Ochratoxin A in Korean food commodities: occurrence and safety evaluation

To evaluate the exposure of Koreans to ochratoxin A, we conducted a survey in 2003 for ochratoxin A in various domestic food commodities: 60 polished rices, 22 barleys, 35 wheat flours, 46 beers, and 14 unstrained rice wine (makkolli) samples. They were analyzed for ochratoxin A using immunoaffinity column and high-performance liquid chromatography (HPLC)-fluorescence detection, and the positive samples were confirmed using HPLC-tandem mass spectrometry. By combining results from different surveys on the levels of ochratoxin A in selected foods and the consumption patterns, we obtained the Korean probable daily intakes (PDI) of ochratoxin A. The polished rice commodity had the highest mean levels of ochratoxin A, which ranged from 0.2 (not detected, i.e., ND = 0) to 1.0 ng/g (ND = limit of detection, i.e., LOD). The estimated PDI for all Koreans fell into the range of 0.8-4.1 ng/kg bw/day, while for heavy consumers the estimates ranged from 1.7 to 9.1 ng/kg bw/day, which did not exceed the PTDI value (14 ng/kg bw/day). Staple rice is the major contributor (>90%) to the Korean dietary intake of ochratoxin A. On the basis of these estimates, it may be concluded that there is at present no considerable risk of ochratoxin A exposure for the average Korean consumer.     

A novel angiotensin I converting enzyme inhibitory peptide from Alaska pollack (Theragra chalcogramma) frame protein hydrolysate

Alaska pollack frame protein, which is normally discarded as an industrial byproduct in the processing of fish in plants, was hydrolyzed with pepsin. This was fractionated into five major types of Alaska pollack frame protein hydrolysates (APH-I, 10-30 kDa; APH-II, 5-10 kDa; APH-III, 3-5 kDa; APH-IV, 1-3 kDa; and APH-V, below 1 kDa) using an ultrafiltration membrane bioreactor system. Angiotensin I converting enzyme (ACE) inhibitory activities of the fractionated hydrolysates were investigated, and the fraction that exhibited the highest ACE inhibitory activity was further purified using consecutive chromatographic methods on SP-Sephadex C-25 column, Sephadex G-25 column, and high-performance liquid chromatography (HPLC) on an octadecylsilane column. Finally, we purified a novel ACE inhibitory peptide with an IC50 value of 14.7 microM, and the sequence of the peptide was Phe-Gly-Ala-Ser-Thr-Arg-Gly-Ala. In addition, the ACE inhibition pattern of the peptide was found to be noncompetitive.     

Changes in orexin-A and neuropeptide y expression in the hypothalamus of obese and lean Zucker Diabetic Fatty rats

This study was carried out to investigate the changes of orexin-A (OXA) and neuropeptide Y (NPY) expression in the hypothalamus of the obese and lean Zucker Diabetic Fatty (ZDF) rats which have a missense mutation in the leptin receptor gene. The mean body weights (MBW) between the obese and lean ZDF rats were significantly different at 28 and 70 postnatal days. However, at 14 postnatal day, there was no significant difference in the MBW between the obese and lean ZDF rats in both male and female. The OXA immunoreactivities were not significantly different between the obese and lean ZDF rats in both sexes at 14, 28, and 70 postnatal days, respectively. The NPY immunoreactivity was higher in the obese than in the lean ZDF rats in both male and female at 28 and 70 postnatal days, whereas there was no significant difference between the obese and lean ZDF rats at 14 postnatal day. These results indicate that both OXA and NPY might halt their roles for food intake in the obese phenotype of the male and female ZDF rats in the preweaning period of 14 postnatal day, whereas NPY might play a main role in the obesity of these rats in the weaning period of 28 and 70 postnatal days.     

Allelic relationships at the pvr1 locus in Capsicum annuum

Diseases caused by members of the Potyviridae family currently threaten pepper crops (Capsicum annuum L.) worldwide. A series of monogenic recessive resistance genes that control potyvirus resistance at the pvr1 locus in Capsicum species are widely known and used in pepper breeding programs, each allele with a differential resistance spectrum affecting a distinct range of viral strains across three viruses, Tobacco etch virus (TEV), Pepper mottle virus and Potato virus Y. In this study, we systematically analyzed the resistant spectra, and the level of the resistance each allele confers using a set of pepper genotypes homozygous or heterozygous for the following alleles; Pvr1 ⁺, pvr1, pvr1 ¹ and pvr1 ². The resistance alleles at the pvr1 locus show recessive inheritance when combined with a susceptible allele in F1 progenies. However, our results show that resistance in this system is, in fact, not always fully recessive and establish a hierarchy of allelic interactions We identified the resistance phenotype in F₁ progenies generated by combining a complete resistant allele with an incomplete resistance allele against TEV strains implying that the resistance alleles at the pvr1 shows dominant inheritance when combined with an incomplete resistance allele. Resistance alleles against TEV-HAT, pvr1 and pvr1 ², show dominance inheritance when each is combined with pvr1 ¹. The resistance allele, pvr1, shows dominance inheritance when combined with pvr1 ² against TEV-N. These results clarify the allelic relationship between resistance alleles at the pvr1 locus displaying different resistance spectra, and will assist breeders to select the preferred combinations of the resistance alleles to obtain durable resistance against multiple potyviral strains.     

Changes in glial fibrillary acidic protein immunoreactivity in the dentate gyrus and hippocampus proper of adult and aged dogs

Astrocytes perform neuron-supportive tasks, repair and scarring process in the central nervous system. In this study, we observed glial fibrillary acidic protein (GFAP), a marker for astrocytes, immunoreactivity in the dentate gyrus and hippocampus proper (CA1-3 region) of adult (2-3 years of age) and aged (10-12 years of age) dogs. In the adult group, GFAP immunoreactive astrocytes were distributed in all layers of the dentate gyrus and CA1-3 region, except in the stratum pyramidale of the CA1-3 region. In the aged group, GFAP immunoreactivity decreased markedly in the molecular layer of the dentate gyrus. However, GFAP immunoreactivity in the CA1-3 region increased in all layers, and the cytoplasm of GFAP immunoreactive astrocytes was hypertrophied. GFAP protein levels in the aged dentate gyrus decreased; however, GFAP levels in the CA1-3 region increased. These results suggest that the morphology of astrocytes and GFAP protein levels in the hippocampal dentate gyrus and CA1 region are changed, respectively, with age.     

Multilocus PCR assay reveals high diversity of vegetative compatibility types in populations of Cryphonectria parasitica in Croatia

The ascomycete fungus Cryphonectria parasitica, causal agent of chestnut blight, is probably one of the best known invasive fungal pathogens in forests of Europe and North America. Mycovirus that reduces virulence of C. parasitica can be used as a biocontrol agent of the chestnut blight. However, anastomosis‐mediated virus transmission is limited by a vegetative (in)compatibility (vc) system involving at least six known diallelic vic genetic loci. This study looked at vegetative compatibility (vc) diversity in two populations of C. parasitica in Croatia. For that purpose, a PCR assay was validated and implemented using already known/published and newly designed primers for amplification of six known vic loci. The vc genotypes determined by PCR for 158 C. parasitica isolates investigated in this study were in complete agreement with the vc genotypes determined by pairwise co‐culturing of the same isolates, revealing the specificity and accuracy of the PCR‐based molecular vic genotyping assay. Twenty‐six unique vc genotypes were found among 158 isolates, and 19 vc types per population, which makes Croatian C. parasitica populations among the most diverse in Europe regarding the number of vc types and genetic diversity. Low values of multilocus linkage disequilibrium suggest sexual reproduction as a major contributor to high C. parasitica genetic diversity in studied populations.     

Chitosan derivatives killed bacteria by disrupting the outer and inner membrane

Six kinds of water-soluble chitosan were prepared by grafting aminofunctionality onto chitosan at the C-6 position, and their antimicrobial activities were investigated against three Gram-negative and three Gram-positive bacteria. Among the derivatives, dimethylaminoethyl-chitosan (DMAEC) has the highest potential to suppress the growth of bacteria. To elucidate detailed antimicrobial modes of action against bacteria, cell integrity, outer membrane (OM), and inner membrane (IM) permeabilization assays were investigated. When treated with DMAEC, the release of 260 nm absorbing materials quickly increased for both Escherichia coli and Staphylococcus aureus, but the absorbance value was different due to the difference in cell structures. In OM and IM permeabilization assays, DMAEC rapidly increased 1-N-phenylnaphthylamine uptake and the release of cytoplasmic beta-galactosidase via an increase in the permeability of OM and IM. Moreover, DMAEC90 prepared from 90% deacetylated chitosan had more activity than DMAEC50 prepared from 50% deacetylated chitosan, and these results revealed that the antimicrobial action of water-soluble chitosans was dependent on the degree of deacetylation and the substituted group.