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71.
Vesicles Cytoplasmic Injection: An Efficient Technique to Produce Porcine Transgene‐Expressing Embryos
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CG Luchetti RJ Bevacqua MS Lorenzo MF Tello M Willis CP Buemo DM Lombardo DF Salamone 《Reproduction in domestic animals》2016,51(4):501-508
The use of vesicles co‐incubated with plasmids showed to improve the efficiency of cytoplasmic injection of transgenes in cattle. Here, this technique was tested as a simplified alternative for transgenes delivery in porcine zygotes. To this aim, cytoplasmic injection of the plasmid alone was compared to the injection with plasmids co‐incubated with vesicles both in diploid parthenogenic and IVF zygotes. The plasmid pcx‐egfp was injected circular (CP) at 3, 30 and 300 ng/μl and linear (LP) at 30 ng/μl. The experimental groups using parthenogenetic zygotes were as follows: CP naked at 3 ng/μl (N = 105), 30 ng/μl (N = 95) and 300 ng/μl (N = 65); Sham (N = 105); control not injected (N = 223); LP naked at 30 ng/μl (N = 78); LP vesicles (N = 115) and Sham vesicles (N = 59). For IVF zygotes: LP naked (N = 44) LP vesicles (N = 94), Sham (N = 59) and control (N = 79). Cleavage, blastocyst and GFP+ rates were analysed by Fisher's test (p < 0.05). The parthenogenic CP naked group showed lower cleavage respect to control (p < 0.05). The highest concentration of plasmids to allow development to blastocyst stage was 30 ng/μl. There were no differences in DNA fragmentation between groups. The parthenogenic LP naked group resulted in high GFP rates (46%) and also allowed the production of GFP blastocysts (33%). The cytoplasmic injection with LP vesicles into parthenogenic zygotes allowed 100% GFP blastocysts. Injected IVF showed higher cleavage rates than control (p < 0.05). In IVF zygotes, only the use of vesicles produced GFP blastocysts. The use of vesicles co‐incubated with plasmids improves the transgene expression efficiency for cytoplasmic injection in porcine zygotes and constitutes a simple technique for easy delivery of plasmids. 相似文献
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73.
Genotypic diversity in field isolates of Babesia bovis from cattle with babesiosis after vaccination
AE LEW RE BOCK JM CROFT CM MINCHIN TG KINGSTON RJ DALGLIESH 《Australian veterinary journal》1997,75(8):575-578
Objective To determine whether particular genotypes of Babesia bovis were common to field isolates obtained from cattle properties in Queensland where the B bovis vaccine had apparently failed.
Design A comparative study of polymerase chain reaction genotypes in different populations of B bovis .
Procedure Two polymerase chain reaction assays were applied to analyse DNA extracts of B bovis vaccine (K, T and Dixie strains) and 27 field isolates from 24 properties where disease outbreaks had occurred despite the use of the vaccine. To evaluate the stability of the genotypes identified, 11 of the field isolates were inoculated into experimental cattle that had either been previously vaccinated with T strain or not vaccinated.
Results No particular genotype of B bovis was responsible for the problems observed in previously vaccinated herds. None of the isolates had genotypes identical to the vaccine strains used. No geographic trends among the genotypes were observed. Isolates that originated from the same property also had different genotypes. Blood passage of the 11 field isolates in either previously vaccinated or nonvaccinated cattle did not alter the original genotype.
Conclusion No particular genotypes identified by the Bv80 and BvVA1 polymerase chain reaction assays could be associated with vaccine failures. 相似文献
Design A comparative study of polymerase chain reaction genotypes in different populations of B bovis .
Procedure Two polymerase chain reaction assays were applied to analyse DNA extracts of B bovis vaccine (K, T and Dixie strains) and 27 field isolates from 24 properties where disease outbreaks had occurred despite the use of the vaccine. To evaluate the stability of the genotypes identified, 11 of the field isolates were inoculated into experimental cattle that had either been previously vaccinated with T strain or not vaccinated.
Results No particular genotype of B bovis was responsible for the problems observed in previously vaccinated herds. None of the isolates had genotypes identical to the vaccine strains used. No geographic trends among the genotypes were observed. Isolates that originated from the same property also had different genotypes. Blood passage of the 11 field isolates in either previously vaccinated or nonvaccinated cattle did not alter the original genotype.
Conclusion No particular genotypes identified by the Bv80 and BvVA1 polymerase chain reaction assays could be associated with vaccine failures. 相似文献
74.
Objective To determine the frequency of damage to the medial palmar intercarpal ligament (MPICL), and the range of sizes of the dorsomedial intercarpal ligament (DMICL) of the midcarpal joint in horses with no history of carpal joint disease.
Materials and methods
Cadaver limbs were collected from 72 horses with no history of carpal joint disease. One hundred and forty-two midcarpal joints were dissected and the MPICL and DMICL were examined. Measurements were made with a digital micrometer.
Results MPICL tearing was present in 88 of 96 joints from horses 2 years and older. Tears were predominantly of the dorsolateral bundle and complete rupture of the dorsolateral and dorsomedial bundles was not observed. Tearing was not present in foals less than 4 months of age and the severity of tearing increased significantly with age (P < 0.0001). Severity of tearing was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01), but there was no significant difference between racing and non-racing horses. The lateromedial thickness of the DMICL ranged from 0.4 mm to 2.6 mm in horses 2 years and older. Lateromedial thickness increased significantly with age, and was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01). There was no significant difference between racing and nonracing horses.
Conclusions Damage confined to the dorsolateral bundle of the MPICL is a common finding in horses over 1 year of age and is probably of little clinical significance. Complete rupture of both dorsolateral and dorsomedial bundles is uncommon in horses with no history of midcarpal joint disease. Variation in size of the DMICL is observed in horses of all ages, but is most marked in 2-year-old horses. 相似文献
Materials and methods
Cadaver limbs were collected from 72 horses with no history of carpal joint disease. One hundred and forty-two midcarpal joints were dissected and the MPICL and DMICL were examined. Measurements were made with a digital micrometer.
Results MPICL tearing was present in 88 of 96 joints from horses 2 years and older. Tears were predominantly of the dorsolateral bundle and complete rupture of the dorsolateral and dorsomedial bundles was not observed. Tearing was not present in foals less than 4 months of age and the severity of tearing increased significantly with age (P < 0.0001). Severity of tearing was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01), but there was no significant difference between racing and non-racing horses. The lateromedial thickness of the DMICL ranged from 0.4 mm to 2.6 mm in horses 2 years and older. Lateromedial thickness increased significantly with age, and was significantly greater in racing Standardbreds than racing Thoroughbreds (P < 0.01). There was no significant difference between racing and nonracing horses.
Conclusions Damage confined to the dorsolateral bundle of the MPICL is a common finding in horses over 1 year of age and is probably of little clinical significance. Complete rupture of both dorsolateral and dorsomedial bundles is uncommon in horses with no history of midcarpal joint disease. Variation in size of the DMICL is observed in horses of all ages, but is most marked in 2-year-old horses. 相似文献
75.
Bressan M Liberatore L d'Alessandro N Tonucci L Belli C Ranalli G 《Journal of agricultural and food chemistry》2004,52(5):1228-1233
A novel system was investigated, finalized to reduce the impact of highly polluting wastewaters, and based on combined actions of catalytic oxidations and microbial biotechnologies. Olive oil mill wastewaters (COD 10,000-100,000 mg O(2)/L) were oxidized up to 80-90% by stoichiometric amounts of dilute hydrogen peroxide (35%) and in the presence of water soluble iron catalysts, either Fe(II) or Fe(III), at concentrations up to 1% w/w and more, i.e., much larger than those reported for conventional Fenton processes. In the combined action, the mineralization activity of a selected microbial consortium was used to degrade residual volatile and nonvolatile organic compounds into CO(2) and biomass. The results of this search could suggest an improved operational methodology capable to reduce the potential impact of wastewater. 相似文献
76.
77.
In a pen feeding trial fresh Echium plantagineum was fed as the sole diet to crossbred sheep with or without a history of previous access to the plant. Control groups received a diet of lucerne chaff and oats. During the trial, sheep on the Echium diet lost weight and deaths occurred with histological evidence of excessive copper accumulation, usually accompanied by pyrrolizidine alkaloid damage, in the liver and biochemical evidence of liver toxicity. It is concluded that E. plantagineum alone is not a suitable fodder for sheep and can be toxic due to its pyrrolizidine alkaloid content and high copper to molybdenum ratio. 相似文献
78.
Major chemical exchange between the crust and mantle occurs in subduction zone environments, profoundly affecting the chemical evolution of Earth. The relative contributions of the subducting slab, mantle wedge, and arc lithosphere to the generation of island arc magmas, and ultimately new continental crust, are controversial. Isotopic data for lavas from a transect of volcanoes in a single arc segment of northern Honshu, Japan, have distinct variations coincident with changes in crustal lithology. These data imply that the relatively thin crustal lithosphere is an active geochemical filter for all traversing magmas and is responsible for significant modification of primary mantle melts. 相似文献
79.
80.
RJ Schoelkopf P Wahlgren AA Kozhevnikov P Delsing DE Prober 《Science (New York, N.Y.)》1998,280(5367):1238-1242
A new type of electrometer is described that uses a single-electron transistor (SET) and that allows large operating speeds and extremely high charge sensitivity. The SET readout was accomplished by measuring the damping of a 1.7-gigahertz resonant circuit in which the device is embedded, and in some ways is the electrostatic "dual" of the well-known radio-frequency superconducting quantum interference device. The device is more than two orders of magnitude faster than previous single-electron devices, with a constant gain from dc to greater than 100 megahertz. For a still-unoptimized device, a charge sensitivity of 1.2 x 10(-5) e/hertz was obtained at a frequency of 1.1 megahertz, which is about an order of magnitude better than a typical, 1/f-noise-limited SET, and corresponds to an energy sensitivity (in joules per hertz) of about 41 Planck's over 2pi. 相似文献