• Journal of agricultural medicine and community health
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• v.13 no.1
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• pp.41-59
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• 1988

In many of the helminthic infections, diagnosis is accomplished by the demonstration of the eggs or, occasionally the adult worms or their parts. Diagnosis can be made by the identification of larval stage obtained from stool or surgically extracted materials too. However some kinds of parasitic disease can not be diagnosed by above mentioned procedure alone. Brain cysticercosis, ectopic paragonimiasis, Capillaria hepatica infection in liver is a good example. In such a case, immunologic method would be helpful for the decision of physician. In this paper, immunologic tools such as indirect hemagglutination test, indirect fluorescent antibody test, circumoval precipitation test, ELISA, western blot were applied for the diagnosis of Clonorchis sinenisis, Cysticercosis and C. hepatica a infection and their efficacy was evaluated. The results obtained were as follows ; 1) In the diagnosis of clonorchiasis, ELISA revealed sensitiveity of 83.3%, but cross reaction against antibody of Paragonimus westermani and Taenia species were observed. For the identification of cross reaction and species specific band of Ag-Ab reaction, western blot was applied. 59Kd relative molecular weight and 21Kd band were identified as a Clonorchis sinensis specific band. OD values of ELISA performed with sera of 18 months after praziquantel treatment decreased to half level compared to that of before treatment. Negative conversion rate of ELISA after 18 months of treatment was 60%. 2) In the diagnosis of cysticercosis, IFAT disclosed 95.8%(23/24) of sensitivity and reaction was most strongly occurred in inner membrane. ELISA revealed 90.0% (36/40) of sensitivity, but cross reaction was observed in both technique. In western blot, 91, 63 and 21Kd Mw bands were identified as a strongly positive band. Among them 63Kd band showed positive reaction against almost all sera of cysticercosis patient. 3) Circumoval precipitation, ELISA, IFAT, showed 85.0% of sensitivity in the diagnosis of C. hepatica infection in rat. The antigenic localities were inner membrane of sectioned egg antigen on the prectipitates around the mucoid plugs which were induced by circumoval precipitation reaction. Sera from rats infected with 2000eggs were collected periodically to observe the changing patterns of antibody titers by IFAT and ELISA, which showed that high titers were detected at weeks 3 and 5, then gradually declined through weeks 9until to negatively converted at weeks 13.

• Journal of the Korean Society of Food Science and Nutrition
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• v.12 no.1
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• pp.46-50
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• 1983

In order to find out the possibility of utilizing red pepper seed as food resources of fats and proteins, a series of studies were conducted. The red pepper seed contained 27.6% of crude fat and 22.2% of crude protein. The lipid fractions obtained by silicic acid column chromatography were mainly composed of 95.4% neutral lipid, where as compound lipid were 4.6%. Among the neutral lipid separated by thin layer chromatography, triglyceride was 85.6%, sterol ester 4.9%, free fatty acids 3.4%, diglyceride 2.5%, sterol 2.2% and monoglyceride 1.1%, respectively. The predominant fatty acids of red pepper seed oil were linoleic acid (57.1-75.4%), palmitic acid (13.9-21.3%) and oleic acid (8.0-15.1%), especially glycolipid contained 1.7% of linolenic acid and small amount of myristic acid and arachidic acid. The salt soluble protein of red pepper seed was highly dispersible in 0.02M sodium phosphate buffer containing 1.0M $MgSO_4$, and the extractability of seed protein was about 25.0%. Glutamic acid and arginine were major amino acids of red pepper seed protein. The electrophoretic analysis showed 6 bands in seed protein, and the collection rate of the main protein fraction purified by sephadex G-100 and G-200 was about 62.2%. Glutamic acid (19.9%) was major amino acid of the main protein, followed by glycine and alanine. The molecular weight of the main protein was estimated to be 93,000.

• Journal of Animal Science and Technology
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• v.46 no.1
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• pp.83-90
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• 2004

Processed meals, such as a ground meat and hamburger patty, are required to ensure that no pathogens remain in the final products. However, there was no rapid method available to verify that the recommended end-point cooking temperature(EPT) was reached. Thus, the objective of this study was to rapidly determine EPT of ground pork hams using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SOS-PAGE), based on the disappearance of sarcoplasmic proteins after cooking. Fresh pork hams were added two levels of salt(0, 2%) and fat(15, 25%) combinations, and stored in refrigerator overnight, and cooked to internal cooking temperatures of $64^{\circ}C$ to $74^{\circ}C$ with $2^{\circ}C$ increments. Cooked pork hams were measured cooking loss(CL, %), protein solubility(PS) and SOS-PAGE. CL(%) was reduced with the addition of 2% salt, as compared to the control, regardless of fat contents. It was also increased with increasing eooking temperature. Protein solubility was affected by the cooking temperature, resulting in reduced PS up to $64^{\circ}C$(P < 0.05), but remained constant higher than $68^{\circ}C$. In SOS-PAGE analysis, protein bands with the molecular weights of 36 and 66 kDa were affected by the addition of salt and fat combinations. regardless of treatments. These protein fractions were decreased gradually with increased cooking temperatures up to $68^{\circ}C$ ${\sim}$ $70^{\circ}C$ and might be good indicators for the determination of EPT in ground pork hams.

• Korean Journal of Food Science and Technology
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• v.14 no.3
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• pp.250-256
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• 1982

A series of studies were conducted to find out the possibility of utilizing grape seed as resources of food fats and proteins, and the results of the studies are as follows: The grape seed contained 25.1%, of crude fat and 12.0% of crude protein. The lipid, fractions obtained by silicic acid column chromatography were mainly composed of about 95.5% neutral lipid, whereas compound lipid was only 4.5% level. Among the neutral lipid by thin layer chromatography, triglyceride was 91.89%, sterol ester, sterol, diglyceride and free fatty acid were 3.24%, 2.87%, 1.20% and 0.80%, respectively The predominant fatty acids of total and neutral lipids were linoleic acid $(69.72{\sim}71.72%)$ and oleic acid $18.09{\sim}19.46%)$, but those of glycolipid and phospolipid were linoleic acid $(31.49{\sim}38.18%)$, oleic acid $(20.20{\sim}35.27%)$ and palmitic acid $(26.80{\sim}39.98%)$. The major fatty acids of triglyceride separated from neutral lipid were oleic acid (43.08%), linoleic acid (38.42%) and palmitic acid (11.60%). The salt soluble protein of grape seed was highly dispersible in 0.02M sodium phosphate buffer containing about 1.0M $MgSO_4$, and the extractability of seed protein was 31%. Glutamic acid was the major amino acid in salt soluble protein, followed by arginine and aspartic acid. The electrophoretic analysis showed 3 bands in grape seed protein, and the collection rate of the main protein fraction purified by Sephadex G-100 and G-200 was 82%. Glutamic acid, aspartic acid and arginine were the major amino acids of the main grape seed protein. The molecular weight for the main protein of the grape seed was estimated to be 81,000.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.1-35
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• 1997

Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

• Parasites, Hosts and Diseases
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• v.32 no.4
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• pp.249-258
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• 1994

Recently, the importance of toxoplasmosis is raised as a complication in immunosuppressed or AIDS patients. Our study focused on the identification of a variety of Toxoplasma antigens by immunoblotting. Rabbits and BALB/c mice were immunized with Toxoplosmo Iysate (RH strain) , frozen tachyzoites (RH strain) or cysts (Beverly and Fukaya strain) . Blood were collected from ear vein, heart or orbital plexus for detecting the serum antibody levels. For excretory-secretory (E.S) antigens, T gondii (RH) tachyzoite were cultured in CHL (Chinese hamster lung) cells with MEM containing of 5% FCS. After 72hrs, culture supernatant was collected. BALB/c mice were inoculated with RH tachyzoite intraperitoneally and peritoneal fluids were extracted three days later. E.S antigens were detected in culture supernatant and infected mouse peritoneal fluid by EITB. Serum IgG levels in rabbit were 1 :512 of 10 days after primary immunization, 1 : 2,048 of 10 days after secondary immunization, 1: 1,024 of 20 days after secondary immunization by IFAT, respectively. Serum IgG levels of immunized mice were 1:128 after 7 weeks. Tachyzoite antigens of the RH strain were detected 25 protein bands ranging 10 kDa-220 kDa of molecular weights with Coomassie blue stain. Toxoplcsma major antigens corresponding to n of 24 kDa, 27 kDa,30 kDa, 35 kDa, 38 kDa were recognized by IgG and IgM antibodies. Excretory-secretory antigens present in culture supernatant with M. W. of 20, 30 kDa and in infected mouse peritoneal fluid with M.W. of 33 (P30), 45 kDa. When RH tachyzoite antigen was probed with different mice sera immunized with 2 strains of T gondii, the IgG antibody bud of Fukaya and Beverly strain (8 week-serum) is identical to those of RH strain. It is considered that the 30 kDa polypeptide detected in excretory- secretory materials and Iysate was important major antigen of T gondii (RH).

• Applied Microscopy
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• v.31 no.1
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• pp.19-35
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• 2001

Outlines for plasma $estradiol-17\beta$, components, electrophoretic patterns, and ultrastructural changes were obtained in female rainbow trout (Oncorhynchus mykiss) during the seasonal reproductive cycles. Plasma $estradiol-17\beta$ under the natural conditions, exhibited distinct seasonal variation, peaking very late in vitellogenic season during September, decreasing gradually the halt of spawning in December, and ultimately falling during the early stages of seasonal ovarian recrudescence in February and March. This change in $estradiol-17\beta$ appeared to stimulate vitellogenin production as evidenced by increases in plasma calcium, phosphorus, glucose, albumin and total protein levels. The electrophoretic patterns of late maturing or spawning oocytes were stained more intensively than those of late perinucleolus oocytes (molecular weights of approximately 70,000 and 200,000). Two protein bands were found in the SDS-PAGE separation, coincident with the $estradiol-17\beta$ hormone peak. Gonadosomatic indices (GSI) significantly increased from October to January, and showed the highest peak in January, coinciding with the numerically abrupt increase of ripe ova in female. A positive correlation (r=0.701, p<0.01) was established between plasma $estradiol-17\beta$ levels and the gonadosomatic index during the prespawning. The highest level of hepatosomatic index (HSI) observed in December. During the breeding season (December), the gonadotropes were large and filled with GTH-containing inclusions such as granules and globules. The vitellogenic phase began as late perinurleolus oocytes became transformed into early maturing oocytes through the accumulation of yolk, and oocytes reached the late maturing stages as the ooplasm was completely packed with yolk. Marked ultrastructural changed in the granulosa cells during nuclear migration involve the dilation of the rough endoplasmic reticulum and the appearance of the rod-shaped mitochondria with tubular cristae. Microvilli (finger-like projections), from the zona radiata and from the oocyte grew, and made contact with each other in the pore canals of the zona radials during vitellogenesis, but were withdrawn as the zona radiata became more compact and devoid of pore canals during oocyte maturation. The zona radiata grew to a tripartite structure such as an outer thin homogeneous layer, and two inner thick helicoidal layers (zona radials interna and zona radiata externa). Under the normal conditions, the ovarian follicle influenced the histological development and periodical secretion of the hormones , sufficient for a oogenesis and gonadal steroid production.

• Journal of Life Science
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• v.22 no.11
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• pp.1487-1492
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• 2012

Peach (Prunus persica) has been recognized as a food allergen for over 20 years. However, there is little information about cross-reactivity with other foods. The aim of this study was to research cross-reactivity of Korean cherry and hot pepper on patients allergic to peach and its stability by digestive enzyme treatment. Peach, Korean cherry, and hot pepper proteins were extracted and separated by Tricine-SDS-PAGE analysis. The protein extracts had a wide range of molecular weight, from 3 kDa to more than 26 kDa, and displayed different patterns of protein bands on Tricine-SDS-PAGE. Peach allergic patients' sera were used to detect the allergenic protein in three samples. Three peach allergic patients' sera reacted strongly with 9 kDa protein of peach, which was the expected lipid transfer protein (LTP) as the major allergen of peach and was detected with anti-LTP1 polyclonal antibody. However, the reactivity of the 23 kDa protein in Korean cherry and hot pepper protein was stronger than that of the 9 kDa protein. The stability of protein extracts on digestive enzyme treatment was examined using simulated gastric fluids (SGF) and simulated intestinal fluids (SIF), in which digestive enzyme stability is one of the characteristics of allergen potentially causing food allergy. Findings confirmed that allergenic proteins in peach, Korean cherry, and hot pepper were not completely digested by SGF and SIF treatments from results of SDS-PAGE analysis. These results confirmed that Korean cherry and hot pepper might cause cross-reactivity in peach allergic patients, and its allergenic proteins have stability against digestive enzymes.

• Tuberculosis and Respiratory Diseases
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• v.44 no.2
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• pp.251-263
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• 1997

Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

• Tuberculosis and Respiratory Diseases
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• v.47 no.3
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• pp.347-355
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• 1999

Background: Phospholipase C(PLC) plays a central role in cellular signal transduction and is important in cellular growth, differentiation and transformation. There are currently ten known mammalian isozymes of PLC reported to this date. Hydrolysis of phosphatidylinositol 4,5-bisphosphate($PIP_2$) by PLC produces two important second messengers, inositol 1,4,5-trisphosphate($IP_3$) and diacylglycerol. PLC-${\gamma}1$, previously, was known to be activated mainly through growth factor receptor tyrosine kinase. Other mechanisms of activating PLC-yl have been reported such as activation through tau protein in the presence of arachidonic acid in bovine brain and activation by $IP_3$, phosphatidic acid, etc. Very recently, another PLC-${\gamma}1$ activator protein such as tau has been found in bovine lung tissue, which now is considered to be AHNAK protein. But there has been no report concerning AHNAK and its associated disease to this date. In this study, we examined the expression of the PLC-${\gamma}1$ activator, AHNAK, in lung cancer specimens and their paired normal. Methods: From surgically resected human lung cancer tissues taken from twenty-eight patients and their paired normal counterparts, we evaluated expression level of AHNAK protein using immunoblot analysis of total tissue extract Immunohistochemical stain was performed with primary antibody against AHNAK protein. Results: Twenty-two among twenty-eight lung cancer tissues showed overexpression of AHNAK protein (eight of fourteen squamous cell lung cancers, all of fourteen adenocarcinomas). The resulting bands were multiple ranging from 70 to 200 kDa in molecular weight and each band was indistinct and formed a smear, reflecting mobility shift mainly due to proteolysis during extraction process. On immunohistochemistry, lung cancer tissues showed a very heavy, dense staining with anti-AHNAK protein antibody as compared to the surrounding normal lung tissue, coresponding well with the results of the western blot Conclusion: The overexpression of PLC-${\gamma}1$ activator protein, AHNAK in lung cancer may provide evidence that the AHNAK protein and PLC-${\gamma}1$ act in concerted manner in carcinogenesis.