Insects or insect remains found in beer are one of major issues in consumer claim. Accurate estimation of inflow time isa critical factor for the settlement of such claims related with beer-contaminating insects but no reliable methods have been developed. In an attempt to establish a molecular marker-based diagnostic method, the degradation rates of 18S rRNA genes in the insectssoaked in 500 ml beer were investigated by quantitative real-time PCR (qPCR) over one month period at room temperature. Among the six insect species tested, the house fly (Musca domestica) and honey bee (Apis mellifera) revealed high correlations (r2=0.974-0.990) between the degradation of 18S rRNA gene and inflow time. In these insects, statistically significant distinction was possible between the samples stored in beer less than 14 days and more than 14 days. Other insects, including the fruit fly, common house mosquito, German cockroach and Indian meal moth, displayed poor correlations, which appeared attributed to the inefficient genomic DNA extraction likely due to small sample size or disintegration of body parts during storage in beer. With proper improvement in DNA extraction, this 18S rRNA-based diagnostic method would be applicable for estimating the inflow time of beer-contaminating insects.

The hemipteran whitefly Bemisia tabaci (Gennadius) is one of the most destructive pests damaging more than 600 agricultural crop species worldwide. The B and Q biotypes are most widely spread in Korea but they are not distinguishable based on morphological characters. In order to search for protein markers that can be employed for rapid and accurate diagnosis of biotypes, two-dimensional PAGE (2DE) in conjunction with mass spectroscopic analysis were conducted. Eleven biotype-specific spots were repeatedly identified during three repetitions of 2DE and analyzed by Q-TOF. One of the B type-specific protein spots was identified as carboxylesterase 2 (Coe2). The transcript level of coe2 was determined to be 6 times higher in B type than in Q type by quantitative real-time PCR. In addition, comparison of genomic DNA sequence of coe2 between B and Q types identified a biotype-specific intron, from which specific primer sets were designed. One-step PCR using these biotype-specific primers successfully distinguished the two biotypes in a high accuracy. Availability of the biotype-specific protein and DNA markers will greatly improve the detection of B. tabaci biotype in the field.

A full genomic DNA microarray technique was employed to investigate the effects of Dongchunghacho on aortal and hepatic gene expression in apolipoprotein E knockout mice fed a high-fat/high-cholesterol diet. Male 8- week - old ApoE-/- mice were randomly divided into two groups, control(high cholesterol group; HC) and supplementation of Dongchunghacho (SD). All of the mice were fed a high-fet/high cholesterol diet with or without Dongchunghacho supplemented by 1% for 6 weeks. At first, lipid profile of the Dongchunghacho was measured by biochemical analysis. No differences were observed in serum triglyceride and total cholesterol levels between the two groups. Antigenotoxic effect of the Dongchunghacho was measured by the single cell gel electrophoresis assay (Comet assay) and quantified as % fluorescence in tail. Dongchunghacho supplementation decreased significantly leukocytic DNA damage and also there was a tendency of reduction in hepatic DNA damage in Dongchunghacho group compared with the control group. In up regulated genes in liver and aorta of the mice, genes with 0 to 2- fold difference in expression level between the two group (HD and SD) was very much more in liver than in aorta, on the contrary, those with 2-fold to 16-flod difference increased greatly rather in aorta than in liver. Also, almost the same results were observed in down regulated genes in liver and aorta between the two groups. These results suggested that supplementation of Dongchunghacho might be helpful in preventing leukocytic DNA damage induced by high fat diet, and has a more crucial roles in aortal gene expression.

Developmental periods of sweet potato whitefly, Bemisia tabaci(Gennaius) Q-biotype, were investigated on three host plants- sweet bell pepper (Capsicum annuum L.), eggplant(Solanum melongena L.) and oriental melon(Cucumis melo L. var. makuwa MAKINO). Egg and nymph development of B. tabaci were studied within temperature ranging of 15℃-35℃ by 2.5℃ under photoperiod 16:8(L:D). Egg period of B. tabaci was the shortest at 32.5℃ and nymphal period was shortest at 27.5℃ on sweet bell pepper. Nymphal period of B. tabaci on eggplant was the shortest at 27.5℃ as well. On the other hand, nymphal period of B. tabaci was shortest at 30℃ and 32.5℃. Lower temperature threshold and effective degree-day for completing egg development on sweet bell pepper were estimated as 13.11, 91.95, respectively. Lower threshold temperature of nymphal stage on sweet bell pepper, eggplant, oriental melon were estimated as 13.01, 13.39, 12.31,respectively. Degree-days required to complete nymphal stage on sweet bell pepper, eggplant, oriental melon were estimated as 191.22, 164.41, 190.34, respectively. The relationships between development rates of egg and nymph were well described by poikilothermal rate function and weibull function. The fitted curves will be used as input for a simulation model of the population dynamics of B. tabaci.

Tomato fruit color, which is the most visible characteristic among the other fruit traits, is considered to have a substantial influence on consumers. The pink-colored tomatoes with high soluble solids content are considerably preferred especially in Asia compared to the other colors. Generally the pink fruit trait of tomatoes is easily determined by visual examination of intact fruit, however, it is technically determined by the characteristic of the fruit peel. The pink trait is regulated by variations of the SlMYB12(y) gene located on chromosome 1, which controls the accumulation of the naringenin chalcone, which comprises a large proportion of flavonoids. In this study, we developed a derived Cleaved Amplified Polymorphic Sequences (dCAPS) marker and a sequence characterized amplified regions (SCAR) marker in order to discriminate of pink/non-pinktomatoes in the domestic breeding lines. Quantitative RT-PCR analysis indicated that the SlMYB gene is highly expressed in non-pink fruit peel, whereas the expression is significantly lowered in the pink fruit peel. These gene based markers are expected to enhance the efficiency and accuracy of selection pink-tomatoes in tomato breeding programs.

The isolated single coronary artery is a rare congenital anomaly, in which both coronary arteries arise from a solitary ostium. Diagnosis of coronary anomalies and identification of the exact anatomy of coronary arteries has significant clinical importance, hence, myocardial ischemia or sudden cardiac death is usually related to its course of anomalous coronary artery. Most patients with a single coronary artery are asymptomatic and have normal electrocardiogram and negative stress tests. However, if the patient has other structural abnormalities, for example, ventricular hypertrophy, the exam is determined. This report describes a case of single coronary artery, where the right coronary artery originated from the distal left circumflex artery in a patient with hypertrophic ardiomyopathy.

The canine major histocompatibility complex (MHC) is referred to dog leukocyte antigens (DLA), which is known to be the most polymorphic genetic system in canine species. Many cloned dogs have been produced since Snuppy, first cloned dog, there was no research about genetic identity of MHC among cloned animals. Recently in Lee’s group, two non-transgenic cloned beagles (BG1, 2) were produced by somatic cell nuclear transfer (SCNT) using fetal fibroblast (BF). Also, four transgenic cloned beagles (Ruppy 1-3, 5) were generated using transgenic BF transfected with Red fluorescent protein (RFP) gene. We hypothesize that non-transgenic (BG1, 2) and transgenic (Ruppy 1-3, 5) cloned beagles derived from identical donor cells have the same immunological genetic characteristic except for RFP gene insertion in the genome. Thus, the aim of this study is to confirm the immunological identity of DLA class II in cloned beagles produced using same nuclear donor cell. Genomic DNA was extracted from blood of BG1, BG2, Ruppy 1, 2, 3 and 5. Genomic DNA of normal two control beagle, no correlation with BF was also investigated for rulling out the possibility that beagles were inbred. Forward and reverse primers used for DLA-DQA1 and DQB1 respectively were DQAF: 5’-TAAGGTTCTTTTCTCCCTCT-3’ and DQAR: 5’-GGACAGATTCAGTGAAGAGA-3’ DQBR:5’-CTCACTGGCCCGGCTGTCTC-3’ and DQBR: 5’-CACCTCGC CGCTGCAACGTG-3’. Polymerase Chain Reaction (PCR) products were purified, sequenced directly using the Big Dye Terminator kit. Sequencing analysis was performed on an automated 3730xl DNA analyzer. In experiment 1, sequence of DLA-DQ alpha 1 (DQA1) and DLA-DQ beta 1 (DQB1) exon 2, hypervariabel region, was compared in BG1 and BG2. Experiment 2 also compared the sequence of DQA1 and DQB1 among Ruppy 1, 2, 3 and 5. Experimental 3 compared sequence of DQA1 and DQB1 among all cloned dogs (BG1, BG2 and Ruppy 1-3, 5). As a result, BG1 and BG2 have same allele for DQA1 and DQB1 as we expected. They share DQA1*00101 and DQB1*02901 in experiment 1. In experiment 2, Ruppy 1, 2, 3 and 5 also have identical DQA1*00101 and DQB1*02901 allele. No discrimination between transgenic dogs and cloned dogs was seen in DQA1 and DQB1 Allele in experiment 3. DQA1, DQB1 allele was identified as *00101 and *02901 in all dogs. We provided the allele identity of DQA1and DQB1 in cloned beagles, which can be used as preliminary data for immunological related studies. In conclusion, transgenic cloned dogs despite of red fluorescent protein genes being inserted in their nuclear DNA were immunologically compatible with non-transgenic cloned dogs. We demonstrated that cloned beagles produced using identical nuclear donor were immunologically compatible.

Barley Yellow Mosaic Virus (BaYMV) caused significant reduction in barley yield and is difficult to control due to alive parasitic soil-borne fungus, Palmyra gamines that transmits the, virus. Previous studies have indicated that a virus-free soil could be infested by using virus-contaminated farming machineneries and implements. For the further confirmation of this finding, different proportions of BaYMV-infested soil were mixed into virus free soil. Three barley varieties (Hordum vulgarae, cv "Olbori", "Baegdong" and "Sacheon 6") were sown in pots treated with different rate of P. graminis-infested soil ranging from 0% to 100% in October 20, 2001. Results showed that BaYMV infection increased as the rate of infested soil increased. Initial symptoms were observed in a pots treated with 10% infested soil in all the 3 varieties of barley. "Olbori" had about 5% infection in 20% infested soil and about 10% infection in 40% or 50% infested soil and about 20% infection in 60% infested soil. In "Baegdong", the trend of BYMV occurrence was similar with "Olbori" but the time of severe infection was earlier than "Olbori". BaYMV infection in "Sacheon 6" was even earlier than "Baegdong" with much more severe symptoms than "Baegdong". The growth rate of barley was affected by about 19-22% when grown in 20% infested soil. As the rate of BaYMV infested soil increased the heading date was delayed but the maturing date was early in "Olbori" and "Sacheon 6". Also, reduction rate of culm length in 3 varieties increased with increase of infested soil content. However, "Olbori" showed the highest reduction. "Sacheon 6", have been characterized with long spike length, however was significantly reduced as the infested soil increased. On the other hand, spike length of "Olbori" was not significantly affected despite of increased of infested soil. The reduction rate of 1000 kernel weight was higher in large kernel size cultivar "Sacheon 6" and "Olbori" than small kernel size "Baegdong" as increase of BaYMV-infested soil content.

The aim of this study was to isolate chicken feather-degrading bacteria with high keratinolytic activity and to investigate cultural conditions affecting keratinolytic enzyme production by a selected isolate. A chicken feather-degrading bacterial strain CH3 was isolated from poultry wastes. Isolate CH3 degraded whole chicken feather completely within 3 days. On the basis of phenotypical and 16S rDNA studies, isolate CH3 was identified as Bacillus thuringiensis CH3. This strain is the first B. thuringiensis described as a feather degrader. The bacterium grew with an optimum at pH 8.0 and 37℃, where maximum keratinolytic activity was also observed. The composition of optimal medium for keratinolytic enzyme production was feather 0.1%, sucrose 0.7%, casein 0.3%, K2HPO4 0.03%, KH2PO4 0.04%, MgCl2 0.01% and NaCl 0.05%, respectively. The keratinolytic enzyme had a pH and temperature optima 9.0 and 45℃, respectively. The keratinolytic activity was inhibited ethylenediaminetetraacetic acid, phenylmethylsulfonyl fluoride, and metal ions like Hg2+, Cu2+ and Zn2+. The enzyme activated by Fe2+, dithiothreitol and 2-mercaptoethanol.