The mixed kidney cell-lymphocyte reaction in rats

Optimal conditions for obtaining the proliverative response in mixed kidney cell-lymphocyte culture (MKLC) have been determined in rats. The addition of 2-mercaptoethanol to the culture medium was essential for successful stimulation of lymphocytes by allogeneic kidney cells. Proliferative responses by standard MLC and MKLC were compared in rats with the same or a different Ag-B locus. In most strain combinations, there was a significant correlation between MLC and MKLC reactions, However, the DA-ACI combination, compatible at the Ag-B locus, consistently showed negative MLC and positive MKLC reactions. This suggested the possibility that this technique allowed the expression of kidney specific antigens.     

The MHD effect and its relaxation process on electric current in the electrolysis of ferricyanide reduction and ferrocyanide oxidation

The electrolytic reduction of ferricyanide and the electrolytic oxidation of ferrocyanide have been carried out with exposing the magnetic field of 1000 or 1800 gauss. The current clearly increased after the magnetic exposure. The maximum current was obtained when the magnetic flux directed in parallel with the surface of electrode. These are speculated in terms of magnetohydrodynamic mechanism. The current decrease caused by relaxation process was observed after the removal of magnetic flux. The relaxation time obtained was temperature-dependent. Therefore the values of apparent transition energy, E_trans, were determined from the Arrhenius' plots of relaxation time against temperature. The magnitude of E_trans was dependent on the concentration of ferricyanide or ferrocyanide, and the viscosity and the conductivity of electrolyte solution. The activation energy of viscosity of electrolyte solution was compared with E_trans. As a conclusion, it was suggested that E_trans may be influenced by the velocity of magnetohydrodynamic flow, which was controlled by the diffusion current in electrolysis and the viscosity of electrolyte solution.     

Applicability of DNA barcode for identification of fish species

DNA barcoding is a species identification technique, which uses a very short DNA sequence from a region of approximately 650 base-pairs in the 5'-end of the mitochondrial cytochrome c oxidase subunit I gene as a marker to identify species of mammals and fishes. The applicability of DNA barcoding for identification of fish species consumed in Japan was studied. Among thirty-one fresh or processed fishes were obtained from the market, two samples could not be identified due to lack of data in the Barcode of Life Data (BOLD) database. However, BLAST-search of 16S rRNA genes in the National Center for Biotechnology Information (NCBI) database and the PCR-RFLP method published by the Food and Agricultural Materials Inspection Center (FAMIC) were found to be applicable to identify these 2 fishes. The results show that the DNA barcoding technique is potentially useful as a tool for confirming the proper labeling of fish species in the Japanese market.     

Immunomodulatory effect of mushrooms on cytotoxic activity and cytokine production of intestinal lamina propria leukocytes does not necessarily depend on β-glucan contents

We evaluated the effects of seven mushroom extracts (Grifola frondosa, Pholiota nameko, Panellus serotinus, Hypsizygus marmoreus, Pleurotus cornucopiae, Armillaria mellea, and Flammulina velutipes) on cytotoxic activity and cytokine production of lamina propria leukocytes (LPLs) isolated from rat small (S) and large (L) intestinal mucosa. Boiling water extracts from seven species of mushrooms showed no direct cytotoxicity against the YAC-1 target cells. However, prominent increases of cytotoxicity were observed in S- and L-LPLs co-cultured with P. serotinus extract. Cytokine production (TNFα, IFNγ, IL-12 p70, and IL-4) of S- and L-LPLs was stimulated in response to P. cornucopiae extract. Mushroom extracts contributed to target cell adhesion and/or cytokine production in the effector cells. The promotion of cytotoxic activity in S- and L-LPLs was not necessarily related to β-glucan content of the mushroom.     

Characterization of Acetylxylan Esterase from White-Rot Fungus Irpex lacteus

The carbohydrate esterase family 1 (CE1) in CAZy contains acetylxylan esterases (AXEs) and feruloyl esterases (FAEs). Here we cloned a gene coding for an AXE belonging to CE1 from Irpex lacteus (IlAXE1). IlAXE1 was heterologously expressed in Pichia pastoris, and the recombinant enzyme was purified and characterized. IlAXE1 hydrolyzed p-nitrophenyl acetate, α-naphthyl acetate and 4-methylumbelliferyl acetate, however, it did not show any activity on ethyl ferulate and methyl p-coumarate. We also examined the activity on partially acetylated and feruloylated xylan extracted from corncob by hydrothermal reaction. Similarly, ferulic and p-coumaric acids were not liberated, and acetic acid was only detected in the reaction mixture. The results indicated that IlAXE1 is an acetylxylan esterase actually reacted to acetyl xylan. However, since IlAXE1 was unable to completely release acetic acid esterifying xylopyranosyl residues, it is assumed that acetyl groups exhibiting resistance to deacetylation by IlAXE1 are present in corn cob xylan.