Antioxidant activity of some protein hydrolysates and their fractions with different isoelectric points

Antioxidant activities of commercially available enzymatic hydrolysates of milk and plant proteins were examined. Among them, soy protein and wheat gluten hydrolysates showed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and antioxidation activity against linoleic acid oxidation in emulsion systems. Peptide fractions with higher antioxidant activities than crude enzymatic hydrolysates of gluten and soy protein were prepared without toxic solvents and reagents. Peptides in these plant protein hydrolysates were fractionated on the basis of the amphoteric nature of sample peptides by preparative isoelectric focusing without adding chemically synthesized carrier ampholytes, which is termed autofocusing. The acidic fractions from both protein hydrolysates showed stronger DPPH radical scavenging activities than the basic fractions, while the basic fractions strongly suppressed 2,2'-azobis (2-amidinopropane) dihydrochloride-induced oxidation of linoleic acid in an emulsion system. These acidic and basic peptide fractions would be useful to examine the mechanism underlying the antioxidant activities of peptides in food.     

Pepsinogens and pepsins from mandarin fish (Siniperca chuatsi)

Four pepsinogens (PG-I, PG-II, PG-III(a), and PG-III(b)) were highly purified from the stomach of the freshwater fish mandarin fish (Siniperca chuatsi) by ammonium sulfate fractionation, anion exchange, and gel filtration. The molecular masses of the four purified PGs were 36, 35, 38, and 35 kDa, respectively. All the pepsinogens converted into their active form pepsins within a few minutes under pH 2.0. The optimum pH and temperature of the four enzymes were 3.0-3.5 and 45-50 degrees C, using hemoglobin as a substrate. The N-terminal amino acid sequences of PG-I and PG-II were determined to the 12th and 17th amino acid residues, respectively. Western blot analysis using antisea bream polyclonal antibodies cross reacted with PG-I, PG-II, and PG-III(b) while no cross reaction with PG-III(a) was detected, suggesting the diversity of pepsinogens in fish.     

Purification and characterization of parvalbumins, the major allergens in red stingray (Dasyatis akajei)

Fish has received increasing attention because it induces IgE-mediated food allergy. Parvalbumin (PV) represents the major allergen of fish, and IgE cross-reactivity to PV in various teleost fish species has been shown, while little information is available about allergens in elasmobranch fish. In this study, two PV isoforms (named as PV-I and PV-II) from red stingray (Dasyatis akajei) were purified to homogeneity by a series of procedures including ammonium sulfate precipitation and column chromatographies of DEAE-Sepharose and Sephacryl S-200. Purified PVs revealed a single band on tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular masses of PV-I and PV-II were 12.29 and 11.95 kDa, respectively, as determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot using antifrog PV monoclonal antibody (PARV-19) showed positive reactions to the two proteins, confirming that they were PVs, although their immunological reactivities were weaker than those of PV from silver carp. The N-terminal amino acid sequence of PV-I was determined, and comparison with PVs from other fish species showed low homology between teleost and elasmobranch fish. The isoelectric points of PV-I and PV-II were 5.4 and 5.0, respectively, as determined by two-dimensional electrophoresis (2-DE), suggesting that both isoforms belong to the α-group. IgE immunoblotting analysis showed that sera from fish-allergic patients reacted to both PV-I and PV-II from red stingray. Thermal stability revealed that PV-I easily formed oligomers than PV-II, which might contribute to the maintenance of its allerginicity during heat processing.     

Safety of reduced-dosage ketoprofen for long-term oral administration in healthy dogs

OBJECTIVE: To evaluate the safety of reduced-dosage ketoprofen (RDKET) for long-term oral administration in healthy dogs. ANIMALS: 14 healthy Beagles. PROCEDURES: Racemic ketoprofen (0.25 mg/kg, PO) and gelatin capsules, as a drug-free placebo, were each administered to 7 dogs for 30 days. Dogs were periodically monitored via physical examination, blood analyses, endoscopic examinations, fecal occult blood tests (tetramethylbenzidine and guaiac methods), renal function tests (effective renal plasma flow and glomerular filtration rate), urinalyses, urinary enzyme indices (N-acetyl-beta-D-glucosaminidase and gamma-glutamyl-transferase), and hemostatic function tests (buccal mucosa bleeding time, cuticle bleeding time, prothrombin time, activated partial thromboplastin time, and fibrinogen concentration). RESULTS: Pyloric antrum lesion grade was significantly higher in the RDKET group on day 28, compared with the pretreatment and control group grades. Fecal occult blood grade measured by use of the tetramethylbenzidine method was significantly higher in the RDKET group on day 30, compared with the pretreatment grade. No other significant differences were detected between treatment groups. CONCLUSIONS AND CLINICAL RELEVANCE: RDKET induced mild to moderate gastric mucosal injuries especially in the pyloric antrum in healthy Beagles, whereas no adverse effects were observed in renal function or hemostasis. Fecal occult blood tests may be useful as screening tests for adverse gastrointestinal effects induced by RDKET in dogs.     

Collembolan community dynamics during deciduous forests regeneration in Japan

The structure and feeding group composition of collembolan communities were studied in secondary deciduous forests of different ages to investigate the collembolan community response to environmental changes associated with forest cycles. The study was carried out at eight sites forming a chronosequence (1, 4, 12, 24, 51, 54, 71 and 128 years after clear cutting) of deciduous forest stands in northern Ibaraki (Japan). Total collembolan density and species richness was low at the 1-year-old site, and there was little difference in density among sites over 4 years of age. The density of sucking feeders was especially low at the 1-year-old site. Species richness of trees of a diameter at breast height (DBH)<5 cm positively correlated with the density of fungal feeders. Species richness of total Collembolans and of sucking feeders correlated positively with the water content of the organic layer. Ordination of the collembolan community with Canonical Correspondence Analysis suggested that species richness of larger trees (DBH 5 cm) contributed to the differences in species composition of fungal feeders and sucking feeders. We conclude that total abundance and species richness of collembolans recovered within 4 years after clear-cutting, but species composition of fungal feeders and sucking feeders took longer to recover.     

Neuronal pathway from the liver modulates energy expenditure and systemic insulin sensitivity

Coordinated control of energy metabolism and glucose homeostasis requires communication between organs and tissues. We identified a neuronal pathway that participates in the cross talk between the liver and adipose tissue. By studying a mouse model, we showed that adenovirus-mediated expression of peroxisome proliferator-activated receptor (PPAR)-g2 in the liver induces acute hepatic steatosis while markedly decreasing peripheral adiposity. These changes were accompanied by increased energy expenditure and improved systemic insulin sensitivity. Hepatic vagotomy and selective afferent blockage of the hepatic vagus revealed that the effects on peripheral tissues involve the afferent vagal nerve. Furthermore, an antidiabetic thiazolidinedione, a PPARg agonist, enhanced this pathway. This neuronal pathway from the liver may function to protect against metabolic perturbation induced by excessive energy storage.     

Soy protein concentrate as an alternative in replacement of fish meal in the feeds of hybrid grouper,brown‐marbled grouper (Epinephelus fuscoguttatus) × giant grouper (E. lanceolatus) juvenile

Hybrid grouper juveniles (body weight, 6.1 ± 0.7 g) (brown‐marbled grouper, Epinephelus fuscoguttatus × giant grouper, E. lanceolatus) were fed with six isoproteic (50% crude protein) and isolipidic (12% crude lipid) feeds containing different levels of soy protein concentrate (SPC) in replacement of fish meal (SPC at 20%, 30%, 40%, 50% and 60% protein) and control feed (SPC0) for 6 weeks. Hybrid grouper juveniles were cultured in 100‐L fibreglass tank equipped with flow‐through water system and fed twice a day to apparent satiation level. The highest and lowest growth was recorded in fish fed SPC20 and SPC60 respectively. However, growth of SPC20 was not significantly higher than those fed SPC0, SPC30, SPC40 and SPC50 (p > .05). A decreasing growth trend was observed with the increasing level of SPC from feed SPC40 to SPC60. A noticeable better feed utilization was also observed in fish fed SPC0, SPC20, SPC30, SPC40 and SPC50 compared to fish fed SPC60 (p < .05). The fish condition factor, hepatosomatic index, viscerosomatic index and whole body proximate content of the fish were not affected by the graded levels of SPC. However, the body lipid content was significantly lower in fish fed SPC40 to SPC60 (p < .05). The apparent digestibility coefficient (ADC) of protein and lipid was significantly higher in fish fed SPC0 and SPC20 compared to other dietary treatments (p < .05). Based on the regression analysis on specific growth rate, the study suggests that the hybrid grouper grow best at 21.4% and can utilize up to 50% inclusion level of SPC in protein without significantly affect their growth and its body condition.     

IMAGING DIAGNOSIS—ECTOPIC SPLEEN MIMICKING HEPATIC TUMOR WITH INTRA‐ABDOMINAL METASTASES INVESTIGATED VIA TRIPLE‐PHASE HELICAL COMPUTED TOMOGRAPHY IN A DOG

A 10‐year‐old castrated male miniature dachshund was presented with an abdominal mass. The dog had a history of splenectomy. Triple‐phase helical computed tomography was utilized, revealing a hepatic mass and multiple intra‐abdominal solid masses. In triple‐phase helical computed tomography the images, hepatic mass and two of four intra‐abdominal masses were heterogenous in all phases. Therefore, we diagnosed a malignant hepatic tumor and presumed intra‐abdominal metastases. The masses were surgically removed and were histologically composed of normal spleen tissues, findings which were consistent with ectopic spleen.     

Effect of supplemented sericin on the development,cell number,cryosurvival and number of lipid droplets in cultured bovine embryos

Sericin was investigated as an alternative to fetal bovine serum (FBS) for bovine embryo culture. In vitro matured oocytes were developed using 0.05%, 0.1% or 0.15% sericin. The developmental rate, cryosurvival rate and blastulation time of these embryos were compared with those of embryos developed using 5% FBS. The number of lipid droplets was compared among the blastocysts developed using 5% FBS, using 0.05% sericin and in vivo. The rate of cleavage and blastocyst formation was similar among all groups. Blastulation occurred significantly earlier in the embryos developed using 5% FBS than in those developed using sericin at any concentration (P < 0.05). At 72 h after thawing, the cryosurvival rate of the blastocysts developed using 5% FBS and 0.05% sericin were significantly higher compared with those developed using 0.1% and 0.15% sericin (P < 0.05). The blastocysts developed using 0.05% sericin and in vivo produced a significantly fewer number of medium and large lipid droplets than those developed using 5% FBS. These results suggest that the blastocysts developed using 0.05% sericin show characteristics similar to those of the blastocysts developed in vivo and that the use of sericin as an alternative to FBS is feasible.