Preparation, characterization, and antioxidative effects of oligomeric proanthocyanidin-L-cysteine complexes

Controlled acid-catalyzed degradation of proanthocyanidin polymers in grape seeds together with L-cysteine led to oligomeric proanthocyanidin-L-cysteine complexes along with monomeric flavan-3-ol derivatives being isolated, and their structures were confirmed on the basis of spectroscopic data and by chemical means. In addition, comparative studies on the antioxidative and survival effects of oligomeric proanthocyanidin-L-cysteine complexes and proanthocyanidin polymers were performed. The oligomeric proanthocyanidin-L-cysteine complexes showed higher bioavailability and antioxidant capacity and enhanced survival time in the animal test groups. In addition, it is suggested that the oligomeric complexes may help to prevent oxidative stress and may reduce free radical production.     

Advanced broodstock diets for the mangrove red snapper and a potential importance of arachidonic acid in eggs and fry

Mangrove red snapper fed advanced broodstock diets containing squid meal and squid oil exhibited higher hatching rates, cumulative survival and survival activity index than those fed a basal diet or a basal diet supplemented with mixture of antioxidants. On the other hand, fatty acid analyses of ovaries and fry of wild fish and eggs and larvae of broodstock fed raw fish revealed high arachidonic acid (ARA) and docosahexaenoic acid (DHA) levels and relatively lower eicosapentaenoic acid (EPA) levels consequently showing high ARA/EPA and DHA/EPA ratios compared to cold water species. This suggests that ARA may be nutritionally more important for egg and larval development and survival in tropical marine fish and its supplementation in broodstock diets may enhance reproductive performance of mangrove red snapper.     

In vivo and in vitro effects of Rhizopus extract on body growth and steroid production in masu salmon, Oncorhynchus masou Brevoort

This study investigated the in vivo and in vitro effects of Rhizopus (filamentous fungus) extract (RU) in masu salmon Oncorhynchus masou Brevoort. Underyearling fish were fed with RU for 16 months. Monthly changes in body growth, gonadal maturation and serum levels of sex steroids were monitored. Gonads were also incubated at 0, 1, 10, 100 and 1000 μg RU mL⁻¹ Leibovitz's L‐15 medium for 18 h. The levels of steroids in serum and cultured medium were measured. It was determined that RU‐fed immature and mature males, when compared with control groups, showed significantly higher body growth during spring, summer and the spawning period. Similarly, immature RU‐fed females showed significantly higher fork length and body weight in autumn, spring and summer. Furthermore, RU‐fed males showed significantly higher levels of serum testosterone (T) and 11‐ketotestosterone (11‐KT) levels in the pre‐spawning season, and 17α, 20β‐dihydroxy‐4‐pregnen‐3‐one (DHP) in the spawning season. In vitro RU incubation of gonads showed a dose‐dependent and significant increase in T, 11‐KT, oestradiol‐17β and DHP release in the medium. It appears that the causes of enhanced body growth and increased steroid production herein observed in salmonids are the physiologically active substances that may be present in the mycelium of the fungus.     

EMG telemetry studies on upstream migration of chum salmon in the Toyohira river,Hokkaido, Japan

The movements of 28 adult chum salmon, Oncorhynchus keta (Walbaum) tagged with electromyogram (EMG) transmitters were tracked along the Toyohira river, Hokkaido, Japan, in October of 2007 and 2008 to investigate and evaluate the upstream migratory behavior through the protection bed and fishway of ground sills. The approach time of fish that ascended successfully through the protection bed and fishway was shorter than that of unsuccessful fish. The unsuccessful fish were observed to swim in currents with high water velocity and shallow water depth at swimming speeds that exceeded their critical swimming speed (U _crit) during the approach to these structures. In consequence, unsuccessful fish frequently alternated between burst and maximum sustained speeds without ever ascending the fishway, and eventually became exhausted. It is important that fishway are constructed to enable chum salmon to find a passage way easily, so that they can migrate upstream rapidly without wasting excessive energy.     

Identification of weakly beta-hemolytic porcine spirochetes by biochemical reactions, PCR-based restriction fragment length polymorphism analysis and species-specific PCR

We examined the usefulness of PCR-based restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR combined with a newly devised rapid biochemical test using microplates for identifying weakly beta-hemolytic intestinal spirochetes (WBHIS) isolated from pigs. WBHIS strains showing atypical biochemical characteristics were decisively identified at the species level by PCR-RFLP and species-specific PCR. Identification of WBHIS at the species level in routine diagnostic work will certainly contribute to clarifying the pathogenicity of WBHIS.     

Dominant effects of litter substrate quality on the difference between leaf and root decomposition process above- and belowground

Initial decomposition rates, changes in organic chemical components (acid-insoluble fraction, holocellulose, polyphenols, soluble carbohydrates) and nutrient dynamics (K, Mg, Ca, P, N) were examined for fine roots and leaves of Japanese cypress (Chamaecyparis obtusa). Litterbag experiments designed to evaluate the relative effects of litter type and position of litter supply in the soil were carried out, considering that root and leaf litter typically occupy different locations and have different substrate qualities. Litterbags of roots and leaves were placed at two positions (on the soil surface and in the humus layer), and collected every 3 months over one year. The mass loss rate and N release were slower during root decomposition in the humus layer than during leaf decomposition on the soil surface. These differences between root and leaf decomposition were mainly caused by the litter type, and the effect of the position on decomposition was relatively small. Root litter was less influenced by position related effects, such as differences in humidity, than leaf litter, and this recalcitrant trait to environmental effects may be responsible for the slower mass loss rate and N release in root decomposition. The results of the present study suggest that fine roots are persistent in the soil and serve an important role in N retention in forest ecosystems because of their litter substrate quality.     

Effects of heating on the interaction of lipid and zein in a dry powder system

The effects of heat treatment on the interaction of lipid and zein in a dry powder system were investigated. Linolenic acid ethyl ester (LAE) was mixed with the zein powder. The glass transition temperature (T(g)) for the dry powder zein was shown to be approximately 107 degrees C by differential scanning calorimetry. The thermogram of the zein-LAE mixed powder showed an exothermic transition near the T(g) of zein. Fourier transform infrared (FT-IR) spectroscopy was used for detecting the structural changes in zein by heat treatment, that is, elevating the temperature from 25 to 160 degrees C. The heat treatment of the powdery zein with and without LAE caused increases in the alpha-helix, beta-turn, and beta-sheet, concomitant with decreases in the intermolecular hydrogen-bonded beta-sheet and random coil. Such changes in the secondary structure were more drastic for the powder with LAE. The heating of the zein-LAE mixed powder also caused decreases in the peaks originating from LAE in the FT-IR spectra. These results suggest that the heat treatment induced the interaction of the zein and LAE in the powdery system. The influence of heating on the antioxidative activity of dry powder zein was studied by measurements of the peroxide value. When zein-LAE mixed powder was heated before storage, the oxidation of LAE was inhibited for 7 days, whereas LAE was oxidized within 1 day in the absence of heat treatment.     

Interaction of grammistins with lipids and their antibacterial activity

ABSTRACT: Grammistins are hemolytic and ichthyotoxic peptides in the skin secretion of soapfishes and are structurally characterized by their abundance in amphiphilic α-helicity. In the present study, their interaction with lipids and lipid vesicles as well as antibacterial activity were examined using four grammistins (Gs 1 and Gs 2 from Grammistes sexlineatus and Pp 1 and Pp 3 from Pogonoperca punctata ). The hemolytic activity of grammistins was inhibited by phospholipids but not by cholesterol. Moreover, grammistins released carboxyfluorescein entrapped within liposomes made of phosphatidylcholine. In contrast, grammistins were found to have antibacterial activity with a broad spectrum against nine species of bacteria, including both Gram-negative and Gram-positive groups. The potency of their antibacterial activity was not related to that of hemolytic activity, suggesting that grammistins bind to membrane phospholipids but lyse erythrocyte and bacterial membranes via different mechanisms. Conclusively, grammistins are new members of the family of cell non-selective membrane-lytic peptides with amphiphilic α-helices, being similar to pardaxins, which are secreted from the skin of soles, and to melittin, which is derived from bee venom.     

Comparison of dendritic cell-mediated immune responses among canine malignant cells

Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials.