Protein Hydrolysates and Ultrafiltration Fractions Obtained from Krill (Euphausia superba): Nutritional,Functional, Antioxidant,and ACE-Inhibitory Characterization

ABSTRACT

Krill (Euphausia superba) was hydrolyzed by proteolytic enzymes in order to produce multifunctional bioactive peptides, and their functional properties were evaluated. Krill protein hydrolysate (KPH) by pepsin with 4-h hydrolysis showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and angiotensin I converting enzyme (ACE) inhibitory activities. The solubility and foaming properties of KPH were higher than those of the unhydrolyzed krill protein at a wide range of pHs. KPH was further fractionated based on molecular weight. The 1- to 3-kDa peptide fraction exhibited the highest DPPH scavenging activity (IC₅₀ value of 0.5 mg/mL), oxygen radical absorbance capacity (497.39 ± 4.31 µM TE/mg fraction), 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid cation radical scavenging activity (48.41 ± 0.23 µM TE/mg fraction), and reducing power (110.40 ± 2.07 µM TE/mg fraction). However, the < 1-kDa peptide fraction exhibited a higher ACE inhibitory activity than that of other fractions. The 1- to 3- and < 1-kDa peptide fractions are rich in aromatic and hydrophobic amino acids, respectively.     

A Comparison of Ultrastructural Changes of Barley Cells Infected with Mild and Aggressive Isolates of Barley stripe mosaic virus

Journal of Plant Diseases and Protection - Electron microscopy studies were carried out to investigate the association of visible disease symptoms with ultrastructural changes in barley plants...     

Mitigative Effects of PFF-A Isolated from Ecklonia cava on Pigmentation in a Zebrafish Model and Melanogenesis in B16F10 Cells

Melanin synthesis is a defense mechanism that prevents skin damage, but excessive accumulation of melanin occurs in the skin in various reactions such as pigmentation, lentigines, and freckles. Although anti-melanogenic effects have been demonstrated for various naturally occurring marine products that inhibit and control tyrosinase activity, most studies have not been extended to in vivo applications. Phlorofucofuroeckol-A (PFF-A, 12.5–100 µM) isolated from Ecklonia cava has previously been shown to have tyrosinase-mitigative effects in B16F10 cells, but it has not been evaluated in an in vivo model, and its underlying mechanism for anti-melanogenic effects has not been studied. In the present study, we evaluated the safety and efficacy of PFF-A for anti-melanogenic effects in an in vivo model. We selected low doses of PFF-A (1.5–15 nM) and investigated their mitigative effects on pigmentation stimulated by α-MSH in vivo and their related-mechanism in an in vitro model. The findings suggest that low-dose PFF-A derived from E. cava suppresses pigmentation in vivo and melanogenesis in vitro. Therefore, this study presents the possibility that PFF-A could be utilized as a new anti-melanogenic agent in the cosmeceutical industries.     

Changes in cyclooxygenase-2 immunoreactivity in the hippocampus in a model of streptozotocin-induced type 1 diabetic rats

In this study, we investigated diabetic stage dependent cyclooxygenase-2 (COX-2) immunoreactivity in the dentate gyrus in streptozotocin (STZ)-induced type 1 diabetic rats. The animals were sacrificed at 2, 3 and 4 weeks after STZ treatment. Blood glucose levels were increased after STZ treatment. COX-2 immunoreactivity in dentate gyrus was significantly increased in these regions 3 weeks after STZ treatment and restored to its basal level to 4 weeks after STZ treatment. In contrast, COX-2 immunoreactivity was not changed in CA3 region in all groups. These results suggest that STZ-induced type 1 diabetes transiently, but not permanently, decreased synaptic transmission and plasticity 3 weeks after STZ treatment in the dentate gyrus.     

Antihypertensive effect of angiotensin i converting enzyme-inhibitory peptide from hydrolysates of Bigeye tuna dark muscle, Thunnus obesus

Angiotensin I converting enzyme (ACE) inhibitory peptide was isolated from tuna dark muscle hydrolysate prepared by alcalase, neutrase, pepsin, papain, alpha-chymotrypsin, and trypsin, respectively. Among hydrolysates, the pepsin-derived hydrolysate exhibited the highest ACE I inhibitory activity versus those of other enzyme hydrolysates. The structure of the peptide was identified to be Trp-Pro-Glu-Ala-Ala-Glu-Leu-Met-Met-Glu-Val-Asp-Pro (molecular weight 1581 Da) by time of flight mass spectrometry/mass spectrometry analysis, and the IC 50 value of the peptide was 21.6 microM. The Lineweaver-Burk plots revealed that the peptide acts as a noncompetitive inhibitor, and the inhibitor constant ( K i) was calculated as 26.6 microM using the secondary plots. The peptide had an antihypertensive effect according to the time-course measurement after oral administration to spontaneously hypertensive rats. Maximal reduction was detected 3 h after oral administration at a dose of 10 mg/kg of body weight. These results suggest that the peptide derived from tuna dark muscle would be a beneficial ingredient for functional food or pharmaceuticals against hypertension and its related diseases.     

Effect of pressure cooking on aflatoxin B1 in rice

The effect of pressure cooking on aflatoxin residues in polished rice was conducted to determine reduction of aflatoxin and mutagenic potentials. Three rice lots consisting of naturally contaminated, A. parasiticus-infested, and aflatoxin-spiked rice were steamed by ordinary and pressure cookers after they were washed with water. They were chemically analyzed for aflatoxins using a silica solid phase extraction tube and high-performance liquid chromatography (HPLC)-fluorescence detection (FD), and the presence of aflatoxin residues was confirmed using HPLC-electrospray ionization (ESI)-mass spectrometry (MS). An in vitro mutagenicity test with Salmonella typhimurium TA100 was employed to verify the results based on chemical analyses. The aflatoxin loss (78-88%) was notable after pressure cooking, and the reduction of aflatoxin-induced mutagenic potential (68-78%) was in good agreement with the HPLC results. It can be concluded that Koreans are safe from the aflatoxin-related risk if a pressure cooker is employed for cooking rice. The average Korean daily intake of aflatoxin through the consumption of staple rice would fall to 0.15 ng/kg bw/day, which would not exceed the established tolerable daily intake (0.40 ng/kg bw/day).     

Serosurveillance for Japanese encephalitis virus in wild birds captured in Korea

Climate change induced by recent global warming may have a significant impact on vector-borne and zoonotic diseases. For example, the distribution of Japanese encephalitis virus (JEV) has expanded into new regions. We surveyed the levels of hemagglutination-inhibition (HI) antibodies against JEV (Family Flaviviridae, genus Flavivirus) in wild birds captured in Korea. Blood samples were collected from 1,316 wild birds including the following migratory birds: Oceanodroma castro (n = 4), Anas formosa (n = 7), Anas penelope (n = 20), Fulica atra (n = 30), Anas acuta (n = 89), Anas crecca (n = 154), Anas platyrhynchos (n = 214), Aix galericulata (n = 310), and Anas poecilorhyncha (n = 488). All were captured in 16 locations in several Korea provinces between April 2007 and December 2009. Out of the 1,316 serum samples tested, 1,141 (86.7%) were positive for JEV. Wild birds captured in 2009 had a higher seroprevalence of ant-JEV antibodies than those captured in 2007. Wild birds with an HI antibody titer of 1 : 1,280 or higher accounted for 21.2% (280/1,316) of the animals tested. These findings indicated that wild birds from the region examined in our study have been exposed to JEV and may pose a high risk for introducing a new JEV genotype into Korea.     

Improved sero-monitoring assay for classical swine fever (CSF) using the recombinant E2 protein of a recent Korean isolate

Classical swine fever (CSF) is a highly contagious disease of pigs that causes fever, diarrhea and paralysis, often resulting in death. E2 is the major structural protein of the CSF virus (CSFV) and mediates the entrance of the virus, subsequently inducing a neutralizing immune response. In this study, the E2 gene of a recent Korean isolate of CSF, SW03, was cloned and the DNA sequence was compared to other strains via phylogenetic analysis. With the purified E2 protein, an enzyme-linked immunosorbent assay (ELISA) was developed for the serodiagnosis of CSFV infection. The sensitivity and specificity of the E2-ELISA were 96.1% and 94.8%, respectively. A total of 17 out of 485 field-collected pig sera tested demonstrated conflicting results between two ELISA methods, a commercial kit and the E2-ELISA. Of these sera, 60% were determined to be CSFV positive by a virus neutralization test (VNT), suggesting involvement of different immune responses in the cases of CSFV infection. As the E2-ELISA was developed using a recent Korean isolate, SW03, this assay is capable of rapidly identifying newly emerging CSFV strains.     

Natural occurrence of grape poisoning in two dogs

Clinical grape poisoning in two dogs (a 1.6-year-old male Shih Tzu and a 5-year-old female Yorkshire Terrier) was described in the present study. Clinical signs included decreased urine output in the Shih Tzu and ataxia in the Yorkshire Terrier after grape ingestion. The Shih Tzu died 5 days post-grape ingestion, while the Yorkshire Terrier died 3 days post-grape ingestion. Erythematous serosae and mucosae, multifocal red small intestinal foci, and blood and grape seeds were identified in the intestinal lumen. Brownish-yellow crystals were bilaterally identified in the renal pelvis. The primary histological findings were acute tubular necrosis of the proximal convoluted tubules, severe necrosis, and mineralization in the renal cortical tubules. Blood urea nitrogen, creatinine, and alanine aminotransferase were increased in the dogs. Many Korean veterinary clinicians have suspected clinical grape poisoning. However, to our knowledge, grape poisoning has not been identified by pathologic and clinicopathologic basis until this writing in Korea. Education and knowledge about the risks of grape poisoning is necessary for the prevention of accidental exposures.     

N and P losses from a paddy field plot in central Korea

Abstract

The study was carried out to investigate the water balance and runoff and infiltration losses of nutrients in a paddy field plot located in southern Korea. Field monitoring was carried out during the cropping season from May 1, 1999 to September 30, 2000. The soil of the experimental paddy field belonged to the Jisan series (SiL; fine loam, mixed, mesic Fluventic Haplaquepts) covering on area of 5,000 m² (100 m × 50 m). The measured input quantities of N and P into the paddy field were as follows: 122 and 140 kg N ha^?1 and 29 and 30 kg P₂O₅ kg ha^?1 from chemical fertilizer, 20 and 28 kg N ha^?1 and 0.35 and 0.36 kg P ha^?1 from precipitation, and 26 and 35 kg N ha^?1 and 0.57 and 0.72 kg P ha^?1 from irrigation water, respectively. The measured outputs of N and P during the study period were as follows: 48 and 52 kg N ha^?1 and 1.1 and 1.6 kg P ha^?1 from runoff water, and 9 and 12 kg N ha^?1 and 0.04 and 0.05 kg P ha^?1 from infiltration. The runoff loading was the highest in June, presumably because of the higher concentrations of chemical components associated with chemical fertilizer application. The runoff losses of nutrients were compared to the amounts of nutrients supplied by chemical fertilizers. It was found that the losses of N accounted for 34.3 and 42.6% of the chemical fertilizer applied, while those of P accounted for 3.8 and 5.3%. The ratio between nutrient losses by infiltration and the chemical fertilizer applied was 6.4 and 9.8% for N and 0.1 and 0.2% for P, respectively.