Feline Ubiquitin Fusion Protein Genes

Using cDNA from a CRFK cell line as a template, PCR amplification was performed with the Ub1S and poly(dT) primers to isolate feline ubiquitin genes. Sequencing of the 495 bp PCR fragment revealed that the putative amino acids induced by this fragment gave a fusion protein consisting of a ubiquitin polypeptide (76 amino acids) and an extension protein of ribosomal proteins L40 (52 amino acids). The putative amino acid sequence of ubiquitin was identical to those of humans, rats and pigs.The recombinant glutathione S-transferase (GST)–feline ubiquitin fusion proteins were produced in Escherichia coli and purified. The fusion proteins had a molecular weight of about 42 kDa and were detected by immunoblot assay with rabbit anti-ubiquitin antiserum.The mRNAs from heat-shocked and non-heat-shocked cells were subjected to RT-PCR (Ub1S and poly(dT) primers) analysis. The molecular weights of the ubiquitinated proteins in heat-shocked CFRK cells were between 18 kDa and 24 kDa by immunoblot assay.These results suggested that there were more ubiquinated proteins in the heat-shocked CRFK cells than in the pre-heat-shocked cells.     

Measurement of nitrate leaching losses from arable plots under different nitrogen input regimes

Abstract. Leaching losses of nitrate-nitrogen were measured from a set of eight hydrologically isolated plots on a clay loam soil over the period from September 1987 to February 1990. Variable drainflow recovery from the plots hampered accurate estimation of nitrate loading, but results suggest that, when inorganic nitrogen fertilizer is applied up to the recommended amount, there is little influence of the amount applied on the amount leached. We did, however, observe the following effects on nitrate leaching: leguminous green manure incorporated in autumn increased leaching of nitrate-nitrogen by 10–15 kg per hectare during the winter; autumn cultivation caused some increase in leaching compared with no cultivation in one year; some systematic variations in nitrate leaching occurred between years and between plots, but were unrelated to treatments.
From the results we conclude that green manuring does not provide sufficient nitrogen for organically grown crops on this soil but contributes significantly to nitrate leaching, and that growing spring cereals, with the land remaining in stubble as long as possible in autumn, may be the best strategy to minimize nitrate leaching.     

Investigations on the Occurrence of Hereditary Diseases in the Danish Cattle Population 1989-1991

Agerholm, J.S., A. Basse and K. Christensen: Investigations on the occurrence of hereditary diseases in the Danish cattle population 1989-1991. Acta vet. scand. 1993, 34, 245-253.– The methods of the Danish Bovine Genetic Disease Programme are outlined, and the results obtained during the first 3 years in function are described. The most common disease reported was spinal muscular atrophy in calves of the Red Danish Dairy breed with 312 reports. Necropsy was performed on 162 cases, and spinal muscular atrophy was diagnosed in 82 of these. Bovine progressive degenerative mye-loencephalopathy, rectovaginal constriction, syndrome of arthrogryposis and palatoschisis, hereditary chondrodysplasia (2 different types), syndactylism, epitheliogenesis imperfecta, and osteogenesis imperfecta was diagnosed with 1 case each. Lethal trait A46 was diagnosed in 4 calves. Some of these diseases have not previously been described in Denmark, and epitheliogenesis imperfecta was for the first time diagnosed in the Hereford breed. Chromosome translocation 1/29 was detected in the Blonde d’Aquitaine (BAQ), Limousine, and Red Danish Dairy breed. The aberration occurred frequently in BAQ. Furthermore, a complex chromosome translocation t(l;8;9)(q45;ql3;q26) was detected in the Red Danish Dairy breed.     

Feline leukemia virus detection by immunohistochemistry and polymerase chain reaction in formalin-fixed, paraffin-embedded tumor tissue from cats with lymphosarcoma.

The prevalence of feline leukemia virus (FeLV) antigen and DNA was assessed in formalin-fixed, paraffin-embedded tumor tissues from 70 cats with lymphosarcoma (LSA). Tissue sections were tested for FeLV gp70 antigen using avidinbiotin complex (ABC) immunohistochemistry (IHC); DNA was extracted and purified from the same tissue blocks for polymerase chain reaction (PCR) amplification of a 166 base pair region of the FeLV long terminal repeat (LTR). Results were related to antemortem FeLV enzyme-linked immunosorbent assay (ELISA) for serum p27 antigen, anatomic site of LSA, and patient age. Viral DNA was detected by PCR in 80% of cases and viral antigen by IHC in 57% of cases. Seventeen cases were PCR-positive and IHC-negative; one case was PCR-negative and IHC-positive. Clinical records included FeLV ELISA results for 30 of 70 cats. All 19 ELISA-positive cats were positive by PCR and IHC; of the 11 ELISA-negative cats that were negative by IHC, seven were positive by PCR. When evaluated according to anatomic site, FeLV DNA and antigen were detected less frequently in intestinal LSAs than in multicentric and mediastinal tumors. Lymphosarcoma tissues from cats < 7 yr were several fold more likely to be positive for FeLV antigen by IHC than were tumors from cats > or = 7 yr. However, there was no significant difference in PCR detection of FeLV provirus between LSAs from cats < 7 yr and those > or = 7 yr.(ABSTRACT TRUNCATED AT 250 WORDS)     

THE INSENSITIVITY OF 99mTc PERTECHNETATE FOR DETECTING METASTASES OF A FUNCTIONAL THYROID CARCINOMA IN A DOG

This report describes the use of ^99mtechnetium pertechnetate (^99mTcO₄⁻) and ¹³¹[for imaging of a metastatic thyroid carcinoma in a dog. The ¹³¹] imaging showed metastatic lesions undetected by the ^99mTcO₄⁻ imaging on 2 separate occasions. The possible mechanisms for the discrepancies between ¹³¹I and ^99mTcO₄⁻ imaging of thyroid carcinomas are discussed. The use of ¹³¹I for the imaging of functional thyroid carcinomas in the dog is recommended.     

Purification and characterization of hepatic triacylglycerol lipase isolated from rainbow trout,Oncorhynchus mykiss

Trialcylglycerol (TG) lipase was isolated and partially purified from rainbow trout liver. Triacylglycerol lipase activity was assayed by measuring¹⁴C-oleic acid release from¹⁴C-triolein.¹⁴C-oleic acid release was linear for up to two hours. Optimal activity occurred at pH 7.0 and 15°C. Most of the lipase activity was recovered in the cytosolic fraction. A 27,000-fold purification was achieved after Sepharose (Bio-gel A 0.5 M, 200–400 mesh) chromatography of a resuspended 20% ammonium sulfate fraction. The molecular weight of the trout hepatic lipase as determined by size-exclusion chromatography and by SDS-polyacrylamide gel electrophoresis was 40–43 kD. Lipase-mediated hydrolysis of TG resulted in the production of diacylglycerols, monoacylglycerols, and fatty acids. Kinetic analysis indicated that V_max=0.016 nmol/h/mg protein and that K_m=0.28 mM triolein. Lipolytic activity was enhanced in the presence of cAMP/ATP-Mg²⁺. These results suggest that the liver of trout possesses a neutral TG lipase that is responsible for mobilizing stored TG and is catalytically activated by phosphorylation.A part of this work was presented at the Annual Meeting of the American Society of Zoologists, December 26–30, 1990, San Antonio, TX.     

Plasma cholesterol and lipoprotein concentrations in the dog: The effects of age, breed, gender and endocrine disease

A combined ultracentrifugation and precipitation technique was used to quantify the plasma lipoprotein concentrations of control dogs (n=33) and dogs with diabetes mellitus (n=11), hyper-adrenocorticism (n=14), hypothyroidism (n=10) and obesity (n=20). In addition, the effect of breed type, age and gender on the lipoprotein phenotype was assessed. Breed type and age were found to have no effect upon cholesterol and lipoprotein concentrations but the high density lipoprotein cholesterol (HDL-C) concentration was greater in intact females than intact males. Cholesterol concentrations were significantly higher than those of the control group in dogs with diabetes mellitus (P<0·01), hyper-adrenocorticism (P<0·01) and hypothyroidism (P<0·001). In dogs with diabetes mellitus this was due to increased concentrations of very low density lipoprotein cholesterol (VLDL-C) (P<0·01) and HDL-C (P<0·05). The concentrations of low density lipoprotein cholesterol (LDL-C) (P<0·05) were significantly increased in dogs with hyperadrenocorticism, while in the hypothyroid dogs, VLDL-C (P<0·05), LDL-C (P<0·001) and HDL-C (P<0·05) were significantly higher than the control group. The cholesterol and lipoprotein concentrations in the obese population were not significantly different from control dogs.     

Influence of Low Light Irradiance on Grain Filling in Rice (Oryza saliva L.) cultivars

Time course of grain filling pattern in two rice (Oryza sativa L.) cultivars, grown under normal and low light intensities, was studied. The number of spikelets, partially filled grains and high density grains were monitored at a 5 day interval during grain filling period from uniform panicles tagged at anthesis. The low light tolerant cultivar Swarnaprabha had more high density grains and less partially filled grains throughout the grain filling period and at harvest compared to cultivar Ratna under both normal and low light intensities. Further, the opening of spikelets in two flushes in Swarnaprabha seemed to result in a more efficient resource allocation and hence better yield as compared to Ratna, in which the opening of spikelets was in a single flush on day 10.