A novel HPLC method for glutathione-insulin transhydrogenase assay using fluorescence labeled insulin

The activity of rat liver glutathione-insulin transhydrogenase (GIT) was measured by HPLC. The degradation of fluorescein isothiocyanate-I (FITC-I)-labeled insulin is separated into several peaks, which are bound different amount of FITC-I. We selected mono-fluorescein-thiocarbamylated insulin to estimate the decrease of insulin content and it became possible to assay GIT activity. This novel method was time-saving and simple, and this system could utilize instead of previous method.     

Changes in the concentration of mRNAs for the inhibin subunits in ovarian follicles after administration of gonadotropins to progestin treated ewes.

RNA was extracted from single or small groups of ovine ovarian follicles after treatment of ewes with FSH and/or LH. The content of mRNA for the alpha-inhibin and beta A-inhibin subunits was analyzed by hybridization with specific cDNA probes. All ewes were treated with progestin vaginal pessaries to suppress spontaneous preovulatory follicle maturation and ewes were given three intramuscular injections of gonadotropins at 8-hr intervals starting 24 hr prior to collection of ovaries. In experiment I, both Schering-FSH and NIDDK-oFSH-17 (oFSH) significantly increased alpha- and beta A-inhibin mRNA per ewe in 2-5 mm follicles and tended to increase alpha- and beta A-inhibin mRNA in large (greater than 5 mm) follicles. In experiment II, oFSH and NIDDK-oLH-25 (oLH) were administered in a 2X2 factorial arrangement. Separate administration of oFSH or oLH increased (P less than .05) the alpha-inhibin mRNA concentration in large follicles. alpha-inhibin mRNA concentration in 4-5 mm follicles was also increased by oFSH but was decreased by oLH. Concomitant treatment with oFSH and oLH did not change alpha-inhibin mRNA concentrations from those measured in oFSH treated ewes. In experiment II, beta A mRNA concentrations followed a pattern similar to that of alpha A mRNA, but the differences were not statistically significant. We conclude that, in the ewe, exogenous FSH increases the concentration of inhibin mRNA in the whole follicle. The ability of exogenous oLH to alter expression of the inhibin subunit genes may depend upon the stage of follicle maturation.     

In situ hybridization analysis of ovarian prostaglandin endoperoxide synthase mRNA throughout the periovulatory period of the ewe.

Cellular alterations in level of expression of mRNA encoding for prostaglandin endoperoxide synthase were quantified within ovarian tissues of sheep obtained before, during and after induction of the preovulatory surge of LH and ovulation with LHRH. This was accomplished by isotopic in situ hybridization using a selective cRNA probe to ovine prostaglandin endoperoxide synthase mRNA. A significant elevation in mRNA was detected within the theca interna of the preovulatory follicle at 8, 16 and 24 hr following administration of LHRH. Very close to the time of ovulation (ie., at 24 hr post-LHRH) a marked rise in mRNA was observed in association with epithelial cells covering the apical surface of the follicle. Ovarian cyclooxygenase metabolites of arachidonic acid produced during the ovulatory process in the ewe originate within the thecal layer and germinal epithelium of the follicle destined to ovulate.     

N_(98s)×特青(亚种间杂交稻)抽穗后物质生产及分配特点研究初报

对N×特青示范田中试区与肥料试验田各处理小区稻株分析结果表明:抽穗后,亚种间杂交稻N×特青比常规稻特青干物质生产能力强,灌浆速度快;地上部各器官占植株干重比例,前者叶片比重大于茎,后者茎比重大于叶。亚种间杂交稻对肥料反应敏感,抽穗后,干物质累积,植株高度,每穗总粒数和粒重均随施肥量增加而增加,在高肥区有利于进一步提高产量,但增施氮肥后,灌浆速度明显滞后。     

Trigger Points in 48 Dogs with Myofascial Pain Syndromes

Seven foci of pain (trigger points) were identified in the triceps brachii, infraspinatus, adductor-pectineus, peroneus longus, gluteus medius, ileocostorum lumborum, and quadriceps femoris muscles in 48 lame dogs. The dogs had been lame for 1 day to 150 weeks (mean, 24 weeks). Thirty-one dogs had been treated unsuccessfully with corticosteroids, nonsteroidal anti-inflammatory drugs, analgesics, or acupuncture. Palpating the trigger points induced severe pain. Treatment consisted of weekly stimulation of the trigger points by needling or injection of a local anesthetic. The mean treatment period was 2.8 weeks. Excellent results and complete recovery were observed in 34 dogs (60%).     

Importance of Some Biochemical Parameters in the Quality and Yield of Rapeseed (Brassica napus L.)

Samples of rapeseed from three Italian growing environments (Bologna, Perugia and Palermo) were analysed for glucose content and dry weight of 1000 seeds every three or four days starting from the end of flowering until complete ripening. In addition, the content of oil, soluble and total proteins, glucosinolates and myrosinase activity was determined in samples of mature seeds. The cultivars used were jet Neuf and Lingot (type 0) and Tandem, Jade and Santana (type 00). From the results it emerged that the point of intersection of the two branches of the linear regression plots for different glucose-consumption kinetics found during seed filling, in addition to being strongly affected by the climate of the test environment, is correlated with quantitative and qualitative production, independently of the genotype.     

Humoral immune responses of black-footed penguins (Spheniscus demersus) after DNA-mediated immunization with a beta-galactosidase reporter gene.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a beta-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-beta, containing the beta-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-beta-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to beta-galactosidase were detected in the sera of pCMV-beta-inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding beta-galactosidase and will mount a humoral response against the in vivo-expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.