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N L Hubbert

Publications and source records attributed to N L Hubbert.

24 records · Page 2Linked to original sources

Harvey murine sarcoma virus p21 ras protein: biological and biochemical significance of the cysteine nearest the carboxy terminus.

Previous studies of premature chain termination mutants and in frame deletion mutants of the p21 ras transforming protein encoded by the transforming gene of Harvey murine sarcoma virus (Ha-MuSV) have suggested that the C terminus is required for cellular transformation, lipid binding, and membrane localization. We have now further characterized the post-translational processing of these mutants and have also studied two C-terminal v-rasH point mutants: one encodes serine in place of cysteine-186, the other threonine for valine-187. The Thr-187 mutant was transformation-competent, and its p21 protein was processed normally, as was the p21 encoded by a transformation-competent deletion mutant from which amino acids 166-175 had been deleted. The Ser-186 mutant was defective for transformation. The p21s encoded by the Ser-186 mutant and by the previously described transformation-defective mutants did not undergo the posttranslational processing common to biologically active ras proteins: their electrophoretic migration rate did not change, they remained in the cytosol, and they failed to bind lipid. Since the cell-encoded ras proteins also contain this cysteine, we conclude that this amino acid residue is required for all ras proteins.

Animals↗

Serum proteins in healthy Basenjis and Basenjis with chronic diarrhea.

A serologic survey was conducted to characterize the electrophoretic patterns of serum proteins in healthy Basenji dogs (n = 137) and Basenjis with chronic diarrhea (n = 32). Serum protein electrophoresis values for Basenjis were similar to previously reported values from dogs of other breeds. Dogs with histologically confirmed lymphocytic-plasmacytic enteritis had a statistically significant (P less than 0.05) decrease in total protein, albumin, and albumin/globulin ratio. alpha 2-Globulin and gamma-globulin values were significantly increased in old dogs (9 years of age or older) with lymphocytic-plasmacytic enteritis. Although not statistically significant, gamma-globulin values were generally increased in Basenjis with lymphocytic-plasmacytic enteritis when compared with the values in age-matched clinically healthy Basenjis.

Alpha-Globulins↗

Counterimmunoelectrophoresis (immunoelectroosmosis) and serum electrophoretic pattern in serologic diagnosis of canine blastomycosis.

Counterimmunoelectrophoresis (CIEP) with blastomyces and histoplasma antigens was used in a serologic study of 181 dogs clinically suspected of having blastomycosis and of 8 dogs with confirmed blastomycosis or histoplasmosis. Thirteen of the 181 dogs, positive by CIEP, were euthanatized, and the diagnosis was confirmed by cultivation and/or microscopic detection of Blastomyces dermatitidis. Additional CIEP-positive dogs were confirmed by staining of aspirates collected in vivo. Radiographic support for the diagnosis was reported in 4 other dogs in which histoplasmosis was excluded by a negative CIEP with histoplasma antigen. The precipitating antibody may disappear during the course of the disease, as it did in 1 dog treated with amphotericin B, but not cured. This dog reverted from CIEP-positive to CIEP-negative within 17 months of treatment (with a weak reaction after 10 months of treatment). The CIEP-detectable antibody was present only in 1 dog without a confirmation by histopathologic findings or cultivation among 24 well-documented cases and 181 total tested sera. The CIEP was more sensitive and specific than was the gel-diffusion precipitin test, eliminated the problems of anticomplementarity that often affected the results of complement-fixation tests with canine sera, and served well in detecting dogs with blastomycosis. Electrophoretic pattern of sera from CIEP-positive dogs with blastomycosis showed a decrease in albumin and an increase in alpha 2- and often in beta- and gamma-globulins, with a substantial decrease of the albumin/globulin ratio.

Animals↗

Canine brucellosis: comparison of clinical manifestations with serologic test results.

Slide agglutination and mercaptoethanol tube agglutination tests for canine brucellosis were performed on 158 dogs. Clinical status was compared with the serologic test results. Sera were from 56 clinically normal dogs, 63 dogs with reproductive disorders, and 39 dogs with various nonreproductive disorders that could be associated with canine brucellosis. Ten of 21 (48%) aborting bitches and 2 of 9 (22%) bitches with other reproductive disorders were seropositive for brucellosis. Enlarged testicles, orchitis, and epididymitis were the main clinical disorders associated with positive (33%) or suspect (20%) serologic reactions in 15 male dogs. In 13 dogs of both sexes, diskospondylitis and osteomyelitis were the most common nonreproductive disorders associated with seropositive status for canine brucellosis (46%). Of 138 stray dogs, 17 were seropositive for canine brucellosis. Treatment of seropositive animals with antibiotics gave variable results. Of the 296 serum samples tested, 43 (14.5%) gave a positive reaction by the slide agglutination test but were negative by the mercaptoethanol tube agglutination test. Correlation was not found between serologic results and sex or breed.

Abortion, Veterinary↗

Testing of hemolytic complement components in domestic animals.

Total complement (C) and its components were assayed in the serum of 8 species of domestic animals, using commercially prepared cellular intermediates of sheep erythrocytes and functionally pure guinea pig and human components of the C system. Testing was done according to methods recommended by the producer for testing human C components. The late-acting components (C6 throug C9) and C1 were detected in carnivorous (dog and cat) and omnivorous (swine) animals. Undetectable or low titers of C4, C2, C3, and C5 were present in large herbivorous animals (cattle, horse, sheep, and goat), indicating major differences in comparison with human or guinea pig components of C. Porcine serum contained an inhibiting substance which interfered with testing C2 and later-acting components at serum dilutions up to 1:100. All components except C2 were detected in chicken serum. The binding or activation (or both) of C4, C2, C3, and C5 is more species specific than is the binding or activation (or both) of other components. Requirements for species specificity between antibody and C1 were not detected. Presence of C1 inactivator was detected in bovine, caprine, equine, and ovine sera. The CH50 (50% hemolysis) titers of C components tested in pooled serum samples from the 8 species of clinically healthy domestic animals are presented.

Animals↗

The p21 ras C-terminus is required for transformation and membrane association.

The Harvey murine sarcoma virus (Ha-MuSV) transforming gene, v-rasH, encodes a 21,000 molecular weight protein (p21) that is closely related to the p21 proteins encoded by the cellular transforming genes of the ras gene family. The primary translation product (prop21), which is found in the cytosol, undergoes posttranslational modification and the mature protein subsequently becomes associated with the inner surface of the plasma membrane and binds lipid tightly. The p21 proteins have the capacity to bind guanine nucleotides non-covalently in vitro. To assess the biological relevance of these biochemical features of the protein, we have now studied a series of deletion mutants located at or near the C-terminus of the viral p21 protein. Our tissue culture studies indicate that amino acids located at or near the C-terminus are required for cellular transformation, membrane association and lipid binding.

Animals↗