Purification and anti-inflammatory action of tripeptide from salmon pectoral fin byproduct protein hydrolysate.

In this study, the anti-inflammatory peptide from salmon pectoral fin byproduct protein hydrolysate by pepsin hydrolysis, was purified and identified using Sephadex G-25 gel permeation chromatography, high performance liquid chromatography and time-of-flight liquid chromatography/tandem mass spectrometry (TOF LC/MS/MS). The purified anti-inflammatory peptide was identified to be a tripeptide (PAY). Lipopolysaccharide treatment significantly (p<0.05) stimulated the production of nitric oxide (NO) and prostaglandin E2 (PGE2) in RAW264.7 cells. However, PAY treatment significantly (p<0.05) inhibited the production of NO by 63.80% and PGE2 by 45.33%. Western blotting analysis revealed that PAY significantly (p<0.05) suppressed the protein expression of inducible nitric oxide synthase and cyclooxygenase-2, which are responsible for the production of NO and PGE2. Additionally, PAY treatment also significantly (p<0.05) attenuated the production of pro-inflammatory cytokines, including tumour necrosis factor-α, interleukin-6 and -1ß.

Dipeptide Phe-Cys derived from in silico thermolysin-hydrolysed RuBisCO large subunit suppresses oxidative stress in cultured human hepatocytes.

A dipeptide (Phe-Cys) was predicted to be bioactive following bioinformatics analysis of the large subunit of plant and microalgae ribulose-1,5-bisphosphate carboxylase (RuBisCO), which was hydrolysed in silico with thermolysin. The peptide was synthesised and found to possess in vitro reducing potential and inhibitory activity against lipid peroxidation, comparable to the activity of glutathione. In cultured Chang human hepatocytes, 2.5-10 µM Phe-Cys was found to induce the suppression of reactive oxygen species formation and membrane lipid peroxidation in oxidative stressed cells. Intracellular glutathione levels were found to increase in the peptide-treated cells under normal condition, which can potentially contribute in protecting the cells from oxidative damage. Furthermore, Western blot analysis showed that the levels of antioxidant enzymes, catalase and superoxide dismutase-1, increased in the hepatic cells when treated with Phe-Cys in the presence of the oxidant. The results show that this peptide has great potential to be used against oxidative stress-induced health conditions.

Effect of steaming, blanching, and high temperature/high pressure processing on the amino Acid contents of commonly consumed korean vegetables and pulses.

In the present report, the effects of blanching, steaming, and high temperature/high pressure processing (HTHP) on the amino acid contents of commonly consumed Korean root vegetables, leaf vegetables, and pulses were evaluated using an Automatic Amino Acid Analyzer. The total amino acid content of the samples tested was between 3.38 g/100 g dry weight (DW) and 21.32 g/100 g DW in raw vegetables and between 29.36 g/100 g DW and 30.55 g/100 g DW in raw pulses. With HTHP, we observed significant decreases in the lysine and arginine contents of vegetables and the lysine, arginine, and cysteine contents of pulses. Moreover, the amino acid contents of blanched vegetables and steamed pulses were more similar than the amino acid contents of the HTHP vegetables and HTHP pulses. Interestingly, lysine, arginine, and cysteine were more sensitive to HTHP than the other amino acids. Partial Least Squares-Discriminate Analyses were also performed to discriminate the clusters and patterns of amino acids.

Almond protein hydrolysate fraction modulates the expression of proinflammatory cytokines and enzymes in activated macrophages.

Simulated gastrointestinal treatment of almond proteins with pepsin and pancreatic proteases resulting in 16.6% degree of hydrolysis or 1.33 milliequivalent leucine per g protein yielded a hydrolysate that modulated excessive nitric oxide production in lipopolysaccharide-activated RAW264.7 macrophages. After fractionation, a resulting fraction of molecular size > 5 kDa retained the nitric oxide modulatory effect observed initially in the crude hydrolysate. The high molecular size fraction was found to modulate levels of proinflammatory cytokines, interleukin (IL)-6, IL-1ß, and tumour necrosis factor (TNF)-α in the activated cells. Immunoblotting analysis indicated that the hydrolysate fraction decreased the expression levels of inflammatory enzyme indicators, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 in the activated cells. RT-PCR analysis showed that treatment of the activated cells with the hydrolysate fraction resulted in the inhibition of relative gene expressions of proinflammatory IL-6, IL-1ß, TNF-α, iNOS and COX-2. These results indicate a potential application of almond protein hydrolysates against inflammatory conditions, and will contribute to delineating the possible contributions of proteins to health benefits attributed to almond consumption.

Antioxidant activities and determination of phenolic compounds isolated from oriental plums (Soldam, Oishiwase and Formosa).

The purposes of this study were to determine phenolic compounds and to evaluate antioxidant activities of plums (Soldam, Oishiwase and Formosa). Soldam contains the highest amount of total phenolics among cultivars (Formosa: 4.0%, Oishiwase: 3.3%, Soldam: 6.4% for total phenolic) as well as the total flavonoids of which constituents were mainly myricetin and anthocyanidin. The antioxidant activities were measured by DPPH, ABTS radical scavenging, and SOD-like activities. The DPPH radical scavenging activity of Korean plum extracts (200 µg/mL) showed more than 43%, and the Soldam turned out to be the highest : ID(50) value: 160-177 µg/mL for Formosa and Oishiwase; 58-64 µg/mL for Soldam. The ABTS radical scavenging activity of Korean plum extracts (200 µg/mL) was found to be more than 50%. The SOD-like activity of Korean plum extracts (200 µg/mL) showed more than 70%. Among three kinds of cultivars, Soldam had the highest antioxidant activity. The nitrite scavenging activity of Soldam was 61.5%, which is the highest, compared with that of the other cultivars, about 50%. From these results, Korean plums turned out to be phytochemical rich fruit as well as to show high antioxidant activities.

Chitosan gallate as potential antioxidant biomaterial.

Chitosan gallate were synthesized using a free radical-induced grafting reaction. Chitosan gallate showed enhanced water-solubility compared to plain chitosan, and exhibited good thermal stability. The IC(50) value of chitosan gallate against 2,2-diphenyl-1-picrylhydrazyl (DPPH) was 17.86 µg/mL. In addition, chitosan gallate effectively inhibited the generation of intracellular reactive oxygen species (ROS), and also suppressed lipid peroxidation in RAW264.7 macrophage cells. Chitosan gallate also exhibited the protection effect on genomic DNA damage by induced hydroxyl radical, and up-regulated the protein expression of antioxidant enzymes including superoxide dismutase-1 and glutathione reductase under H(2)O(2)-mediated oxidative stress in RAW264.7 macrophage cells. These results indicate that chitosan gallate might be potential antioxidant biomaterials.

Absence of association between two HECTD2 polymorphisms and sporadic Creutzfeldt-Jakob disease.

BACKGROUND: HECT (homologous to E6-AP carboxyl terminus) E3 ubiquitin ligases are fundamental components of the eukaryotic ubiquitin-proteasome system and are involved in the pathogenesis of several human diseases, including polyglutamine diseases. HECTD2, an E3 ubiquitin ligase, has been linked to the incubation time of prion disease in mice, and its polymorphisms have been associated with sporadic Creutzfeldt-Jakob disease (CJD) in the British population. OBJECTIVE: To investigate whether 2 HECTD2 polymorphisms, -247G→A (rs7081363) and +16066T→A (rs12249854), are associated with sporadic CJD in the Korean population. METHODS: We compared the genotype, allele and haplotype frequencies of the 2 HECTD2 polymorphisms in 205 sporadic CJD patients to those of 208 healthy Koreans. RESULTS AND CONCLUSION: Our study does not show significant differences in the genotype and allele frequencies of these 2 polymorphisms between sporadic CJD and normal controls. Significant differences in the haplotype frequencies of these 2 polymorphisms were not observed between sporadic CJD and normal controls either. Our results indicate that these 2 HECTD2 polymorphisms are not associated with genetic susceptibility to sporadic CJD in a Korean population. This is the first genetic association study of HECTD2 with sporadic CJD in an Asian population.

Portion sizes of foods frequently consumed by the Korean elderly: Data from KNHANES IV-2.

The purpose of this study was to define a one-portion size of food frequently consumed by the Koreans aged 65 years or over. From the original 8,631 people who took part in the Forth Korea National Health and Nutrition Examination Survey (KNHANES IV-2) 2008, we analyzed the data on 1,458 persons (16.9%) aged 65 and over, and selected food items consumed based on the intake frequency of 30 or more by all participant. A total of 158 varieties of food items were selected. The portion size of food items was set on the basis of the median amount (50 percentile) in a single intake by a single person. In the cereals category, 13 items were selected, of which the most frequently consumed item was well-polished rice with portion size of 75 g. Among legumes, 7 items were selected, of which the most frequent item was dried black soybean with a portion size of 6 g. Among the 16 groups, the most varied food group (49 items) was vegetables, and among these the most frequently occurring item was garlic (5 g), while among the fruit group, only 11 items were selected, as their intake frequency was low. Fish and shellfish were more frequently consumed by the elderly than meats. The most frequently consumed meat was pork loin, with a portion size of 30 g. In fish and shellfish, the most frequently consumed item was dried and boiled large anchovy with a portion size of 2 g. Portion sizes for food items consumed regularly by the elderly may be conveniently and effectively used in dietary planning and in nutritional education programs, and in assessing the diet intake status of the elderly.

Genetic association of a cathepsin D polymorphism and sporadic Creutzfeldt-Jakob disease.

BACKGROUND: Cathepsin D is the most abundant lysosomal and endosomal aspartyl protease; it shows beta and gamma secretase activity in vitro by cleaving the amyloid precursor protein into amyloid beta protein. In recent studies, cathepsin D was co-localized with PrP(Sc), the disease-associated form of the prion disease, and abnormal expression of cathepsin D correlated with tissue damage in brains of sporadic Creutzfeldt-Jakob disease (CJD). OBJECTIVE: To investigate whether a polymorphism at position 224, C224T, on exon 2 of the cathepsin D gene (CTSD) is associated with sporadic CJD in the Korean population. METHODS: We compared the genotype and allele frequencies at this polymorphism site in 172 sporadic CJD patients with those in 197 healthy Koreans. RESULTS AND CONCLUSION: Our study does not show a significant difference in genotype (p = 0.901) and allele (p = 0.509) frequencies of CTSD C224T between sporadic CJD patients and normal controls. This was the first genetic association study of CTSD in a sporadic CJD population.

Lack of association between PRNP 1368 polymorphism and Alzheimer's disease or vascular dementia.

BACKGROUND: Polymorphisms of the prion protein gene (PRNP) at codons 129 and 219 play an important role in the susceptibility to Creutzfeldt-Jakob disease (CJD), and might be associated with other neurodegenerative disorders. Several recent reports indicate that polymorphisms outside the coding region of PRNP modulate the expression of prion protein and are associated with sporadic CJD, although other studies failed to show an association. These reports involved the polymorphism PRNP 1368 which is located upstream from PRNP exon 1. In a case-controlled protocol, we assessed the possible association between the PRNP 1368 polymorphism and either Alzheimer's disease (AD) or vascular dementia (VaD). METHODS: To investigate whether the PRNP 1368 polymorphism is associated with the occurrence of AD or VaD in the Korean population, we compared the genotype, allele, and haplotype frequencies of the PRNP 1368 polymorphism in 152 AD patients and 192 VaD patients with frequencies in 268 healthy Koreans. RESULTS AND CONCLUSION: Significant differences in genotype, allele and haplotype frequencies of PRNP 1368 polymorphism were not observed between AD and normal controls. There were no significant differences in the genotype and allele frequencies of the PRNP 1368 polymorphism between Korean VaD patients and normal controls. However, in the haplotype analysis, haplotype Ht5 was significantly over-represented in Korean VaD patients. This was the first genetic association study of a polymorphism outside the coding region of PRNP in relation to AD and VaD.

Serologic screening of pregnant Korean women for primary human cytomegalovirus infection using IgG avidity test.

BACKGROUND: Primary human cytomegalovirus (CMV) infection during pregnancy is a major cause of congenital malformation. We detected primary CMV infection in pregnant Korean women by using an algorithm that comprises CMV IgG, IgM, and IgG avidity tests. METHODS: During a 2-month period, 744 pregnant women who were at 10-19 weeks of gestation were consecutively enrolled in this study. Human anti-CMV IgG and IgM levels in their sera were determined by chemiluminescence immunoassays. Serum samples from the women who were positive for CMV IgG and IgM were assayed by the ARCHITECT CMV IgG avidity test in order to distinguish primary from non-primary CMV infection. Gross examination of the neonates of the women who were positive for CMV IgM was conducted. RESULTS: The seroprevalence of CMV IgG and IgM was estimated to be 98.1% and 1.7%, respectively. The samples from all the women who were positive for CMV IgM or with grey zone results contained high avidity CMV IgG. Seven women with positive CMV IgG and IgM results who completed follow-up up to delivery showed no gross evidence of in utero CMV transmission. CONCLUSIONS: Maternal primary CMV infection was not detected in any of the pregnant women included in this study cohort. CMV IgG avidity test enabled the identification of women who were at a low risk of transmitting CMV infection and provided informative for subsequent pregnancy outcomes. Compared to previous studies, the seroprevalence of CMV IgG antibody across pregnant Korean women remained unchanged.

Characterization of (aminoethyl)chitin/DNA nanoparticle for gene delivery.

Nonviral gene delivery systems have been increasingly proposed as a safer alternative to viral vehicles. In the present study, we synthesized water-soluble chitin by aminoalkylating onto chitin at the C-6 position, and its transfection efficiency was investigated. Aminoethyl-chitin (AEC) was complexed with DNA, and AEC/DNA nanoparticles were characterized. AEC/DNA nanoparticles showed good DNA binding ability, high protection of DNA from nuclease and serum, and low cytotoxicity. Mean particle size decreased from 367 to 290 nm and zeta potential increased from -4.58 to 22.87 mV when the AEC/DNA charge ratio (N/P) increased from 1.15 to 18.5. The transfection efficiency of AEC/DNA nanoparticles was investigated in a human embryonic kidney cell line (HEK293), and the results showed that AEC/DNA nanoparticles were much enhanced compare with naked DNA.

Cytotoxic activities of water-soluble chitosan derivatives with different degree of deacetylation.

Chitosans with different degree of deacetylation (DD) (90% and 50% deacetylated chitosan) were prepared by N-deacetylation followed by grafted onto chitosan to form water-soluble aminoethyl-chitosan (AE-chitosan), and dimetylaminoethyl-chitosan (DMAE-chitosan), diethylaminoethyl-chitosan (DEAE-chitosan). In the present study, cytotoxic activities of the chitosan derivatives were evaluated using three tumor cell lines and two normal cell lines, and structure-activity relationship was suggested. The cytotoxic activity was dependent on their DD and substituted group.