Effects of water activity and lipid addition on secondary structure of zein in powder systems

The effects of water activity (A(w)) and lipid addition on the secondary structure of powdery zein were investigated using Fourier transform infrared spectroscopy. Two fatty acid esters, i.e., the linolenic and eicosapentaenoic acid ethyl esters (LAE and EPE), were mixed with the zein powder. The powders were stored in the "dry" state (with silica gel) and the "humid" state (A(w) = 0.9). The powdery zein without the lipids was shown to have a high content of the intermolecular hydrogen-bonded beta-sheet in the "dry" state, indicating the presence of protein aggregates. An increase in A(w) induced a decrease in this beta-sheet, concomitant with increases in the alpha-helix and beta-turn structures. The addition of LAE caused decreases in the alpha-helix and intermolecular hydrogen-bonded beta-sheet of zein when the powder was stored in the "humid" state, suggesting the strong interaction of LAE and zein molecules. However, LAE did not affect the secondary structure of zein in the "dry" state. The addition of EPE hardly influenced the secondary structure of zein, irrespective of A(w). These results are discussed in relation to the antioxidative activity of zein in the powder system, which had studied previously.     

Oogenesis and relevant changes in egg quality of abalone Haliotis discus hannai during a single spawning season

Oogenesis of abalone Haliotis discus hannai was examined histologically during a single spawning season using broodstock of various maturation conditions, which were controlled by effective accumulative temperature (EAT). The quality of eggs spawned was determined in relation to oogenesis. For histological examinations, three to five females were sacrificed at 300, 600, 850, 1050, and 1150 °C days EAT, without induction of artificial spawning. Other females were successfully induced to spawn at 700 °C days EAT and were reared following spawning. Three of these females were then sacrificed every 200 °C days EAT until 1300 °C days EAT. Gonad histology showed that two oocyte cohorts matured in H. discus hannai ovaries during a single spawning season. One mature oocyte cohort could be spawned in multiple times. The second oocyte cohort started developing after the first oocyte cohort had been spawned or reabsorbed, and became fully mature 400 °C days EAT after the first cohort was depleted. For egg quality measurements, three to five females were successfully induced to spawn at 850, 1050, 1150, 1900, and 2350 °C days EAT (Experiment 1). Three females were induced to spawn twice, at 700 and 1500 °C days EAT, resulting in two batches of eggs from the same individuals (Experiment 2). Total lipid and protein content of eggs were measured and were greater in eggs from the second cohort than in eggs from the first cohort. No carbohydrates were detected in eggs and there was no difference in cytoplasm volume between the two cohorts. In hatcheries producing H. discus hannai, it is important to increase post-larval starvation tolerance by increasing the quality of eggs, to yield higher and more consistent survival. The results of this study suggest that H. discus hannai hatcheries should use eggs from the second oocyte cohort, which are of higher quality, rather than eggs from the first oocyte cohort.     

Effects of enzymatic deamidation by protein-glutaminase on structure and functional properties of alpha-zein

The performance of novel protein-glutaminase (PG) purified from Chryseobacterium proteolyticumon alpha-zein was investigated. Highly insoluble alpha-zein was able to be deamidated to the extent of deamidation degree 62% by using 50 mM potassium phosphate (pH 8) containing 11.7% ethanol, at 40 degrees C for 137 h. Analysis by sodium dodecyl sulfate polyacrylamide-gel electrophoresis showed that deamidated and non-deamidated zeins have different mobilities. Results of circular dichroism spectra revealed the decline in alpha-helix contents of alpha-zein by deamidation. Besides, Fourier transform infrared spectroscopy analysis demonstrated alterations in the secondary structure of alpha-zein by deamidation. The assignment of the amide I region showed a remarkable decrease in antiparallel intermolecular beta-sheets (around 1690 cm(-1)) as an indication of the weakening aggregation ability of the deamidated molecules. Solubility and emulsification properties of alpha-zein, particularly at pH 7, were remarkably improved after the deamidation by PG. Gas chromatography and peroxide value studies pointed out that deamidated alpha-zein in powder form exhibited an inferior antioxidative property as compared with the non-deamidated one.     

Effects of ovariectomy on bone metabolism and bone mineral density in spontaneously diabetic Torii-Lepr(fa) rats

The Spontaneously Diabetic Torii-Lepr(fa) (SDT- fa/fa) rat is a new model of obese type 2 diabetes. The female SDT-fa/fa rat shows obesity, hyperglycemia and hyperlipidemia from a young age. However, it is not known whether diabetes and estrogen deficiency can lead to bone abnormalities in the female SDT-fa/fa rat. The objective of the present study was to investigate the effects of ovariectomy (OVX) on bone metabolism and bone mineral density (BMD) in the female SDT-fa/fa rat. Female Sprague-Dawley rats were used as control animals. The BMDs of the whole tibia and fifth lumbar (L5) vertebral body were analyzed at 30 weeks after OVX. Serum osteocalcin, a bone formation marker, and urine deoxypyridinoline (DPD), a bone resorption marker, were sequentially analyzed before and at 5, 15 and 30 weeks after OVX. Serum osteocalcin and urine DPD levels were lower in SDT-fa/fa rats than in control rats before OVX. Both serum osteocalcin and urine DPD levels were elevated in control rats 5-30 weeks after OVX, but only the urine DPD levels were elevated in SDT-fa/fa rats 5-30 weeks after OVX. SDT-fa/fa rats showed a decrease in the BMDs of the whole tibia and L5 vertebral body compared with control rats. OVX decreased the BMDs of the whole tibia and L5 vertebral body in control rats, but not in SDT-fa/fa rats. These data suggest that estrogen deficiency is not a risk factor for bone loss in type 2 diabetes mellitus.     

Rapid activation of Mac-1(CD11b/CD18) molecules on macrophages by a new chemotactic factor 'Gasserokine' produced by Lactobacillus gasseri JCM1131T

The chemoattractant activity of a new chemotactic factor, 'Gasserokine' produced by Lactobacillus gasseri JCM1131^T, has been proposed as a novel immunological function of probiotic lactic acid bacteria. The focus of the present study was to understand the mechanism of the chemotaxis induced by Gasserokine, using activation of an adhesion molecule, Mac-1 (CD11b/CD18) on macrophages. The macrophage chemotaxis to Gasserokine was abolished by preincubation of macrophages with the anti-Mac-1 mAb. Gasserokine induced rapid serine phosphorylation of CD18 molecules within 1 min of stimulation, but the effect was short-lived. Substantial tyrosine phosphorylation was observed in CD18-associated protein of macrophages stimulated by Gasserokine. The tyrosine phosphorylation was confirmed in macrophages stimulated with Gasserokine and also serine/threonine phosphorylation was detected on CD18 molecules by laser microscopy using a double immunostaining method. These results suggest that selective activation of intracellular signaling cascades, such as the mitogen-activated protein kinase pathway, are related to the macrophage chemotaxis induced by Gasserokine.     

Predation by neogastropods on <Emphasis Type="Italic">Turbo cornutus</Emphasis> juveniles and other small gastropods inhabiting coralline algal turfs

To understand the effects of predation by gastropods on juveniles of the Japanese spiny turban snail Turbo cornutus, a field survey and laboratory experiments were conducted. The species compositions of the order Neogastropoda inside turfs of articulated coralline algae (ACA) on the east coast of Sagami Bay were surveyed monthly, and the dominant species inside the ACA turfs were identified. Laboratory experiments were conducted to examine the effects of predation by the dominant neogastropods on small gastropods in ACA turfs. Turban snails, which inhabit ACA turfs during their juvenile stages, were predated by two muricid gastropods, Ergalatax contractus and Thais bronni, with the predation rate of E. contractus being significantly higher than that of T. bronni. While E. contractus could also predate two other species of gastropods, Anachis misera and Cantharidus japonicus, the predation rates on these two snails were significantly lower than on T. cornutus. The observed defensive behaviors were different among the three prey species, and the defensive strategy related to the calcified operculum of T. cornutus was not efficient against predation by E. contractus. It is concluded that the predation by E. contractus represents an important factor in the mortality of juvenile turban snails inside ACA turfs.     

Identification of juvenile abalone <Emphasis Type="Italic">Haliotis diversicolor</Emphasis> based on number of open and sealed respiratory pores

ABSTRACT:   Changes in the number of respiratory pores (open pores) and their imprints (sealed pores) in post-larvae and juveniles were observed and compared for four abalone species Haliotis diversicolor , H. discus discus , H. madaka , and H. gigantea . The first open pore was evident at a shell length (SL) of 1.5 mm in H. diversicolor , 1.9–2.0 mm SL in H. discus discus , and 2.3–2.4 mm SL in H. madaka and H. gigantea . The number of open pores in H. diversicolor gradually increased with growth, with four to five pores at 2.5–18.0 mm SL and five to six pores at 18.0–27.0 mm SL. The other three species maintained four to five open pores after they reached 3.4–4.5 mm SL. The total number of open and sealed pores (TNP) was greater in H. diversicolor than in the other species at the same SL. Juvenile H. diversicolor were identified among field-caught abalone by the difference in the relationship between SL and TNP (SL–TNP relationship) and also by the monoclonal antibody reaction method. The results of the two methods were in perfect agreement, indicating that our method using the SL–TNP relationship is reliable for the identification of H. diversicolor .     

Maturation and spawning of <Emphasis Type="Italic">Spratelloides gracilis</Emphasis> Clupeidae in temperate waters off Cape Shionomisaki,central Japan

ABSTRACT:   The reproductive ecology of Spratelloides gracilis was investigated in the temperate waters off Cape Shionomisaki, central Japan. Cape Shionomisaki is located at the northern margin of the distribution range of this species. Females of S. gracilis with gonadosomatic index (GSI) ≥ 4.0 were defined as mature based on the relationship between GSI and the histological maturity phases of their ovaries. More than 50% of females greater than 60 mm standard length (SL) were mature. Hatch dates of larvae and juveniles collected in the study area were determined by otolith daily ring counts and found to extend from April to November. The size at maturity of females (60 mm SL) and the spawning season of S. gracilis in the temperate waters off Cape Shionomisaki (April–November) was larger and shorter, respectively, than those in tropical waters in the western Pacific. The reproductive traits observed for S. gracilis off Cape Shionomisaki appear to be adaptive to northern temperate waters with seasonal changes in environmental conditions.     

Intracerebroventricular administration of chicken glucagon‐like peptide‐2 potently suppresses food intake in chicks

Glucagon‐related peptides, such as glucagon‐like peptide (GLP)‐1, GLP‐2 and oxyntomodulin (OXM), are processed from an identical precursor proglucagon. In mammals, all of these peptides are suggested to be involved in the central regulation of food intake. We previously showed that intracerebroventricular administration of chicken OXM and GLP‐1 significantly suppressed food intake in chicks. Here, we show that central administration of chicken GLP‐2 potently suppresses food intake in chicks. Male 8‐day‐old chicks (Gallus gallus domesticus) were used in all experiments. Intracerebroventricular administration of chicken GLP‐2 significantly suppressed food intake in chicks. Plasma glucose concentration was significantly decreased by chicken GLP‐2, whereas plasma nonesterified fatty acid concentration was significantly increased. Intracerebroventricular administration of chicken GLP‐2 did not affect plasma corticosterone concentration. In addition, the anorexigenic effect of GLP‐2 was not reversed by the corticotropin‐releasing factor (CRF) receptor antagonist α‐helical CRF, suggesting that CRF is not a downstream mediator of the anorexigenic pathway of GLP‐2 in chicks. Intracerebroventricular administration of an equimolar amount of GLP‐1 and GLP‐2, but not OXM, significantly suppressed food intake in both broiler and layer chicks. All our findings suggest that GLP‐2 functions as a potent anorexigenic peptide in the brain, as well as GLP‐1, in chicks.     

Induction of interferon-gamma (IFN-gamma) and T helper 1 (Th1) immune response by bitter gourd extract

Mice were inoculated intraperitoneally wih 34 different types of vegetable juices, and interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) were measured as markers for the induction of Th1 and Th2 cells, respectively. Serum IFN-gamma level was markedly increased in mice inoculated with bitter gourd (Momordica charantia) juice, but IL-4 levels were not increased with any of the 34 vegetable juices. Testing of the various components of bitter gourd, including peel, pulp, and seed, showed that the pulp induced the highest levels of IFN-gamma. Trial immunogen including the heat extract of the pulp induced specific IgG(2a) antibody of the mice serum inoculated with this immunogen. These results demonstrate that bitter gourd pulp induced IFN-gamma production and show its promise as a means of effective immunostimulatory therapy specific for Th1 cells and IFN-gamma production.