• Journal of Life Science
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• v.31 no.11
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• pp.1028-1036
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• 2021

Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.

• Journal of Microbiology and Biotechnology
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• v.9 no.1
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• pp.106-112
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• 1999

The therapeutic potential of phosphodiesterase 4(PDE4) inhibitors in inflammatory diseases including some autoimmune diseases has been explored recently with some hopeful results. These PDE4 inhibitors are thought to show their anti-inflammatory effect by down-regulating tumor necrosis factor-a (TNF-$\alpha$) production in lymphocytes and macrophages. A high concentration of TNF-$\alpha$has been found in rheumatoid arthritis (RA) synovium and reducing TNF-$\alpha$using biological agents was proven to be an effective RA treatment. To test the possibility of using PDE4 inhibitors for RA treatment, the effects of a newly synthesized PDE4 inhibitor, DWP205505, on TNF-$\alpha$ and IL-10 production was tested in cells isolated from normal peripheral blood and rheumatoid arthritis synovial fluid. Cytokine production was assayed at the protein level by sandwich enzyme-linked immunosorbent assay (ELISA) and at the mRNA expression level by semi-quantitative RT-PCR. Another PDE4 inhibitor, RP73401, was used for comparison. DWP205505 and RP73401 had no harmful effect on cell viability up to 10 $\mu$M concentration during the 24 h culture period. DWP205505 as well as RP73401 significantly reduced TNF-$\alpha$ secretion from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (pBMC) and synovial fluid mononuclear cells (SFMC). The effect of DWP205505 or RP73401 treatment on the mRNA expression of TNF-$\alpha$ was also studied in LPS-stimulated PBMC and SFMC. TNF-$\alpha$ mRNA expression was increased by LPS stimulation and both of the PDE4 inhibitors suppressed TNF-$\alpha$ mRNA expression. For interleukin-l0 (IL-l0), a little different results were obtained from PBMC and SFMC; IL-l0 secretion was unaffected by LPS stimulation and only minimally affected by both of the PDE4 inhibitors in PBMC. In unstimulated SFMC, DWP205505 and RP73401 slightly enhanced IL-10 secretion, while they reduced IL-l0 secretion from LPS-stimulated SFMC where IL-l0 secretion was a lot higher than unstimulated SFMC. These results suggest that the newly synthesized PDE4 inhibitor DWP205505 may have anti-rheumatoid arthritis activity.

• Tuberculosis and Respiratory Diseases
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• v.63 no.3
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• pp.242-250
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• 2007

Background: Abnormal angiogenesis can induce hypoxia within a highly proliferating tumor mass, and these hypoxic conditions can in turn create clinical problems, such as resistance to chemotherapy. However, the mechanism by which hypoxia induces these changes has not yet been determined. Therefore, this study was conducted to determine how hypoxia induces changes in cell viability and extracellular microenvironments in an in vitro culture system using non-small cell lung cancer cells. Methods: The non-small cell lung cancer cell line, A549 was cultured in DMEM or RPMI-1640 media that contained fetal bovine serum. A decrease in the oxygen tension of the media that contained the culture was then induced in a hypoxia microchamber using a $CO_2-N_2$ gas mixture. A gas analysis and an MTT assay were then conducted. Results: (1) The decrease in oxygen tension was checked the anaerobic gas mixture for 30 min and then reoxygenation was induced by adding a 5% $CO_2-room$ air gas mixture to the chamber. (2) Purging with the anaerobic gas mixture was found to decrease the further oxygen tension of cell culture media. (3) The low oxygen tension resulted in a low pH, lactic acidosis and a decreased glucose concentration in the media. (4) The decrease in glucose concentration that was observed as a result of hypoxia was markedly different when different types of media were evaluated. (5) The decrease in oxygen tension inhibited proliferation of A549 cells. Conclusion: These data suggests that tumor hypoxia is associated with acidosis and hypoglycemia, which have been implicated in the development of resistance to chemotherapy and radiotherapy.

• The Research Journal of the Costume Culture
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• v.3 no.2
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• pp.241-260
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• 1995

The Maori's traditional clothing materials, basic forms of dress, and the pattern and technique of tatoo were examined in the present study in order to deepen the appreciation of the cultural heritage of the Maori. The research method employed was the analysis of written materials. And a fild-trip was also made for the study. The study was limitted to the traditional culture of body adornment of the Maori including the clothing which is preserved and practicing by them at the present day, and the origin and the process of the historical development of those are not included in the scope of the present study. Followings are the results of the study: (1) By far the most widely used fiber for Maori clothing is abtained from what is commonly called New Zealand Flax. The fiber of kiekie(Freycinetia baueriana) and cabbage trees(Cordyline spp.) may also be used. The strong, long-lasting fiber of toi(cordyline indivisa) is used for a prestige warrior's cloak. Flat strips of ti kauka(Cordyline australi) are also used as thatch on rain cloaks. (2) Regardless of technique used, Maori weaving is always worked horizontally from left to right. Traditionally the work was suspended between two upright turuturu or weaving sticks. As the work progressed a second pair of uprights was used to keep the work off the ground. These uprights were moved forward as required. Because the weaver sat on the ground, the working edge was kept at a height that was comfortable to reach. No weaving tools are used, the wefts(aho) being manipulated by the fingers. The two main Maori weaving techniques are whatu aho patahi(single-pair twining) and whatu aho rua(double-pair twining). (3) The Maori wore two basic garments - a waist met and a cloak. The cloth of commoners were of plain manufacture, while those of people of rank were superior, sometimes being decorated with feather or dyed tags and decorated borders. Children ran more-or-less naked until puberty, being dressed only for special events. Some working dress consisted of nothing more than belts with leaves thrust under them. Chiefs and commoners usually went barefoot, using rough sandals on journeys over rough country (4) The adornment of men and women of rank was an important matter of tribal concern as it was in chiefly persons that prestige of the group was centred, The durable items of Maori persons adornment were either worn or carried. Ornaments of various kinds were draped about the neck or suspended from pierced earlobes. Combs decorated the head. Personal decorations not only enhanced the appearance of men and women, but many had protective magical function. The most evident personal ornament was the hei-tiki made of jade or other material. Maori weapons were treasured by their owners. They served on bottle and were also personal regalia. A man of rank was not fully dressed without a weapon in hand. Also weapons were essential to effective oratory. (5) No man or woman of rank went without some tattoo adornment except in extremely rare instances when a person was too sacred to have any blood shed. The untattooed were marked as beeing commoners of no social standing. This indelible mark of rank was begun, with appropriate rite and ritual, at puberty. And tattoo marked the person as being of a marriageable age. Maori tattoo was unlike most traditional tattoo in that its main line were 'engraved' on the face with deep cuts made by miniature bone chisels. The fill-in areas were not tattooed with cuts but with the multiple pricks of small bone 'combs' that only lightly penetrated the skin surface. The instrument of tattoo consisted of small pots of pumice or wood into which was placed a wetted black pigment made from burnt kauri gum, burnt vegetable caterpillars or other sooty materials. A bird bone chisel or comb set at right angles on a short wooden handle was dipped into the gigment, that a rod or stick was used to tap head of this miniature adze, causing penetration of the skin surface. Black pigment lodged under the skin took on a bluish tinge. A full made facial tattoo consisted of major spirals with smaller spirals on each side of the nose and sweeping curved lines radiating out from between the brows over the forehead and from the nose to the chin. The major patterns were cut deep, while the secondary koru patterns were lightly pricked into the skin.

• Clinical and Experimental Pediatrics
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• v.49 no.1
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• pp.56-63
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• 2006

Purpose : This study was designed to document the etiologies and the characteristics of parapneumonic effusion in children. Methods : During a 17-year period from 1987 to 2004, parapneumonic effusion was confirmed in 86 children at Gyeongsang National University Hospital. The clinical records of these children were reviewed and radiological findings and laboratory data, especially results of thoracentesis, were analyzed retrospectively. Results : M. pneumoniae(34 subjects) was the most common pathogen at all over age, especially above 1-years-old. There were diagnosed with clinical characteristics and serologic tests. The $2^{nd}$ most common pathogen revealed non tuberculous bacteria(14 subjects). A species of bacteria at no tuberculous bacteria revealed S. aureus(5), S. pneumoniae(3), P. aeroginosa(3), other staphylococcus (2), and K. pneumoniae(1). There were confirmed with sputum culture or pleural fluid culture or blood culture. S. aureus was most common pathogen in infants. The $3^{rd}$ common pathogen was M. tuberculosis(7). There were confirmed with skin tuberculin tests and AFB stains. Another that was classified as a non bacteria was adenovirus(2). Complications of parapneumonic effusion such as pleural thickness occurred on M. tuberculosis(1). Non tuberculous bacteria, especially S. aureus revealed a serious predominance of polymorphocyte at pleural fluid, and lowest pleural pH and glucose, and highest pleural protein and LDH. Tuberculosis revealed high pleural protein and LDH. Conclusion : Age and chemistries of pleural fluid might be helpful in differentiating various etiologies of parapneumonic effusion. If there were suspicious of tuberculosis and non-tuberculous bacteria, more aggressive approaches were needed to prevent complication.

• Tuberculosis and Respiratory Diseases
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• v.55 no.1
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• pp.59-68
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• 2003

Background : In intubated patients, cultures of endotracheal aspirates (EA) are apt to contamination throughout the endotracheal tube. Therefore, the identification of etiologic agents via conventional EA cultures is not always reliable. In order to differentiate a pulmonary infection from a non-infectious disease, and to identify the true etiologic agent of acute pulmonary infection, blinded protected specimen brushing (PSB) was used, and its efficacy evaluated. Methods : In 51 intubated patients, with suspected pneumonia, blind PSB were performed, and the results compared with blood and EA cultures. A protected specimen brush was introduced through the endotracheal tube, and settled at the affected large bronchus. A specimen brush was introduced to the expected region using the blind method. The tip of the brush was introduced with an aseptic technique after vigorously mixed for 1 minute in $1cm^3$ of Ringer's lactate solution. The specimens were submitted for quantitative culture within 15 minutes, with a culture being regarded as positive if the colony forming units were above $10^3/ml$. Results : Of the 51 patients, 15 (29.4%) had community-acquired pneumonia (CAP), 27 (52.9%) hospital-acquired pneumonia (HAP) and 9 (17.6%) non-infectious diseases. The sensitivity and specificity of the quantitative PSB culture for the diagnosis of pneumonia were 52.4 and 88.9%, respectively. The sensitivity and specificity of EA were 78.6 and 77.8%, respectively. The blind PSB was superior to the EA for the identification of true etiologic agents. Of 53 episodes of 27 HAP patients, MRSA (Methicillin-resistant staphylococcus aureus) (41.5%) was the most common causative agent followed by Pseudomonas aeruginosa (15.1%), Klebsiella sp. (7.5%) and Acinetobacter sp. (7.5%). Conclusions : As a simple, non-invasive diagnostic modality, the blind PSB is a useful method for the differentiation of a pulmonary infection from non-infectious diseases and to identify the etiologic agents in intubated patients. A blind PSB can be performed without bronchoscopy, so is safer, more convenient and cost-effectiveness for patients where bronchoscopy can not be performed.

• 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).

• Food Science of Animal Resources
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• v.28 no.1
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• pp.51-58
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• 2008

This study was performed to evaluate the physico-chemical properties of fermented sausages containing probiotic starter cultures (LK-30 plus, Lactobacillus plantarum 155 and 167, and Pediococcus damnosus L12) with reduced fat levels, and to determine the optimum condition for the manufacture of these products. Although low-fat fermented sausages were reduced fat content at the amount of 90% and the ripening time by 1-2 weeks, as compared to regular-fat counterpart, they became harder and had many winkles outside due to the extreme drying. In addition, fat level in fermented sausages affected the composition and shear force values. During ripening, pH, lightness and yellowness values tended to decrease, however, microbial counts of inoculated lactic acid bacteria were increased up to $10^8-10^9cfu/g$ within 3 days and remained constant thereafter. Low-fat fermented sausages had higher microbial counts than regular-fat ones. Although the inoculated probiotic starter cultures alone had the functional properties, such as cholesterol reduction, anti-high blood pressure and antimicrobial activity, they did not have distinctive characteristics in the fermented sausages. Based on these results, the low-fat fermented sausages were successfully manufactured, but a little bit increased fat level and improved functional properties in the fermented sausages would be required to have better quality as compared to regular-fat counterparts.

• Journal of Biomedical Engineering Research
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• v.16 no.4
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• pp.463-470
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• 1995

Complete prelining of artificial vascular grafts with autologous endothelial cells may be one of the ideal solutions to obtain a nonthrombogenlc blood-contacting surface. To establish an intact endothelial cell monolayer on a prosthetic surface at the time of implantation,a sufficient number of endothelial cells and adequate propagation condition In cell culture are prerequisites. In this experimental study, endothelial cells from microvessels of adult human oriental adipose tissue were enzymatically harvested, and optimal culture conditions for proliferation of the endothelial cells in cell culture were examined. Human oriental adipose tissue was digested with collagenase and endothelial cells were separated from other stromal elements by mesh filtration method. Cultured cells were identified as endothelial cells by immunofluorescent staining for factor VIII-related antigen. Proliferation in usual 20% fetal bovine serum (FBS) medium or medium containing endothelial cell growth factor (ECGF)(5 ng/ml) and heparin (HEP)(1,000 units/ml) were compared,and the effects of adding compounds that increase intracellular cyclic adenosine monophosphate levels, that is,cholera toxin (CT)(1 $\mu\textrm{g}$/ml) and isobutylmethylxanthine (IBMX)(0.2 ml),were also analyzed. In total,following eight media groups were examined. 1) FBS medium + ECGF + HEP, 2) FBS medium + ECGF + HEP+CT, 3) FBS medium+ECGF+HEP+lBMX, 4) FBS medium+ECGF+HEP+CT+ IBMX, 5) FBSmedium, 6) FBS medium +CT, 7) FBS medium + IBMX, 8) FBS medium + CT + IBMX. It was shown that the medium containing ECGF + HEP with or without cholera toxin was most efficient in Stimulating cell proliferation. IBMX was considered to have antagonistic effect to ECGF. Among experimental groups without ECGF and HEP, the addition of cholera toxin and IBMX was shown to significantly potentiate cell proliferation. This results could provide a practical method for use of cultured human endothelial cells for endothelial cell seeding of cardiovascular prosthetic device, particularly in small-diameter vascular grafts.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.7
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• pp.959-967
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• 2005

Lead is a ubiquitous environmental and industrial pollutant that causes a major health concerns. It is known to induce a broad range of physiological, biochemical, and behavioral dysfunctions in laboratory and humans, including hematopoietic system, kidneys, liver, and reproductive system. This study was conducted to investigate the effect of Saengshik supplementation on the lead-induced toxicity in rats. Five week old male Sprague­Dawley rats were randomly assigned to five groups for six weeks as followings: control group (CT), lead acetate treated group (PT), and lead acetate groups administered with three different dosages of Saengshik $(SI2.5-12.5\%,\;S25-25\%,\;and\;S50-50\%).$ Lead acetate (12 mg/rat) was intragastrically administered daily for 6 weeks. The results were summarized as follows; Weight gain and food efficiency ratio were significantly lower (p<0.05) in lead administered group compared with those of the control group. Also, significant lead-induced alteration in blood hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and reticulocyte distribution width (RDW) were observed. In the liver of lead-exposed animals, there was an increase in the lipid peroxidation (MDA) and the level of glutathione (GSH), but superoxiede dismutase (SOD) activity did not change. Lead-exposed animals with $25\%\;and\;50\%$ Saengshik supplementation showed marked improvements in the values of MCH, MCV, and RDW. Also, the level of HCT was significantly increased by $50\%$ Saengshik supplementation. The levels of liver MDA in $12.5\%\;and\;50\%$ Saengshik administered groups and GSH level in $50\%$ Saengshik administered group were significantly decreased compared to the lead administered group. Also, hepatic SOD activity tended to increase in the presence of Saengshik supplementation. Furthermore, the accumulation of lead in liver and kidney was reduced by presence of Saneghshik supplementation. Liver lead concentration was significantly reduced by both $25\%\;and\;50\%$ Saengshik supplementations and kidney lead concentration was significantly reduced by the $25\%$ Saengshik supplementation. These results show that Saengshik may have a protective effect against lead intoxication but the mechanism of their effects remains unclear.