Permuted tRNA genes expressed via a circular RNA intermediate in Cyanidioschyzon merolae

A computational analysis of the nuclear genome of a red alga, Cyanidioschyzon merolae, identified 11 transfer RNA (tRNA) genes in which the 3' half of the tRNA lies upstream of the 5' half in the genome. We verified that these genes are expressed and produce mature tRNAs that are aminoacylated. Analysis of tRNA-processing intermediates for these genes indicates an unusual processing pathway in which the termini of the tRNA precursor are ligated, resulting in formation of a characteristic circular RNA intermediate that is then processed at the acceptor stem to generate the correct termini.     

Detection of the EGFP sequence in breast muscle of 3‐year‐old chicken after transfection using sonoporation

In our continuing effort to generate transgenic chickens, sonoporation was chosen to insert an exogenous gene into the chicken genome. An EGFP expression vector (pCAG‐EGFPac) and microbubbles were injected into the central disc of stage‐X blastoderm or the germinal crescent of stage‐4 embryos, followed by ultrasonic vibration. Nineteen chicks out of 108 treated embryos hatched, six females and six males out of these 19 chicks grew to sexual maturity and two females and three males lived for 3 years. Genomic DNA from 17 out of 35 gonads from embryos and chicks that died before sexual maturity was EGFP‐positive by PCR. No EGFP sequence was detected in the genomic DNA of 322 embryos from six sexually mature females and the semen from four sexually mature males by PCR. When genomic DNA was obtained from various tissues of five 3‐year‐old chickens, the EGFP sequence was amplified from the genomic DNA of the breast muscle of a female (No. 85). The above sequence was subjected to DNA sequencing and verified to be the EGFP sequence. These results showed that sonoporation is an effective tool for the transduction of exogenous genes into chicken embryos for the generation of transgenic chickens.     

Precise, simple screening for resistance in potato varieties to common scab using paper pots

Resistance to common scab pathogen Streptomyces turgidiscabies of seven potato varieties was compared in the field with a newly developed paper pot method. Seedlings raised in soil in paper pots containing inocula at 1 × 10³ to 10⁷cfu/g soil were transplanted into a scab-free field and grown for 3 months. The disease severity of the seven varieties in the field trials differed in iteration and from year to year, even though their resistance levels were approximately similar at the expected levels. With the paper pot method, the seven varieties had different resistance levels, which were almost completely consistent with the results of the field trials, at more than 1 × 10⁵cfu/g soil. Significant differences in disease severity between resistant and susceptible varieties were observed (P = 0.05) for 2 years, and the resistance level of the varieties was elucidated.     

Chicken peripheral blood CD3+CD4-CD8- cells are regulated by endocrine and nerve systems

The existence of CD3(+)CD4(-)CD8(-) T cells in thymus and spleen has already been known. However, because of the presence of large amounts of thrombocytes in peripheral blood (PB), the proportion of CD3(+)CD4(-)CD8(-) T cells in PB has yet to be investigated. Therefore, the proportion of peripheral T cell-subsets was investigated in 6-week-old chickens. The percentage of CD3(+) cells, CD4(+) cells, CD8 alpha(+) cells, CD8 beta(+), and CD3(+)CD4(-)CD8(-) cells was 76%, 41%, 14%, 5%, and 15%, respectively. The proportion of CD3(+)CD4(-)CD8(-) cells in PB increased during egg-laying periods and in chickens treated with an analog of estrogen, while it decreased with age and in response to restraint stress. All of the CD3(+)CD4(-)CD8(-) cells expressed TCR1, and did not have NK activity. CD3(+)CD4(-)CD8(-) cells represent about 60% of peripheral TCR1(+) cells. These findings indicate that the proportion of CD3(+)CD4(-)CD8(-) cells is regulated by the endocrine and nerve systems.     

Usefulness of MRI compared with CT for diagnosis of mesenteric lymphoma in a dog

We evaluated the usefulness of MRI and compared it with CT for diagnosis of mesenteric lymphoma in a dog. The results in the plain CT, dynamic CT and plain MR (T1WI and T2WI) images suggested that the mass was a large single nodular lesion with abundant blood perfusion. On enhanced MRI(T1WI) , the mass was depicted as a tumor with adhesion to the gut wall. Exploratory laparotomy confirmed the mass was consistent with the findings on enhanced MRI. We think that MRI might be a useful imaging tool for diagnosis of canine mesenteric lymphoma.     

Evaluation of bovine spongiform encephalopathy (BSE) infection risk of cattle via sewage sludge from wastewater treatment facilities in slaughterhouses in Japan

Scattered SRM residues from BSE-infected cattle are possible to contaminate sewage during the slaughtering process in slaughterhouses. A proportion of the sludge discharged from wastewater treatment facilities at slaughterhouses has historically been processed into fertilizer. We therefore investigated the associated risk of BSE infection to cattle via sludge-derived fertilizer. Each stage of the process associated with BSE exposure was qualitatively evaluated and quantitative evaluations were subsequently performed using infectious dose as a unit of concern. Results of these qualitative evaluations indicated that installation of filter(s) at the drains to the wastewater treatment facilities has been undertaken by many slaughterhouses and has decreased the likelihood of SRM contamination of sewage. The level of sludge-derived fertilizer ingested by cattle was considered to be very low since the fertilizer is mixed with the ground soil, and the amount of soil ingested by cattle is likely to be small. Results from the quantitative analysis indicated the total infectious dose ingested by cattle in Japan from an infected cow has been estimated to be 5.5 x 10(-3) ID(50). Preventing scattering of SRM during the slaughtering process, installing filters to the drains with the removal of residues from the drain water and preventing the application of sludge-derived fertilizer to pasturelands would be effective to reduce the risk. Although the limited extent of available information, this study should provide useful indication for the development of an inclusive risk assessment for slaughterhouse sludge in the future.     

Establishment and biological characterization of canine histiocytic sarcoma cell lines

Seven novel cell lines from canine histiocytic sarcoma (HS), three of which were disseminated cutaneous HS and four of which were synovial HS, were established. All of the established cell lines had the same morphological (by light and electron microscopic findings), cytochemical (alpha-naphthyl butyrate esterase-positive), and immunohistochemical (vimentin- and lysozyme-positive, and cyto-keratin-negative) characteristics as the original HS tumor cells. All of the established cell lines injected into nude mice subcutaneously produced solid tumors. Because the established cell lines also showed phagocytic and processing activities, the HS tumor cells appear to originate from the mononuclear phagocytic system cells, despite their differences in locations or organs.     

The influence of age and body mass index on relative accuracy of energy intake among Japanese adults

OBJECTIVE: To examine relationships between the ratio of energy intake to basal metabolic rate (EI/BMR) and age and body mass index (BMI) among Japanese adults. DESIGN: Energy intake was assessed by 4-day semi-weighed diet records in each of four seasons (16 days in total). The EI/BMR ratio was calculated from reported energy intake and estimated basal metabolic rate as an indicator of reporting accuracy. SETTING: Residents in three areas in Japan, namely Osaka (urban), Nagano (rural inland) and Tottori (rural coastal). SUBJECTS: One hundred and eighty-three healthy Japanese men and women aged > or =30 years. RESULTS: The oldest age group (> or =60 years) had higher EI/BMR values than the youngest age group (30-39 years) in both sexes (1.74 vs. 1.37 for men; 1.65 vs. 1.43 for women). In multiple regression analyses, age correlated positively (partial correlation coefficient, beta = 0.012, P < 0.001 for men; beta = 0.011, P < 0.001 for women) and BMI correlated negatively (beta = -0.031, P < 0.001 for men; beta = -0.025, P < 0.01 for women) with EI/BMR. CONCLUSION: Age and BMI may influence the relative accuracy of energy intake among Japanese adults.     

Age Difference in Morphology and Immunohistology inthe Thymus and Spleen in Crl:CD (SD) Rats

We investigated chronological changes in immunohistochemical phenotyping in the thymus and spleen in Crl:CD rats up to the age of about one year. In the thymus, T cells increased markedly from 3 to 4 weeks of age. Proliferating cells also increased markedly at these points. B cells tended towards an increase with age. In the spleen, white pulp increased until 9 weeks of age and remained fairly stable thereafter. In the periarteriolar lymphoid sheath and marginal zone, T cells gradually increased until 9 weeks of age and became almost flat thereafter. In the lymph follicle, T cells increased with age. B cells tended towards an increase with age in all areas of the spleen. It was concluded that development of the thymus was most marked from 3 to 4 weeks of age and that both the thymus and spleen had matured by 9 weeks of age.