A recombinant-based feline immunodeficiency virus antibody enzyme-linked immunosorbent assay.

We have developed an antibody detection enzyme-linked immunosorbent assay (ELISA) for the identification of animals infected by feline immunodeficiency virus (FIV). The ELISA solid-phase antigen consists of recombinant FIV gag proteins expressed in bacteria. The proteins are purified from bacterial lysates as insoluble inclusion bodies. In the case of bacterially expressed p24gag, it is shown that all of the linear, sequential epitopes presented by viral p24 during infection are retained. Purified preparations can be substituted for solid-phase whole virus in the IDEXX PetChektm immunoassay. The antibody ELISA duplicates the sensitivity and specificity of the whole virus based PetChek plate assay.     

Artificial Insemination with Frozen Semen in Ewes at Different Times of the Breeding Season

Totally 13575 ewes of two different breeds, Dala and Spel, were inseminated with semen, frozen in straws and thawed at 70°C for 8 sec. An insemination dose of 0.2 ml containing approx. 150 × 10⁶ spermatozoa with at least 45 to 50% progressive motility was imerted 5 to 12 mm into the cervix. The insemination was performed once between 12 and 30 h after the onset of heat. The NR rate of the Dala ewes increased significantly during the season. The NR rate of the ewes inseminated before 15. November was 44.3%, from 15. to 20. November 52.2%. from 20. to 25. November 55.3% and from 25. November and later 61.4%. The corresponding values for the ewes of the Spel breed were 57.3, 58.7, 61.5 and 71.0% respectively, and only the difference between the two last values was statistically significant. The difference between the fertility of the two breeds was significant within each of the periods .     

Cross reaction between bovine enterovirus and South African Territories I5 foot-and-mouth disease virus.

A bovine enterovirus (E76T) isolated from a 2-year-old bull produced serologic cross reactions to South African Territories (SAT) I5 foot-and-mouth disease virus when inoculated into guinea pigs and cattle. Cross-reacting serum titers to SAT I5 virus of 1:320 by the plaque-reduction neutralization test and 1:20 by the radial immunodiffusion test occurred in 2 steers after they were inoculated with the E76T virus. In 1 steer, maximal cross-reacting titers appeared related to a 2nd exposure to the viruses or to a hyperimmune state. Ultracentrifugation and 2-mercaptoethanol studies indicated that the cross reactions were due to immunoglobulin M antibody. Sera from guinea pigs immunized with the E76T or the SAT I5 virus cross reacted with the heterologous virus by postinoculation day 7. Cross-reacting titers had decreased markedly by postinoculation day 35, whereas the homologous virus titer remained constant. Cross reactivity of the E76T virus was primarily with the SAT I5 virus, and to a lesser degree with SAT II3. Cross reactions did not occur with representatives of the 5 other antigenic types of foot-and-mouth disease virus.     

Cross reactions of normal bovine sera with foot-and-mouth disease virus: incidence, duration, and effect of shipping stress.

Serum samples were obtained from 30 Hereford steers in an area known to be free of foot-and-mouth disease (FMD) viruses as follows: before shipment and 4 times during a 70-day period after shipment; the sera were tested for the presence of cross-reacting antibody to various viruses. Percentages of sera containing cross-reacting antibody to FMD virus detected by the plaque-reduction neutralization and the radial immunodiffusion techniques were higher for the FMD viruses Asia and SAT I5 than for the FMD viruses A5, O1, and C1. Cross-reacting antibody was usually of low titer and usually present in only 1 or 2 consecutive serum samples. The incidence of cross reactions increased after stress of shipping and thus an infective agent may be responsible. These results were compared with results from sera collected from Herefords and Holstein-Friesians in a 2nd area; results did not indicate that Herefords have an excess of cross reactions with FMD viruses.     

High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

五种鬼伞过氧化物酶和酯酶的同工酶研究

应用垂直板聚丙烯酰胺凝胶电泳对五种野生鬼伞 (Coprinus)真菌进行了过氧化物酶 (POD)和酯酶(EST)的同工酶分析 ,结果表明 :五种鬼伞的POD和EST同工酶酶谱比较稳定清晰 ,且分别有一条共同的酶带 ,可能是鬼伞属的POD和EST同工酶特征酶带 ;POD和EST同工酶酶谱均表明 ,家园鬼伞 (C .domesticus)和瓦鳞鬼伞 (C .clavatus)间有较近的亲缘关系 ;不同种鬼伞的POD和EST同工酶之间既有共同的特征 ,又各自有本物种的特有特征 ,POD和EST同工酶酶谱可以作为鬼伞属种类鉴定、亲缘关系比较的重要依据。     

Roles of the Kv1.2, Kv1.5, Kv2.1 potassium channels in hypoxia pulmonary vasoconstriction

AIM: To determine the role of Kv1.2, Kv1.5, Kv2.1 in the hypoxia pulmonary vasoconstriction (HPV). METHODS: Male Wistar rats were divided into two groups: normoxic group and hypoxic group. The single smooth muscle cell was obtained from pulmonary artery of Wistar rats with acute enzymatic digestion method. The conventional whole-cell patch clamp technique was used to record the resting membrane potential (Em) and the potassium currents of voltage-gated potassium channel (IKv) in rat pulmonary arterial smooth muscle cells (PASMC). Intracellular application of Kv1.2/Kv1.5/Kv2.1 antibodies (1∶125) was conducted through the whole-cell patch clamp system. RESULTS: ① Em of PASMC was depolarized after 24 h hypoxia compared with that of control cells . IKv of PASMC was decreased after 24 h hypoxia, . ② The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies depolarized Em and inhibited IKv in PASMC from normoxic rat, whereas the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on them. ③ The mixture of Kv1.2/Kv1.5/Kv2.1 antibodies and the mixture of Kir2.1/Kir2.3/Kir4.1 antibodies had no effects on IKv and Em from rats hypoxic for 24 h. CONCLUSION: Kv1.2, Kv1.5, Kv2.1 might be oxygen sensitive potassium channels which mediated HPV.     

Identification of Chlamydophila pneumoniae in an emerald tree boa, Corallus caninus.

Tissues were evaluated from emerald tree boas, Corallus caninus, from a collection in which chlamydiosis was diagnosed. To determine the strain of chlamydia infecting these snakes, tissue samples from 5 frozen snakes were tested by a quantitative TaqMan polymerase chain reaction (PCR) test and a PCR sequence analysis test. Of the 22 samples tested, 9 were categorized as either positive or weakly positive with the TaqMan test, and 6 yielded an amplicon using a serial PCR test that amplified a portion of the 23S ribosomal RNA gene. A PCR product suitable for sequencing was obtained from the heart of one of the snakes. Sequence analysis showed that the snake had been infected with Chlamydophila pneumoniae. These findings show that C. pneumoniae can infect emerald tree boas, broadening the range of reptiles known to be infected by this primarily human pathogen.     

In vitro studies of Norwegian Red bovine semen immobilized and cryopreserved in alginate solid gel network

Development of new semen cryopreservation techniques improving sperm survival and ensuring availability of viable spermatozoa for a prolonged time‐period after AI is promising tools to reduce sensitivity of timing of AI and enhance overall fertility. The SpermVital^® technology utilizes immobilization of bull spermatozoa in a solid network of alginate gel prior to freezing, which will provide a gradual release of spermatozoa after AI. The objective of this study was to compare post‐thaw sperm quality and in vitro sperm survival over time of Norwegian Red bull semen processed by the SpermVital^® (SV) technology, the first commercialized production line of SpermVital^® (C) and by conventional procedure applying Biladyl^® extender (B). Post‐thaw sperm motility was not significantly different between SV, C and B semen (p > .05). However, sperm viability and acrosome intactness were higher for SV than C and B semen (p < .05). Small differences in DNA quality were observed (p < .05). Sperm viability after storage in uterus ex vivo was higher for SV than for C semen (p < .05). Furthermore, sperm survival in vitro over time at physiological temperature was significantly higher for SV semen than C semen as well as B semen during the incubation period of 48 hr (p < .05). In conclusion, the SpermVital^® technology is improved and is more efficient in conserving post‐thaw sperm quality and results in higher sperm viability over time in vitro for SV than for C and B semen.     

Biochar Mitigates Salinity Stress in Potato

A pot experiment was conducted in a climate‐controlled greenhouse to investigate the growth, physiology and yield of potato in response to salinity stress under biochar amendment. It was hypothesized that addition of biochar may improve plant growth and yield by mitigating the negative effect of salinity through its high sorption ability. From tuber bulking to harvesting, the plants were exposed to three saline irrigations, that is 0, 25 and 50 mm NaCl solutions, respectively, and two levels of biochar (0 % and 5 % W/W) treatments. An adsorption study was also conducted to study the Na⁺ adsorption capability of biochar. Results indicated that biochar was capable to ameliorate salinity stress by adsorbing Na⁺. Increasing salinity level resulted in significant reductions of shoot biomass, root length and volume, tuber yield, photosynthetic rate (A_n), stomatal conductance (g_s), midday leaf water potential, but increased abscisic acid (ABA) concentration in both leaf and xylem sap. At each salinity level, incorporation of biochar increased shoot biomass, root length and volume, tuber yield, A_n, g_s, midday leaf water potential, and decreased ABA concentration in the leaf and xylem sap as compared with the respective non‐biochar control. Decreased Na⁺, Na⁺/K⁺ ratio and increased K⁺ content in xylem with biochar amendment also indicated its ameliorative effects on potato plants in response to salinity stress. The results suggested that incorporation of biochar might be a promising approach for enhancing crop productivity in salt‐affected soils.