Relationship between Clostridium septicum alpha-toxin activity and binding to erythrocyte membranes

The activity of Clostridium septicum alpha-toxin was determined in erythrocytes of various animals, with sensitivities observed in the order of mouse, rat, canine, equine, rabbit, chicken, bovine, swine and ovine. Temperature and protease treatment affected the sensitivity of erythrocytes to alpha-toxin. Proteinase K treatment decreased the sensitivity of murine, canine, equine and bovine erythrocytes, but ovine erythrocytes did not change the sensitivity to alpha-toxin activity. On the other hand, the activity of alpha-toxin on swine erythrocytes increased after treatment with proteinase K, trypsin, chymotrypsin or lysyl endopeptidase. Toxin overlay assay showed that alpha-toxin bound to erythrocyte membrane proteins with a molecular mass of 30 to 45-kDa in mouse, equine, bovine, swine and chicken, whereas in rat erythrocyte membranes the toxin reacted with 100-kDa protein. The treatment of murine and swine erythrocyte membranes with phosphatidylinositol-specific phospholipase C resulted in liberation of the toxin-binding protein from the individual membranes in a native state. These results show that alpha-toxin associates with specific erythrocyte membrane proteins in any animal species, and are subsets of glycosylphosphatidylinositol-anchored proteins in various animal species. These results may reflect distinct characteristics of the hemolytic activity of alpha-toxin in response to various erythrocytes.     

Seasonal potential fishing ground prediction of neon flying squid (Ommastrephes bartramii) in the western and central North Pacific

We explored the seasonal potential fishing grounds of neon flying squid (Ommastrephes bartramii) in the western and central North Pacific using maximum entropy (MaxEnt) models fitted with squid fishery data as response and environmental factors from remotely sensed [sea surface temperature (SST), sea surface height (SSH), eddy kinetic energy (EKE), wind stress curl (WSC) and numerical model‐derived sea surface salinity (SSS)] covariates. The potential squid fishing grounds from January–February (winter) and June–July (summer) 2001–2004 were simulated separately and covered the near‐coast (winter) and offshore (summer) forage areas off the Kuroshio–Oyashio transition and subarctic frontal zones. The oceanographic conditions differed between regions and were regulated by the inherent seasonal variability and prevailing basin dynamics. The seasonal and spatial extents of potential squid fishing grounds were largely explained by SST (7–17°C in the winter and 11–18°C in the summer) and SSS (33.8–34.8 in the winter and 33.7–34.3 in the summer). These ocean properties are water mass tracers and define the boundaries of the North Pacific hydrographic provinces. Mesoscale variability in the upper ocean inferred from SSH and EKE were also influential to squid potential fishing grounds and are presumably linked to the augmented primary productivity from nutrient enhancement and entrainment of passive plankton. WSC, however, has the least model contribution to squid potential fishing habitat relative to the other environmental factors examined. Findings of this work underpin the importance of SST and SSS as robust predictors of the seasonal squid potential fishing grounds in the western and central North Pacific and highlight MaxEnt's potential for operational fishery application.     

Identification and kinetic study of tyrosinase inhibitors found in sake lees

The present study found that the n-hexane extract of freeze-dried sake lees inhibits tyrosinase activity and showed that the constituents isolated from the n-hexane extract are the mixture of triacylglycerols. The inhibitory effects of triolein and trilinolein found as the triacylglycerols were examined using tyrosinases from mushroom and Streptomyces castaneoglobisporus. The IC50 values of the triacylglycerol mixture for the oxidase activity on mushroom and Streptomyces tyrosinases were 20 and 0.14 microg/mL, respectively. The IC50 values of trilinolein for the oxidase activity on mushroom and Streptomyces tyrosinases were 8.4 and 0.1 microM, respectively. However, the inhibitory effect of triolein (IC50=30 microM) was lower than that of trilinolein, even when the Streptomyces tyrosinase was used for the assay. Kinetic analyses indicate that both trilinolein and triolein inhibit the tyrosinase activity noncompetitively. When transformed with a plasmid carrying the Streptomyces tyrosinase gene, the melanin-synthesizing ability of the transformed Escherichia coli host was dose-dependently interfered with by trilinolein.     

In vitro metabolism of tyrosine by rumen bacteria, protozoa and their mixture

An anaerobic in vitro study of tyrosine (Tyr) metabolism by rumen bacteria (B), protozoa (P) and their mixture (BP) was conducted using an HPLC analytical method to obtain some basic and systematic information on aromatic amino acid metabolism in the rumen. Most of the Tyr (> 96%) disappeared after 12 h incubation in B and BP, but only 58% in P. Tyr was converted mainly to p‐hydroxyphenylacetic acid (HPA) in all microbial suspensions. About 45% of disappeared Tyr in B and P, and about 35% in BP were converted to HPA. An appreciable amount of phenylalanine (Phe), 13 and 3% of disappeared Tyr, and a small amount of tryptophan (Trp), 8 and 1% of disappeared Tyr, were also produced from Tyr by rumen bacteria and protozoa, respectively. Small amounts of p‐hydroxyphenylpyruvic acid (about 4 and 6% of disappeared Tyr) were produced from Tyr in B and P, respectively. A moderately large amount of phenylacetic acid (14% of disappeared Tyr) was produced from Tyr in P which was 1.9 times higher than that in B. Phenylpropionic acid and p‐hydroxybenzoic acid were produced only in B and BP. It was concluded that the Tyr degradation ability of rumen bacteria was about 1.5 times higher than that of rumen protozoa. Degraded Tyr mainly produces HPA and then two other aromatic amino acids, Phe and Trp, which are considered essential amino acids for ruminants. Therefore, it is speculated that the requirement for Phe and Trp in ruminants may be partially fulfilled if Tyr is sufficiently supplied in rations.     

Impact of paralarvae and juveniles feeding environment on the neon flying squid (Ommastrephes bartramii) winter–spring cohort stock

In this study, we found that there were significant positive correlations between the catch per unit effort (CPUE, a squid abundance index) for the neon flying squid (Ommastrephes bartramii) winter–spring cohort and the satellite‐derived chlorophyll a concentrations in their spawning grounds located at 140–160°E where 21°C < sea surface temperature < 25°C from February to May. The spawning grounds of the winter–spring cohort are located in a quiet stream region, and a particle tracking experiment, based on the velocity field obtained from an ocean data assimilation system, showed that paralarvae and juveniles aged <90 days remained in their spawning grounds and the chlorophyll a concentration in their habitat had a significant positive correlation with the CPUE. A backward particle tracking experiment also showed that the chlorophyll a concentration in the spawning grounds had a significant positive correlation with the autumn–winter mixed layer depth. Based on these results, we hypothesize that the CPUE interannual variability is caused by variations in the feeding environment of the paralarvae and juveniles, which may be linked to autumn–winter mixed layer depth variations.     

Follicle-stimulating hormone (FSH) stimulates the expression of Pin1, a peptidyl-prolyl isomerase, in the bovine granulosa cells

A peptidyl-prolyl isomerase, Pin 1, has been shown to play a role in the regulation of cell cycle progression, both in vitro and in vivo. However, the involvement of Pin 1 during follicular development is not well understood. The aim of this study was first to investigate the expression of Pin 1 mRNA in the granulosa and theca cells of the follicle at different developmental stages of follicles in the bovine ovary, and second, to examine the effects of follicle-stimulating hormone (FSH) and estradiol (E2) on the expression of Pin 1 in the cultured bovine granulosa cells. Follicles were classified into four groups based on the diameter (dominant follicles >8.5mm in diameter, subordinate follicles <8.5mm in diameter) and the relative levels of E2 and progesterone (P4) (E2:P4>1, estrogen active; E2:P4<1, estrogen inactive): i.e. preovulatory dominant follicles (POFs); E2 active dominant follicles (EADs); E2 inactive dominant follicles (EIDs); small follicles (SFs). The expression of the Pin 1 gene was significantly increased in the granulosa cells of EADs as compared with those of other follicles, whereas its expression in theca cells did not differ among follicles at different developmental stages. The concentration of 5 ng/ml FSH alone and the combination of 1 ng/ml E2 and 5 ng/ml FSH stimulated the expression of the Pin 1 gene in bovine granulosa cells. Our data provide the first evidence that Pin 1 expression in the granulosa cells but not the theca cells changes during follicular development, and that FSH stimulate the expression of the Pin 1 gene. These results suggest that Pin 1 regulates the timing of cell proliferation and may act as an intracellular signal responder in the granulosa cells during bovine follicle development.     

Tetrodotoxin accumulation conflicts with low salinity tolerance in juvenile tiger puffer Takifugu rubripes

We evaluated whether bearing tetrodotoxin (TTX) affects salinity stress in the juvenile tiger puffer Takifugu rubripes. Juveniles of hatchery-reared non-toxic T. rubripes [body weight (BW): 1.7?±?0.2 g, n?=?120] were divided into six tanks and acclimatized to salinity (8.5 ppt) that is equivalent to blood osmolality. Fish in three tanks were fed non-toxic diet, and those in the other three tanks were fed a TTX-containing diet (356 ng/g diet) three times a day until satiation. In each diet treatment, salinity of one tank was kept at 8.5 ppt, and the other two tanks were adjusted to either 1.7 or 34.0 ppt, and fish were reared for another 33 days. Then, we compared survival, growth, TTX accumulation, plasma osmolality, plasma cortisol, and glucose levels among treatments. We detected TTX only in the fish in the TTX-diet groups. Survival was highest at 8.5 ppt (70%) and lowest at 1.7 ppt in the TTX-diet group (20%). The BW was greater at 8.5 ppt, and plasma osmolality was significantly higher at 34.0 ppt than at any other salinities. Plasma cortisol level was significantly higher but glucose level was lower at 1.7 ppt. Possessing TTX at a low salinity may be lethal to tiger puffer juveniles.

     

A comparison of the immunological effects of propofol and isoflurane for maintenance of anesthesia in healthy dogs

Most anesthetics have an immuno-suppressive effect on cellular and neurohumoral immunity, and research shows that total intravenous anesthesia (TIVA) with propofol has a greater immuno-protective effect than inhalational anesthesia in human medicine. However, in veterinary clinics, these effects remain ambiguous. To clarify the details, we focused on propofol and isoflurane, investigating clinical blood hematology and immunological profiles drawn from healthy dogs under and after two anesthesia techniques. Twelve healthy adult beagles were included in this study, randomly assigned to the propofol anesthesia group (group P: n=6) or the isoflurane anesthesia group (group I: n=6). In both groups, the number of lymphocytes in peripheral blood decreased after 2 hr of anesthesia (2 hr), but group P showed significantly less decrease than group I. For T-lymphocyte subsets examined by flowcytometry, the ratio of CD3+, CD4+ and CD8+ lymphocytes in the peripheral blood mononuclear cell (PBMC) of group P at 2 hr also exhibited a high level compared to group I. Moreover, for mRNA expression of cytokines measured by real-time PCR, the IL2 (pro-inflammatory cytokine) of group P showed no decrease like group I. The IL10 (anti-inflammatory cytokine) of group P also showed no increase like group I, while both cytokines maintained nearly the same level until 2 hr. These results suggest that, compared to propofol, isoflurane had more strongly immuno-suppression caused by anesthesia, and propofol itself might have some immuno-protective effects. Thus, TIVA with propofol might benefit immunological support in the perioperative period of dogs.     

Effects of hypoxia on principal prey and growth of flathead flounder Hippoglossoides dubius in Funka Bay, Japan

ABSTRACT:   Effects of hypoxia on the principal prey and growth of flathead flounder Hippoglossoides dubius were studied in Funka Bay. Of the three dominant year-classes that occurred in recent years, the 1995 year-class was small in total length at age ≥ 3 and low in condition factor at age ≥ 2. Ophiuroids (almost Ophiura sarsi ), which were the dominant prey in the 1980s, were of little importance and instead, crustaceans such as mysids, natant decapods and pelagic amphipods, bivalves and fish were important prey items for H. dubius in 2000–2001. In addition, the feeding intensity of H. dubius in 2000–2001 was lower than that in the 1980s. These facts are closely related to a reduction of prey abundance, particularly ophiuroids. It seems that the hypoxia that occurred in the central part of the bay during the summer and autumn of 1995–1997 caused the poor food supply and low growth rate at ages 2–4 of the 1995 year-class.     

Synthesis of citrulline from ornithine by the small intestinal mucosa of cattle

Three segments of cattle small intestine (duodenum, upper jejuno‐ileum and lower jejuno‐ileum) were examined in an in vitro system for activity of ornithine carbamoyltransferase (OCT; EC 2.1.3.3) which is involved in the synthesis of citrulline (Cit) from ornithine (Orn). The mucosa of the three segments of small intestine was collected from Japanese black cattle, homogenized and then centrifuged. The supernatant fraction was used as the crude OCT enzyme solution. The OCT activity was assayed by the production of Cit from Orn determined directly by HPLC. The optimal pH and temperature for OCT activities in the duodenum, upper jejuno‐ileum and lower jejuno‐ileum of cattle small intestine were 7.47 and 39°C. Little difference was observed between the three segments. The OCT activity in cattle kidney was also examined for comparison, and almost no activity was found. The OCT activities in crude enzyme solutions of the three segments of small intestine were stable for up to one month of storage at ?20°C in Tris HCl buffer solution. Finally, the role of the small intestine in supplying Cit as a precursor for arginine synthesis in cattle kidney was discussed.