate the physiological roles of <it>OsBADH2</it>, sequencing approach and RNA interference (RNAi) technique were employed to analyze allelic variation and functions of <it>OsBADH2 </it>gene in aroma production. Semi-quantitative, real-time reverse transcription-polymerase chain reaction (RT-PCR), as well as gas chromatography-mass spectrometry (GC-MS) were conducted to determine the expression levels of <it>OsBADH2 </it>and the fragrant compound in wild type and transgenic <it>OsBADH2</it>-RNAi repression lines, respectively.</p> <p>Results</p> <p>The results showed that multiple mutations identical to <it>fgr </it>allele occur in the 13 fragrant rice accessions across China; <it>OsBADH2 </it>is expressed constitutively, with less expression abundance in mature roots; the disrupted <it>OsBADH2 </it>by RNA interference leads to significantly increased 2-acetyl-1-pyrroline production.</p> <p>Conclusion</p> <p>We have found that the altered expression levels of <it>OsBADH2 </it>gene influence aroma accumulation, and the prevalent aromatic allele probably has a single evolutionary origin.</p

Structural specializations of the sperm tail

Sperm motility is crucial to reproductive success in sexually reproducing organisms. Impaired sperm movement causes male infertility, which is increasing globally. Sperm are powered by a microtubule-based molecular machine-the axoneme-but it is unclear how axonemal microtubules are ornamented to support motility in diverse fertilization environments. Here, we present high-resolution structures of native axonemal doublet microtubules (DMTs) from sea urchin and bovine sperm, representing external and internal fertilizers. We identify \u3e60 proteins decorating sperm DMTs; at least 15 are sperm associated and 16 are linked to infertility. By comparing DMTs across species and cell types, we define core microtubule inner proteins (MIPs) and analyze evolution of the tektin bundle. We identify conserved axonemal microtubule-associated proteins (MAPs) with unique tubulin-binding modes. Additionally, we identify a testis-specific serine/threonine kinase that links DMTs to outer dense fibers in mammalian sperm. Our study provides structural foundations for understanding sperm evolution, motility, and dysfunction at a molecular level

Structural specializations of the sperm tail

Sperm motility is crucial to reproductive success in sexually reproducing organisms. Impaired sperm movement causes male infertility, which is increasing globally. Sperm are powered by a microtubule-based molecular machine-the axoneme-but it is unclear how axonemal microtubules are ornamented to support motility in diverse fertilization environments. Here, we present high-resolution structures of native axonemal doublet microtubules (DMTs) from sea urchin and bovine sperm, representing external and internal fertilizers. We identify >60 proteins decorating sperm DMTs; at least 15 are sperm associated and 16 are linked to infertility. By comparing DMTs across species and cell types, we define core microtubule inner proteins (MIPs) and analyze evolution of the tektin bundle. We identify conserved axonemal microtubule-associated proteins (MAPs) with unique tubulin-binding modes. Additionally, we identify a testis-specific serine/threonine kinase that links DMTs to outer dense fibers in mammalian sperm. Our study provides structural foundations for understanding sperm evolution, motility, and dysfunction at a molecular level