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.     

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.     

Genetic diagnosis of Rehmannia species micropropagated by tip tissue culture and an F1 hybrid by RAPD analysis

Shoot formation was induced from the tip tissue of Rehmannia glutinosa f. hueichingensis on hormone-free Murashige-Skoog medium within 4 weeks of culture. Shoots were propagated on Murashige-Skoog medium supplemented with l.0mg/l benzy0ladenine for 4 weeks. Propagated shoots rooted on the hormone-free Murashige-Skoog medium during 4 weeks of culture. Total DNA was extracted from the leaves of a F₁ hybrid and its parents. R. glulinosa f. huekliingensis and R. glutinosa var, purpurea. Analysis of random-amplified polymorphic DNA (RAPD) using 10 arbitrary oligonucleotide 10-mers. showed the genetic homogeneity ofthe above three species. The F₁ hybrid was genetically intermediate between both parental plants, compared with the genetic distance between the F₁ hybrid and individual parents. Furthermore, the comparison of the band patterns between the F₁ hybrid, obtained from the crossing clearly showed that parts of the bands of both parents. R. glutinosa f. hueichingensis and R. glutinosa var. purpurea, were introduced into the F₁ hybrid.     

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.     

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.     

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.     

Availability of NMR microscopic observation of mouse embryo disorder: examination in malformations induced by maternal administration of retinoic acid

Nuclear magnetic resonance (NMR) microscopy is a magnetic resonance imaging method with enhanced spatial resolution due to the use of a high static magnetic field and high magnetic field gradients. It is considered to be a useful tool for non-invasive and continuous investigation of tissue and organs at the histological level. In this study, we applied NMR microscopy to assessment of morphology in mouse embryos using a developmental disorder model induced by retinoic acid administration. Pregnant mice were given 50 mg/kg all-trans retinoic acid at 8.5 dpc. Embryos were collected at several time points after treatment and examined by NMR microscopy after fixation. Two-dimensional and three-dimensional spin echo sequences were used. Tissue contrast on two-dimensional images changed according to length of repetition time and echo time, and also to developmental stage of embryos. Two-dimensional and three-dimensional images nondestructively demonstrated defects in development of the skeleton and soft tissue, e.g. hypoplasia of vertebrae in the lumbar and tail regions and dysplasia of the spinal cord, in embryos exposed to retinoic acid. These morphological abnormalities were confirmed by conventional assessment after imaging. Although further improvements are required, NMR microscopy will provide a new approach for multi-parameter assessment of embryonic development under physiological and pathological conditions.     

Post-mortem evaluation of expanded polytetrafluoroethylene (ePTFE) used in mitral valve repair in dogs

Mitral valve repair is one of the treatment options for mitral regurgitation. Expanded polytetrafluoroethylene (ePTFE) is a polymer that has been widely used in cardiovascular surgery. In this case series, we report the autopsy and histological findings in 6 dogs that underwent cardiopulmonary bypass for mitral annuloplasty using ePTFE sheets and chordoplasty using ePTFE sutures.From May 2005 to October 2009, 3 female and 3 male dogs with severe mitral regurgitation underwent mitral valve repair. This case series included 3 Cavalier King Charles spaniels, 2 Maltese, and 1 Shih Tzu. The survival period after surgery was 19–72 (35 ± 19) months.In all the cases, autopsy revealed that the ePTFE sheets and sutures were not damaged and well integrated into the surrounding highly differentiated, connective tissues. Low-power microscopy revealed that in all cases, the tissues surrounding the ePTFE sheet in the mitral valve annulus had almost completely been covered by granulation tissue. No inflammatory infiltrate or thrombogenesis was observed around the ePTFE in any of the cases.There was no evidence of reactive changes in the region surrounding the ePTFE. These results suggest that ePTFE has excellent tissue compatibility and durability and can be effectively used for canine mitral valve repair.