Changes of lymphocyte subpopulations and natural killer cells in mice sensitized with Toxoplasma lysate antigen before and after Babesia infection

When 8-week-old BALB/c mice were sensitized with two intramuscular injections of Toxoplasma lysate antigen (TLA) at 2 week interval, the numbers of sIg(+), Thy-1,2(+), Lyt-1,2(+) Lyt-2,2(+), and Asialo GM1(ASGM1)(+) cells in the spleen, liver and peripheral blood increased by 2 to 4 times over those found in unsensitized mice of the same age. When TLA-sensitized and unsensitized mice were infected with Babesia, 4 of 10 (40%) of the TLA-sensitized mice survived infection, while none of the unsensitized control mice lived longer than 14 days after Babesia infection. By contrast, sensitization of nude mice with TLA had no effect on survival, and mice did not live more than 12 days. The number of thymic Thy-1,2(+) cells decreased in TLA-sensitized and unsensitized BALB/c mice by almost 80% within 10 days after infection (AI). During the same time, the numbers of B cells, T cells, and NK cells increased in the spleen, liver and peripheral blood of both sensitized and unsensitized mice. Especially notable were increases in numbers of Lyt-2,2(+) cells in the spleen and blood and increases in numbers of NK cells in the spleen, liver and blood in both TLA-sensitized and unsensitized mice. When spleen cells from TLA-sensitized and unsensitized mice were cultured in the presence or absence of TLA for 6 days, assays for cytotoxicity using NK-insensitive P-815 target cells and NK-sensitive YAC-1 target cells demonstrated higher rates of cytotoxicity in cultures of TLA-sensitized spleen cells.     

Narthecium asiaticum Maxim. Poisoning of grazing cattle: observations on spontaneous and experimental cases

A total of 39 Holstein cattle were grazed in tracts of wild grassland on account of shortage in pasture grass. Twenty-nine cattle were affected and 26 of them died during a 21-day period. The main signs were depression, anorexia, ascites, and oliguria. There was elevated serum urea nitrogen and sugar and protein in the urine. Pathological examination revealed turbid swelling of the kidney, an increase in the amount of fluid in the body cavity, edema in the perirenal adipose tissue and hemorrhage in various visceral organs and tissues. Histologically, acute tubular necrosis in the kidney, hypoplasia of the erythroblast series in the bone marrow, atrophy and degeneration of the lymphatic tissue and focal necrosis of the liver were observed in many of the cattle. Among cows experimentally fed Narthecium asiaticum Maxim., Polygonum sachalinense Fr. Schum., and Vitis coignetiae Pulliat which were presumed to have been ingested in large amounts by grazing cattle in the field, the cows fed N. asiaticum revealed the clinical, biochemical and pathological changes similar to those noticed in naturally affected cattle. Cows fed P. sachalinense and V. coignetiae showed no distinct systemic symptoms except transient anorexia and hypothermia.     

Correlation between the Cell Number and Diameter in Bovine Embryos Produced In Vitro

This study was designed to examine the effects of age and developmental stage of in vitro‐produced bovine embryos on the cell number of the embryos and to investigate the correlation between the cell number and diameter in the embryos. The diameter and cell number in blastocysts and expanded blastocysts collected on days 7–9 after in vitro fertilization (IVF) were examined. Although the diameters of the blastocysts collected on days 7 and 8 after IVF were smaller than those of the expanded blastocysts collected on day 9, the cell number in both types of embryos was similar. The cell numbers of the blastocysts and expanded blastocysts decreased with increasing embryo age. There were positive correlations between the cell number and diameter in bovine embryos at each stage collected on each day after IVF. However, the value of the correlation coefficient in the day‐9 expanded blastocyst group tended to be higher than that in the other groups. These results indicate that the cell number of in vitro‐produced embryos is affected by the embryonic stage and age. The diameter of the embryo may be potentially used for the viability testing of the expanded blastocysts collected on day 9 after IVF.     

Flow cytometric enumeration of reticulocyte in the peripheral blood from canine infected with Babesia gibsoni

Peripheral blood samples from dogs infected with Babesia gibsoni were analyzed by a flow cytometer for the percentage of reticulocytes after staining with a membrane-permeable fluorochrome, thiazole orange. Though thiazole orange has been reported to stain human reticulocytes and Plasmodium-infected erythrocytes, number of positive cells determined by the flow cytometry did not include that of erythrocytes infected with Babesia gibsoni. Analysis of 51 samples revealed a correlation coefficient of 0.96 as compared to the conventional determination by light microscopy. Separation of reticulocytes from Babesia gibsoni-infected erythrocytes by flow cytometry with or without the stain remained unresolved.     

Inactivation of staphylococcal enterotoxin-A with an electrolyzed anodic solution.

Electrolyzed anodic NaCl solutions [EW+], prepared by the electrolysis of 0.1% NaCl, have been shown to instantly inactivate most pathogens that cause food-borne disease. Elimination of food-borne pathogens does not necessarily guarantee food safety because enterotoxins produced by pathogens may remain active. We have tested whether EW+ can inactivate Staphylococcal enterotoxin A (SEA), one of the major enterotoxins responsible for food poisoning. Fixed quantities of SEA were mixed with increasing molar ratios of EW+, and SEA was evaluated by reversed-phase passive latex agglutination (RPLA) test, immunoassay, native polyacrylamide gel electrophoresis (PAGE), and amino acid analysis after 30 min incubations. Exposure of 70 ng, or 2.6 pmol, of SEA in 25 microL of PBS to a 10-fold volume of EW+, or ca. 64.6 x 10(3)-fold molar excess of HOCl in EW+, caused a loss of immuno-reactivity between SEA and a specific anti-SEA antibody. Native PAGE indicated that EW+ caused fragmentation of SEA, and amino acid analysis indicated a loss in amino acid content, in particular Met, Tyr, Ile, Asn, and Asp. Staphylococcal enterotoxin-A excreted into culture broth was also inactivated by exposure to an excess molar ratio of EW+. Thus, EW+ may be a useful management tool to ensure food hygiene by food processing industries.     

Improvement of the bitter taste of amino acids through the transpeptidation reaction of bacterial gamma-glutamyltranspeptidase.

The tastes of several bitter amino acids and their gamma-glutamyl derivatives were compared. The bitterness of Phe, Val, Leu, and His was reduced, sourness was produced, and preferences were increased by gamma-glutamylization. Because the effect of gamma-glutamylization of bitter amino acids was most obvious for Phe, which is an atypical bitter amino acid, an enzymatic method for the synthesis of gamma-glutamylphenylalanine (gamma-Glu-Phe) involving bacterial gamma-glutamyltranspeptidase was developed. The optimum reaction conditions were 200 mM Gln, 200 mM Phe, and 0.5 unit/mL GGT, pH 10.4. After 1.5-h of incubation at 37 degrees C, 140 mM gamma-Glu-Phe was obtained, the yield being 70%. gamma-Glu-Phe was purified on a Dowex 1x8 column and then identified by NMR.     

Solute effects on the interaction between water and ethanol in aged whiskey

The hydrogen-bonding structure of water-ethanol in whiskey was examined on the basis of (1)H NMR chemical shifts of the OH of water and ethanol. Phenolic acids and aldehydes (gallic, vanillic, and syringic acids; vanillin and syringaldehyde) exhibited their structure-making effects regardless of the presence or absence of 0.1 or 0.2 mol dm(-3) acetic acid. The OH-proton chemical shifts were measured for 32 malt whiskey samples of a distillery, aged for 0-23 years in five different types of casks. The OH-proton chemical shift values of the whiskies shifted toward the lower field in proportion to their contents of total phenols. It can be concluded that the strength of the hydrogen bonding in aged whiskies is directly predominated by acidic and phenolic components gained in oak wood casks and not dependent on just the aging time.     

Evaluating the durability of wood-based panels using internal bond strength results from accelerated aging treatments

In this study, the durability of wood-based panels was evaluated by comparing the internal bond (IB) strength retention after five different laboratory-based accelerated aging tests with the IB retention after 5 years of outdoor exposure in Shizuoka City. In each accelerated aging test, the IB retention of MDI-bonded panels showed high retention compared to other panels. Outdoor exposure in Shizuoka City resulted in an IB retention value for particleboard (PF) and oriented strandboard (aspen) of less than 10% after the 5-year exposure period. Medium-density fiberboards maintained their initial IB strength over the same period. Calculation of the mean IB retention for all board types allowed comparison of the severity of aging between the accelerated test methods and outdoor exposure. The ASTM six-cycle test method was the most severe among the standard treatment cycles applied.