Comparison of connective tissue structure and muscle toughness of spotted mackerel <Emphasis Type="Italic">Scomber australasicus</Emphasis> and Pacific mackerel <Emphasis Type="Italic">S. japonicus</Emphasis> during chilled and frozen storage

To characterize post-mortem changes in fish meat quality during chilled and frozen storage, muscle toughness and the connective tissue structure of muscle were compared between two mackerel species, spotted mackerel Scomber australasicus and Pacific mackerel S. japonicus. Measurement of the breaking strength of meat slices from both non-frozen fresh fish and frozen-thawed fish revealed that spotted mackerel meat was softer than Pacific mackerel meat. Under scanning electron microscopic observation, the connective tissue of non-frozen meat of spotted mackerel was thinner than that of Pacific mackerel. In addition, the collagen content in the spotted mackerel meat was lower than that of Pacific mackerel. These findings suggest that connective tissue structure might influence muscle toughness in mackerel.     

Extracellular matrix of <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis> may have a role in host adhesion during fungal penetration and is digested by matrix metalloproteinases

Spores and infection structures such as germ tubes and appressoria of Magnaporthe oryzae, the fungus causing blast disease of wheat, produced an extracellular matrix (ECM) on the surfaces of host leaves during fungal differentiation. The chemical components and function of the ECM were studied to understand the pathological roles using two immunological techniques and ECM-digesting enzymes. The ECM was characterized by fibrous and amorphous materials, located in the spaces between fungal cell walls and plant cuticles. Immunohistochemical and immunoelectron microscopy suggested that ECM includes components positively reacted with antibodies of four animal cell adhesion factors (collagen VI, vitronectin, fibronectin and laminin) and an animal integrin α3. ECM, incubated on a cellulose membrane, was rapidly digested by matrix metalloproteinases (collagenase and gelatinase B), resulting in the detachment of most infection structures from membrane surfaces. Both ultrastructural observation and immunological responses showed that more ECM was located at the appressoria than at the spores and germ tubes. This result suggested that appressoria needed a powerful adhesion force for aggressive action of penetration pegs into plant cuticles. An erratum to this article can be found at     

Upregulation of the genes for ferritin,RNase, and DnaJ in leaves of rice plants in response to sulfur deficiency

Microarray analysis was carried out with mRNAs isolated from aerial portions of rice plants subjected to sulfur deficiency. Among the candidate clones regulated by sulfur deficiency, three genes were identified by northern analysis that showed altered levels of mRNA accumulation. Genes similar to those for ferritin, S-like ribonuclease, and DnaJ-like protein were down-, down-, and upregulated by sulfate deficiency, respectively. These genes were found for the first time to be responsive to sulfur nutrition in the present study.     

Relationships between ammonia oxidizers and N2O and CH4 fluxes in agricultural fields with different soil types

Nitrous oxide (N₂O) is a greenhouse gas that contributes to the destruction of stratospheric ozone, and agricultural soil is an important source of N₂O. Aerobic soils are sinks for atmospheric methane (CH₄), a greenhouse gas. Ammonia monooxygenase (AMO) can oxidize CH₄, but CH₄ is mostly oxidized by methane monooxygenase (MMO), and CH₄ oxidation by AMO is generally negligible in the soil. We monitored the N₂O and CH₄ fluxes after urea application in fields containing different soils using an automated sampling system to determine the effects of environmental and microbial factors on the N₂O and CH₄ fluxes. The soil types were Low-humic Andosol (Gleyic Haplic Andosol), yellow soil (Gleyic Haplic Alisol) and gray lowland soil (Entric Fluvisol). Cumulative N₂O emissions from the yellow soil were higher than those from other soil types, although the difference was not significant. The CH₄ uptake level by Andosol was one order of magnitude higher than that by other soils. There were significant relationships between the ammonia oxidation potential, AOB and AOA amoA copy numbers, and the CH₄ uptake. In contrast, the gene copy numbers of methane-oxidizing bacteria (MOB) pmoA were below the detection limit. Our results suggested that the AMOs of AOB and AOA may have more important roles than those previously considered during CH₄ oxidation in agricultural soils treated with N fertilizers.     

Developmental competence of oocytes grown in vitro: Has it peaked already?

In vitro growth of immature oocytes provides opportunities to increase gametic resources and to understand the mechanisms underlying oocyte development. Many studies on the in vitro growth of oocytes have been reported thus far; however, only a few cases have been reported, which demonstrated that oocytes can support full-term development after in vitro fertilization. Our research group recently found that culture of mouse neonatal primordial follicles increased the birthrate; however, the establishment of an in vitro system that can completely mimic follicle or oocyte growth in vivo and control oogenesis remains an ongoing challenge.     

Accumulation of phosphorylated repressor for gibberellin signaling in an F-box mutant

Gibberellin (GA) regulates growth and development in plants. We isolated and characterized a rice GA-insensitive dwarf mutant, gid2. The GID2 gene encodes a putative F-box protein, which interacted with the rice Skp1 homolog in a yeast two-hybrid assay. In gid2, a repressor for GA signaling, SLR1, was highly accumulated in a phosphorylated form and GA increased its concentration, whereas SLR1 was rapidly degraded by GA through ubiquitination in the wild type. We conclude that GID2 is a positive regulator of GA signaling and that regulated degradation of SLR1 is initiated through GA-dependent phosphorylation and finalized by an SCF(GID2)-proteasome pathway.     

Effects of Intermittent Positive Pressure Ventilation on Cardiopulmonary Function in Horses Anesthetized with Total Intravenous Anesthesia Using Combination of Medetomidine,Lidocaine, Butorphanol and Propofol (MLBP-TIVA)

Effects of intermittent positive pressure ventilation (IPPV) on cardiopulmonary function were evaluated in horses anesthetized with total intravenous anesthesia using constant rate infusions of medetomidine (3.5 µg/kg/hr), lidocaine (3 mg/kg/hr), butorphanol (24 µg/kg/hr) and propofol (0.1 mg/kg/min) (MLBP-TIVA). Five horses were anesthetized twice using MLBP-TIVA with or without IPPV at 4-week interval (crossover study). In each occasion, the horses breathed 100% oxygen with spontaneous ventilation (SB-group, n=5) or with IPPV (CV-group, n=5), and changes in cardiopulmonary parameters were observed for 120 min. In the SB-group, cardiovascular parameters were maintained within acceptable ranges (heart rate: 33–35 beats/min, cardiac output: 27–30 l/min, mean arterial blood pressure [MABP]: 114–123 mmHg, mean pulmonary arterial pressure [MPAP]: 28–29 mmHg and mean right atrial pressure [MRAP]: 19–21 mmHg), but severe hypercapnea and insufficient oxygenation were observed (arterial CO₂ pressure [PaCO₂]: 84–103 mmHg and arterial O₂ pressure [PaO₂]: 155–172 mmHg). In the CV-group, normocapnea (PaCO₂: 42–50 mmHg) and good oxygenation (PaO₂: 395–419 mmHg) were achieved by the IPPV without apparent cardiovascular depression (heart rate: 29–31 beats/min, cardiac output: 17–21 l /min, MABP: 111–123 mmHg, MPAP: 27–30 mmHg and MRAP: 15–16 mmHg). MLBP-TIVA preserved cardiovascular function even in horses artificially ventilated.     

Identification of novel genetic markers and evaluation of genetic structure in a population of Japanese crested ibis

Japanese population of the Japanese crested ibis Nipponia nippon was founded by five individuals gifted from the People's Republic of China. In order to exactly evaluate genetic structure, we first performed development of novel genetic makers using 89 microsatellite primer pairs of related species for cross‐amplification. Of these, only three primer pairs were useful for the genetic markers. Additionally, we sequenced allelic PCR products of these three markers together with 10 markers previously identified. Most markers showed typical microsatellite repeat units, but two markers were not simple microsatellites. Moreover, over half of the markers did not have the same repeat units as those of the original species. These results suggested that development of novel genetic markers in this population by cross‐amplification is not efficient, partly because of low genetic diversity. Furthermore, the cluster analysis by STRUCTURE program using 17 markers showed that the five founders were divided into two clusters. However, the genetic relationships among the founders indicated by the clustering seemed to be questionable, because the analysis relied largely on a small number of triallelic markers, in spite of the addition of the three useful markers. Therefore, more efficient methods for identifying large numbers of single nucleotide polymorphisms are desirable.     

Synthesis of new glycosides by transglycosylation of N-acetylhexosaminidase from Serratia marcescens YS-1

Serratia marcescens YS-1, a chitin-degrading microorganism, produced mainly N-acetylhexosaminidase. The purified enzyme had an optimal pH of approximately 8-9 and remained stable at 40 degrees C for 60 min at pH 6-8. The optimum temperature was around 50 degrees C, and enzyme activity was relatively stable below 50 degrees C. YS-1 N-acetylhexosaminidase hydrolyzed p-nitrophenyl beta-N-acetylgalactosamide by 28.1% relative to p-nitrophenyl beta-N-acetylglucosamide. The N-acetylchitooligosaccharides were hydrolyzed more rapidly, but the cellobiose and chitobiose of disaccharides that had the same beta-1,4 glycosidic bond as di-N-acetylchitobiose were not hydrolyzed. YS-1 N-acetylhexosaminidase efficiently transferred the N-acetylglucosamine residue from di-N-acetylchitobiose (substrate) to alcohols (acceptor). The ratio of transfer to methanol increased to 86% in a reaction with 32% methanol. N-Acetylglucosamine was transferred to the hydroxyl group at C1 of monoalcohols. A dialcohol was used as an acceptor when the carbon number was more than 4 and a hydroxyl group existed on each of the two outside carbons. Sugar alcohols with hydroxyl groups in all carbon positions were not proper acceptors.