Effect of supplementation of Agaricus mushroom meal extracts on enzyme activities in peripheral leukocytes of calves.

To investigate the effect of Agaricus mushroom meal on the energy metabolism in animal tissues; plasma glucose, triglyceride, cholesterol and immunoreactive insulin (IRI) concentrations and activities of enzymes related to energy metabolism in plasma and peripheral leukocytes were measured in Japanese Black WagyuxHolstein F1 calves supplemented with Agaricus blazei Murill (A. blazei) extract in milk-replacer at the dose of 60g/head/day for 4 weeks. Activities of malate dehydrogenase and aspartate aminotransferase in cytosol and glutamate dehydrogenase in mitochondria, and the malate dehydrogenase/lactate dehydrogenase ratio in cytosol in peripheral leukocytes of calves with A. blazei were significantly higher than those in control calves without A. blazei. It was concluded that supplementation of Agaricus mushroom meal extract was effective in activation of enzymes related to energy metabolism in peripheral leukocytes of calves.

Serine acetyltransferase involved in cysteine biosynthesis from spinach: molecular cloning, characterization and expression analysis of cDNA encoding a plastidic isoform.

A cDNA clone that encodes a chloroplast-localizing isoform of serine acetyltransferase (SATase) (EC 2.3.1.30) was isolated from spinach (Spinacia oleracea L.). The cDNA encodes a polypeptide of 347 amino acids containing a putative transit peptide of ca. 60-70 amino acids at the N-terminal. Deduced amino acid sequence of SATase from spinach exhibited homology with other SATases from plants. DNA blot hybridization analysis showed the presence of 2-3 copies of Sat gene in the genome of spinach. RNA blot hybridization analysis indicated the constitutive expression of Sat gene in green and etiolated seedlings of spinach. Bacterial expression of the cDNA could directly rescue the cysteine auxotrophy of Escherchia coli caused by a lack of SATase locus (cysE). Catalytically active SATase protein was produced in E. coli cells. L-Cysteine, an end product of the cysteine biosynthetic pathway, inhibited the activity of recombinant spinach SATase, indicating the regulatory function of SATase in this metabolic pathway. A chloroplastic localization of this spinach SATase was revealed by the analyses of transgenic plant expressing transit peptide of SATase-beta-glucuronidase (GUS) fusion protein, and transient expression using the transit peptide-green fluorescent protein (GFP) fusion protein. The result from in vitro translation analysis suggests that this cDNA may encode both plastidic and cytosolic SATases.

Concentrations of trace elements in sweat during sauna bathing.

Trace elements in sweat during sauna bathing were assessed. Sweat collected by the whole body method was compared with that collected by the arm bag method. The sweat samples were collected from ten healthy male adults aged 22-26 years, by heat exposure in dry sauna bathing (60 degrees C, 30 minutes). Concentrations of major (Na, Cl, K, Ca, P and Mg) and trace (Zn, Cu, Fe, Ni, Cr and Mn) elements in sweat tended to be lower in the arm bag method than in the whole body method. It was found that Ca, Mg, Fe and Mn concentrations in the arm bag method were significantly lower than those in the whole body method. The amount of trace elements in sweat measured by the arm bag method was less than that by the whole body method; significant differences were observed in Fe and Mn amounts. These observations suggest that excretion of trace elements by sweating induces trace element decrease. Therefore, athletes and workers who work in a hot environment and sweat much habitually should ingest adequate amounts of trace elements.

[Basic study of Beutin's washed sheep blood agar plate used for selective screening of verocytotoxin-producing/enterohemorrhagic Escherichia coli (VTEC/EHEC)].

Enterohemolysin production of Beutin's washed sheep blood agar plate has been used as an epidemiological marker for selective screening of Verocytotoxin-producing/Enterohemorrhagic Escherichia coli (VTEC/EHEC). In this study, we examined about the component of Beutin's washed sheep blood agar for further improvement of hemolysin production media. The following items were studied: the numbers of washings of sheep blood with phosphate buffered saline (PBS) (pH 7.2), concentration of sheep blood, kind and concentration of divalent metal ion (Ca2+, Mg2+) and basal medium. Twenty-seven strains of VT-producing E. coli of 7 different O-serotypes, 74 strains of VT-nonproducing E. coli of 24 different O-serotypes and one strain of O157 coded Escherichia hermanii were used for this basic study. In comparison of washing times of sheep blood with PBS, 5 times washing was better than 3 times, the original. In sheep blood concentration, supplement with 4% sheep blood was best for hemolysis observation. In experiment of addition of 2 divalent metal ions, Ca2+ and Mg2+, supplement with Ca2+ was more suitable than Mg2+ for hemolysis, and the supplement with 10 mM CaCl2, the original, was the best concentration. On the basal medium used in Beutin's sheep washed blood agar, 4 kinds of media were compared. In addition to Soybean-Casein Digest (SCD) agar, the original, Nutrient agar, Heart Infusion (HI) agar and Brain Heart Infusion (BHI) agar were examined, HI agar was the best blood agar base in the four media. As the above experimental results, the composition of the better Beutin's washed sheep blood agar may be summarize as follows: Heart Infusion agar 'Eiken' used for blood agar base, supplemented with 10 mM CaCl2 and 4% defibrinated sheep blood (Japan biosupp. Center) washed five times in phosphate-buffered saline, pH 7.2.

Entactin-2: a new member of basement membrane protein with high homology to entactin/nidogen.

Using the new signal sequence trap (SST) method, we isolated several clones encoding secreted and transmembrane proteins from KUSA cells, a murine osteoblast-like cell line. One isolated novel clone, termed entactin-2, exhibited a high similarity to mouse entactin/nidogen, a basement membrane protein. Although deduction of the amino acid sequence of entactin-2 revealed only 27.4% homology to entactin, many structural similarities were seen between both proteins. Entactin-2 contains five EGF-like and two thyroglobulin-like motifs, which are both cysteine-rich. Comparison of both proteins clearly revealed that entactin-2 also contains related domain structures. The rod-like domain of entactin-2, containing the RGD integrin recognition sequence, fused to glutathione-S transferase (GST), revealed a cell surface-binding activity similar to that of entactin. In addition, the tissue distribution of entactin-2 mRNA resembled that of entactin. Furthermore, mRNA expression of both genes decreased as osteoblastic differentiation progressed. These results suggest that entactin-2 is a member of the entactin gene family, may have entactin-related functions, and might act as a basement membrane component.

[Portal embolization using lipiodol-gelatin sponge].

Gelatin sponge moistened with lipiodol was prepared for use in portal embolization to improve percutaneous transhepatic portal embolization as preoperative management of patients undergoing extensive liver resection. For one sheet of gelatin sponge (20 x 60 x 7mm)0.5ml of lipiodol was used. Five sheets cut to 2mm square pieces and mixed with physiological saline solution were injected into the right portal vein. This embolic material proved safer and more reliable than gelatin sponge mixed with water-soluble contrast medium, because it provided sufficient opacification to grasp the extent of embolization and prevented back flow.

[A case of cardiomyoplasty with synchronous stimulated latissimus dorsi muscle].

In December, 1988, a 57-year-old man sustained an acute myocardial infarction. He suffered from progressive cardiac failure, despite bed rest and intensive medical treatment. Finally, he developed hepatic and renal failure. Subsequent angiographic studies revealed the total occluded LAD (Seg. 6) and a large LV aneurysm. In January 1989, a left ventricular aneurysmectomy and cardiomyoplasty using the latissimus dorsi muscle were performed successfully. Two years after the operation, he is alive and well.