Development of enzyme-linked immunosorbent assays for conglutinin, mannan-binding protein, and serum amyloid-P component in bovine sera.

Sandwich ELISA were developed to quantitatively determine conglutinin (CG), mannan-binding protein (MBP), and serum amyloid-P component (SAP) in the sera of cattle. The ELISA system was found to have high repeatability for quantitation of these serum proteins at concentration as low as 5 ng/ml. From results obtained for 10 healthy cows aged 2 to 7 years, mean +/- SD serum concentrations were 56.5 +/- 14.4 micrograms of CG/ml, 2.37 +/- 0.87 micrograms of MBP/ml, and 11.14 +/- 3.92 micrograms of SAP/ml, respectively. Values in 6 healthy heifer calves aged 6 months were 3.45 +/- 1.22 micrograms/ml for CG, 1.71 +/- 0.96 micrograms/ml for MBP, and 5.45 +/- 2.75 micrograms/ml for SAP, respectively. Concentrations in 9 healthy bullocks aged 6 months were 1.83 +/- 0.66 micrograms/ml for CG, 1.04 +/- 0.63 micrograms/ml for MBP, and 4.9 +/- 1.13 micrograms/ml for SAP, respectively.     

Conglutinin, mannan-binding protein, and serum amyloid P component concentrations in sera from cows: changes associated with mastitis.

To study the roles of conglutinin (Kg), mannan-binding protein (MBP), and serum amyloid P component (SAP) in the protection of cattle against infections, the concentrations of these proteins in the sera from cows with mastitis were determined by sandwich enzyme-linked immunosorbent assays. The Kg and MBP concentrations in the sera from cows with mastitis were much lower than those from uninfected (or clinically healthy) ones, and increased after recovery. No significant difference in the SAP concentrations was found between healthy and infected cows. With the sera from cows experimentally infected with bovine leukemia virus, the Kg concentrations were also lower than those from uninfected ones, whereas the MBP concentrations were not. From these findings, the Kg concentration is suggested to be serologically a possible indicator for clinical diagnosis of treatment for mastitis although both Kg and MBP concentrations were found to decrease in cows with mastitis and to increase after recovery.     

The influence of age and body mass index on relative accuracy of energy intake among Japanese adults

OBJECTIVE: To examine relationships between the ratio of energy intake to basal metabolic rate (EI/BMR) and age and body mass index (BMI) among Japanese adults. DESIGN: Energy intake was assessed by 4-day semi-weighed diet records in each of four seasons (16 days in total). The EI/BMR ratio was calculated from reported energy intake and estimated basal metabolic rate as an indicator of reporting accuracy. SETTING: Residents in three areas in Japan, namely Osaka (urban), Nagano (rural inland) and Tottori (rural coastal). SUBJECTS: One hundred and eighty-three healthy Japanese men and women aged > or =30 years. RESULTS: The oldest age group (> or =60 years) had higher EI/BMR values than the youngest age group (30-39 years) in both sexes (1.74 vs. 1.37 for men; 1.65 vs. 1.43 for women). In multiple regression analyses, age correlated positively (partial correlation coefficient, beta = 0.012, P < 0.001 for men; beta = 0.011, P < 0.001 for women) and BMI correlated negatively (beta = -0.031, P < 0.001 for men; beta = -0.025, P < 0.01 for women) with EI/BMR. CONCLUSION: Age and BMI may influence the relative accuracy of energy intake among Japanese adults.     

Interaction between ascorbic acid and chlorogenic acid during the formation of nitric oxide in acidified saliva

When saliva and gastric juice are mixed, salivary nitrite is transformed to nitrous acid to produce nitric oxide (NO). The NO formation in acidified saliva was enhanced by ascorbic acid and chlorogenic acid. Thiocyanate ion (SCN(-)) also enhanced the transformation of nitrous acid to NO. During the NO formation in the presence of both ascorbic acid and chlorogenic acid, ascorbic acid was preferentially oxidized. Chlorogenic acid was oxidized after ascorbic acid had been oxidized. Ascorbyl radical was detected during the oxidation of ascorbic acid, and the radical intensity was decreased by chlorogenic acid. The decrease is discussed to be due to the reduction of the oxidation intermediate or product of chlorogenic acid by ascorbyl radical. The result obtained in this study suggests that ascorbic acid was preferentially oxidized and that not only ascorbic acid but also ascorbyl radical could interact with the oxidation intermediate or product of chlorogenic acid when chlorogenic acid was added to the mixture of saliva and gastric juice that contained ascorbic acid.     

Reduction of nitrous Acid to nitric oxide by coffee melanoidins and enhancement of the reduction by thiocyanate: possibility of its occurrence in the stomach

Reactions of nitrous acid with freeze-dried instant coffee and its methanol-insoluble melanoidin fractions were studied at pH 2 in the presence and absence of thiocyanate (SCN (-)), simulating the mixture of coffee, saliva, and gastric juice. Coffee contained stable radicals, and the radical concentration increased by ferricyanide and decreased by ascorbic acid. This result indicates that the radical concentration was affected by the redox state of coffee and that the nature of the radical was due to quinhydrone structure that might be included in coffee melanoidins. Nitrite also increased the electron spin resonance (ESR) signal intensity at pH 2, suggesting that nitrite oxidized melanoidins producing nitric oxide (NO). The formation of NO could be detected by oxygen uptake due to the autoxidation of NO and using an NO-trapping agent. SCN (-) largely enhanced NO formation in coffee and methanol-insoluble melanoidin fractions but only slightly in a methanol-soluble fraction, and the enhancement accompanied the consumption of SCN (-) but did not accompany the formation of a stable ESR signal. The enhancement was explained by the reduction of NOSCN by melanoidins in methanol-insoluble fractions and that the consumption was due to binding of SCN (-) to melanoidins during their oxidation by nitrous acid. The result obtained in this study suggests that when coffee is ingested, in addition to chlorogenic acid and its isomers, melanoidins can also react with salivary nitrite and SCN (-) in the gastric lumen, producing NO.     

Effects of the food additive sulfite on nitrite-dependent nitric oxide production under conditions simulating the mixture of saliva and gastric juice

The food additive sulfite is mixed with saliva, which contains nitrite, in the oral cavity, and the mixture is mixed with gastric juice in the stomach. In the stomach, salivary nitrite can be transformed to nitric oxide (NO). In this study, the effects of sulfite on nitrite-dependent NO production were investigated using acidified saliva (pH 2.6) and acidic buffer solutions (pH 2.0). Sulfite enhanced NO production in acidified saliva and acidic buffer solutions, and the enhancement increased with the increase in sulfite concentration from 0 to 0.1 mM, whereas suppressed NO production and the suppression increased as the concentration was increased over 0.2 mM. The enhancement was due to the increase in reaction rate between nitrous acid and nitrososulfonate (ONSO(3)(-)) that was formed by the reaction of nitrous acid with hydrogen sulfite, and the suppression was due to the increase in hydrogen sulfite-dependent consumption rate of ONSO(3)(-). A salivary component SCN(-) (1 mM) enhanced and suppressed NO production induced by 1 mM nitrite when sulfite concentrations were lower and higher than 1 mM, respectively. ONSO(3)(-) formed from hydrogen sulfite and nitrosyl thiocyanate (ONSCN), which was produced by the reaction of nitrous acid with SCN(-), seemed to contribute to the enhancement and suppression. NO production induced by nitrite/ascorbic acid systems was suppressed by sulfite, and the suppressive effects were decreased by SCN(-), whereas sulfite-induced suppression of NO production in nitrite/rutin systems was increased by SCN(-). During reactions of nitrite with sulfite in the presence and absence of SCN(-), oxygen was taken up. The oxygen uptake is discussed to be due to autoxidation of NO and radical chain reactions initiated by hydrogen sulfite radicals. The results of the present study suggest that sulfite can enhance and suppress nitrite-dependent NO production. It is discussed that radicals including hydrogen sulfite radicals can be formed through the reactions of nitrite and sulfite in the stomach.     

Inhibition of buckwheat starch digestion by the formation of starch/bile salt complexes: possibility of its occurrence in the intestine

During the digestion of starch in foods, starch is mixed with bile in the duodenum. Because fatty acids and some kinds of polyphenols could bind to starch, it was postulated that bile salts might also bind to starch. The purpose of this paper is to study the effects of bile and bile salts on starch/iodine complex formation and pancreatin-induced starch digestion. Bile suppressed starch/iodine complex formation and inhibited pancreatin-induced starch digestion slightly in control buckwheat starch, but did so significantly in buckwheat starch from which fatty acids and polyphenols had been extracted. Such significant suppression and inhibition by bile were also observed in a reagent soluble starch. The effects of cholate and taurocholate on the starch/iodine complex formation and the pancreatin-induced starch digestion were essentially the same as those of bile. Bile, cholate, and taurocholate suppressed amylose/iodine complex formation more significantly than amylopectin/iodine complex formation and inhibited pancreatin-induced amylose digestion more effectively than the digestion of amylopectin. It is concluded from the results that bile salts could bind to starch, especially amylose, the helical structures of which were not occupied by other molecules such as fatty acids and polyphenols, and that the binding resulted in the inhibition of starch digestion by pancreatin. The conclusion suggests that the function of bile salts can be discussed from the point of not only lipid digestion but also starch digestion.     

Protective effects of sugar cane extracts (SCE) on Eimeria tenella infection in chickens

The effects of oral administration of sugar cane extracts (SCE) on Eimeria tenella oocysts infection in chickens were studied with 2 different experiments. In Experiment 1, 3-week-old inbred chickens (MHC; H.B15) were inoculated into the crop with SCE (500 mg/kg/day) for 1 day or 3 consecutive days, and then challenged with E. tenella sporulated oocysts (2 x 10(4) cells/chicken). In Experiment 2, 1-week-old chickens were orally administered SCE at the same dose for 3 consecutive days, and then initially infected with E. tenella sporulated oocysts (2 x 10(3) cells/chicken). At 2 and 3 weeks of age, these chickens were immunized intravenously with the mixed antigens of sheep red blood cells (SRBC) and Brucella abortus (BA). At 4 weeks of age, chickens were challenged with E. tenella sporulated oocysts (1 x 10(5)/chicken). Challenged chickens with E. tenella oocysts showed markedly decreased body weight gain/day, severe hemorrhage and great number of shedding oocysts in feces and high lesion scores. Oral administration of SCE and initial infection with oocysts (2 x 10 (3)/chicken) resulted in a remarkable improvement in body weight gain/day, hemorrhage, the number of shedding oocysts and lesion score, compare to other infected groups. In addition, SCE-inoculated chickens with the initial infection showed a significant increase in antibody responses against SRBC and BA and also improvement in decreased relative proportions of Bu-1a(+) and CD4( )cells in cecal tonsil lymphocytes of E. tenella-challenged chickens. Cecal tissues of chickens administered SCE and initially infected with E. tenella oocysts showed lower numbers of schizonts, gametocytes and oocysts than those of infected control chickens. These results suggest that SCE have immunostimulating and protective effects against E. tenella infection in chickens.     

Immunostimulating and growth-promoting effects of sugar cane extract (SCE) in chickens

Polymorphonuclear cells of the peripheral blood in the chicken significantly increased their phagocytosis when cultured with sugar cane extract (SCE; 250-1,000 microg/ml) for 24 hr. Chickens orally administered SCE (500 mg/kg/day) for 3 or 6 consecutive days at 1 week of age showed significantly higher body weight and gain in body weight/day and a lower food conversion ratio within the growing period of 6 weeks than physiological saline-administered control chickens. Furthermore, oral administration of SCE also resulted in significantly higher immune responses against sheep red blood cells and Brucella abortus. These results suggest that SCE has immunostimulating and growth promoting effects in chickens.