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Effects of Pronuclear Age in Freezing of Mouse Embryos on Survival and Development in Vitro after Cryopreservation (동결보존시 생쥐 전핵배아의 시기에 따른 생존율과 발생율의 비교)

  • Kim, H.S.;Ryu, B.Y.;Oh, S.K.;Suh, C.S.;Kim, S.H.;Choi, Y.M.;Kim, J.G.;Moon, S.Y.;Lee, J.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.59-64
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    • 1998
  • This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from $F_1$ hybrid mice and classified into 4 groups according to the pronuclear stage (6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower (p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower (p<0.05) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei (9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).

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Enhancement of Hybridoma Cell Growth and Anti-Hepatitis B Surface Antigen Monoclonal Antibody Production in Enriched Media with Low Serum (저혈청농축배지에서 세포성장 및 간염표면항원에 대한 단일클론항체 생산의 증가)

  • 전복환;조의철김동일백승복
    • KSBB Journal
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    • v.5 no.1
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    • pp.87-94
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    • 1990
  • Enhancement of hybridoma cell growth and monoclonal antibody(MAb) production by the addition of a small amount of serum into both serum-free medium and enriched medium was studied. The enriched medium was constructed by mixing a basal serum-free medium and a nutrient-fortified RPMI 1640 medium. It was supplemented with human serum albumin, insulin, transferrin, and monoethanolamine. It was found that addition of low concentration of serum with other serum-free supplements was favorable for growth of a mouse hybridoma 2c3.1 cells. The concentration of serum was determined to 0.5%. The maximum cell concentration obtained in this enriched medium supplemented with 0.5% fetal bovine serum (FBS) was $3.06{\times}10^6$ cells/ml and the concentration of secreted anti-Hepatitis surface antigen (antiHBsAg) MAb was $159.7{\mu\textrm{g}}\;/\;ml$ compared to $43{\mu\textrm{g}}\;/\;ml$ in RPMI 1640 medium with 10% FBS and $50{\mu\textrm{g}}\;/\;ml$ in previously-developed serum-free medium. The 2c3.1 cell growth and MAb production could be enhanced considerably by using the enriched medium supplemented with 0.5% FBS and serum-free supplements instead of RPMI 1640 medium or serum-free medium. The enhancement in MAb production in the enriched medium was more noticeable.

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A Survey of Nutrition and Blood Contents of Rural Women in Korea (일부 농촌지역 부인의 영양실태 및 혈액성상에 관한 연구)

  • Lim, Hyeon-Sook;Hwang, Geum-Hee
    • Journal of Nutrition and Health
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    • v.15 no.3
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    • pp.171-180
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    • 1982
  • The nutritional survey and blood sampling was conducted from May 23 to 27, 1971, in order to investigate the state of the nutritional intake in relation to blood contents of rural women. This survey covered a total of 47 women residing in Youngam-gun, Cholla-namdo, Korea. The daily diet of these subjects was observed by 24-hour recall method and weighing method for 3 days and hemoglobin (Hb), hematocrit(Hct), mean corpuscular hemoglobin concentration (MCHC), serum protein, serum albumin, A/G ratio, serum iron, total iron binding capacity (TIBC), transferrin saturation (TS), serum cholesterol and blood specific gravity (SG) were measured. The results were summarized as follows ; The rate of intake of vegetable foods was high, especially cereals, and that of animal foods was low, especially eggs and milk and milk products. The daily intake of niacin was sufficient, and the intakes of energy and iron were also tend to be adequate. On the contrary, the intakes of vitamin A, $B_2$ and C, calcium and protein were insufficient. The mean red blood cell number was $410.8{\pm}26.4{\times}10^{4}/mm^{3},$ the mean values of Hb, Hct and MCHC were $12.5{\pm}0.9gm%$, $37.5{\pm}2.4%$ and $33.2{\pm}0.5%$ thus the anemic prevalences were 21.3%, 12.8% and 93.6% comparing with the anemic criteria of WHO. The levels of serum protein and serum albumin were $7.1{\pm}0.4gm\;%$ and $4.3{\pm}0.3gm\;%$so the A/G ratio was indicated $1.6{\pm}0.2.$. The levels of serum iron and TIBC were $91.9{\pm}27.5{\mu}g/100ml and $297.5{\pm}31.4{\mu}g/100ml,$ so TS was calculated $31.2{\pm}9.7\;%$. The mean value of SG was $1.044{\pm}0.002,$ and the level of serum cholesterol was $164.3{\pm}25.4mg/dl$. In the relation between each of bloood content, Hb showed positive correlation not only with Hct but also protein at the level of significance of 1 per cent and 5 per cent, and protein also showed a positive correlation with serum cholesterol at the 1 per cent level of significance. In the relation between the nutrient intake and blood contents, vitamin C intake showed a positive correlation with Hb, serum protein and serum iron at the level of significance of 1 per cent, iron intake also showed a positive correlation with serum iron and serum cholesterol at the same significant level, and protein intake showed a positive correlation with serum iron at the at the level of significance of 5 per cent.

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Study of sweat content analysis and latent fingerprint developing (땀의 성분 분석과 잠재지문 현출에 관한 연구)

  • Choi, Mi-Jung;Sun, Yale-Shik;Kim, Chang-Seong;Choi, Man-Sik;Sung, Nack-Do;Park, Sung-Woo
    • Analytical Science and Technology
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    • v.20 no.2
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    • pp.147-154
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    • 2007
  • Sweat contents were investigated for using those data to forensic purpose. The experiments of identifying sweat contents were as follow: 1) measurement of amino acids (aspartic acid, serine, glycine etc) by HPLC, 2) anions ($Cl^-$, $F^-$, and $SO{_4}^{2-}$) by IC and 3) trace elements (Cu, Zn, Li, B, etc.) by ICP-MS. Amino acid contents in sweat are varied with individual samples and glycine, threonine, alanine, valine and histidine are detected as the prime one. The detected anions are $Cl^-$(2167~4073 ppm) and $F^-$(454~582 ppm) mostly. Trace elements of Rb, Zn and Cu are detected and those concentrations are relatively very high. The compositions of sweat can be influenced by various factors (diet, anthropometric, characteristics, physical fitness, age, gender and the state of the health).

Qualities and Anti-inflammatory Activity of Kyungokgos Sold in Local Markets (국내 시판 경옥고 제품의 품질 특성 및 항염증 활성)

  • Lee, Ka-Soon;Kim, Gwan-Hou;Kim, Hyun-Ho;Seong, Bong-Jae;Kim, Sun-Ick;Han, Seung-Ho;Kang, Eun Ju;Yoo, Yung Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.335-341
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    • 2013
  • Kyungokgos purchased in local markets in Korea vary in their combination and mixing ratios during processing. This study was investigated qualities of Kyungokgos manufactured traditionally to evaluating its qualities. The general components of Kyungokgos were moisture (18.62~49.78%), ash (0.198~1.211%), protein (0.89~3.58%), lipid (0.16~1.14%) and carbohydrates (47.95~77.08%). The color values of L, a, and b were 26.49~73.87, 16.51~38.64, and 45.41~88.94, respectively. The viscosity was classified into three non-Newtonian type groups: high, medium, and non-dilatant, according to the increase of loop execution times. Three extracts (KOG-1, -7, and -8, in a 30-fold dilution) showed no cytotoxicity toward RAW 264.7 cells, while the extracts of KOG-2, -4, and -5 showed a low cytotoxic effect. KOG-1 and -2 extracts with low cytotoxicity markedly inhibited the production of the inflammatory mediators-nitric oxide (NO) and tumor necrosis factor-alpha (TNF-${\alpha}$) in LPS-stimulated RAW 264.7 cells. These results indicate that KOG-1 and -2 extracts have anti-inflammatory activity in LPS-stimulated RAW 264.7 macrophages.

Production of hTPO Transgenic Chickens using Tetracycline-Inducible Expression System (Tetracycline-Inducible Expression System을 이용한 Human Thrombopoietin (hTPO) 형질전환 닭의 생산)

  • Kwon, M.S.;Koo, B.C.;Kim, D.H.;Kim, M.J.;Kim, T.
    • Korean Journal of Poultry Science
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    • v.36 no.2
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    • pp.177-186
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    • 2009
  • It is well-known that unregulated over-expression of foreign gene may have unwanted physiological or toxic effects in transgenic animals. To circumvent these problems, we constructed retrovirus vector designed to express the foreign gene under the control of the tetracycline-inducible promoter. However, gene expressions in the tetracycline-inducible expression system (Tet system) are not completely regulated but a little leaky due to the inherent defects in conventional Tet-based systems. A more tightly controllable regulatory system can be achieved when the advanced versions ($rtTA2^SM2$) of rtTA and a minimal promoter in responsive components (pTRE-tight) are used in combination therein. In this study, we tried to produce human thrombopoietin (hTPO) from various target cells and transgenic chickens using the retrovirus vector combined with Tet system. hTPO is the primary regulator of platelet production and has an important role in the survival and expansion of hematopoietic stem cells. In a preliminary experiment in vitro, higher hTPO expression and tighter expression control were observed in chicken embryonic fibroblast (CEF) cells. We also measured the biological activity of the hTPO using Mo7e cells whose proliferation is dependant on hTPO. The biological activity of the recombinant hTPO from CEF was higher than both its commercial counterpart and hTPO from other target cells. The recombinant retrovirus was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 138 injected eggs, 15 chicks hatched after 21 days of incubation. Among them, 8 hatched chicks were hTPO positive. When the Go transgenic chicken was fed doxycycline (0.5 mg per 1 gram of feed), a tetracycline derivative, hTPO concentration of the transgenic chicken blood was 200 ng/mL. Germline transmission of the transgene was confirmed in sperm of the Go transgenic roosters. These results are informative to establish transgenic chickens as bioreactors for the mass production of commercially valuable and biological active human cytokine proteins.

Influence of Panax Ginseng on Hepatic DNA Synthesis in Mice (고려인삼이 마우스의 간조직 DNA 합성능에 미치는 영향( I ))

  • Chae, Y.B.;Chang, W.S.;Kwon, Y.C.
    • The Korean Journal of Physiology
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    • v.8 no.1
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    • pp.27-30
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    • 1974
  • It was planned to evaluate the influence of Panax Ginseng upon hepatic DNA synthesis in mice by observing incorporation of $[^3H]$ thymidine into the tissue cells. Thirty male mice$(body\;weight:\;18{\sim}20\;g)$ were divided equally into the ginseng and the saline groups. Each animal of the ginseng and the saline groups received every day (subcutaneously) 0.05 m1/10 g body weight of ginseng extract (4mg of ginseng alcohol extract in 1 ml of saline) and the same amount of saline, respectively, for 5 days. On the 5th experimental day, all animals received $1\;{\mu}Ci/g$ body weight of $[^3H]$ thymidine intraperitoneally 2 hours after the last medication. Five animals, at a lime, of each group were sacrificed 1, 10, and 24 hours after thymidine administration, and their hepatic radioactivity was measured autoradiographically in terms of the % number of radioactive cells in 1,000 cell counts (Radioactive Index, R.I.). Following results were obtained: 1. The hepatic radioactive indices obtained from the saline group 1, 10, and 24 hour after $[^3H]$ thymidine administration were $3.23{\pm}0.23,\;5.20{\pm}0.21,\;and\;6.00{\pm}0.30\;(mean{\pm}S.D.)$, respectively. 2. The corresponding values obtained from the ginseng group $(4.22{\pm}0.33,\;6.32{\pm}0.32,\;and\;7.42{\pm}0.35)$ were significant higher than the values of the saline group. The inference from the above results was that the ginseng facilitated hepatic DNA synthesis.

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Studies on the Physical and Chemical Denatures of Cocoon Bave Sericin throughout Silk Filature Processes (제사과정 전후에서의 견사세리신의 물리화학적 성질변화에 관한 연구)

  • 남중희
    • Journal of Sericultural and Entomological Science
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    • v.16 no.1
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    • pp.21-48
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    • 1974
  • The studies were carried out to disclose the physical and chemical properties of sericin fraction obtained from silk cocoon shells and its characteristics of swelling and solubility. The following results were obtained. 1. The physical and chemical properties of sericin fraction. 1) In contrast to the easy water soluble sericin, the hard soluble sericin contains fewer amino acids include of polar side radical while the hard soluble amino acid sach as alanine and leucine were detected. 2) The easy soluble amino acids were found mainly on the outer part of the fibroin, but the hard soluble amino acids were located in the near parts to the fibroin. 3) The swelling and solubility of the sericin could be hardly assayed by the analysis of the amino acid composition, and could be considered to tee closely related to the compound of the sericin crystal and secondary structure. 4) The X-ray patterns of the cocoon filament were ring shape, but they disappeared by the degumming treatment. 5) The sericin of tussah silkworm (A. pernyi), showed stronger circular patterns in the meridian than the regular silkworm (Bombyx mori). 6) There was no pattern difference between Fraction A and B. 7) X-ray diffraction patterns of the Sericin 1, ll and 111 were similar except interference of 8.85A (side chain spacing). 8) The amino acids above 150 in molecular weight such as Cys. Tyr. Phe. His. and Arg. were not found quantitatively by the 60 minutes-hydrolysis (6N-HCI). 9) The X-ray Pattern of 4.6A had a tendency to disappear with hot-water, ether, and alcohol treatment. 10) The partial hydrolysis of sericin showed a cirucular interference (2A) on the meridian. 11) The sericin pellet after hydrolysis was considered to be peptides composed with specific amino acids. 12) The decomposing temperature of Sericin 111 was higher than that of Sericin I and II. 13) Thermogram of the inner portioned sericin of the cocoon shell had double endothermic peaks at 165$^{\circ}C$, and 245$^{\circ}C$, and its decomposing temperature was higher than that of other portioned sericin. 14) The infrared spectroscopic properties among sericin I, II, III and sericin extracted from each layer portion of the cocoon shell were similar. II. The characteristics of seriein swelling and solubility related with silk processing. 1) Fifteen minutes was required to dehydrate the free moisture of cocoon shells with centrifugal force controlled at 13${\times}$10$^4$ dyne/g at 3,000 R.P.M. B) It took 30 minutes for the sericin to show positive reaction with the Folin-Ciocaltue reagent at room temperature. 3) The measurable wave length of the visible radiation was 500-750m${\mu}$, and the highest absorbance was observed at the wave length of 650m${\mu}$. 4) The colorimetric analysis should be conducted at 650mu for low concentration (10$\mu\textrm{g}$/$m\ell$), and at 500m${\mu}$ for the higher concentration to obtain an exact analysis. 5) The absorbing curves of sericin and egg albumin at different wave lengths were similar, but the absorbance of the former was slightly higher than that of the latter. 6) The quantity of the sericin measured by the colorimetric analysis, turned out to be less than by the Kjeldahl method. 7) Both temperature and duration in the cocoon cooking process has much effect on the swelling and solubility of the cocoon shells, but the temperature was more influential than the duration of the treatment. 8) The factorial relation between the temperature and the duration of treatment of the cocoon cooking to check for siricin swelling and solubility showed that the treatment duration should be gradually increased to reach optimum swelling and solubility of sericin with low temperature(70$^{\circ}C$) . High temperature, however, showed more sharp increase. 9) The more increased temperature in the drying of fresh cocoons, the less the sericin swelling and solubility were obtained. 10) In a specific cooking duration, the heavier the cocoon shell is, the less the swelling and solubility were obtained. 11) It was considered that there are differences in swelling or solubility between the filaments of each cocoon layer. 12) Sericin swelling or solubility in the cocoon filament was decreased by the wax extraction.. 13) The ionic surface active agent accelerated the swelling and solubility of the sericin at the range of pH 6-7. 14) In the same conditions as above, the cation agent was absorbed into the sericin. 15) In case of the increase of Ca ang Mg in the reeling water, its pH value drifted toward the acidity. 16) A buffering action was observed between the sericin and the water hardness constituents in the reeling water. 17) The effect of calcium on the swelling and solubility of the sericin was more moderate than that of magnecium. 18) The solute of the water hardness constituents increased the electric conductivity in the reeling water.

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Sow Transfer of Cultured Freezing Embryos by Open Pulled Straw(OPS) Methods : Preliminary Results (Open Pulled Straw(OPS) 방법에 의한 체외배양 동결수정란의 경산돈 이식 : 예비실험 결과)

  • Kim, I.-D.;Ahn, M.-H.;Hur, T.-Y.;Hong, M.-P.;Seok, H.-B.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.155-163
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    • 2004
  • The aims of this study are 1) to test oocytes and embryos collected from in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to Funahashi et al (1994). Glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$, and 10% fetal bovine serum albumin was added to the culture medium thereafter. Embryos were treated with 7.5 ${\mu}g/ml$ cytochalasin-B for 30 min, centrifuged at 13,000 rpm for 13 min and then exposed sequentially to an ethylene glycol(EG) vitrification solution, aspirated into OPS, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three dornors after AI for control group. Forty-nine embryos were washed 3 times in mPBS + 10% FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients were transferred individually with 100, 100 frozen embryos derived from abattoir and 34 fresh embryos by surgically, and another three recipients were transferred individually with 150, 150 frozen embryos and 100 fresh embryos by nonsurgically, respectively. all recipient sows exhibited delayed returns to estrus. To our knowledge, theses results suggest that required an improved techniques, more vigorous embryos preparation and substitute to gilt with cleaner uterous condition.

The Anti-inflammatory Effect of Skipjack Tuna (Katsuwonus pelamis) Oil in LPS-induced RAW 264.7 Cells and Mouse Models (LPS 유도 RAW 264.7 세포와 마우스 모델에서 참치(Katsuwonus pelamis) 유의 항염증 효과)

  • Kang, Bo-Kyeong;Kim, Min-Ji;Kim, Koth-Bong-Woo-Ri;Ahn, Na-Kyung;Choi, Yeon-Uk;Bark, Si-Woo;Pak, Won-Min;Kim, Bo-Ram;Park, Ji-Hye;Bae, Nan-Young;Ahn, Dong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.45-55
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    • 2015
  • This study was carried out to demonstrate the anti-inflammatory effect of tuna oil (TO) using LPS-induced inflammation responses and mouse models. First, nitric oxide (NO) and pro-inflammatory cytokines levels were suppressed up to 50% with increasing concentrations of TO without causing any cytotoxicity. Also, the expression of a variety of proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and nuclear factor kappa B (NF-κB), was suppressed in a dosedependent manner by treatment with TO. Furthermore, TO also inhibited the phosphorylation of mitogen-activated protein kinases (MAPKs), including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK), and p38 protein kinase (p38). Moreover, in in vivo testing the formation of ear edema was reduced at the highest dose tested compared to that in the control, and a reduction of ear thickness and the number of mast cells was observed in histological analysis. In acute toxicity test, no mortalities occurred in mice administrated 5,000 mg/kg body weight of TO over a two-week observation period. Our results suggest that TO has a considerable anti-inflammatory property through the suppression of inflammatory mediator productions and that it could prove to be useful as a potential anti-inflammatory therapeutic material.