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Antioxidative Effect of Pyruvate, Taurine and Melatonin, and Relationships of the Evaluation Methods on Boar Sperm Viability for In Vitro Storage (돼지 정자의 체외조본에 있어 Pyruvate, Taurine 및 Melatonin의 항산화효과와 평가방법의 상관관계 분석)

  • Jang, H.Y.;Park, C.K.;Cheong, H.T.;Kim, J.T.;Lee, H.K.;Im, S.K.;Yang, B.K.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.729-736
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    • 2007
  • The objective of this study was to investigate the anti-oxidative effects of pyruvate, taurine and melatonin on sperm characteristics(motility, membrane integrity) and lipid peroxidation(LPO) for in vitro storage of boar semen. Semen was treated with various antioxidants such as pyruvate(1mM), taurine(50mM) and melatonin(100nM) with or without 100uM H2O2. Antioxidant treatments were significantly increased the sperm motility when compare to control group in all incubation periods(P≤0.05). Hypoosmotic swelling test(HOST), membrane integrity was similar to the result of motility. In lipid peroxidation measurement by TBA reactions of spermatozoal plasma membrane, malondialdehyde(MDA) level in control and antioxidant treatments were lower than those of antioxidant plus H2O2 or H2O2 treatment for 3 to 6 h incubation period. Relationships of evaluation methods for sperm viability were investigated by motility, membrane integrity and lipid peroxidation. Among evaluation methods, LPO vs motility and membrane integrity vs LPO were negatively correlated(-0.23~-0.92 and -0.68~-0.85), but membrane integrity vs motility was positively correlated (0.53~0.94) in all treatments. These experiments indicate that supplementation of antioxidant to the semen extender can increase the sperm motility and membrane integrity and decrease the lipid peroxidation of spermatozoal plasma membrane. The HOST might be utilized to evaluate the sperm quality instead of lipid peroxidation or motility.

Effect of Feeding High Guality Hay on Performance and Physico-chemical Characteristics of Carcass of Hanwoo Steers (양질의 건초 급여가 한우 거세우의 비육성적 및 도체의 이화학적 특성에 미치는 영향)

  • Kim, S.I.;Jung, K.K.;Kim, J.Y.;Lee, S.W.;Baek, K.H.;Choi, C.B.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.783-800
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    • 2007
  • The current study was conducted to investigate the effect of high quality hay on the performance and carcass characteristics of Hanwoo steers. Twenty(20) Hanwoo(7 months old) were allocated into either Control(rice straw fed) or Treatment(timothy hay and rice straw fed) group(10 animals per group) and fed for 710 days until the animals reached at 30 months old. Concentrates were fed according to the feeding program composed with three(3) phases; growing, fattening, and finishing period. For the overall feeding period, final body weights were 761.3 and 799.6kg for the Control and Treatment groups, respectively, showing 38.3kg heavier body weight in Treatment group. ADG were 0.79 and 0.84kg for Control Treatment groups, respectively. These results might be because of the intake of high quality hay during growing phase and the effects persisted until the end of the experiment. Feed intake tended to be higher in Treatment group whereas feed conversion did not show significant difference between groups. Cold carcass weights were 451.0 and 475.3kg for Control and Treatment, There were no significant difference between groups in both yield and quality grade. There were no remarkable differences in physico-chemical characteristics fatty acid composition of carcasses between groups. In conclusion, it would be beneficial to feed high quality hay such as timothy during growing period of Hanwoo steers to produce high quality beef with heavier live and carcass weight.

NEAR REAL-TIME ESTIMATION OF GEOMAGNETIC LOCAL K INDEX FROM GYEONGZU MAGNETOMETER (경주 지자기관측소 자료를 이용한 준실시간 K 지수 산출에 관한 연구)

  • Choi, K.C.;Cho, K.S.;Moon, Y.J.;Kim, K.H.;Lee, D.Y.;Park, Y.D.;Lim, M.T.;Park, Y.S.;Lim, H.R.
    • Journal of Astronomy and Space Sciences
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    • v.22 no.4
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    • pp.431-440
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    • 2005
  • Local K-index is an indicator representing local geomagnetic activity in every 3 hour. For estimation of the local K-index, a reasonable determination of solar quiet curve (undisturbed daily variation of geomagnetic field) is quiet essential. To derive the solar quiet curve, the FMI method, which is one of representative algorithms, uses horizontal components (H and D) of 3 days magnetometer data from the previous day to the next day for a specific day. However, this method is not applicable to real time forecast since it always requires the next day data. In this study, we have devised a new method to estimate local K-index in near real-time by modifying the FMI method. The new method selects a recent quiet day whose $K_p$ indices, reported by NOAA/SEC are all lower than 3, and replace the previous day and the next day data by the recent quiet day data. We estimated 2,672 local K indices from Gyeongzu magnetometer in 2003, and then compared the indices with those from the conventional FMI method. We also compared the K indices with those from Kakioka observatory. As a result, we found that (1) K indices from the new method are nearly consistent with those of the conventional FMI method with a very high correlation (R=0.96); (2) onr local K indices also have a relatively high correlation (R=0.81) with those from Kakioka station. Our results show that the new method can be used for near real-time estimation of local K indices from Gyeongzu magnetometer.

Variations of Phytoplankton Standing Crops Affecting by Environmental Factors in the Marine Ranching Ground of Tongyeong Coastal Waters from 2000 to 2007 (2000$\sim$2007년 통영바다목장해역에서 환경요인의 영향에 따른 식물플랑크톤 현존량의 변화)

  • Jung, Seung-Won;Kwon, Oh-Youn;Joo, Hyoung-Min;Lee, Jin-Hwan
    • Korean Journal of Environmental Biology
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    • v.25 no.4
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    • pp.303-312
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    • 2007
  • In order to investigate the dynamics of phytoplankton standing crops affecting by environmental factors, biological and environmental factors, this study was examined in the marine ranching ground of Tongyeong coastal waters from 2000 to 2007. During the study, mean water temperature and salinity were 16.7$^{\circ}C$ and 32.9 psu, respectively. pH, DO and SS varied from 7.81$\sim$8.09, 3.02$\sim$8.97 mg $L^{-1}$ and 2.7$\sim$32.2 mg $L^{-1}$, respectively. Mean concentrations of dissolved inorganic nitrogen, phosphate and silicate were 21.75 ${\mu}M$, 0.90 ${\mu}M$ and 14.38 ${\mu}M$, respectively. Chlorophyll a concentrations varied from 0.02 ${\mu}g$ $L^{-1}$ to 25.29 ${\mu}g$ $L^{-1}$ with mean a value of 2.0 ${\mu}g$ $L^{-1}$. These factors did show significant differences on each layer and season, while did not show on the sampling stations. Phytoplankton standing crops varied from $4.21\times10^3$ cells $L^{-1}$ to $1.44\times10^6$ cells $L^{-1}$ with a mean value of $1.92\times10^5$ cells $L^{-1}$. Especially, variations of phytoplankton standing crops had an unimodal pattern as only bloomed in autumn rather than a bimodal pattern as generally bloomed in spring and autumn. In results of stepwise multiple regression analysis, the coefficient of determination $(R^2)$ for total standing crops was 0.35 and the standing crops were affected by water temperature, salinity, phosphate and silicate. The factors affected were different seasonally; water temperature in spring, salinity in summer, water temperature, salinity and silicate in autumn and water temperature, salinity and suspended solids in winter. Therefore, the results from the statistical analysis showed that the environmental factors influencing on the variations of the phytoplankton standing crops were predominantly water temperature and salinity.

Immunoelectron Microscopic Study on the Paneth Cell of Rabbit after the Common Bile Duct Ligation (총담관결찰후 집토끼 Paneth세포의 변화에 대한 면역전자현미경적 연구)

  • Park, Kyung-Ho;Cho, Hwee-Dong;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Kim, Jin-Gook
    • Applied Microscopy
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    • v.24 no.2
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    • pp.78-92
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    • 1994
  • Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.

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Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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Studies on canine babesiosis in Korea I. In vitro isolation and antigenic properties of Babesia gibsoni (개 바베시아병에 관한 연구 I. Babesia gibsoni의 시험관내 분리와 항원성상에 관한 연구)

  • Lee, Ho-kweon;Suh, Myung-deuk
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.681-692
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    • 1996
  • The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

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Nutritional Components and Their Antioxidative Protection of Neuronal Cells of Litchi (Litchi chinensis Sonn.) Fruit Pericarp (리치 과피의 영양화학 성분 및 항산화성 신경세포 보호효과)

  • Jeong, Hee-Rok;Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Kim, Yeon-Su;Jeong, Chang-Ho;Kim, Dae-Ok;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.481-487
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    • 2010
  • The nutritional components, antioxidant, and neuroprotective effects of water and a 50% methanol extract from litchi fruit pericarp were investigated. The most abundant mineral, amino acid, and fatty acid were K, proline, and palmitic acid, respectively. In addition, the total water phenolics and 50% methanol extracts were 8.02 and 12.28 mg/g, respectively. The DPPH, ABTS radical scavenging activities and ferric reducing antioxidant power of the water and 50% methanol extracts showed dose-dependent antioxidant activity. In a cell viability assay using MTT, almost all extracts showed a protective effect against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase leakage was also inhibited by the pericarp extracts. In particular, the 50% methanol extract showed a higher cell membrane protective effect than the water extract at the highest concentration. Consequently, these data suggest that litchi fruit pericarp can be utilized as an effective and safe functional food substances for natural antioxidants and may reduce the risk of neurodegenerative disorders.

Development of Assay Methods for Enterotoxin of Escherichia coli Employing the Hybridoma Technology (잡종세포종기법을 이용한 대장균의 장독소 측정법 개발)

  • Kim, Moon-Kyo;Cho, Myung-Je;Park, Kyung-Hee;Lee, Woo-Kon;Kim, Yoon-Won;Choi, Myung-Sik;Park, Joong-Soo;Cha, Chang-Yong;Chang, Woo-Hyun;Chung, Hong-Keun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.1
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    • pp.151-161
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    • 1986
  • In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

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Studies on the Improvement of the Fishing Efficiency of Purse Seine in the Sea Area of Cheju Island -The Changes of Seine Volume and Tension in the Purseline During Pursing- (제주도 주변해역 선망의 어획성능 향상에 관한 연구 -짐줄 체결 중 선망의 용적과 짐줄의 장력 변화 -)

  • 김석종
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.35 no.2
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    • pp.93-101
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    • 1999
  • A simple experimental method was used in an attempt to realize the elevation of the fishing ability of purse seine in the sea area of Cheju Island, the changes of seine volume and tension in the purseline during pursing. Experiments carried out on the six types simplified reduced model seines which were made of knotless nettings. The nettings were woven in different leg length 4.3, 5.0, 5.5, 6.0, 6.6 and 7.7mm of polyester 28 tex two threads two-ply twine, and each of the seines were named I, II, III, IV, V and Ⅵ seine. Dimension of seine models were 450cm for corkline and 85cmfor seine depth, each seines rigged up 160g of float for a floatline and 50g (underwater weight) of lead for a leadline. These model purse seines were made of the scale of 1/200 of its full scale, a 120 ton in the near sea of Cheju Island. Designing and testing for the model purse seines were based on the Tauti's law. Experiments were measured in the observation channel of a flume tank at the static conditions set up shooting and pursing equipments. Motion of purse seine during purse line was recorded by the two sets video camera for VTR which were placed in top and front of the model seine. The reading coordinate of seine volume carried out by the video digitization system, disk data for the purseline tension. An analysis were performed on the changes seine volume and tension in the purseline during pursing. The results obtained were as follows: 1. The seine volume during pursing was largest for Ⅵ seine with smallest d/l followed by V, IV, III, II and I seines, and tension in the purseline was small. 2. Seine volume during pursing can be expressed by the following equation; CVt=l-EXP[{2.79 (d/l)+0.35}t-33.37 (d/l) + 0.57] Where CVt is volume ratio, d is twine diameter, l is leg length and t is pursing time (sec). 3. Tension in the purse line during pursing can be expressed by the following equation; T= 1- EXP {0.57t + 13.36 (d/l)+2.97} Where T is tension (kg) in the purseline during pursing.

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