ras p21 expression in ovine pulmonary carcinoma

We have adapted an enzyme-linked immunoblot assay (ELIBA) for the detection of a c-ras proto-oncogene and oncogene protein products in human cell lines and tumors of 21,000 daltons molecular weight (p21ras) to studies of tissues derived from sheep. In the ELIBA, a double antibody system is used in which p21ras proteins are initially immunoprecipitated from protein extracts with monoclonal antibodies, and subsequently identified using additional anti-ras antibodies. Binding is identified with a non-radioactive enzyme-linked colorimetric detection system. In the present study, the ELIBA system was used to study twenty-seven ovine lung specimens, representing normal lung, inflammatory, and neoplastic lesions. We detected p21ras protein expression in every tissue examined, but the nature and amount of the protein product varied significantly among the tissues examined. Some tissues expressed multiple ras species. Broncho-alveolar carcinoma specimens were most likely to express c-Ki-ras proteins. Mutant proteins of c-N-ras and c-Ki-ras were detected in several bronchoalveolar carcinoma specimens, based on migrational differences between mutant and normal proteins in 15% polyacrylamide gels. The results of this study demonstrate the utility of the ELIBA system for detection of c-ras expression in ovine lung tissues, and demonstrate the ability of the system to discriminate specific ras protein species. The prognostic significance of ras expression in sheep pulmonary carcinoma has yet to be determined.     

A bioassay method for formulation testing and residue studies of sulfonylurea and sulfonanylide herbicides

A bioassay method using the radicles of pea (Pisum sativum L.) and lupin (Lupinus angustifolius L.) was developed for the assessment of trials on herbicides of common use in the sulfonylurea class (chlorsulfuron, triasulfuron and metsulfuron-methyl) and in the sulfonanylide class (flumetsulam and metosulam). Soils within a range of pH 5.8–8.4 with textures from sand to clay were used in these experiments. The sensitivities of the species were similar in chlorsulfuron and flumetsulam trials and their response range varied with soil type and herbicide, e.g. between 0.75 and 6.0 ng triasulfuron g^?1 in the Wimmera grey clay and between 0.125 and 8.0 ng chlorsulfuron g^?1 soil in the Mallee sand. The method was demonstrated in a wide range of uses, encompassing tests of the initial bioactivity of formulations of chlorsulfuron and flumetsulam, monitoring the field leaching and persistence of triasulfuron and measuring relative potencies between the classes, using metsulfuron-methyl and metosulam. The bioassay response provided a high level of reproducibility and precision, which was measurable by the logistic curve-fitting procedure. In each case, R² values were >0.90 and lack-of-fit tests were clearly non-significant at the 0.05 level. Chi-square tests were used to measure differences between ED₅₀'s. The method does not require the pre-germination and selection of seedlings, daily watering or root-washing and results are obtained 7 days from sowing, providing favourable use for routine analyses and large-scale trials.     

Occurrence of halo spot in barley caused by Pseudoseptoria donacis in Argentina

Barley is an important malting crop extensively cultivated in the province of Buenos Aires (AR). In 1992, an unusual disease was detected with a prevalence of 8% on 36 commercial fields examined in this province. The pattern of the disease, the characteristics of the pathogen and the test for pathogenicity indicated that the causal agent was Pseudoseptoria donacis.     

Bovine Leukocyte Adhesion Deficiency in Danish Holstein-Friesian Cattle: I. PCR Screening and Allele Frequency Estimation

Jergensen, C. B., J. S. Agerholm, J. Pedersen and P. D. Thomsen: Bovine leukocyte adhesion deficiency in Danish Holstein-Friesian Cattle I. PCR screening and allele frequency estimation. Acta vet. Scand. 1993,34,231-236.–A screening program for bovine leukocyte adhesion deficiency (BLAD) in Danish Holstein-Friesian cattle has been initiated. During the first months 1611 animals were tested by a PCR based assay. Of these animals 1256, 346, and 8 were assigned normal, BLAD carriers, and BLAD affected animals, respectively One bull, born as a co-twin, showed weak reaction for the BLAD allele on DNA isolated from leukocytes, but a normal genotype on DNA isolated from semen. Chromosome analysis showed that this bull was a blood chimaera. Estimation of the BLAD allele frequency upon the PCR test results showed that around 450 Danish calves born in 1991 might have been affected with the recessive disorder.     

Multiresidue assay for benzimidazole anthelmintics by liquid chromatography and confirmation by gas chromatography/selected-ion monitoring electron impact mass spectrometry

A multiresidue method utilizing all-disposable labware has been developed for 8 benzimidazole anthelmintics from ovine, bovine, and swine muscle and liver tissues. After an initial extraction with ethyl acetate and subsequent evaporation, a 3-component extraction using hexane, ethanol, and 0.2N HCl was used for final cleanup. Clean extracts were produced for separation and determination by reverse-phase liquid chromatography at 298 nm, using methanol and aqueous buffer as mobile phase. A synthesized internal standard, 2-(n-butylmercapto)benzimidazole, was used for quantitation of all drugs. Results are included along with statistical information verifying the performance of the method. Spiked control tissues and incurred drug tissues were used for an intralaboratory study with a concentration range of 50-1470 ppb. A series of standard curves at 0, 50, 100, and 200 ppb were analyzed. Overall recovery at the 100 ppb level averaged 92% (CV 8%) in liver tissues, across all 3 species and 88% (CV 5%) in muscle tissues across all 3 species. Results were confirmed by gas chromatography/mass spectrometry with acid hydrolysis of the remaining extract in 2N HCl followed by re-extraction of the amine and derivatization to the tert-butyldimethylsilyl derivative. The anthelmintics were identified by gas chromatography/selected ion monitoring electron-impact mass spectrometry. Ion ratio measurements were taken and compared to standard material. CVs averaged 10% or less for all drugs tested.     

Experience with Cyclosporin-A After Renal Allografting in Two Dogs Research Note

After renal allografting, cyclosporin-A was administered to one partially nonmatched dog that was followed for 79 days. Cyclosporin-A and prednisolone were administered to one nonmatched dog that was followed for 805 days. Side effects encountered with cyclosporin-A included lymphocytic dermatitis, papillomatosis, bacterial and fungal infections, and B lymphocyte hyperplasia.     

Serologic response and lesions in goats experimentally infected with Corynebacterium pseudotuberculosis of caprine and equine origins

Fifteen goat kids were experimentally inoculated with Corynebacterium pseudotuberculosis. Five were given a strain of caprine origin (nitrate-negative biotype) intradermally, 5 were given a strain of equine origin (nitrate-positive biotype) intradermally, and 5 were inoculated intranasally with the caprine-origin strain. Animals were monitored for 127 days. The goats given the inocula intradermally developed abscesses; those given caprine-origin strain had multiple lesions both peripherally and in visceral locations (primarily endothoracic abscesses), whereas those given the equine-origin strain had abscesses only at injection sites and draining nodes. The difference in extent of lesions could be due to biotypic bacterial differences or to the individual strains used. Intranasally inoculated goats did not develop abscesses and were essentially no different from controls. The cranial part of the respiratory tract may not be an important portal of entry for C pseudotuberculosis. Serum samples obtained monthly from all animals were subjected to the synergistic hemolysis-inhibition test, which measures antibodies to the exotoxin of C pseudotuberculosis. Animals with abscesses developed titers within 1 month of inoculation. Animals without abscesses remained seronegative. The synergistic hemolysis-inhibition test may be a reliable diagnostic assay for caseous lymphadenitis in goats.     

Pharmacokinetic and Phase I Evaluation of Carboplatin in Dogs

Thirty dogs with spontaneously occurring malignant neoplasms were treated monthly with carboplatin (CBDCA) given as a 30-minute intravenous infusion in a dose escalation study. Twenty-eight dogs were considered evaluable for toxicity. The maximally tolerated dose of CBDCA was conceptually defined as that dose, determined by logistic regression analyses of toxicity data, resulting in a 50% incidence of moderate toxicity (MOD₅₀) or a 5% incidence of severe toxicity (SEV₅). Each designated maximally tolerated dose was calculated for the first course of treatment only and for the first and second courses of treatment combined to estimate cumulative drug toxicity. The MOD₅₀ and SEV₅ for the first treatment course were 340 and 278 mg/M², respectively. MOD₅₀ and SEV₅ values for the first plus second treatment courses were 327 and 231 mg/M², respectively. The nadir of neutrophil and platelet counts occurred approximately 14 days after treatment. The mean neutrophil and platelet values for all dogs experiencing myelosuppression during the first two treatment courses were 1541/μL and 62,600/μL, respectively. Nonparametric pharmacokinetic analysis of plasma CBDCA values suggested that half-life (T_1/2), area-under-the-curve and total body clearance (CL_b) were not dose dependent. Volume of distribution (VD_ss) significantly increased with dose only between 100 and 150 mg/M², not between 150 and 300 mg/M². Dose-independent serum CBDCA pharmacokinetic disposition indicates that detailed investigation of tissue CBDCA distribution would be warranted and may identify novel dosing strategies that could improve the therapeutic index of CBDCA by minimizing toxicity. (Journal of Veterinary Internal Medicine 1993; 7:235–240. Copyright © 1993 by the American College of Veterinary Internal Medicine.)