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Effect of Verapamil on Cellular Uptake of Tc-99m MIBI and Tetrofosmin on Several Cancer Cells (수종의 암세포에서 Verapamil이 Tc-99m MIBI와 Tetrofosmin의 섭취에 미치는 영향)

  • Kim, Dae-Hyun;Yoo, Jung-Ah;Suh, Myung-Rang;Bae, Jin-Ho;Jeong, Shin-Young;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.1
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    • pp.85-98
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    • 2004
  • Purpose: Cellular uptake of $^{99}mTc$-sestamibi (MIBI) and $^{99}mTc$-tetrofosmin (TF) is low in cancer cells expressing multidrug resistance(MDR) by p-glycoprotein(Pgp) or multidrug related protein(MRP). Verapamil is known to increase cellular uptake of MIBI in MDR cancer cells, but is recently reported to have different effects on tracer uptake in certain cancer cells. This study was prepared to evaluate effects of verapamil on cellular uptake of MIBI and TF in several cancer cells. Materials and Methods: Celluar uptakes of Tc-99m MIBI and TF were measured in erythroleukermia K562 cell, breast cancer MCF7 cell, and human ovarian cancer SK-OV-3 cells, and data were compared with those of doxorubicin-resistant K562(Ad) cells. RT-PCR and Western blot analysis were used for the detection of mdr1 mRNA and Pgp expression, and to observe changes in isotypes of PKC enzyme. Effects of verapamil on MIBI and TF uptake were evaluated at different concentrations upto $200{\mu}M\;at\;1{\times}10^6\;cells/ml\;at\;37^{\circ}C$. Radioactivity in supernatant and pellet was measured with gamma counter to calculate cellular uptake ratio. Toxicity of verapamil was measured with MTT assay. Results: Cellular uptakes of MIBI and TF were increased by time in four cancer cells studied. Co-incubation with verapamil resulted in an increase in uptake of MIBI and TF in K562(Adr) cell at a concentration of $100{\mu}M$ and the maximal increase at $50{\mu}M$ was 10-times to baseline. In contrast, uptakes of MIBI and TF in K562, MCF7, SK-OV3 cells were decreased with verapamil treatment at a concentration over $1{\mu}M$. With a concentration of $200{\mu}M$ verapamil, MIBI and TF uptakes un K562 cells were decreased to 1.5 % and 2.7% of those without verapamil, respectively. Cellular uptakes of MIBI and TF in MCF7 and SK-OV-3 cells were not changed with $10{\mu}M$, but were also decreased with verapamil higher than $10{\mu}M$, resulting 40% and 5% of baseline at $50{\mu}M$. MTT assay of four cells revealed that K562, MCF7, SK-OV3 were not damaged with verapamil at $200{\mu}M$. Conclusion: Although verapamil increases uptake of MIBI and TF in MDR cancer cells, cellular uptakes were further decreased with verapamil in certain cancer cells, which is not related to cytotoxicity of drug. These results suggest that cellular uptakes of both tracers might differ among different cells, and interpretation of changes in tracer uptake with verapamil in vitro should be different when different cell lines are used.

Optimum Harvest Stage of Italian Ryegrass 'Kowinearly' According to One and Two Harvests During Spring Season (이탈리안 라이그라스 '코윈어리'의 봄철 1회 및 2회 이용에 따른 수확적기 구명)

  • Seo, Sung;Kim, Meing Jooung;Kim, Won Ho;Lee, Sang Hak;Jung, Min Woong;Kim, Ki Yong;Ji, Hee Chung;Park, Hyung Soo;Kim, Jong Geun;Choi, Gi Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.1
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    • pp.15-20
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    • 2013
  • This study was carried out to determine the optimum harvest stage of Italian ryegrass (Lolium multiflorum Lam., IRG) for maximum forage production during the spring season in Suwon, 2010. The variety of IRG was the early maturity type, 'Kowinearly', and six harvest stages (treatments) were first heading (T1), heading (T2), late heading to early bloom (T3), bloom to late bloom (T4), ripeness (T5), and late ripeness stage (T6). The dates of the first heading and heading of 'Kowinearly' were seen on 4 to 5 May, and 14 May, respectively. Plant length and dry matter (DM) percentage at first harvest were from 69 cm and 14.8% at T1 stage to 103 cm and 35.0% at T6 stage, respectively. The content of crude protein (CP) and in vitro DM digestibility (IVDMD) of T1, T2, T3, T4, T5 and T6 at first harvest were 15.6%, 10.6%, 10.1%, 8.1%, 7.3% and 5.4%, and 81.8%, 72.1%, 64.8%, 63.8%, 61.4% and 59.0%, respectively. The content of neural detergent fiber (NDF) and acid detergent fiber (ADF) were increased continuously with delayed harvest. A significantly higher yield of DM, CP and in vitro digestible DM (IVDDM) were observed for T3, and T4 (p<0.05). DM yield of 3,526 kg, 6,278 kg, 7,842 kg, 8,984 kg, 8,346 kg and 8,008 kg/ha, CP yield of 549 kg, 665 kg, 795 kg, 725 kg, 608 kg and 430 kg/ha, and IVDDM of 2,883 kg. 4,526 kg, 5,083 kg, 5,728 kg, 5,124 kg and 4,722 kg/ha at first harvest were recorded in T1, T2, T3, T4, T5 and T6, respectively. Regrowth yield of DM, CP and IVDDM were shown to be higher at T1 and T2 (p<0.05). However, no significant differences were observed between the two stages. Daily DM and DDM production of regrowth IRG were higher at T2, followed by T1. The total yield (at first and at regrowth) of DM, CP and IVDDM were significant higher for T2, followed by T3, T4 and T1 in order. At T2 stage, the yield was 11,089 kg, 1,254 kg, and 7,669 kg/ha in DM, CP, and IVDDM. In conclusion, the late heading to bloom stage was determined to be the optimum harvest stage for a single harvest, while the heading stage was a suitable stage of first harvest of 'Kowinearly' where two harvests were sought in a single year.

In vitro Antioxidant and Anti-Inflammatory Activities of Ethanol Extract and Sequential Fractions of Flowers of Prunus persica in LPS-Stimulated RAW 264.7 Macrophages (복숭아꽃 에탄올 추출물과 분획물의 in vitro 항산화 효과 및 RAW 264.7 대식세포에서의 항염증 효과)

  • Kwak, Chung Shil;Choi, Hye-In
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.10
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    • pp.1439-1449
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    • 2015
  • Prunus persica Flos (PPF) were investigated for their antioxidant and anti-inflammatory activities to find a natural functional food resource preventing degenerative diseases associated with excessive oxidative stress and chronic inflammation. PPF was extracted using ethanol (EtOH) and then sequentially fractioned by hexane (Hx), dichloromethane (DM), ethyl acetate (EA), n-butanol (BtOH), and water (DW). Contents of total phenolics and flavonoids, as well as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities were measured. Anti-inflammatory effects in terms of nitric oxide (NO), prostaglandin (PG) E2, and pro-inflammatory cytokines such as interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$ production were also measured using LPS-treated RAW 264.7 macrophages. EtOH extract showed relatively high antioxidant activity with high total phenolic (78.1 mg tannic acid/g) and flavonoid contents (55.3 mg rutin/g). EA fraction contained the highest total phenolic and flavonoid contents (394.6 mg tannic acid/g, 253.7 mg rutin/g), followed by BtOH (128.3 mg tannic acid/g, 93.1 mg rutin/g). EA and BtOH fractions and EtOH extract showed higher DPPH radical and ABTS radical scavenging activities than the others (P<0.05). In LPS-treated RAW 264.7 macrophages, EtOH extract ($200{\mu}g/mL$) showed significantly reduced (P<0.05) NO, PGE2, and TNF-${\alpha}$ production levels to 38.5%, 32.3%, and 48.9% of the control, respectively, as well as reduced iNOS and COX-2 protein expression. DM fraction ($50{\mu}g/mL$) showed significantly reduced (P<0.05) NO, PGE2, IL-6, and TNF-${\alpha}$ production levels to 43.5%, 13.3%, 38.7%, and 41.3% of the control, respectively, and EA fraction ($50{\mu}g/mL$) showed significantly reduced NO, PGE2, IL-6, and TNF-${\alpha}$ production levels to 44.8%, 22.4%, 45.7%, and 62.0% of the control, respectively. Taken together, EtOH extract of PPF showed potent antioxidant and anti-inflammatory activities, and EA and BtOH fractions showed comparatively stronger antioxidant activities while DM and EA fractions showed stronger anti-inflammatory activities. It can be concluded that EtOH extract of PPF and its fractions are good candidates as natural resources for the development of anti-oxidative and anti-inflammatory functional food products.

The Effects of Supplemental Bacterial Phytase to the Calcium and Nonphosphorus Levels in Feed of Laying Hens (산란계 사료 내 칼슘 및 무기태 인 수준에 따른 Bacterial Phytase 급여 효과)

  • Kang, H.K.;Park, S.Y.;Yu, D.J.;Kim, J.H.;Kang, G.H.;Na, J.C.;Kim, D.W.;Suh, O.S.;Lee, S.J.;Lee, W.J.;Kim, S.H.
    • Korean Journal of Poultry Science
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    • v.35 no.2
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    • pp.143-151
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    • 2008
  • This study was conducted to identify the correlation of bacterial phytase ($Transphos^{(R)}$) to the calcium level in feed. Of all 21-week-old 720 HyLine brown laying hens, 2 birds of similar weight were placed on each individual cage. The experiment was conducted by $3{\times}2{\times}3$ factorial design with including 3 different levels of phytase (0, 300, and 1,000 DPU/kg), 2 different levels of calcium (3.5% and 4.0%), and 3 different levels of no NPP addition 0% (0.095 NPP), 0.5% (0.185% NPP), and 1.0% (0.275% NPP). The feeding trial maintained the ME level of 2,800 kcal/kg and 16% for crude protein. The diet was fed ad libitum and 17 hours of lighting was provided throughout the experimental period. Egg production seemed to increase, in the 300 DPU of bacterial phytase added group and the cracked egg tended to reduce in Transphos added group. The egg productivity between treatment groups did not show significant difference by dietary calcium level, whereas non NPP added group (0.095% NPP) was found to be low compared to NPP added groups (P<0.05). The highest mean egg weight and the highest daily egg mass were detected in 300 DPU phytase added group. Although the mean egg weight was significantly higher in treatment groups fed with 3.5% calcium containing feeds (P<0.05), daily egg mass was no among treatment groups. The mean egg weight and daily egg mass were the lowest in non NPP added group (0.095% NPP) compared to other treatment groups (P<0.05). The feed intake showed similar pattern regardless of the bacterial phytase and calcium levels in the diet. However, the treatment groups fed diets containing NPP level of 0.275% and 0.165% showed significantly higher feed intake than the group fed with 0.095% NPP (P<0.05). Although the feed conversion was not affected by calcium and NPP levels in the diet, the most improved result was obtained from 300 DPU phytase added group (P<0.05). The eggshell breaking strength and thickness increased as dietary calcium level increase the level of calcium increases in diet. The treatment groups fed diet containing 0.275% and 0.165% NPP revealed to show improvement in eggshell breaking strength and yolk color index compared to the NPP non added (0.095% NPP) treatment group. The result of the present study suggests that the appropriate level of microbial phytase is 300 DPU and at this level, tricalciumphosphate supplementation in feed can be reduced to 40% of NRC recommendation. Higher calcium level in feed fail to show synergistic effect by adding microbial phytase.

The Signal Transduction Mechanisms on the Intestinal Mucosa of Rat Following Irradiation (방사선조사후 백서소장점막에서 발생하는 신호전달체계에 관한 연구)

  • Yoo Jeong Hyun;Kim Sung Sook;Lee Kyung Ja;Rhee Chung Sik
    • Radiation Oncology Journal
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    • v.15 no.2
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    • pp.79-95
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    • 1997
  • Purpose : Phospholipase C(PLC) isozymes play significant roles in signal transduction mechanism. $PLC-\gamma$ 1 is one of the key regulatory enzymes in signal transduction for cellular proliferation and differentiation. Ras oncoprotein, EGFR, and PKC are also known to be involved in cell growth. The exact mechanisms of these signal transduction following irradiation, however, were not clearly documented Thus, this study was Planned to determine the biological significance of PLC, ras oncoprotein, EGFR, and PKC in damage and regeneration of rat intestinal mucosa following irradiation. Material and Method : Sixty Sprague-Dawley rats were irradiated to entire body with a single dose of 8Gy. The rats were divided into S groups according to the sacrifice days after irradiation. The expression of PLC, ras oncoprotein, EGFR and PKC in each group were examined by the immunoblotting and immunohistochemistry. The histopathologic findings were observed using H&I stain, and the mitoses for the evidence of regeneration were counted using the light microscopy & PCNA kit. The Phosphoinositide(PI) hydrolyzing activity assay was also done for the indirect evaluation of $PLC-\gamma$ 1 activity. Results: In the immunohistochemistry , the expression of $PLC-{\beta}$ was negative for all grøups. The expression of $PLC-{\gamma}1$ was highest in the group III followed by group II in the proliferative zone of mucosa. The expression of $PKC-{\delta}1$ was strongly positive in group 1 followed by group II in the damaged surface epithelium. The above findings were also confirttled in the immunoblotting study. In the immunoblotting study, the expressions of $PLC-{\beta}$, $PLC-{\gamma}1$, and $PKC-{\delta}1$ were the same as the results of immunohis-tochemistry. The expression of ras oncoprctein was weakly positive in groups II, III and IV. The of EGFR was the highest in the group II, III, follwed by group IV and the expression of PKC was weakly positive in the group II and III. Conclusion: $PLC-{\gamma}1$ mediated signal transduction including ras oncoprotein, EGFR, and PKC play a significant role in mucosal regeneration after irradiation. $PLC-{\delta}1$ mediated signal transduction might have an important role in mucosal damage after irradiation. Further studies will be necessary to confirm the signal transduction mediating the $PKC-{\delta}1$.

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Neuroprotective Effect of Cyclosporin A on Spinal Cord Ischemic Injury in Rabbits (토끼를 이용한 척수 허혈 손상 모델에서 Cyclosporin A의 척수 손상에 대한 보호 효과)

  • Shin Yoon-Cheol;Choe Ghee-Young;Kim Won-Gon
    • Journal of Chest Surgery
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    • v.39 no.10
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    • pp.739-748
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    • 2006
  • Background: The purpose of this study is to ascertain the neuroprotective effect of cyclosporin A on the 25-min surgical ischemia model in the spinal cords of rabbits with neuropathological correlation and histoimmunochemical analyses, Material and Method: Thirty-two New Zealand white rabbits were randomly divided into four groups: Rabbits were randomly divided into four groups: the control 12 group (n=8), the control 17 group (n=8), the cyclosporin Cs2 group (n=8), and the cyclosporin Cs7 group (n=8). The 12 group underwent a 25-min aortic cross- clamp without intervention and were sacrificed on the 2nd day postoperatively, while the 17 group underwent a 25- min of aortic cross-clamp without intervention and were sacrificed on the 7th day postoperatively. The Cs2 group received cyclosporin A (25 mg/kg) intravenously 15 min after the 25-min cross-clamp and were sacrificed on the End day postoperatively, while the Cs7 group received cyclosporin A (25 mg/kg) intravenously 15 min after the 25-min cross-clamp and were sacrificed on the 7th day postoperatively. The rabbits underwent 25-min surgical aortic cross-clamp. Neurologic functions were evaluated on the 2nd day and 7th postoperative day using Tarlov scoring system. After scoring neurologic function, all rabbits were sacrificed for histopathologic observation. Result: All rabbits survived the experimental procedure. The values of Tarlov score did not show any differences between the control and cyclosporin groups on the 2nd day. The scores of group Cs7 ($2.75{\pm}0.89$) were significantly higher than those of group 17 ($1.25{\pm}1.39$) on the 7th day (p<0,05). On the histologic exanminations, specimens of the spinal cord showed necrosis and apoptosis. The pathologic scores of group Cs7 ($1,0{\pm}0.53$) was less than those of group 17 ($2.13{\pm}1.36$, p<0.05). TUNEL staing showed apoptosis of the specimen in group 12 and Cs2 but there was no stastically significant difference between groups on the score. There were more overexpression of HSP70 and nNOS in cyclosporine group than in control group. Conclusion: We think that cyclosporin A may decrease neuronal cell death with induced upregulation of HSP70 against 25-min ischemia of the spiral cord in the rabbit.

Comparison of Growth Characteristics and Productivity of Summer Forage Crops in Sihwa Reclaimed Land (시화 간척지에서 하계 사료작물의 초종 및 품종에 따른 생육특성 및 생산성 비교)

  • Kim, Jong Geun;Jeong, Eun Chan;Kim, Meing Joong;Li, Yan Fen;Kim, Hak Jin;Lee, Su Hwan
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.41 no.2
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    • pp.110-118
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    • 2021
  • This study was conducted to investigate the effects of species and varieties of summer forage crops on growth characteristics and productivity in Sihwa reclaimed land. The summer forage crops used in the trial were silage corn, sorghum×sudangrass hybrid(SSH), and proso millet. For each forage species, Gwangpyeongok(GPO), P15453, P1952 and P2088 were used for silage corn, and 877F, Green star, Honey chew, and Turbo gold cultivars were used for SSH. For proso millet, Ibaekchal, Geumsilchal and Manhongchal developed by the National Institute of Crop Science were used. Silage corn and SSH were sown on May 21, 2019 and proso millet on June 4, and harvested on September 2. There was no significant difference in plant and ear height of silage corn among varieties. P1543 was the highest and P2088 was the lowest in yield of silage corn, but there was no significant difference among treatments. Among the SSH, the plant height of 877F was the highest and Turbo gold variety had the smallest (p<0.05). As for the dry matter(DM) yields, 877F had the highest at 3,862 kg/ha and Green star had the lowest at 2,669 kg/ha (p<0.05). The fresh matter yield of proso millet was 15,778 kg/ha, which was higher than that of corn or SSH, The average dry matter yield was 4,780 kg/ha, and Ibaekchal variety had the highest DM yield compared to other varieties (p<0.05). P2088 had the highest TDN content and GPO was the lowest (p<0.05). As for the SSH, the TDN content of Green star and Honey chew varieties was significantly higher, and the RFV value was the lowest in Turbo gold. The average crude protein content of proso millet was 7.03%, and the highest TDN and RFV values were 64.36% and 106 in Geumsilchal. In the experiment of the germination rate of summer forage crops according to salt concentration, silage corn showed a germination rate of 83.1% even at 0.4% salinity. In particular, P2088 and P1921 varieties had more than 80% germination rate even at 0.6% salt concentration. As for the SSH, the germination rate of 877F was 93.3% even at 0.8% salinity, and 88.3% with Honey chew, indicating higher resistance to salt concentration compared to other varieties. Proso millet showed a high germination rate of 84.0 to 88.7% even at a salt concentration of 0.6%. Considering the above results, proso millet was recommended as the most suitable forage crop species in the Sihwa reclaimed land with high salt concentration, and the Ibaekchal variety is recommended as a suitable forage crop due to its high yield.

Studies on Dairy Farming Status, Reproductive Efficiencies and Disorders in New Zealand (I) A Survey on Dairy Farming Status and Milk Yield in Palmerston North Area (뉴질랜드 (Palmerston North) 의 낙농 현황과 번식 및 번식장해에 관한 연구(I) Palmerston North 지역의 낙농 현황과 우유 생산량에 관한 조사 연구)

  • 김중계;맥도날드
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.1-18
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    • 2000
  • Eighty dairy farms in Palmers ton North area in New Zealand were surveyed on 1) general characteristics (10 Questions), 2) milk yield and feed supplementary (7 questions), 3) reproductive efficiencies (12 questions) and 4) reproductive disorders (12 questions) by mail questions from February to July, 1998. Among those 4 items from 38 dairy farms (47.5%), especially in items 1) and 2), overall dairy farming situation, supplementary feeding and milk yields were surveyed and analyzed for Korean dairy farmers (especially in Cheju island) to have better understanding or higher economical gains. The results were as follows. 1. In dairy experience, 21 (45%) among 38 dairy farms surveyed were answered that farming less than 15 years, 15~19 year, 20~25 years and over 26 years experience were 3 (7.9%), 7 (18.4%), 6 (15.8%) and 5 (13.2%) which generally showed longer experience compare to Korean dairy farming situation. In survey of labour input and business goal of dairy farming, self-managing farms, sharemilkers, unpaid family manpowering farms, manager running farms, farms with hired worker, farms with part time helper and other type was 21 (55.3%), 10 (26.3%), 2 (3.5%), 3 (5.3%), 18 (31.6%), 2 (3.5%), and 1 (1.8%), respectively. 2. Analyzing pasture and tillable land, pasture according to feeding scale (200, 300 and 400 heads) were 56, 90 and 165.3 ha, and tillable lands were 51, 78 and 165 ha which showed some differences among feeding scale. In recording methods in 38 farms replied, 36 (95%) dairy handbook and 23 (70%) dual methods taking farms were higher than that of 10 (26.3%) computer and 15(39.5%) well-recorder methods. 3. Dairy waste processing facilities in environmental field were almost perfect except of metropolitan area, and so no problem was developed in its control so far. Hence, 26 farm (68.4%) of pond system was higher rather than those in 8 (21.2%) of using as organic manure after storing feces of dairy cattle, 1(2.6%) bunker system and 3 (7.9%) other type farms. 4. In milking facilities, 33 farms (86.9%) of Harringbone types were higher than those in 3 (7.9%) of Walkthrough types, 1 (2.6%) of Rotary system and other types. Although the construction facilities was not enough, this system show the world-leveled dairy country to attempted to elevate economic gains using the advantage of climatic condition. 5. In milking day and yearly yield per head, average 275 milking days and 87 drying days were longer than that of 228 average milking days in New Zealand. Annual total milk yield per head and milk solid (ms) was 3,990 kg and approximately 319 kg. Dairy milk solid (ms) per head, milk yield, fat percentage was 1.2 kg, 15.5 kg and average 4.83% which was much higher than in other country, and milk protein was average 3.75%. 6. In coclusion, Palmerstone North has been a center of dairy farming in New Zealand for the last 21 years. Their dairy farming history is 6~9 year longer than ours and the average number of milking cows per farm is 355, which is much greater than that (35) of Korea. They do not have dairy barn, but only milking parlors. Cows are taken care of by family 0.5 persons), are on a planned calving schedule in spring (93%) and milked for 240~280 days a year, avoiding winter. Cows are dried according to milk yield and body condition score. This management system is quite different from that of Korean dairy farms. Cows are not fed concentrates, relying entirely on pasture forages and the average milk yield per cow is 3,500 kg, which is about 1/2 milk yield of Korean dairy farms. They were bred to produce high fat milk with an average of 4.5%. Their milk production cost is the lowest in the world and the country's economy relies heavily on milk production. We Korean farmers may try to increase farming size, decreasing labor and management costs.

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Effects of Total Mixed Fermentation Feeds Based on Rice-straw and Six Forage Crops on the Productivity of Holstein Cows (청예사료작물과 볏짚 위주의 완전배합발효사료 급여가 Holstein 착유우의 생산성에 미치는 영향)

  • Lee, H. J.;Kim, H. S.;Ki, K. S.;Jeong, H. Y.;Baek, K. S.;Kim, J. S.;Cho, K. K.;Cho, J. S.;Lee, H. G.;Woo, J. H.;Choi, Y. J.
    • Journal of Animal Science and Technology
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    • v.45 no.1
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    • pp.69-78
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    • 2003
  • This experiment was carried out to evaluate the value of total mixed fermentation feeds(TMFF) as completely mixed ration and to observe the effect of various kinds of TMFF on the palatability, feed intake, and milk performance in Holstein cows. The dry matter (DM) content of TMFF used in the experiment was 23.98-28.42% range, and CP, TDN, ADF and NDF were 16.2${\sim}$19.2%, 58.3-65.1%, 34.4-39.6% and 46.9${\sim}$49.9% levels, respectively. The relative feed value (RFV) in rape-, alfalfa-, grass-, oat-, corn-TMFF groups were 138.6, 133.9, 116.5, 111.8, 111.4 and 108.1, respectively. Among these groups, RFV of rye-TMFF group was lowest. Dry matter disappearance(DMD) showed 0.8${\sim}$.9% to the all kinds of TMFF groups. The pH was 3.89${\sim}$.87 and $NH_3$-N concentration was 6.93-8.66 mg/$d\ell$. The acetic acid concentration in the raw material of TMFF showed low level of 0.19${\sim}$0.57%, lactic acid showed high level of 1.17${\sim}$3.21% and butyric acid was very high as 0.03${\sim}$0.32%. Therefore, these results provide evidence that the quality of TMFF was not so bad. In the daily fresh matter intake on the alfalfa-, grass-, rape-, corn-, oats- and rye-TMFF were showed 62.85, 60.48, 58.04, 57.11, 54.61 and 45.74 kg respectively. All TMFF showed high palatability as daily dry matter intake of 1.95 to 2.90% by body weight of experimental cows. Body condition score(BCS) was gradually increased in during 60 days of the experiment term. Average daily gain(ADG) showed about 140.0${\sim}$326.7g. In alfalfa-TMFF group, the ADG was higher than in the other groups (p<0.05). Also, the increase in BCS was observed in grass-TMFF group (3.07 to 3.34) and rye-TMFF group was decreased in 3.07 to 3.34 (p<0.05). The milk yield appropriately showed a range of 16.16${\sim}$18.95 kg in all groups. Among these groups, alfalfa-TMFF group was highest(P<0.05). Average milk fat contents showed high levels of 4.06${\sim}$4.79% and the level was high in order of rape-, grass-, corn-, alfalfa-, rye- and oats-TMFF. Milk protein was highest in forage-TMFF and level of lactose in milk was approximately 4.56% in overall groups. Solid non fat(SNF) and total solid(TS) contents were 8.75% and 12.8%, respectively. However, milk composition was not significantly affected by TMFF.

The Study of anti-inflammatory Mechanism with Cobra Venom on Astrocytes of Rats (뇌(腦) 성상세포(星狀細胞)를 대상으로 한 Cobrotoxin의 염증(炎症) 치료(治療) 기전(機轉) 연구(硏究))

  • Yoo, Jae-ryong;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.155-167
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    • 2005
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.

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