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To characterize the low-molecular-weight glutenin subunit (LMW-GS), we developed specific PCR primer sets to distinguish 12 groups of LMW-GS genes of Norin 61 and to decide their loci with nullisomic-tetrasomic lines of Chinese Spring. Three, two, and ten groups were assigned to Glu-A3, Glu-B3, and Glu-D3 loci, respectively. To identify the proteins containing the corresponding amino acid sequences, we determined the N-terminal amino acid sequence of 12 spots of LMW-GSs of Norin 61 separated by two-dimensional gel electrophoresis (2DE). The N-terminal sequences of the LMW-GS spots showed that 10 of 12 groups of LMW-GSs were expressed as protein products, which included LMW-i, LMW-m, and LMW-s types. Four spots were encoded by Glu-A3 (LMW-i). Three spots were encoded by Glu-B3 (LMW-m and LMW-s). Five spots were encoded by Glu-D3 (LMW-m and LMW-s). A minor spot of LMW-m seemed to be encoded by the same Glu-B3 gene as a major spot of LMW-s, but processed at a different site. Comparing among various cultivars, there were polymorphic and non-polymorphic LMW-GSs. Glu-A3 was highly polymorphic, i.e., the a, b, and c alleles showed one spot, the d allele showed four spots, and the e allele had no spot. Insignia used as one of the Glu-A3 null standard cultivars had a LMW-GS encoded by Glu-A3. We also found that Cheyenne had a new Glu-D3 allele. Classification of LMW-GS by a combination of PCR and 2DE will be useful to identify individual LMW-GSs and to study their contribution to flour quality.

We have identified long-incubation viral agents that share epitopes with hepatitis B virus (HBV). During chimpanzee infectivity studies, these agents may be recognized in the liver since they possess complementary nucleic acid sequences with HBV DNA; the genomic size was found to be 3.2 kilobases, identical to that of HBV. Liver injury was produced and there was antigen expression in hepatocytes. Chimpanzees were not protected by prior immunization with hepatitis B surface antigen; conversely, they were still susceptible to HBV after recovery from infection with such agents. These findings suggest that these hepatitis B virus-related variants appear to be immunologically distinct from HBV.Previous studies on sera using monoclonal anti-hepatitis B surface antigen antibody (anti-HBsAg) radioimmunoassays (M-RIAs) have established that there are individuals with acute and chronic hepatitis infection and hepatocellular carcinoma whose serum demonstrates positive binding activity only in these tests (1-8). Such patients were found to be negative for hepatitis B surface antigen (HBsAg) by polyclonal anti-HBsAg RIA (P-RIA) and some lacked evidence of recent or past exposure to hepatitis B virus (HBV) as shown by the absence of antibodies to the core (antiHBcAg) and to surface antigens (anti-HBsAg) in the blood (4-8). HBV DNA hybridizable sequences were detected in the liver and serum ofa significant number of such individuals and this finding correlated with the detection of antigen in serum by 6). However, very little is known regarding the biologic properties and characteristics of the M-RIA HBV DNA-positive agents in serum. We report here the identification, transmission, and partial characterization of some viral agents during chimpanzee infectivity studies. These hepatitis viral variants appear to be antigenically related to and share nucleic acid homology with HBV. His serum was negative for HBsAg, anti-HBsAg and antiHBcAg antibodies by P-RIAs but was found to be reactive at a S/N ratio of 24.0 by M-RIA, and HBV DNA complementary sequences were detected by dot blot analysis. METHODSTwo chimpanzees (Dolf and lanthe) were previously immunized with a yeast recombinant HBV DNA-derived HBsAg vaccine. These chimpanzees had been subsequently challenged with a standard National Institutes ofHealth HBV inoculum known to produce infection in unimmunized chimpanzees; both animals were found to be protected from HBV infection. The details of these experiments have been reported elsewhere (9). Two other chimpanzees (Sjef and Jimmie) had not been previously exposed to HBV or to the non-A, non-B hepatitis agents, and their serum was negative for HBsAg, anti-HBsAg, and anti-HBcAg antibodies. In contrast, Dolf and Ianthe had high titer anti-HBsAg antibodies as the result of past vaccination with HBsAg; anti-HBcAg antibodies, however, were not present. lanthe and Sjef were inoculated intravenously with 3 ml of serum M, and Jimmie and Dolf were inoculated with 3 ml of serum G. After injection of serum, the chimpanzees were s...

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Abstract:In the present survey, the historical data for spontaneous tumors observed in our laboratory was compared in relation to the time-related changes between recent (2000)(2001)(2002)(2003)(2004) and past (1990-1999) samples from Fischer 344 rats used in carcinogenicity studies. In the recent samples, there were statistically increased incidences of islet cell adenomas in males and uterine adenocarcinomas in females. On the other hand, there were decreased incidences of pheochromocytomas, prostatic adenomas, pituitary anterior adenomas, large granular lymphocytic (LGL) leukemias and Leydig cell tumors in males and pituitary anterior adenomas in females. Furthermore, there were decreases in the body weights of both sexes at 58 weeks of age in the recent samples, and this may have been related to the decreases in the incidences of pheochromocytomas, Leydig cell tumors, prostatic adenomas and LGL leukemias. Moreover, the decreased incidence of pheochromocytoma in males revealed a positive correlation with a decreased percentage of severe chronic progressive nephropathy (CPN). On the other hand, there were no distinct factors responsible for increased incidences of tumors observed in the recent samples, suggesting a possible genetic drift. In conclusion, the incidences of spontaneous tumors obtained in our laboratory have been changed with time. Smaller body weights in both sexes and reduction in the number of male cases with severe CPN might be related to the reduced incidence of certain types of tumors in recent animals. (J Toxicol Pathol 2008; 21: 53-60)