• Journal of the Korean Society of Food Science and Nutrition
• /
• v.21 no.4
• /
• pp.353-366
• /
• 1992

This study was devised to purify the compound from tuna that have cytotoxic activities against various cancer cell lines and to observe its immunopotentiating activities. The cytotoxic compound was partially purified 277 fold, from petroleum ehter extract (crude extract) of tuna by silicic acid column chromatography (fraction D) and thin layer chromatography (Spot I). Cytotoxic activity was monitored using human colon cancer cell, HCT-48. The active compound (Spot I) was composed of seven materials which are fatty acids of four kinds ($C_{14:0},\;C_{16:0},\;C_{17:1},\;and\;C_{18:0}$) and unknown three fat materials. The active compound has cytotoxic activities against various cancer cell lines, that is, murine leukemic lymphocytes (L1210, P388) and human rectal (HRT-18) and colon cancer cells (HCT-48, HT-29). The patterns of size distribution of HCT-48 cells in the medium containing tuna extract were shifted to direction of the small size region. Also, the microscopic shape of HCT-48 cells were shrinked and distracted. The number of plaque forming cell and immunoglobin fraction of serum protein obtained from tuna-treated mice were increased, but natural killer cell activity was not affected.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.28 no.5
• /
• pp.375-395
• /
• 2006

Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

• Korean Journal of Occupational Health Nursing
• /
• v.8 no.2
• /
• pp.193-211
• /
• 1999

This study was carried out to investigate the management and support system affecting to the occupational health nursing services(OHNS) provided in group occupational health agencies(GOHA). Questionnaire was developed and distributed to 82 nurses who were working in GOHA and who agreed to participate in the survey. The results were as follow: 1. OH nurses responded were mostly in the age of twenty to thirties(89%), married(73.7%), technical college graduates(88.9%), worked in hospital(85.4%) and participated more than 1 year in group occupational health services (96.3%). 2. Fifty eight point four percent of the OH nurses worked in number of workplace more than 30 to less than 60 in the OHNS form. The figure of workplaces undertaken by nurses was ranged greatly from 9 to more than 100. Number of employees who cared by nurses were mostly under 5,000 peoples in 93.3%. The types of industry was mostly manufacturing and located in the order of factory complex area, suburban, urban and others. 3. Most OH nurses(87.8%) were fully involved in the OHNS for the SSE. Their working days to visit SSE was 5 days per week(77.8%) and one day in the GOHA at 41.3%. 4. The OH documents using by nurses were found in more than 23 different types. However, they were largely summarized in the types of 'Workplace Health Management Card', 'Personal Health Counselling Card', 'Daily Health Management Report', 'Visiting List of Workplace' and 'Sick Employee List'. 5. The items of laboratory test provided by GOHA were mostly achieved in the purpose of basic health examination. They were used to be the blood pressure check(98.8%), blood sugar test (98.8%), urine sugar and protein(91.4%), SGOT and SGPT(85.3% each), cholesterol (82.9%), hepa vaccine immunization(82.9%), r-GPT(81.7%), hemoglobin(79.3%) and triglyceride(75.5%). 6. The OH nurses(92.7%) followed the work pattern to visit the GOHA before and after small-scale enterprises(SSE) visit by car driven by nurses in 74.3%. They were payed by GOHA for transportation fees in certain amounts. However, nurse is the main person(75.0%) who covers up in case of traffic accident. If the GOHA has no transportation regulation for the formal workplace visit, data showed that nurses had been responsible to take charge(31.7%). 7. The personnel manager who takes in charge for nursing services was 'nurse' in 61.7% and 41.2% worked as the final decision maker related to nursing work. The OH nurses' opinions about factors affecting to the management were classified in the four areas such as 'Nature(Quality) of health professional'. 'Content of OHNS', 'Delivery system of the GOHS', and 'Others'. The factors were indicated highly in 'Authority as health professional', 'Level of perception of director on the OH' and 'Physical work condition for OHNS'. The things that this study suggests in the recommendation would be summarized in such as the management and supporting system working for SSE in the OHNS is necessary to reform thoroughly. The reconsidered aspects might be in the matters of number of workplaces undertaken by nurses, development of effectively practical health documents, preparation for guideline of the laboratory test in the workpleces, establishment of convenient and encouraging support system and cooperation between other health professionals with respect and skill.

• Clinical and Experimental Reproductive Medicine
• /
• v.26 no.3
• /
• pp.407-417
• /
• 1999

Objective: Mammalian follicle cells are the most important somatic cells which help oocytes grow, mature and ovulate and thus are believed to provide oocytes with various functional and structural components. In the present study we have examined whether cumulus or granulosa cells might playa role in establishing the plasma membrane structure of mouse oocytes during meiotic maturation. Design: In particular the differential resistances of mouse oocytes against chymotrypsin treatment were examined following culture with or without cumulus or granulosa cells, or in these cell-conditioned media. Results: When mouse denuded oocytes, freed from their surrounding cumulus cells, were cultured in vitro for $17{\sim}18hr$ and then treated with 1% chymotrypsin, half of the oocytes underwent degeneration within 37.5 min ($t_{50}=37.5{\pm}7.5min$) after the treatment. In contrast cumulus-enclosed oocytes showed $t_{50}=207.0$. Similarly, when oocytes were co-cultured with cumulus cells which were not associated with the oocytes but present in the same medium, the $t_{50}$ of co-cultured oocytes was $177.5{\pm}13.1min$. Furthermore, when oocytes were cultured in the cumulus cell-conditioned medium, $t_{50}$ of these oocytes was $190.0{\pm}10.8min$ whereas $t_{50}$ of the oocytes cultured in M16 alone was $25.5{\pm}2.9min$. Granulosa cell-conditioned medium also increased the resistance of oocytes against chymotrypsin treatment such that $t_{50}$ of oocytes cultured in granulosa cell-conditioned medium was $152.5{\pm}19.0min$ while that of oocytes cultured in M16 alone was $70.0{\pm}8.2min$. To see what molecular components of follicle cell-conditioned medium are involved in the above effects, the granulosa cell-conditioned medium was separated into two fractions by using Microcon-10 membrane filter having a 10 kDa cut-off range. When denuded oocytes were cultured in medium containing the retentate, $t_{50}$ of the oocytes was $70.0{\pm}10.5min$. In contrast, $t_{50}$ of the denuded oocytes cultured in medium containing the filtrate was $142.0{\pm}26.5min$. $T_{50}$ of denuded oocytes cultured in medium containing both retentate and filtrate was $188.0{\pm}13.6min$. However, $t_{50}$ of denuded oocytes cultured in M16 alone was $70.0{\pm}11.0min$ and that of oocytes cultured in whole granulosa cell-conditioned medium was $156.0{\pm}27.9min$. When surface membrane proteins of oocytes were electrophoretically analyzed, no difference was found between the protein profiles of oocytes cultured in M16 alone and of those cultured in the filtrate. Conclusions: Based upon these results, it is concluded that mouse follicle cells secrete a factor(s) which enhance the resistance of mouse oocytes against a proteolytic enzyme treatment. The factor appears to be a small molecules having a molecular weight less than 10 kDa.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.30 no.3 s.47
• /
• pp.393-397
• /
• 2004

L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

• Tuberculosis and Respiratory Diseases
• /
• v.39 no.6
• /
• pp.517-525
• /
• 1992

Background: Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent, and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis. Method: To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB-1, -2), and 123 base pair DNA, a part of IS6110 fragment, which multiple copies are known to exsist PCR one genome (the primers are Sal I-1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results. Results: 1) Using TB-1, -2 primers, PCR was positive in 73.1% (19/26) of culture positive sputa, in 12.5% (1/8) of culture negative. but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology. 2) Using Sal I-I, -2 primers, PCR was positive in 94.1% (32/34) of culture positive sputa, in 23.1% (6/26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14/16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology. Conclusion: PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method.

• Tuberculosis and Respiratory Diseases
• /
• v.44 no.6
• /
• pp.1353-1365
• /
• 1997

Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majority of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to host. In the previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF, and the probability was shown that the acquired resistance to TNF might be associated with synthesis of some protective protein. Understanding the mechanisms of TNF -resistance in TNF-$\alpha$ cDNA transfected cancer cells would be. an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of MnSOD, an antioxidant enzyme, in the acquired resistance to TNF of TNF-$\alpha$ cDN A transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164(murine fibrosarcoma cell line), NCI-H2058(human mesothelioma cell line), A549(human non-small cell lung cancer cell line), ME180(human cervix cancer cell line) cells using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF-$\alpha$ cDNA transfected cells(WEHI164-TNF, NCIH2058-TNF, A549-TNF, ME180-TNF) and the changes of MnSOD mRNA expressions with Northern blot analysis. Results : The MnSOD mRNA expressions of parental cells and genetically modified cells of WEHI164 and ME180 cells(both are naturally TNF sensitive) were not significantly different The MnSOD mRNA expressions of genetically modified cells of NCI-H2058 and A549(both are naturally TNF resistant) were higher than those of the parental cells, while those of parental cells with exogenous TNF were also elevated. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ cDNA transfection may not be associated with the change in the MnSOD expression, but the difference in natural TNF sensitivity of each cell may be associated with the level of the MnSOD expression.

• Food Science and Preservation
• /
• v.16 no.5
• /
• pp.705-713
• /
• 2009

We prepared salmon patties and compared the quality characteristics thereof with those of commercial tuna and pork patties. The moisture and crude ash contents of salmon patty were lower, whereas the crude protein content was higher, than those of commercial patties. The crude lipid content of salmon patty was higher than that of tuna patty, but lower than that of pork patty. The pH value and the volatile basic nitrogen content of salmon patty were lower than those of the commercial patties. Hunter color values (L, a, b) in a cross-section of cooked salmon patty were higher, whereas the ${\Delta}E$ value was lower, than those of the two commercial patties. The lipophilic browning index (0.397) of salmon patty was higher, whereas the hydrophilic browning index (0.047) was lower, than those of commercial patties. Trichloroacetic acid-soluble N content (272 mg/100 g) of salmon patty was lower than of commercial patties. The major fatty acids of salmon patty were palmitic acid (11.9%), oleic acid (27.6%), and linoleic acid (30.1%), whereas small amounts of eicosapentaenoic acid (EPA, 3.7%) and docosahexaenoic acid (DHA, 8.4%) were also found. The predominant amino acids of all patties were arginine, glutamic acid, aspartic acid, leucine, threonine, and proline, and the contents of these amino acids in salmon patty were higher than in the two commercial patties. The Fe, Ca, K, P, and Mg contents of salmon patty were 2.4 mg/100g, 42.6 mg/100g, 207.5 mg/100g, 211.6 mg/100 g, and 29.9 mg/100 g, respectively. The sensory quality of salmon patty was higher than that of pork patty. These results indicate that salmon patty may have good quality characteristics, comparable to those of the two commercial patties.

• Journal of Dairy Science and Biotechnology
• /
• v.14 no.2
• /
• pp.157-164
• /
• 1996

This study aimed increase the quality during ripening of Cheddar cheese made with proteinase-negative mutant of Streptococcus lactis KCTC 1913 selected by curing. The degradation of protein during cheese ripening were investigated by electrophoresis and chromatography. The results were summarized as follow ; 1. The number of lactic acid bacteria decreased with the ripening stage, and that of the control cheese decreased faster than that of the cheese made with mutant. 2. Polyacrylamide gel electrophoretic analysis of cheese caseins revealed no difference between the cheese made with mutant and the control cheese, but differences along with the ripening stage were evident. 3. On Sephadex G-25 column chromatography, the extracts of bitter components from the green cheese and 3 month ripended cheese were fractionated into 3 fractions. With the progress of ripening, bitter peptides were degraded to rather small peptides or free amino acids. 4. Sensory evaluation of the 3 month ripended Cheddar cheese found no significant differences in color but the cheese made with mutant evidenced higher palatability in flavor and better texture than the control cheese. 5. The yields of the cheddar cheese made with mutant was 0.14% higher than that of the control cheese.

• Korean Journal of Poultry Science
• /
• v.42 no.3
• /
• pp.223-230
• /
• 2015

Eggs are an important source of protein for the human diet. Consumers want fresher, more delicious and more sanitary eggs. In Korea, the Shell Egg Grading system (EGS) was employed in 2001. The portion of graded shell eggs has increased every year, but graded shell eggs account for only 6% of all eggs. The EGS should satisfy producers, distributors and consumers. However, the EGS does not have an official function because of many problems. Consumers cannot select various graded shell eggs in the market, and producers do not receive enough profit even though they produce top-quality graded shell eggs. There are few studies on the EGS, Therefore, this study was performed to improve the EGS. We surveyed the EGS, GP Centers and farmers. Large companies (farmers) are more satisfied than small companies with the EGS. There was a high tendency for the companies (farmers) that are not involved with the EGS to think that graded and ungraded shell eggs are similar, in contrast to the companies (farmers) connected to the EGS. We should have to change the grading system of grade shell eggs, establish of the cold chain system, change of the law for the school meals, minimize payment for the grading shell eggs for developing EGS. Based on this study, the egg industry can benefit through the improvement of the EGS.