Induction of reactive oxygen generation and functional changes in mitochondria and their involvement in the development of bacterial rot in lettuce caused by Pseudomonas cichorii

Pseudomonas cichorii is the major causal agent of bacterial rot disease in lettuce, and apoptosis-like programmed cell death is closely associated with disease development. Depletion of cellular ATP and expression of the alternative oxidase gene was observed in lettuce leaves inoculated with P. cichorii suggesting mitochondrial dysfunction. Cytochemical observation showed production of reactive oxygen species in mitochondria of P. cichorii-inoculated lettuce leaf cells. Release of cytochrome c into the cytosol was also induced by inoculation with the bacterium. Superoxide production was observed in the mitochondrial fraction isolated from P. cichorii-infected leaves much more intense than water-treated leaves. Loss of swelling ability was also observed in the mitochondrial fractions following inoculation with P. cichorii. Intriguingly, inhibitors of the mitochondrial respiratory complex III prevented loss of swelling ability, whereas superoxide generation was scarcely affected by inhibitors of mitochondrial permeability transition in the mitochondrial fractions. Inhibitors of the mitochondrial respiratory complex III and mitochondrial permeability transition delayed not only P. cichorii-induced cell death, but also disease development. In contrast, P. cichorii-induced DNA fragmentation was not inhibited in the presence of either type of inhibitor. The findings suggest that mitochondria may play a crucial role in DNA fragmentation-independent cell death pathway(s).     

Spatiotemporal catch distribution of age-0 Pacific bluefin tuna Thunnus orientalis caught by the Japanese troll fishery in relation to surface sea temperature and seasonal migration

This study used a delta-lognormal model to analyze monthly catches of age-0 Pacific bluefin tuna by the troll fishery. The model included fixed effects of month, area, and month–area interaction, and random effects of port, year and port–year interaction. The catch patterns by month and area predicted by the statistical model (standardized catch) revealed that main fishing grounds along the Tsushima Warm Current generally shifted from north to south as the season turned from autumn to winter. In contrast, the standardized catch along the Kuroshio Current did not show such clear spatiotemporal patterns. The standardized catch along the Tsushima Warm Current is significantly associated with average monthly sea surface temperatures in the fishing grounds and consistent with migration routes revealed by tagging experiments in previous studies. These associations indicate the spatiotemporal catch pattern in the Tsushima Warm Current region partly reflects seasonal migration. Knowledge of the possible associations among fish migration, environmental factors and spatiotemporal distribution of the catch will contribute to future management of this species.     

Nickel-catalyzed carbonization of wood for coproduction of functional carbon and fluid fuels II: improved fuel quality of oil fraction and increased heating value of gas fraction

The yields and properties of oil and gas fractions coproduced during carbonization of larch wood loaded with Ni 2%, Ni 2%+Ca 1%, and Ni 4% and without catalyst (None) at 700°–900°C were examined to clarify the catalytic effect in terms of conversion into fluid fuels. The net calorific value of oil occurred mainly below 500°C and increased in the order None < Ni 2% < Ni 4% < Ni 2%+Ca 1%, while the yield decreased in this order. The same order held for the production of gases enriched with hydrogen at 500°–700°C. Even above 800°C, markedly promoted evolution of hydrogen took place for all catalyst systems. These observations confirmed the effectiveness of nickel-catalyzed carbonization at 900°C, particularly Ni 2%+Ca 1%, for both upgrading of oil and gaseous fractions, although the quality of oil was not satisfactory. The catalysis of nickel with and without calcium is discussed on the basis of the modified Broid-Shafizadeh scheme, and the scheme was altered to adapt to the high temperature region where oil was no longer produced. Part of this study was presented at the 14th Annual Meeting of the Japan Institute of Energy, Suita, August 2005, and at the 18th Symposium, Session D, of the Materials Research Society of Japan, Tokyo, December 2007     

A piglet with concurrent polioencephalomyelitis due to porcine teschovirus and postweaning multisystemic wasting syndrome

A piglet developed respiratory distress followed by difficulty in standing and unsteady gait. The lesions were characterized by polioencephalomyelitis with the predominant distribution in the brain stem, as well as lymphocyte depletion and histiocyte infiltration with cytoplasmic inclusion bodies in the lymphoid tissues throughout the body and interstitial pneumonia. Porcine teschovirus (PTV) antigens were found in the former lesions and porcine circovirus 2 (PCV2) in the latter two lesions. PTV genes were detected from the diencephalon. The results suggest that the piglet was concurrently affected with polioencephalomyelitis due to PTV and postweaning multisystemic wasting syndrome (PMWS) associated with PCV2. They also suggest that the immunosuppressive condition developing in PMWS may have facilitated the infection of the brain with PTV.     

Multiplication of low and high cell culture passaged strains of transmissible gastroenteritis virus in organs of newborn piglets

Distribution and persistence of four different strains of transmissible gastroenteritis (TGE) virus in newborn piglets were compared.The piglets inoculated with high-passaged TO-163 strain did not show any clinical signs of TGE on any days postinoculation (DPI), but the piglets inoculated with one of the other three strains, SH-14, SH-164 or TO-16, had soft feces or diarrhea. In the latter cases, the virus was isolated mainly from respiratory organs, lymph nodes, and digestive tract on any DPI, but was rarely detected in the digestive tract of piglets inoculated with the TO-163 strain. The frequency of virus recovery from the tissues was the highest till 4 DPI in all of the piglets inoculated with one of the four virus strains, and it was markedly reduced thereafter in the piglets inoculated with high-passaged strains.The TO-163 strain was subjected to serial passage in newborn piglets for seven passages. There was no evidence of regained pathogenicity with advance in passage, and detection of virus was restricted to lymph nodes and lung of these piglets.In gnotobiotic piglets inoculated with the TO-163 strain, frequent virus recovery and high titers of virus from the tissues were obtained on up to the 4th DPI. The viruses in high titer were found in the digestive tract of some of the piglets; however, none of them showed any clinical signs of TGE.     

Transport stress increases somatic cell counts in milk, and enhances the migration capacity of peripheral blood neutrophils of dairy cows

The present study was designed to determine the effects of physiological stress on milk-somatic cell counts (SCC) and function of bovine peripheral blood leukocytes (PBL). Nine healthy lactating cows were used in the examination. Five cows were transported 100 km for 4 hr (transported group; TG), and 4 cows were penned (non-transported group; NTG). Blood and milk samples were collected at 0, 2, and 4 hr after loading, and at 2 hr, and 1, 2, 3, and 6 days after unloading. The following activities were measured: adhesion receptor (CD 18 and L-selectin) expression of neutrophils and monocytes, migration capacity and percentage of apoptotic cells of neutrophils, serum soluble L-selectin (sL-selectin), plasma cortisol, and SCC. A significant increase in plasma cortisol and milk SCC was observed in TG. Leukocytosis, derived from neutrophils was recorded in TG, and was indicated by apoptotic measurement as an increase of young cells from the marginal pool. Increased migration and decreased surface expression of both L-selectin and CD 18 in neutrophils were observed after transportation. Elevated serum sL-selectin was also noted as a result of transportation. The present study indicated that transport stress modulates peripheral blood neutrophil function, particularly enhancing migration capacity, and causes diapedesis across the mammary epithelium. Increased milk SCC in transported cattle might be due to these phenomena, and severe physiological stress may bring about an increase in SCC in milk.     

Effects of nuclear transfer procedures on ES cell cloning efficiency in the mouse

Enucleated oocytes receiving mouse embryonic stem (ES) cells develop into fertile young. The developmental potential to young is low, however, and the rate of postnatal death is high. We examined the effect of various nuclear transfer procedures on the in vitro and in vivo developmental potential of nuclear-transferred oocytes. The potential of oocytes receiving ES cells at M phase to develop into blastocysts after fusion by Sendai virus was high compared with that after direct injection (67% vs. 30%). The developmental potential of oocytes receiving ES cells at the M phase is higher than that of oocytes receiving ES cells at the G(1) phase (30-67% vs. 2-5%). Developmental ability to live young was low in all groups (0-4%). Different activation protocols affected the potential to develop into blastocysts to a different extent (27-62%), but did not affect the potential to develop into live young (0-3%). The present study demonstrated that the various conditions examined did not affect the potential of nuclear-transferred oocytes receiving ES cells to develop into live young or the incidence of postnatal death.     

Prominent expression of spinocerebellar ataxia type-1 (SCA1) gene encoding ataxin-1 in LH-producing cells, LbetaT2

The LH-producing cell line, LbetaT2, and non LH-producing cell line, alphaT3-1 cells, established from a pituitary tumor, were employed for cDNA subtraction cloning to identify genes with expression unique to LH producing cells. Several cDNAs that code for known as well as for many unidentified clones were discovered. Most clones were the spinocerebellar ataxia type-1 (SCA1) gene encoding ataxin-1, the abnormality of which causes neurodegeneration and loss of cerebellar Purkinje cells. We examined whether the expression of SCA1 gene in LbetaT2 cells is related to hormone production. We also compared the expression of SCA1 with that in various other pituitary tumor derived cell lines, and confirmed the prominent expression of SCA1 in LbetaT2 cells. The effect of gonadal factor(s) for SCA1 gene expression was examined. The expression level in female rats was low and did not change during the estrus cycle, but increased significantly after ovariectomy and did not return to the normal level under low and high doses of estrogen. In the male pituitary SCA1 gene expression increased markedly after castration and was not decreased by estrogen or testosterone. The Ontogeny of SCA1 gene expression was investigated in porcine fetal and postnatal pituitaries and revealed biphasic and sexually dimorphic expression. Transient expression of SCA1 gene was observed at fetal day 50 and 65 in males and day 40 in females, followed by a decline and increased expression before birth in both genders. Thus the expression of SCA1 gene is prominent in LH-producing cells and is not under direct control of gonadal factor(s) in both genders. In addition to the variable expression of SCA1 gene during the fetus period, the present results provide a novel aspect to the understanding of Boucher-Neuhauser syndrome (Ataxia Hypogonadism Choroidal Dystrophy).