Background: Endotoxin plays an important role in the initiation and aggravation of alcoholic liver disease. In this study, we evaluated plasma endotoxin levels and serum concentrations of cytokines and lipopolysaccharide binding protein (LBP) during the acute and recovery phase of patients with alcoholic hepatitis; we also explored the prognostic factors associated with a fatal outcome.Methods: Fourteen patients, consisting of eight patients with alcoholic hepatitis (AH), five cirrhotics with superimposed AH (LC+AH), and one patient with severe alcoholic hepatitis (SAH), were studied. Among these, two with LC+AH died of hepatic failure.Results: Plasma endotoxin levels in the acute phase were higher in patients with AH (184.4 ± 159.4 pg/ml) and LC+AH (206.9 ± 174.9 pg/ml) than in healthy subjects (10.4 ± 5.5 pg/ml, p < 0.001). In particular, in one patient with SAH and one of two nonsurvivors, plasma endotoxin levels were markedly high relative to the other cases. In most survivors, plasma endotoxin levels decreased in the recovery phase, whereas they further increased at the terminal stage in one of two nonsurvivors. Serum interleukin (IL)‐6 and IL‐8 levels in the acute phase were significantly higher in patients with AH and LC+AH as compared with healthy subjects. These levels were especially high in nonsurvivors and in one patient with SAH. IL‐10 increased in two nonsurvivors, one patient with SAH, and one with LC+AH. In the recovery phase, these cytokine levels in survivors tended to decrease, but in nonsurvivors, IL‐6 remained high, and IL‐8 and IL‐10 further increased. Tumor necrosis factor‐α levels were below the detection limit throughout the course in all patients. Serum lipopolysaccharide binding protein (LBP) generally was elevated in the acute phase and decreased in the recovery phase in all survivors, but in one of the nonsurvivors, LBP was elevated markedly at the terminal stage. In the acute phase, plasma endotoxin levels were correlated positively with white blood cell counts, neutrophil counts, and serum IL‐8. IL‐8 was correlated positively with neutrophil counts and negatively with serum Cholinesterase, hepaplastin test, and serum albumin levels. IL‐6 was correlated positively with white blood cell and neutrophil counts, C‐reactive protein, and serum total bilirubin and negatively with hepaplastin test and serum total protein levels. Serum LBP was correlated positively with white blood cell and neutrophil counts.Conclusions: Endotoxemia and related elevation of IL‐8 may play an important role in the activation and migration of neutrophils in patients with alcoholic hepatitis. Marked elevation of inflammatory cytokines, IL‐6 and IL‐8, are related to severity and poor prognosis of alcoholic hepatitis. Serum LBP may serve as an index of inflammatory reaction in alcoholics.

Gas chromatography-mass spectrometry was employed to identify the two volatile amines in male mouse urine. These amines were much less concentrated in urine of castrated males. The identified amines, isobutylamine and isoamylamine, were assayed for the potential of puberty acceleration in postweaning female mice. A total of 105 young female mice were exposed to one of the following five odors: distilled water (control), 0.1 M isobutylamine, 0.1 M isoamylamine, a mixture of 0.05 M isobutylamine and 0.05 M isoamylamine, or fresh male mouse urine. The mixture of these amines accelerated the vaginal opening of young females. Except for the control, all experimental odors accelerated the first vaginal estrus in ICR strain mice.

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We examined the participation of the superoxide anion radical (O(2)(-)) in the beneficial effects of L-ascorbic acid on heat-induced fish gel (Kamaboko). The generation of a thiyl radical (S.) in glutathione, ovalbumin, and actomyosin was examined by electron spin resonance spectroscopy coupled with spin trapping. O(2)(-) was provided by the photoactivation of riboflavin. The typical line shape for S. was observed with the glutathione and ovalbumin samples. A signal different from that for S. was detected with the actomyosin sample, and its intensity markedly decreased when the SH groups of actomyosin had been modified. The signal was eliminated when superoxide dismutase was added, but unaffected when catalase or an equivalent amount of heat-inactivated superoxide dismutase or catalase were added. These results suggest that S. in actomyosin was produced by the reaction with O(2)(-) and that the beneficial effects of L-ascorbic acid are due to a different mechanism in Kamaboko from that in bread.

There is evidence that mucus of the female bovine genital tract contains pheromones that induce physiological and behavioral responses in other animals. To study these pheromones, vaginal mucus was collected from heifers either at estrus or during diestrus. The mucus was then applied to the hindquarters of the same animal during diestrus or to the hindquarters of herdmates during diestrus. The behaviors of the treated animal and its herdmates were then observed. To attempt to isolate the mounting-inducing substance, mucus was dialyzed or separated on ion-exchange resins. Diestrous heifers to which their own estrual mucus has been applied were nearly always mounted by herdmates (P less than .01). But, heifers to which another's estrous mucus had been applied were not mounted. This suggests that vaginal mucus contains not only estrus-related pheromones, but also individual distinctive odors. The dialyzable fraction of vaginal mucus and the neutral fraction, prepared by ion-exchange chromatography of the dialyzable solution of vaginal mucus, had a mounting-inducing activity on the herdmates, as did the application of an animal's own vaginal mucus. These findings suggest that mounting-inducing pheromones are relatively low molecular weight, neutral substances.

The effects of endogenous protein disulfide isomerase (PDI) family proteins on the properties of gluten proteins in dough during breadmaking were determined using bacitracin, an inhibitor of PDI. Bread loaf volume in the presence of bacitracin was increased to 118% of that in the absence of bacitracin. The addition of bacitracin caused a decrease in the extension tolerance of the dough. The amount of sodium dodecyl sulfate (SDS)-insoluble glutenin macropolymer (GMP) in dough decreased to approximately 70% of that in flour during the 20 min of mixing for doughmaking. The addition of bacitracin to dough caused a dramatic GMP decrease, corresponding to ∼20-30% of that in flour during the 20 min of mixing. The decrease in GMP was compensated by an increase in SDS-soluble glutenin polymer. Taken together, these results suggest that the endogenous PDI family proteins in flour suppress the depolymerization of GMP during dough mixing.

Cereal Chem. 83(5):472-477

The rate of autolysis after thawing in intact frozen krill (whole body, cephalothorax, and abdomen) and in its homogenates was studied by determining both the increase of TCA-soluble nitrogen and the change of protein pattern in SDS-polyacrylamide gel electrophoresis. Autolysis in the intact state proceeded rapidly in the separated cephalothorax, but very slowly in separated abdomen. The pH dependence in autolysis differed between cephalothorax and abdomen homogenates, indicating differences in the proteolytic enzymes involved. Autolysis of intact whole body proceeded to an extent similar to that of intact cephalothorax, liberating about 30% of total nitrogen in 24 hr. The rapid autolysis of abdomen proteins in the intact whole body was attributed to the action of cephalothorax proteases which penetrate into abdomen. Amongmuscle proteins, the myosin heavy chain (MW=206,000) was degraded very rapidly