Purification and properties of β-galactosidase from Tilapia intestine: Digestive enzyme of Tilapia-X

ABSTRACT:   β-galactosidase of the intestine of Tilapia nilotica was purified by ammonium sulfate precipitation, followed by PAPTG-Sepharose 4B affinity chromatography, ethylenediamineetetraacetic acid ion-exchange chromatography, polyexchanger PBE 94 chromatofocusing, and Sephadex G-100 gel filtration. β-galactosidase was found to be a single band when examined by poly-acrylamide gel electrophoresis. The purifications of β-galactosidase were 27-fold from the crude extract. β-galactosidase showed optimum activity at pH 5.0 at 40°C, and was specifically found to be able to hydrolyze p -nitrophenyl β-galactopyranoside. It degrades galactan and agarose, and produces galactose. β-galactosidase was strongly inhibited by Hg²⁺ and PCMB. β-galactosidase is considered to be secreted by the upper and middle parts of the intestine and most of the activity was detected in the intestinal juice.     

Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.     

Comparative external morphology of cetacean spermatozoa

SUMMARY: To compare size and morphology of spermatozoa in cetaceans, sperm and epididymis samples were collected from 10 species in four families and spermatozoa were observed with phase-contrast and scanning electron microscopes. According to the average total length of the spermatozoa, the 10 species examined were classified into the following four groups in order of increasing size: (i) Baird's beaked (Ziphiidae) and Bryde's whales (Balaenopteridae); (ii) Dall's and finless porpoises (Phocoenidae); (iii) common, bottlenose, and Pacific white-sided dolphins (Delphinidae); and (iv) killer and short-finned pilot whales, and Risso's dolphin (Delphinidae). Spermatozoa head length of Bryde's whale and the finless porpoise were shorter than those of the other species. Spermatozoa head width was widest in the killer whale and thinnest in the Baird's beaked whale. The lateral aspects of sperm heads from the 10 species were characterized as the 'anterior region of the sperm head is thin and flat while the posterior region is thick.' The dorsal aspects of sperm heads were 'paddle-shaped' in Bryde's whales, 'bowling pin-shaped' in Baird's beaked whales, 'Japanese fan-shaped' in killer whales, an 'elongated ellipsoid shape' in Delphinidae except for killer whales, and 'ellipsoid shaped' in Phocoenidae. Size and morphology of the spermatozoa showed interspecific differences among the 10 species examined, which correspond to cetacean taxonomic classification.     

Comparison of environmental conditions in two representative oyster farming areas: Hiroshima Bay,western Japan and Oginoham a Bay (a branch of Ishinomaki Bay), northern Japan

ABSTRACT:   Sea water environmental conditions over annual cycles were investigated and compared between two oyster farming areas, western Hiroshima Bay and Oginohama Bay (a branch of Ishinomaki Bay) in Miyagi Prefecture, to appropriately manage oyster culture or more efficiently utilize farming areas. The environmental parameters of temperature, salinity, nutrient concentrations (NO₂–N, NO₃–N, NH₄–N, PO₄–P, and SiO₂–Si) and size-fractionated chlorophyll- a (<0.2, 2–20, >20µm), and abundances of microzooplankton were measured in each bay at the surface, and 2 and 5 m depth layers. Differences in the annual mean values and results with monthly paired Student's t -tests showed that salinity was lower, and temperature, nutrient (especially PO₄–P) and chlorophyll- a concentrations, and abundance of microzooplankton, were higher in Hiroshima Bay than in Oginohama Bay. Differences in environmental conditions between inshore and offshore areas of each bay suggest that inflows of river water in western Hiroshima Bay and sea water from offshore had the most significant effects on the environmental conditions. It is concluded that such oceanographic and biological differences strongly affect the oyster farming system, especially regarding the optimum usage of offshore areas in Summer under clean, cold and stable seawater conditions, rather than food quantity in Hiroshima Bay, and under more abundant food conditions in Oginohama Bay.     

氢能作为内燃机燃料的研究

该文论述了氢发动机运转中所存在的问题，提出了氢发动机的燃烧改进方案。研究表明，采用高压喷射、火花点火，并优化调整喷射正时和点火正时，就可控制异常燃烧，获得良好的性能     

Oxygen isotopic compositions of asteroidal materials returned from Itokawa by the Hayabusa mission

Meteorite studies suggest that each solar system object has a unique oxygen isotopic composition. Chondrites, the most primitive of meteorites, have been believed to be derived from asteroids, but oxygen isotopic compositions of asteroids themselves have not been established. We measured, using secondary ion mass spectrometry, oxygen isotopic compositions of rock particles from asteroid 25143 Itokawa returned by the Hayabusa spacecraft. Compositions of the particles are depleted in (16)O relative to terrestrial materials and indicate that Itokawa, an S-type asteroid, is one of the sources of the LL or L group of equilibrated ordinary chondrites. This is a direct oxygen-isotope link between chondrites and their parent asteroid.     

Regulation of muscular glutamate metabolism by high‐protein diet in broiler chicks

The major taste active component, glutamate (Glu), improves the taste of meat. In this study, we investigated the effect of a short‐term high‐protein (HCP) diet on the intramuscular free Glu content to improve the taste of meat. Furthermore, we elucidated how the muscle free Glu content was controlled by the HCP diet. Chicks (14 days old) were fed the control diet or HCP diet for 10 days. Plasma and muscle free amino acid concentrations, and activity and messenger RNA (mRNA) expression of muscle enzymes related to Glu metabolism were determined. Muscle free Glu content was increased (P < 0.01) by 51%. Activity and mRNA expression of glutaminase (GA), which is one of the major Glu‐related enzymes, were significantly decreased (P < 0.05) in the HCP group because of feedback inhibition. The mRNA expression of lysine α‐ketoglutarate reductase (LKR), which is the enzyme involved in lysine (Lys) degradation and Glu production, was significantly increased (P < 0.001) in the HCP group. These results suggest that short‐term dietary HCP feeding is an effective treatment for improving the taste of meat. Furthermore, our results suggest that the free Glu content in muscle is regulated by GA and LKR.