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21.
J Li K Villemoes Y Zhang Y Du PM Kragh S Purup Q Xue AM Pedersen AL Jørgensen JE Jakobsen L Bolund H Yang G Vajta 《Reproduction in domestic animals》2009,44(1):122-127
The purpose of our work was to establish an efficient-oriented enucleation method to produce transgenic embryos with handmade cloning (HMC). After 41–42 h oocytes maturation, the oocytes were further cultured with or without 0.4 μg/ml demecolcine for 45 min [chemically assisted handmade enucleation (CAHE) group vs polar body (PB) oriented handmade enucleation (OHE) group respectively]. After removal of the cumulus cells and partial digestion of the zona pellucida, oocytes with visible extrusion cones and/or polar bodies attached to the surface were subjected to oriented bisection. Putative cytoplasts without extrusion cones or PB were selected as recipients. Two cytoplasts were electrofused with one transgenic fibroblasts expressing green fluorescent protein (GFP), while non-transgenic fibroblasts were used as controls. Reconstructed embryos were cultured in Well of Wells (WOWs) with porcine zygote medium 3 (PZM-3) after activation. Cleavage and blastocyst rates were registered on day 2 and day 7 of in vitro culture respectively. Meanwhile, the total blastocyst cell number was counted on day 7. We found that the difference was only observed between blastocyst rates (38.6 ± 2% vs 48.1 ± 3%) of cloned embryos with GFP transgenic fibroblast cells after CAHE vs OHE. With adjusted time-lapse for zonae-free cloned embryos cultured in WOWs with PZM-3, it was obvious that in vitro developmental competence after CAHE was compromised when compared with the OHE method. OHE enucleation method seems to be a potential superior alternative method used for somatic cell nuclear transfer (SCNT) with transgenic fibroblast cells. 相似文献
22.
Studies have demonstrated that oviductal fluid (ODF) proteins associate with eggs of numerous species including the bovine. In this study, the association of three ODF proteins, the bovine oestrus‐associated protein, osteopontin (OPN), lipocalin‐type prostaglandin D synthase (L‐PGDS), with the bovine zona pellucida (ZP) was demonstrated by immunohistochemistry and western blot. The biological function of ODF derived egg‐associated OPN and L‐PGDS in sperm binding, fertilization and embryonic development was also explored. In vitro matured bovine oocytes were pre‐incubated with ODF collected by cannula from cows in oestrus, or ODF with antibodies to OPN, L‐PGDS and bovine serum albumin (BSA). Following incubation, oocytes were inseminated with 1 × 105 frozen‐thawed spermatozoa, and they were evaluated for sperm binding, fertilization and embryonic development in vitro. Pre‐treatment of ODF with antibodies to all of proteins reduced sperm binding to the ZP and fertilization in vitro. Cleavage rates were not significantly different among incubations, but rates of embryo development were significantly decreased. We conclude that antibodies to OPN, L‐PGDS and BSA react with oocytes incubated with ODF and inhibit sperm binding, fertilization and embryonic development in vitro, suggesting a potential role of these proteins in these events. 相似文献
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鹿蹄草等中草药对油桃采后褐腐病抑制效果研究 总被引:5,自引:2,他引:5
在离体和活体条件下,测定鹿蹄草、厚朴等中草药单剂及鹿蹄草为主的各中草药复配组合对油桃采后褐腐病菌的抑制效果。结果表明:11号(鹿蹄草+厚朴+大黄),14号(鹿蹄草+高良姜+虎杖)复配组合在离体试验中抑菌效果最好,10 mg/mL浓度的抑制率分别达到89.77%,85.50%;两种组合的最小抑菌浓度(MIC)分别是1.25,2.5 mg/mL;最小杀菌浓度(MBC)分别是2.5,5.0 mg/mL。活体抑菌测定结果表明,11号和14号复配组合对油桃褐腐病菌有不同程度延缓侵染和抑制作用,但不能完全防治;0.05 mg/mL扑海因与11号组合20 mg/mL,0.10 mg/mL扑海因与11号组合10 mg/mL均能完全抑制褐腐病菌在油桃果实上的发生和蔓延。 相似文献
24.
G Scamarcio F Capasso C Sirtori J Faist AL Hutchinson DL Sivco AY Cho 《Science (New York, N.Y.)》1997,276(5313):773-776
A quantum-cascade long-wavelength infrared laser based on superlattice active regions has been demonstrated. In this source, electrons injected by tunneling emit photons corresponding to the energy gap (minigap) between two superlattice conduction bands (minibands). A distinctive design feature is the high oscillator strength of the optical transition. Pulsed operation at a wavelength of about 8 micrometers with peak powers ranging from approximately 0.80 watt at 80 kelvin to 0.2 watt at 200 kelvin has been demonstrated in a superlattice with 1-nanometer-thick AlInAs barriers and 4.3-nanometer-thick GaInAs quantum wells grown by molecular beam epitaxy. These results demonstrate the potential of strongly coupled superlattices as infrared laser materials for high-power sources in which the wavelength can be tailored by design. 相似文献
25.
MELISSA A. JACKSON ANDREW L. VIZARD GARRY A. ANDERSON JOHN S. MATTOON ROGER B. LAVELLE BRYAN T. SMITHENSON NOLA V. LESTER ANDREW F. CLARKE ROBERT C. WHITTON 《Veterinary radiology & ultrasound》2011,52(3):262-269
We aimed to identify common mistakes made when radiographing yearling sale horses. Radiographic examinations from repositories at eight yearling sales held in Australia in 2003 were assessed by one of four veterinary radiology specialists. Each radiographic examination consisted of a maximum of 34 radiographs. Each radiograph was assessed for errors associated with movement, exposure, positioning, labeling or marker placement, and processing, and categorized as either ideal, less than ideal or nondiagnostic. In addition, from the first 800 sets catalogued, 167 were selected randomly and read twice by the four radiologists for agreement analysis. A total of 81,297 radiographs were examined for errors affecting quality. Positioning errors were the most common reason for radiographs to be considered nondiagnostic (2432/81,297; 3%), with the flexed lateromedial (LM) metacarpophalangeal joint, LM metatarsophalangeal joint, and the dorsomedial palmarolateral (DMPaLO) carpal views being the most frequently involved. Overexposure (14,357/81,297; 17.7%) was the most common reason for radiographs being categorized as less than ideal with the LM stifle view the most represented. Agreement within and between radiologists for reporting errors in positioning of the flexed LM metacarpophalangeal joint, LM metatarsophalangeal joint, and DMPaLO carpal views varied from slight to almost perfect. The low repeatability within radiologists on some views suggests that before declaring a radiograph nondiagnostic it is worth considering rereading it at another time. Care should be taken in positioning of the flexed LM metacarpophalangeal, LM metatarsophalangeal, and DMPaLO carpus views to maximize radiograph quality. 相似文献
26.
SUMMARY: Linear regression analysis was used to describe the decline in numbers of Culicoides brevitarsis Kieffer into winter with monthly maximum, average and minimum temperatures at the southern limits to its distribution in New South Wales. From this, low temperature thresholds were derived when C brevitarsis would be absent from the field. The low minimum threshold ± 2 SE (95% Confidence Interval) of 8.1 ± 0.3°C was used with historical temperature data to estimate the last month that the species should occur (March to June) and the mean number of months (2 to 6.5) below the threshold at 17 selected sites. Probability for survival during winter at these sites was estimated from years when the number of consecutive months below the threshold was ≤ 2 months. This varied from zero to 51% depending on the location of the site. Last occurrence was 0.7 months later on average and absolute probabilities for survival ranged from zero to 100% when the temperatures were increased by an arbitrary 2°C. 相似文献
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