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.     

Exposure of Ankole and crossbred cattle to theileriosis in Rwanda

Summary Susceptible Ankole (Sanga:Bos indicus/Bos taurus) and crossbred Ankole x Jersey (B. taurus) and Ankole x Sahiwal (B. indicus) cattle derived from a farm in Rwanda with no recent history of theileriosis, were infected withTheileria parva stocks from Rwanda either by feeding infectedRhipicephalus appendiculatus ticks on the ears, inoculation of tick derived stabilate or natural exposure to tick challenge. The Ankole cattle originated from local stock born and bred in East Coast fever (ECF) endemic areas of Rwanda. Disease, followed by spontaneous recovery, was observed in 49 of the 72 Ankole cattle after infection withT. parva (68%); the other 23 animals (32%) died of ECF. In contrast 21 of the 33 infected crossbred cattle (64%) died of ECF. It is concluded that the partialTheileria tolerance of the Ankole is, to a great extent, genetic. The basis of this partial tolerance seems to be their ability to limit the explosive multiplication of macroschizonts during the acute phase of the disease.

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Resumen Se infectó ganado Ankole susceptible (Sanga:Bos indicus/Bos taurus) y ganado cruzado Ankole x Jersey (Bos taurus) y Ankole x Sahiwal (Bos indicus) proveniente de una finca en Rwanda sin historia reciente de theileriosis, conTheileria parva procedente de Rwanda, mediante la adhesión en la oreja deRhipicephalus appendiculatus infectado, inoculación de estabilados derivados de garrapatas, o exposición natural a la enfermedad a través del vector. El ganado Ankole era originario de un área endémica de theileriosis en Rwanda. Se observó la enfermedad seguida de recuperación, en 49 de 72 animales Ankole infectados conT. parva (68%); los otros 23 animales murieron (32%). En contraste, 21 de los 33 animales cruzados infectados (64%), murieron. Se concluye, que la tolerancia parcial del ganado Ankole es de origen genético. Las bases de ésta tolerancia genética, parece debida a la habilidad para limitar en cierto modo la multiplicación explosiva de macroesquizontes, durante la fase aguda de la enfermedad.

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Résumé On a infecté du bétail sensible, de race Ankolé (Sanga:Bos indicus/Bos taurus) et des croisés Ankolé x Jersey (B. taurus) et Ankolé x Sahiwal (B. indicus), provenant d'une ferme au Rwanda sans cas récents de theilériose, avec des souches deTheileria parva du Rwanda, par l'application de tiques (Rhipicephalus appendiculatus) infectées sur les oreilles, par inoculation de stabilat de tiques infectées ou par l'exposition à un challenge naturel de tiques. La souche d'Ankolé était originaire du cheptel local Rwandais, né et élevé dans les régions où la theilériose est endémique. La maladie suivie par la guérison spontanée fut observée dans 49 des 72 Ankolés (68%), les autres 23 animaux (32%) ont succombé à la theilériose. Par contre 21 des 33 croisées infectées (64%) ont succombé à la theilériose. Nous concluons que la tolérance partielle des Ankolés contre la theilériose, est génétique. La base de cette tolérance semble être leur capacité de limiter la multiplication explosive des macroschizonts pendant la phase aigu de la maladie.

     

Advances in diagnostic ultrasonography

A wide variety of ultrasonographic equipment currently is available for use in equine practice, but no one machine is optimal for every type of imaging. Image quality is the most important factor in equipment selection once the needs of the practitioner are ascertained. The transducer frequencies available, transducer footprints, depth of field displayed, frame rate, gray scale, simultaneous electrocardiography, Doppler, and functions to modify the image are all important considerations. The ability to make measurements off of videocassette recorder playback and future upgradability should be evaluated. Linear array and sector technology are the backbone of equine ultrasonography today. Linear array technology is most useful for a high-volume broodmare practice, whereas sector technology is ideal for a more general equine practice. The curved or convex linear scanner has more applications than the standard linear array and is equipped with the linear array rectal probe, which provides the equine practitioner with a more versatile unit for equine ultrasonographic evaluations. The annular array and phased array systems have improved image quality, but each has its own limitations. The new sector scanners still provide the most versatile affordable equipment for equine general practice.     

Postovulatory Effect of Intravenous Administration of Lipopolysaccharide (Escherichia coli, O55:B5) on the Endocrine Status of Recently Ovulated Sows

The effects of lipopolysaccharide ( Escherichia coli , O55:B5), administered 18 h after ovulation in the second oestrus after weaning on the hormonal profiles in 14 Swedish cross-bred (Landrace × Yorkshire) multiparous sows were studied. The endotoxin group (E-group) sows were administered with 300 ng/kg of lipopolysaccharide (LPS) whereas the control group (C-group) sows were administered 5 ml of saline intravenously via an indwelling jugular cannula. Blood samples for hormonal analyses were collected from all sows until slaughter. In the E-group, progesterone, cortisol and prostaglandin F_2α metabolite levels increased significantly (p < 0.05) following LPS compared with the C-group. It can be concluded from this study that apart from elevating cortisol and prostaglandin F_2α metabolite, LPS also elevates progesterone levels.