To clarify the changes of serum haptoglobin (HP) and α1-acid glycoprotein (α1-AGP) concentrations and the effects of serum in new born calves with diarrhea on lymphocyte blastogenesis, eight healthy Japanese Black calves and 24 Japanese Black calves during the 12-24 h after the onset of diarrhea with Rota virus infection, 3-35d of age, were studied. Though serum HP was not detected in healthy calves, it did appear in the serum of all calves with diarrhea (599±414μg/ml). On the other hand, there was no significant difference between normal calves (398±100μg/ml) and calves with diarrhea (441±104μg/ml) in serum α1-AGP concentration. The lymphocyte blastogenesis induced by concanavalin A in peripheral mononuclear cells, isolated from one clinically healthy 13-month-old Japanese Black castrated male steer, was significantly suppressed compared to those from normal calves and the steer by adding heat-inactivated (56°C for 30min) sera from calves with diarrhea (P<0.01). Furthermore, a significant negative correlation between the serum HP concentrations and serum-lymphocyte blastogenesis reaction was obtained in calves with diarrhea (R²=0.693, P<0.01). However, no significant correlation was observed between the serum α1-AGP concentrations and the serum-lymphocyte blastogenesis reaction (R²=0.001). These results indicate that HP appears in the serum of newborn calves with Rota virus infection within 12-24h after the onset of diarrhea and that lymphocyte blastogenesis induced by T-cell mitogen is inhibited by serum from calves with Rota virus infection in a dosedependent manner of serum HP.

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The effects of glucose, a non-essential amino acid mixture and oleic acid on cytosolic calcium concentration ([Ca²+]_i) in individual cells were investigated with a confocal laser scanning microscope in reference to growth hormone (GH) release in caprine anterior pituitary cells cultured for 3d in Dulbecco's modified Eagle's medium. An increase in glucose concentration from 5.5 to 11.0mmol/l in HEPES-buffered saline solution (HBS) caused a slight, but not significant, increase in basal [Ca²+]_i and GH release. Administration of amino acids (NEAA; consisting of 7 non-essential amino acids; L-Asp, Gly, L-Ala, L-Ser, L-Pro, L-Asn, and L-Glu; the total concentration was 1.4mmol/l) significantly raised basal [Ca²+]_i (86.7% of the total cells) (P<0.05) and GH release (P<0.05). In contrast, oleic acid (10μmol/l) significantly reduced basal [Ca²+]_i (83.3% of the total cells)(P<0.05) and GH release (P<0.001). From these findings we conclude that some nutrients such as amino acids and oleic acid have a direct action on caprine anterior pituitary cells to alter the [Ca²+]_i level consequently modifying hormone release.

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The metabolic fates of carbon skePetans of lysine, threonine and leucine were examined in growing rats fed on the zein diets supplemented with graded levels of lysine (0.2, 0.4, 0.6, 0.8, 1.0 and 1.2%). The body weight gain and feed efficiency increased with an increment in lysine level up to 0.6% of the diet, and thereafter remained constant. The concentration of plasma free lysine and the expired ¹⁴CO₂ production from ¹⁴C-lysine in ratsremained a talow and cons tant level up to the lysinelevel of O.6% of thediet, at which point they started to increase inearlyas the dietary lysine level increased. However, the out put of ¹⁴CO₂ from labeled threonine and leucine decreased with the increasing dietary lysine level from 0.2 to 0.6 or 0.4%, respectively, and thereafter remained constant. The incorporation of radioactivity from ¹⁴C-lysine into the body protein was constant at the two low levels of lysine and at more than 0.6%, it gradually decreased. The incorporation of labeled amino acids into the body protein was inversely correlated to the ¹⁴CO₂ production from labeled amino acids. Under these experimental conditions, the calculated requlrement for lysine with body weight gain, plasma lysine and lysine oxidation as response criteria was 0.6% of the diet. These results showed that;1) when the zein diet was supplemented with mare lysine than its requirement, ¹⁴CO₂ release from the ¹⁴C-lysine increased and that from the labeled threonine and leucine remained constant, 2) addition of lysine up to the optimum level increased the amino acids used for the body protein synthesis and decreased the oxidation of amino acids.

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Diaminopimelate decarboxylase (EC4.1.1.20) from a cell free extract of a rumen protozoon Entodinium caudatum was purified 430-folds by ammonium sulphate precipitation and column chromatography with hydroxylapatite, DEAE-cellulose and sephadex G-150. The molecular weight of the enzyme was estimated to be 54, 000 by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The optimal pH for catalytic activity of the purified enzyme from E. caudatum was found to be 6.3 and catalytic activities were retained between 5.0 and 6.8. The optimal temperature was 50°C. The enzyme was stable at below 60°C and the activity was completely lost at 65°C. For the stability of the enzyme the presence of pyridoxal 5'-phosphate (PLP) and thiol group such as 2-mercaptoethanol and dithiothreitol were necessary. PLP was the only pyridoxine derivative which acted as a cofactor. The Michaelis constant (Km) of this enzyme for 2, 6-diaminopimelate was 0.8mM.

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A rapid new method for the simultaneous determination of tryptophan (Trp) and its related compounds such as indolelactic acid (ILA), tryptophol (TPP), tryptamine (TPM), indole-3-acetaldehyde (ICA), indoleacetic acid (IAA), indole-3-aldehyde (ILD), skatole (SKT) and indole (IND), and p-hydroxyphenylacetic acid (HPA) and trans-cinnamic acid (CNM) which appeared within the retention time of final compound, SKT, in the chromatogram was established by highperformance liquid chromatography (HPLC). LiChrospher 100 RP-18 column and methanol-50 mM s odium acetate buffer (pH 4.2) (30: 70, v/v) as a mobile phase were used for the analysis. The compounds were monitored at 280nm with a UV absorbance detector. Prior to the analysis, sample was mixed with an equal volume of 4% (w/v) sulfosalicylic acid (SSA) for deproteinization. A 5μl portion of deproteinized and filtered sample was injected into the HPLC and the final peak of SKT appeared at about 60min. The recovery of every compound was more than 95%. The minimum detectable limits (pmol/5μl) of the secompounds were as follows: Trp, ILA, IAA, ILD and TPP, 19; ICA, HPA, IND and CNM, 39; SKT, 156 and TPM, 625. When this method was applied to the analysis of goat rumen fluid, HPA, ICA, Trp, ILA, IAA, ILD and TPP were detected. Among them ICA, HPA and IAA showed relatively high amounts. The concentrations of these compounds varied with the time after feeding.

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This study comparatively evaluated the influence of quail or chicken lysozyme and bovine or human α-lactalbumin at concentrations ranging from 0 to 100μg/ml on proliferative responses of mouse spleen lymphocytes stimulated by lipopolysaccharide, pokeweed mitogen, phytohemagglutinin and concanavalin A. The lysozymes and α-lactalbumins inhibited the mitogen-induced proliferations in a dose-dependent manner, although the magnitude of inhibition depended on the kind of protein or mitogen. The α-lactalbumins revealed a cytotoxicity toward lymphocytes while the lysozymes were inert. The cytotoxicity of bovine α-lactalbumin diminished completely when the protein was reduced S-carboxymethylated, while some inhibitory effects on the PHA-or ConA-induced proliferation remained after the modification. No mitogen combined with the lysozymes and α-lactalbumins, and the mitogen could bind to lymphocytes previously incubated with the lysozymes or α-lactalbumins. Bovine α-lactalbumin linked to murine CD⁴⁺ T-lymphocytes, CD⁸⁺ T-lymphocytes and monacytes/macrophages, while quail lysozyme bound to only monocytes/macrophages. These results indicate that both the lysozyme and α-lactalbumin suppress the proliferation of mouse spleen lymphocytes via a different specific interaction of lymphocytes and/or macrophages with each protein.

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Selenium (Se) balance trials were undertaken to determine the amount of Se accumulation in the late pregnancy and lactation period, and the effects of fetus number and feeding level on the Se balance and blood Se concentration were investigated.(Experiment 1) Holstein dairy cows carrying single (n=5) or twin (n=4) Japanese Black fetuses were used for the Se balance trials at 210 and 266 day of pregnancy. Feeding of 0.106ppm Se containing diet resulted in 973.3μg/day of the Se intake and 136.9μg/day of the Se accumulation on average. The Se accumulation of dam fed the amount of diet for twin was significantly larger than for single. Although there were no significant differences in the Se accumulation between single and twin pregnant cattle, the plasma and whole blood Se concentration of twin calf was significantly lower than that of single. The plasma Se concentration of all calves was not in sufficient level regardless of the dam's treatment condition or fetus number.(Experiment 2) Se balance trials were undertaken at mean 67 day of the lactation by using 4 Holstein milking cattle. The Se intake and accumulation in the lactation increased 4.7 times and 2.2 times to those of the late pregnancy, respectively, because of an increase of feed intake and of average Se concentration of the diet (0.226ppm). The blood Se concentration was in the sufficient level. These results suggest that dairy cattle are short of Se not in the lactation but the late pregnancy.

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Ten weeks old wister strain pregnant rats were fed ad libitum CE-2 rat chow (Clea Japan) and water. On the 15th days of pregnancy, 4 groups of 5 rats were each orally administered, 2ml (i) water (control), (ii) Trp-P1 solution (25mg/kg in 5ml distilled water), (iii) lyophilised Leuconostoc paramesenteroides R62 cells (350mg/kg) suspended in 5ml Trp-P-1 solution or (iv) lyophilised R62 cells (350mg/kg) suspended in 5ml distilled water. The same administration was continued for three consecutive days. The brain weight of the fetus was significantly decreased by the administration of Trp-P1 (P<0.05). The administration of Trp-P1 along with R62 cells also showed decrease in brain weight of the fetus, but the decrease was comparatively less than that of the Trp-P1 group (P<0.05). The ratio between brain weight and body weight was higher in lactic acid bacteria with Trp-P1 group than the Trp-P1 group (P<0.05).

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