• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.93-100
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• 2009

In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.18 no.3
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• pp.500-514
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• 1996

The purpose of this study was to observe the tissue response in applying staples and various suture materials to both scalp and buccal mucosa in rabbits. 18 rabbits were divided into 6 groups. The incised wounds of both scalp and buccal mucosa were sutured with staples, polyglactin 910, chromic catgut, mer silk and nylon. The experimental animals were sacrificed after 1, 3, 5, 7, 10, 14 days posto peratively 3 animals at one time. The tissue was stained with Hematoxylin and eosin, and Masson's Trichrom. In light microscopic examinations, the sutured sites were examined histologi cally according to 6 degrees about inflammation and collagen deposit. The results were obtained as follows, 1. The chromic catgut, an absorbable suture material, was absorbed by 7 days, whereas polyglactin 910 and mersilk began to get absorbed after 7 days. 2. Mersilk manifested a broad range of inflammation in the scalp, and both staple and nylon showed a severe inflammatory reaction in the buccal mucosa. 3. With polyglactin 910, both tissue samples showed only minor foreign body reaction, however in the scalp, the process of fibrosis took place compara tively slowly, whereas in the buccal mucosa, it occurred promptly and manifested active fibrosis by 7 days. 4. Mersilk showed widespread a matrix formation in both scalp and buccal mucosa, and showed the most severe inflammatory reaction by 3 days, which did not seem to decrease even after 7 days. 5. Both staple and nylon showed relatively a severe inflammatory reaction, however fibrosis took place rather promptly compared to the other groups. 6. Generally, in the buccal mucosa fibrosis occurred more promptly than in the scalp in both control and experimental groups. 7. Retention of the suture material and stability of the knot were the best with the staple, and better stability was manifested by the multi-stranded poly glactin 910 and mersilk than singlestranded chromic catgut and nylon. From above results, in the buccal mucosa absorbable suture materials especially polyglactin 910 showed better response in the aspect of inflammatory reaction, while in the scalp monofilament suture materials such as staple and nylon manifested a early fibrosis and collagen formation.

• Journal of Periodontal and Implant Science
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• v.38 no.4
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• pp.565-578
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• 2008

Purpose: Porphyromonas gingivalis(P. gingivalis) heat shock protein (HSP)60 may play a role in the immunopathogenesis of periodontitis as well as atherosclerosis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. The purpose of this study was to identify immunodomiant epitope of P. gingivalis HSP60 that is reactive exclusively to the homologous bacteria without reacting with human HSP. Materials and methods: The present study was performed to identify the peptide specifically recognized by anti-P. gingivalis HSP60 monoclonal antibodies mono-reactive to P. gingivalis HSP60. Results: Four different hybridomas were cloned producing monoclonal IgG antibodies exclusively to P. gingivalis HSP60. Thirty seven synthetic peptides (20-mer with 5-amino acid overlapping) were synthesized. All of these peptide were subject to SDS-PAGE for immunblot analysis. One peptide (TVPGGGTTYIRAIAALEGLK) and the other peptide (TLVVNRLRGSLKICAVKAPG) were recognized by all and one of the four monoclonal antibodies, respectively, that reacted solely with P. gingivalis HSP60. Immunohistochemistry to identify the localization of the HSP60 in the diseased gingival tissues revealed that all of the four monoclonal antibodies were highly reacted with the diseased gingival tissue than normal gingival tissue. Conclusion: The P. gingivalis HSP60 peptides (TVPGGGTTYIRAIAALEGLK and TLVVNRLRGSLKICAVKAPG, respectively) are positively involved in the immunopathologic process of periodontal disease. The peptide may potentially be developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P. gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.871-884
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• 2009

Gene expression profiling is a useful tool for identifying critical genes and pathways in metabolism. The objective of this study was to determine the major differences in the expression of genes associated with metabolism and metabolic regulation in liver and mammary tissues of lactating cows. We used the Michigan State University bovine metabolism (BMET) microarray; previously, we have designed a bovine metabolism-focused microarray containing known genes of metabolic interest using publicly available genomic internet database resources. This is a high-density array of 70mer oligonucleotides representing 2,349 bovine genes. The expression of 922 genes was different at p<0.05, and 398 genes (17%) were differentially expressed by two-fold or more with 222 higher in liver and 176 higher in mammary tissue. Gene ontology categories with a high percentage of genes more highly expressed in liver than mammary tissues included carbohydrate metabolism (glycolysis, glucoenogenesis, propanoate metabolism, butanoate metabolism, electron carrier and donor activity), lipid metabolism (fatty acid oxidation, chylomicron/lipid transport, bile acid metabolism, cholesterol metabolism, steroid metabolism, ketone body formation), and amino acid/nitrogen metabolism (amino acid biosynthetic process, amino acid catabolic process, urea cycle, and glutathione metabolic process). Categories with more genes highly expressed in mammary than liver tissue included amino acid and sugar transporters and MAPK, Wnt, and JAK-STAT signaling pathways. Real-time PCR analysis showed consistent results with those of microarray analysis for all 12 genes tested. In conclusion, microarray analyses clearly identified differential gene expression profiles between hepatic and mammary tissues that are consistent with the differences in metabolism of these two tissues. This study enables understanding of the molecular basis of metabolic adaptation of the liver and mammary gland during lactation in bovine species.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.52 no.1
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• pp.51-54
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• 2007

Soybean has a morphological type with a broadened and flattened stem. Fasciation has been suggested as a new gene for soybean research. SSR marker linked to the $\Large f$ locus that controls fasciation phenotype has not identified within 10 cM. A mapping population consisting of 94 $F_2$ progenies was derived from a cross between wild type Clark (FF) and fasciation mutant C32 (${\Large f}{\Large f}$). The phenotype of $F_2$ individual plants was recorded at R2 and R3 growth stage from field. One-thousand 10-mer oligonucleotide RAPD primers and 29 SSR primers selected from the D1b+W of the soybean molecular linkage map were used. A genetic map was constructed from the segregating 35 RAPD, four SSR markers and one phenotypic(wild type/fasciation) marker. The segregation ratios of 3 : 1 observed in the $F_2$ population and the Chi-square values strongly suggest that the fasciation trait is controlled by a single recessive gene. Satt537 marker was linked to $\Large f$ locus at a distance of 9.6 cM. Assignment of the $\Large f$ locus to linkage group D1b+W and identification of markers can be used as an initial step for fine mapping of the $\Large f$ gene.

• Horticultural Science & Technology
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• v.17 no.3
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• pp.352-354
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• 1999

Randomly and specifically amplified polymorphic DNA band patterns based on polymerase chain reaction (PCR) analysis were used to assess genetic variation of somaclonal variants obtained from tissue culture of lisianthus (Eustoma grandiflorum). Five different types of variant were classified by morphological characters such as leaflet number, leaf shape, caulicle length, plant height, and leaf area. Five primers out of 20 primers (10 mer) resulted in 34 random amplified DNA fragments with polymorphisms (64.7%) in all tested plants. The dissimilarity coefficient was from 0.71 to 0.91 by UPGMA cluster analysis. Based on the presence of polymorphic bands, normal plant and five somaclonal variants were divided into two groups at the similarity coefficient value of 0.79.

• Journal of fish pathology
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• v.20 no.3
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• pp.307-313
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• 2007

In this study, a QCM biosensor was made to detect Edwardsiella tarda (E. tarda) using a specific antibody. A 9 MHz AT-cut piezoelectric wafer layered with two gold electrodes of 5mm diameter had a reproducibility of 0.1 Hz in frequency response and was used as the transducer of the QCM biosensor. Self assembled layer (SAM) was conformed on a quartz crystal by treating with 3-mer-captopropionic acid (MPA) and activated with N-ethyl-N'-(3-dimethyl-aminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). The resulting NHS group was further converted to hydrazide by the reaction with hydrazine. Aldehyde group was introduced into the carbohydrate moiety of anti-E. tarda antibody by the reaction with periodic acid and was used to immobilise the antibody through the reaction with hydrazide group on the electrode surface. A baseline was established in the presence of phosphate-buffered saline (PBS) and a resonant frequency (F1) was measured. Sample was added to the sensor surface and second resonant frequency (F2) was measured after unbound substances were washed out with PBS several times. Finally, the frequency shift (ΔF) representing the mass change was calculated by subtracting F2 from F1. After adding the oxidized anti-E. tarda antibody to the electrode surface containing hydrazide group, frequency shift of 288.811.4 Hz (mean S.E) was observed, thus proving that considerable amount of antibody was immobilized. In the immunoassay test, the frequency shift of 1877.75 Hz, 580.67 Hz, 221.39 Hz, 7.671.83 Hz (mean S.E) were observed at doses of 1000, 500, 100, 50 g of bacterial cells, respectively. It was also demonstrated that the prepared sensor chip was stable enough to withstand repeated surface regeneration with 0.2 M Tris-glycine and 1 % DMSO, pH 2.3 more than ten times.

• International Journal of Industrial Entomology and Biomaterials
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• v.8 no.2
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• pp.161-167
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• 2004

Silkworms sex determination drew high attention from researchers. Sex chromosomes on the silkworm are of ZW type for females and ZZ type for males. Chromosome W plays an important role in sex determination. Although several molecular linkage maps have been constructed for silkworm, very few markers are discovered on the W chromosome. In order to look for molecular markers and to further locate the Fern gene on chromosome W, we used genomic DNA from both female and male larvae of a silkworm strain named 937 as PCR templates for RAPD amplification with 200 arbitrary 10-mer primers. The amplification results showed three female-specific bands, namely ${OPG-07_496}, {OPC-15_1,660} and {OPE-18_1,279}$. Further verification, however, revealed no band from OPG-07 and OPC-15 in either sex in the strain 798, but OPE-18 provided female-specific band in the strains Suluan7 and C108, and absent in both males and strain 798. This indicates that the bands from ${OPG-07_496} and {OPC-15_1,660}$ are probably female-specific in strain 937, and the band from OPE-18 was probably amplified from a common segment shared by most strains. The genomic DNAs from OPG-07 and OPC-15 were cloned and sequenced. Sequence analysis showed that the DNAs from OPG-07 and OPC-15 have high identities with the retrotransposable elements, and DNA from OPC-15 contains a portion of sequence which probably encodes an eukaryotic translation initiation factor 4E binding protein (eIF4EBP).

• Proceedings of the Korea Inteligent Information System Society Conference
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• 2001.01a
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• pp.73-77
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• 2001

In this paper, we propose a virtual action system based on information agents, We call the system the MultiHammer, MultiHammer can be used for studying and analyzing online actions. MuiltiHammer provides functions of implement-ing a meta online action site and an experiment environ-ment. We have been using MultiHammer as an experiment as an experiment environment for BiddinBot. BiddingBot aims at assisting users to bid simultaneously in multiple online auctions. In order to bid simultaneously in multiple online auctions. In order to analyze the behavior of BiddngBot, we need to pur-chase a lot of items. It is hard for us to prepare a lot of fund to show usability and advantage of BiddingBot. MultiHam-mer enables us to effectively analyze the behavior of BiddingBot. MultiHammer consists of three types of agents for information collecting data storing and auctioning. Agents for information wrappers. To make agent work as wrarp-pers, we heed to realize software modules for each online action site. Implementing these modules reguires a lot of time and patience. To address this problem, we designed a support mechanism for developing the modules. Agents for data storing record the data gathered by agents for informa-tion collecting. Agents for auctioning provide online services using data recorded by agents for data storing. By recording the activities in auction sites. MultiHammer can recreate any situation and trace auction for experimentation, Users can participate in virtual using the same information in real online auctions. Users also participate in real auc-tions via wrapper agents for information collecting

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.4
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• pp.687-694
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• 1996

Myrosinase was purified from Korean mustard seed(Brassica juncea) by a sequential process of DEAE-cellulose, concanavalin A-sepharose, and Superose 6 chromatography. The molecular weight of puri-fied myrosinase(II-2) determined by SDS-polyacrylamide electrophoresis was 67KD. About a 248-fold purification for myrosinase II-2 was obtained after Superose 6 chromatography. Optimum pH of the myrosinase was 7.0 and optimum temperature of the enzyme was $3^{\circ}C.$ The enzyme was stable at pH 7.0, and below $30^{\circ}C.$ Cu, Hg and Fe ion significantly inhibited the enzyme activity, but ascorbic acid enhanced, resulting in a maximum activity by 1mM ascorbic acid. Among tile ascorbic acid ana-logues, dehydroascorbic acid inhibited the enzyme activity, whereas others showed a little effect. Reducing agents such as 2-mercaptoethanol and dithiothreitol inhibited the enzyme activity, but the reducing agents with ascorbic acid was enhanced enzyme activity.