Comparison of cytokine mRNA expression in peripheral CD4+, CD8+ and γδ T cells between healthy Holstein and Japanese Black calves

Japanese Black (JB) calves are more susceptible to infectious diseases compared to Holstein (Hol) calves. To clarify the immunological differences between JB and Hol calves, expression of cytokine messenger RNA (mRNA) was examined using peripheral CD4⁺, CD8⁺ and γδ T cells. Healthy calves, 24 from each species, were examined. Blood samples were obtained from calves at 1 week, 1 month and 3 months old, eight calves for each age of each species. Peripheral blood mononuclear cells were stimulated with phytohemagglutin (PHA), and T cell subsets were isolated by positive selection using magnetic cell sorting (MACS). Levels of interlekin (IL)‐2, IL‐4, IL‐10 and interferon (IFN)‐γ mRNA in three T cell subsets were analyzed. WC1‐N1⁺ γδ T cell percentages were significantly lower in JB calves at 1 week and 1 month of age compared to Hol calves. In addition JB calves had significantly lower IL‐2, IL‐10 and IFN‐γ mRNA in WC1‐N1⁺ γδ T cells at 1 and 3 months of age, whereas there were no significant differences in cytokine mRNA of CD4⁺ and CD8⁺ cells between the two groups. Decreased cytokine mRNA and cell number of peripheral γδ T cells may affect negatively on the immune system of JB calves.     

Anatomical features of ossa cordis in the Steller sea lion

Irregular triangular cartilage or bone fragments are sometimes found in the fibrous triangle of the heart. Ossa cordis and/or cartilago cordis has been demonstrated in various terrestrial animal species. Regarding marine mammals, sperm whales lack heart bones, and there have been no studies on bones or cartilage in pinniped hearts. Therefore, we examined the ossa cordis and/or cartilago cordis of the Steller sea lion. Eleven Steller sea lion hearts were examined morphologically and histologically. Before dissection, some hearts were imaged by CT to confirm the presence of ossa cordis or cartilago cordis. As a result, ossa cordis-like fragments were confirmed in four adults and one pup. All of the fragments were found at the right fiber triangle, and one adult had ossified tissue, including adipose tissue in the bone marrow cavity. The ossa cordis probably support the aorta because they surround the aorta as in other terrestrial animals. Steller sea lions can dive to a few hundred meters, but they need to rest on land frequently. Hence, their ossa cordis help maintain heart function during the tachycardia that occurs upon repeated surfacing and movements on land after diving in water.     

不同地下水位胡杨蒸腾速率与叶水势的变化分析

本文研究了不同地下水位胡杨蒸腾速率、叶水势及蒸腾速率的季节变化等水分生理特性,结果表明:地下水位影响胡杨蒸腾作用,同时土壤水分状况也影响蒸腾速率,当地下水位在2.28~2.43 m时,胡杨蒸腾速率变化幅度大,蒸腾速率的日最大值为7.50 mmol.m-2s-1,日均值为4.73 mmol.m-2s-1,对应叶水势的日最低值为-3.55(Mpa)和日均值为-1.94(Mpa);地下水位2.91~3.33 m时,叶水势的日最低值为-2.61(Mpa)和日均值为-1.62(Mpa)。胡杨蒸腾速率季节变化在7月份达到最大值,靠近河岸胡杨蒸腾速率变化较远离河岸胡杨大,表明地下水位低时,胡杨保持较高的水势就能从环境中吸收到水分来满足其正常生长。     

Micropores and mesopores in the cell wall of dry wood

To investigate micropores and mesopores in the cell walls of dry wood, CO₂ gas and N₂ gas adsorption onto dry wood were measured at ice-water temperature (273 K) and liquid nitrogen temperature (77 K). CO₂ gas adsorption isotherms obtained were used for determining micropore volumes smaller than 0.6 nm by the HK method (Horvath-Kawazoe method), and N₂ gas adsorption isotherms obtained were used for determining the mesopore volume between 2 nm and 50 nm by the Barrett-Joyner-Halenda (BJH) method. Micropores and mesopores existed in cell walls of dry wood, and the cumulative pore volume was much larger for micropores than for mesopores. Micropores in the cell wall of dry wood decreased with elevating heat treatment temperature, and the decreased micropore was reproducible by wetting and drying. Mesopores did not decrease so much with elevating heat treatment temperature. Micropore volumes for the softwood Hinoki and the hardwood Buna were compared. A larger amount of micropores existed in hardwood Buna than in softwood Hinoki, and this relationship was considered to correspond to the difference in thermal softening properties for lignin in water-swollen Hinoki and Buna. This result probably indicates that micropores in the cell walls of dry wood relate to the structure of lignin.     

Molecular Monitoring of hrpB-disrupted Mutant of Ralstonia solanacearum in Tomato Plants

A nonpathogenic mutant of Ralstonia solanacearum was produced by the insertion of transposon Tn4431. The mutagenized gene was then cloned from a genomic DNA library by the gene tagging method, using the labeled lux operon located on Tn4431 of pUCD623 as a hybridization probe. From nucleotide sequence analysis of the transposon-inserted genomic clone, the hrpB gene was shown to be disrupted by the inserted transposon. Tomato plants were inoculated with the hrpB-disrupted mutant bacteria, for which multiplication and translocation were then monitored using the colony hybridization method. In addition, the original pathogenic bacteria in which the lux operon had been functionally ligated with the genomic promoter were also used for inoculation and traced by their bioluminescence. Multiplication of the hrpB-disrupted mutant was suppressed initially in the invaded root tissues and then in upper hypocotyl after translocation, suggesting that the pathogenic strain of R. solanacearum overcomes at least two steps of host responses expressed in root and hypocotyl tissues. Thus, our approach for molecular monitoring of the bacteria enabled us to precisely analyze the infection behavior of the pathogenic bacteria in planta. Received 16 April 1999/ Accepted in revised form 10 August 1999     

Gene families encoding isoforms of two major sesame seed storage proteins, 11S globulin and 2S albumin

Sesame (Sesamum indicum L.) seed has been recognized as a nutritional protein source owing to its richness in methionine. Storage proteins have been implicated in allergenic responses to sesame consumption. Two abundant storage proteins, 11S globulin and 2S albumin, constitute 60-70 and 15-25% of total sesame proteins, respectively. Two gene families separately encoding four 11S globulin and three 2S albumin isoforms were identified in a database search of 3328 expressed sequence tag (EST) sequences from maturing sesame seeds. Full-length cDNA sequences derived from these two gene families were completed by PCR using a maturing sesame cDNA library as the template. The amino acid compositions of these deduced storage proteins revealed that the richness in methionine is attributed mainly to two 2S albumin isoforms and partly to one 11S globulin isoform. The presence of four 11S globulin and three 2S albumin isoforms resolved in SDS-PAGE was confirmed by MALDI-MS analyses. The abundance of these isoforms was in accord with the occurrence frequency of their EST sequences in the database. A comprehensive understanding of these storage proteins at the molecular level may also facilitate the identification of allergens in crude sesame products that have caused severe allergic reactions increasingly reported in the past decade.     

A case of feline sporotrichosis

We excised surgically a feline granulomatous lesion and performed histopathological, mycological and molecular examinations. As a result, it was diagnosed as sporotrichosis, which was the second recorded case of a cat so afflicted in Japan. After the operation, we recognized another nodule on the lymph node. Histopathological examination was therefore performed, but no fungi were detected. To prevent recurrence, the cat was administered a antimycotic drug, itraconazole. As a result, no recurrence was found. Excision of the lesion is the treatment of choice for feline sporotrichosis.     

Phytotoxic and antiphytopathogenic compounds from Thai Alpinia galanga (L.) Willd. rhizomes

The management of weeds and diseases that are caused by phytopathogenic fungi is important for preventing the loss of agricultural products. The aim of the present study was to identify phytotoxic and antiphytopathogenic agents from the Thai Alpinia galanga rhizome. Extracts of the dried rhizomes of A. galanga (Zingiberaceae) were separated and tested for phytotoxic activity against lettuce (Lactuca sativa L. cv. Great Lakes) and Italian ryegrass (Lolium multiflorum Lam. cv. Wasefudou) and for antiphytopathogenic activity against Alternaria porri, Colletotrichum gloeosporioides, Fusarium oxysporum and Phytophthora nicotianae. 1′‐Acetoxychavicol acetate ( 1 ) was identified as one of the main components, together with trans‐p‐coumaryl acetate ( 3 ) and trans‐p‐acetoxycinnamyl acetate ( 2 ). 1′‐Acetoxychavicol acetate ( 1 ) was solvolyzed with 2% EtOH to yield trans‐p‐coumaryl ethyl ether ( 6 ), trans‐p‐coumaryl acetate ( 3 ) and trans‐p‐coumaryl alcohol ( 5 ). 1′‐Acetoxychavicol acetate ( 1 ) completely inhibited the root growth of the lettuce seedlings at 50 μg mL^–1, but had a weaker inhibitory effect on the growth of Italian ryegrass. 1′‐Acetoxychavicol acetate also inhibited the growth of P. nicotianae and A. porri, with minimum inhibition concentration values of 15.6 and 31.5 μg mL^–1, respectively. The plant growth‐inhibitory activity and fungal growth‐inhibitory activity of trans‐p‐coumaryl acetate ( 3 ), trans‐p‐coumaryl ethyl ether, trans‐p‐coumaryl alcohol ( 5 ) and trans‐p‐acetoxycinnamyl acetate ( 2 ) were lower than those of 1′‐acetoxychavicol acetate. A structure–activity relationship suggested that the strong phytotoxic and antiphytopathogenic activity of 1′‐acetoxychavicol acetate relied on the 1′‐acetoxyl group.     

A plastidic glucose-6-phosphate dehydrogenase is responsible for hypersensitive response cell death and reactive oxygen species production

Glucose-6-phosphate dehydrogenase (G6PDH) has been implicated in the supply of reduced nicotine amide cofactors for resistance to biotic and abiotic stresses. Here, we show participation of the plastidic P2 isoform of G6PDH in plant immunity. A cytosolic isoform (NbG6PDH-Cyto) and two plastidic isoforms (NbG6PDH-P1 and NbG6PDH-P2) cloned from Nicotiana benthamiana were localized in cytosol and chloroplasts, respectively. Hypersensitive response (HR) cell death and NADPH oxidase (RBOH; respiratory burst oxidase homolog)-dependent reactive oxygen species (ROS) production after recognition of INF1 elicitin, secreted by oomycete Phytophthora infestans, decreased in NbG6PDH-P2-silenced plants, but not in NbG6PDH-Cyto- and NbG6PDH-P1-silenced plants. Silencing of the cytosolic NAD kinase NbNADK1, which phosphorylates NADH to form NADPH, compromised HR cell death and ROS production, and concomitant silencing with NbG6PDH-P2 reduced HR cell death and ROS to levels near those in NbG6PDH-P2-silenced plants. Similarly, silencing NbG6PDH-P2 and NbNADK1 resulted in high susceptibility to P. infestans. These results suggest that NADPH produced by the P2 isoform of G6PDH in chloroplasts is responsible for HR cell death and ROS production mediated by RBOH and that NbNADK1 is involved in this pathway.