Watercore in Japanese pear. III. Changes in the activities of some enzymes relating to the degradation of cell walls and the accumulation of sugar

The relationship between cellular breakdown and the activities of some cell-wall degrading enzymes (endocellulase, exocellulase, polygalacturonase, pectin methylesterase and β-galactosidase) was investigated in the watercored tissue of Japanese pear fruit, Pyrus serotina Rehder var. ‘Culta’ Rehder, cultivar ‘93-3’. In watercored tissue at an early stage, endocellulase activity, especially neutral form, was higher than that in healthy tissue. As the symptom of the watercore developed, polygalacturonase and β-galactosidase activities in the watercored tissue increased compared with those in healthy tissue. This indicated that in tissues at an early stage of watercore, these cell-wall degrading enzymes were activated, and operative in the breakdown of the cell wall. However, exocellulase and pectin methylesterase were not activated in the watercored tissue. The breakdown of cellular tissue was also supported by the fact that the DNA and protein contents decreased in the watercored tissue. Invertase activity decreased in the watercored tissue, compared with healthy tissue, and it was suggested that its decrease could be one of the causes of sucrose accumulation in the watercored tissue.     

Effects of extracellular lactate on production of reactive oxygen species by equine polymorphonuclear leukocytes in vitro

Objective-To evaluate effects of extracellular lactate on viability, shape change, lactate metabolism, and reactive oxygen species (ROS) production in equine polymorphonuclear leukocytes (PMNs). Sample-PMNs isolated from equine venous blood samples. Procedures-PMNs were incubated with 0 to 300mM lactate for 30 minutes before each experiment. Viability was assessed via trypan blue exclusion. Shape change was assessed via flow cytometry and light microscopy. Relative quantification of monocarboxylic acid transporter and lactate dehydrogenase lactate dehydrogenase (LDH) isotype mRNAs was performed with a real-time PCR assay. Effects of lactate at a pH of 7.4 to 6.0 on ROS production in response to phorbol 12-myristate 13-acetate, opsonized zymosan, or N-formyl-methionyl-leucyl-phenylalanine was assessed by luminol-dependent chemiluminescence. Results-Lactate had no effect on viability of PMNs but did alter their size and density. Monocarboxylic acid transporter 1 and lactate dehydrogenase B mRNA values were not altered. Monocarboxylic acid transporter 4 and lactate dehydrogenase A mRNA values were significantly decreased. Lactate incubation of cells significantly decreased PMN-derived luminol-dependent chemiluminescence and induced different sensitivities to stimulants (phorbol 12-myristate 13-acetate, opsonized zymosan, and N-formyl-methionyl-leucyl-phenylalanine). The response ratio to N-formyl-methionyl-leucyl-phenylalanine revealed that PMNs were primed by incubation with up to 50mM lactate, significantly increasing the production of ROS. Incubation with lactate and acidic pH caused a synergistic effect on ROS production. Conclusions and Clinical Relevance-Extracellular lactate potentially has a direct effect on the capacity to produce ROS by equine PMNs, which may be associated with alterations in innate immune functions within a short period after high-intensity exercise.     

Proteome analysis of cerebrospinal fluid in healthy beagles and canine encephalitis

We performed proteomics analysis of the cerebrospinal fluid (CSF) of healthy dogs and dogs with meningoencephalitis of unknown etiology (MUE). By comparing two-dimensional electrophoreses (2DE), an upregulated spot was found in MUE dogs. This protein was identified as a neuron-specific enolase (NSE) by analysis with MALDI-TOF mass spectrometry. In comparing dot blots using an antibody against NSE, the NSE levels in the CSF of MUE dogs was significantly higher than that of the controls. NSE is a diagnostic marker of neuroendocrine tumors, brain injury and spinal cord trauma in humans. It seems that the NSE concentration in the CSF is increased by cellular destruction in canine encephalitis. Though elevation of NSE may not be specific in canine encephalitis because the NSE level was increased in other CNS diseases, further study including measurement with serum is necessary.     

Vertical soil–air CO2 dynamics at the Takayama deciduous broadleaved forest AsiaFlux site

At the Takayama deciduous broadleaved forest Asiaflux site in Japan, the ecosystem carbon dynamics have been studied for more than two decades. In 2005, we installed non-dispersive infrared CO₂ sensors in the soil below the site’s flux tower to systematically study vertical soil–air CO₂ dynamics and explain the behavior of soil surface CO₂ efflux. Soil–air CO₂ concentrations measured from June 2005 through May 2006 showed sinusoidal variation, with maxima in July and minima in winter, similar to the soil CO₂ effluxes measured simultaneously using open-flow chambers. Soil–air CO₂ concentrations increased with soil depth from 5 to 50 cm: from 2,000 to 8,000 ppm in the summer and from 2,000 to 3,000 ppm in the winter under snow. Summer soil–air CO₂ concentrations were positively correlated with soil moisture on daily and weekly scales, indicating that the Oi, Oe, and A horizons, where decomposition is accelerated by high-moisture conditions, contributed substantially to CO₂ emissions. This result is consistent with the short residence time (about 2 h) of CO₂ in the soil and larger emissions in shallow soil layers based on our diffusion model. We revealed for the first time that soil–air CO₂ concentrations in winter were correlated with both snow depth and wind speed. CO₂ transfer through the snow was hundreds of times the gas diffusion rates in the soil. Our estimate of the CO₂ efflux during the snow-cover season was larger than previous estimates at TKY, and confirmed the important contribution of the snow-cover season to the site’s carbon dynamics.     

Breed differentiation among Japanese native chickens by specific skull features determined by direct measurements and computer vision techniques

1. Inter-breed morphological comparisons were made among 11 breeds of Japanese native chickens (Gifujidori, Hinaidori, Shokoku, Totenko, Tomaru, Satsumadori, Shamo, Koshamo, Koeyoshi, Chabo and Nagoya), White Leghorn, broiler chickens (Chunky) and red junglefowl collected in the Philippines, based on results of direct measurements and analysis by computer vision techniques of the skull. 2. Analysis of direct measurements identified two groups of chicken: a small type that included the Chabo, Koshamo, red junglefowl, Gifujidori and Shokoku and a large type that included the remaining breeds studied. These groupings were made based on size determined both in the first (PC1) and second principal component (PC2). The greatest length of the cranium and condylobasal length greatly contributed to the morphological differences between these two groups. 3. Analysis by computer vision techniques, however, identified three groups: the Bantam group (which includes red junglefowl), Shokoku group and Shamo group. White Leghorn clustered within the Shokoku group while the broiler chicken belonged to the Shamo group. The region around the junction of the neural cranium and the visceral cranium contributed greatly to the morphological differences among breeds, both in the PC1 and PC2.     

Effect of dietary fish oil intake on ubiquitin ligase expression during muscle atrophy induced by sciatic nerve denervation in mice

Dietary fish oil intake improves muscle atrophy in several atrophy models however the effect on denervation‐induced muscle atrophy is not clear. Thus, the aim of this study was to investigate the effects of dietary fish oil intake on muscle atrophy and the expression of muscle atrophy markers induced by sciatic nerve denervation in mice. We performed histological and quantitative mRNA expression analysis of muscle atrophy markers in mice fed with fish oil with sciatic nerve denervation. Histological analysis indicated that dietary fish oil intake slightly prevented the decrease of muscle fiber diameter induced by denervation treatment. In addition, dietary fish oil intake suppressed the MuRF1 (tripartite motif‐containing 63) expression up‐regulated by denervation treatment, and this was due to decreased tumor necrosis factor‐alpha (TNF‐α) production in skeletal muscle. We concluded that dietary fish oil intake suppressed MuRF1 expression by decreasing TNF‐α production during muscle atrophy induced by sciatic nerve denervation in mice.