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Leukocyte-Endothelial Cell Adhesion Induced by Ischemia and Reperfusion Observed with in vivo Videomicroscopy (허혈-재관류에 의해 유도된 백혈구-혈관내피세포 유착에 대한 Videomicroscopy 영상소견)

  • Lee, Young Bae;Kang, Han Sug;Park, Shin Byung
    • Journal of Korean Neurosurgical Society
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    • v.29 no.10
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    • pp.1289-1295
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    • 2000
  • Purpose : Recent evidence suggests a possible role for leukocytes in brain injury following ischemia and reperfusion. This study examined the temporal profile of ischemic tissue damage and leukocyte response after transient middle cerebral artery occlusion(MCAO) with reperfusion in the mouse. Methods : Focal cerebral ischemia was made by temporary occluding of the stem of the proximal MCA. Two groups of the mouse were investigated : (1) sham operation(n=10), and (2)those having the arterial occlusion released after 90 minute(n=20). By 4 hours(n=10) and 24 hours(n=10) after the onset of ischemia-reperfusion, fluorescein videoimages were under-taken in the pial venules of the mouse using a closed cranial window technique. Rhodamine 6G was administered as a $80-100{\mu}l/min$ i.v. loading dose and a $30-40{\mu}l/min$ i.v. maintenance dose in saline to selectively label circulating leukocytes. Neuropathologic evaluation for brain injury was accomplished using the histochemical stain 2,3,5-triphen-yltetrazolium chloride(TTC) and hematoxylin and eosin(H & E) stain. Results : The mean number of adherent leukocytes to cerebral venules in the 90 minutes MCAO and 24 hours reperfusion group were $306{\pm}24$ compared with $72{\pm}8$ in the sham operation group. In the TTC staining method, the cortical infarct affecting 34.8% of hemispheric volume were created in all of animals (n=10) undergoing 90 minute MCAO with 24 hours reperfusion, but the infarcted area were not found in the other(sham operation and 90 minute MCAO with 4 hours reperfusion)groups. In the H & E stain, the brain tissue following 90 minute MCAO with 4 hours reperfusion revealed only a pyknosis of the nuclei with shrunken cytoplasm, but infiltrated leukocytes were not observed. After 24 hours of reperfusion, a many leukocytes were infiltrated within parenchyma and blood vessles. Conclusions : These findings demonstrate the feasiblity of continous in vivo monitoring of leukocyte adherence in cerebral venules and suggest that reperfusion induced leukocyte adherence to venular endothelium may contribute to tissue injury following focal cerebral ischemia.

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Bacterial Community Dynamics during Swine In vitro Fermentation Using Starch as a Substrate with Different Feed Additives for Odor Reduction

  • Alam, Md.J.;Jeong, C.D.;Mamuad, L.L.;Sung, H.G.;Kim, D.W.;Cho, S.B.;Lee, K.;Jeon, C.O.;Lee, Sang-S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.690-700
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    • 2012
  • The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen ($NH_3$-N) and $H_2S$ gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, $NH_3$-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as $NO_2$, $NO_3$ and $SO_4$ concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that $NH_3$-N, $H_2S$ and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds.

Influence of Human Follicular Fluid and Gonadotropins in the Culture Medium on the In Vitro Maturation, Fertilization and Development of Human Immature Oocytes (배양액 내 인간 난포액 및 성선자극호르몬 첨가가 인간 미성숙 난자의 체외성숙, 수정 및 체외 배발달에 미치는 영향)

  • Kim, Eun-Kuk;Kim, Dong-Won;Jeong, Byung-Jun
    • Journal of Embryo Transfer
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    • v.24 no.3
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    • pp.145-150
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    • 2009
  • This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) ${\sim}4$ h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to $6{\sim}8$ cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.

Effects of Bromelain and Double Emulsion on the Physicochemical Properties of Pork Loin

  • Shin, Hyerin;Kim, Hyo Tae;Choi, Mi-Jung;Ko, Eun-Young
    • Food Science of Animal Resources
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    • v.39 no.6
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    • pp.888-902
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    • 2019
  • Our aim was to investigate the effects of bromelain embedded in double emulsion (DE) on physicochemical properties of pork loin. We evaluated DE characteristics such as size, zeta potential, and microscopy after fabrication. We marinated meat with distilled water (DW), 1% (w/v) bromelain solution, blank DE, and 1% (w/v) bromelain loaded in double emulsion (DE E) for 0, 24, 48, and 72 h at 4℃, and prepared raw meat for control. The marinated samples were assessed for color, water holding capacity, cooking loss, moisture content, pH, protein solubility, Warner-Bratzler shear force (WBSF) and gel electrophoresis. The droplet size of 1% (w/v) bromelain embedded in DE was increased compared with blank DE (p<0.05) and values of zeta potential decreased. The increase in lightness and color difference range of the DE-treated group was lower than that of the DW-treated group (p<0.05). Moreover, treatment by immersion in 1% (w/v) DE E resulted in the highest water holding capacity values (p<0.05) and lower cooking loss values than water base treatment (p<0.05). Results for myofibrillar protein solubility and WBSF showed a similar trend. 1% (w/v) DE E showed degradation of myosin heavy chain after 48 h in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Thus, bromelain-loaded DE is useful for controlling and handling enzyme activity in food industry.

Disinfection efficacy of slightly acidic electrolyzed water (SlAEW) against some fresh vegetables (미산성 차아염소산수의 신선 채소류에 대한 살균 유효성)

  • Park, Kee-Jai;Lim, Jeong-Ho;Jung, Heeyong;Jeong, Mooncheol
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.312-319
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    • 2017
  • In the present study, disinfection efficacy of slightly acidic electrolyzed water [SlAEW, 30 ppm of effective chlorine at $20{\pm}1^{\circ}C$, oxidation-reduction potential (ORP) $562{\pm}23mV$, pH 6.4] on 4 kinds of vegetables (lettuce leaf, endive leaf, perilla leaf and kale leaf) was evaluated to obtain a microbial reduction characteristics which are necessary to design a process control for non-thermal sterilization of fresh vegetables. Active chlorine, residual chlorine, microbial counts and residual microbial counts, which are the key factors in the non-thermal sterilization process were measured by dipping them in SlAEW three times for 30 minutes in order to analyze the relationship between factors. Total microbial count was decreased mostly during the first 10 minutes of washing, and the limit value that can be reduced by immersion treatment was 3 log CFU/g for the total microbial count surviving in 4 kinds of vegetables. The total number of microorganism that can be reduced by washing in SIAEW for 10 min was found to be about 2 log CFU/g on average. In addition, the active chlorine decreased in the initial 10 minutes in 2.2 ppm, 2.0 ppm, 1.7 ppm and 2.5 ppm in lettuce, perilla leaf, endive leaf and kale leaf, respectively, and about 50-80% of the chlorine was reduced in the initial 10 min appear.

Induction of Apoptotic Cell Death in Human Jurkat T Cells by a Chlorophyll Derivative (Cp-D) Isolated from Actinidia arguta Planchon

  • Park, Youn-Hee;Chun, En-Mi;Bae, Myung-Ae;Seu, Young-Bae;Song, Kyung-Sik;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.27-34
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    • 2000
  • The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

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Early Aneurysm Surgery using Eyebrow Incision for Poor Grade Patients

  • Lee, Chul-Jae;Jeon, Byung-Chan;Kim, Young-Soo;Chun, Tae-Sang;Kim, Nam-Kyu
    • Journal of Korean Neurosurgical Society
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    • v.39 no.2
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    • pp.120-124
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    • 2006
  • Objective : This study is aimed to assess the clinical outcome in early and minimally invasive surgery using incision for the patients with poor grade aneurysm. Methods : The authors retrospectively reviewed all 46 poor grade patients of Hunt and Hess[H-H] grade IV and V who suffered aneurysmal subarachnoid hemorrhage[SAH] between 1999 and 2004. All 35 patients harboring 43 aneurysms who underwent early surgery within 72 hours were included in this study. Clinical outcome was assessed by Glasgow outcome scale[GOS] and compared with that of conventional pterional approach. Results : Twenty four patients were operated with conventional pterional approach and 11 with eyebrow approach within 72 hours after SAH. Seven multiple aneurysm patients harbor 15 aneurysms, Forty one aneurysms were treated with clippings. All 11 patients of eyebrow surgery group[ESG] were in H-H grade IV, 3 in Fisher grade III and 8 in Fisher grade IV. Among 24 patients of pterional approach group[PAG]. 20 were in H-H grade IV and 4 in H-H grade V, 3 were in Fisher grade III and 21 in Fisher grade IV. Overall favorable outcome was achieved in 41.7% and 54.5% in PAG and ESG, respectively. Favorable outcome of H-H grade IV in PAG showed 45.0%. Overall mortality rate was 14.3%. Conclusion : It is concluded that the clinical outcome of early and minimally invasive aneurysmal surgery using eyebrow incision in the selected poor grade aneurysm patients can be compatible with that of conventional pterional surgery.

Evaluation of Mixtures of Certain Market Wastes as Silage

  • Ozkul, H.;Kilic, A.;Polat, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1243-1248
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    • 2011
  • The aim of this study was to determine the usefulness of vegetable wastes as silage for ruminants. Varying amounts of wheat straw (WS), wheat bran (WB) and salt (S) were combined with minced vegetable wastes (VW) during ensilage. Seven different ingredient combinations were investigated viz: 100% VW (Group I, control), 90% VW+9% WS+1% S (Group II), 80% VW+15% WS+4% WB+1% S (Group III), 70% VW+20% WS+9% WB+1% S (Group IV), 90% VW+9% WB+1% S (Group V), 80% VW+15% WB+4% WS+1% S (Group VI) and 70% VW+20% WB+9% WS+1% S (Group VII). The inclusion of straw and bran increased (p<0.01) the DM content of silage. The highest contents of the pure silage were CP (p<0.001), EE (p<0.01) and NFE (p<0.05). NDF contents of VW silage and group V were significantly lower and especially the VW silage was found to have the lowest ADL content (p<0.01). The in vitro ME values of VW silage and bran added silage were higher than other groups (p<0.01). pH, lactic acid and acetic acid values of silage groups were changed between 4.09-4.20, 2.43-3.46% and 0.60-0.86%, respectively. In conclusion, different mixtures of VW have a high ensilage capacity and can serve as an alternative roughage source for ruminants. The addition of 9% bran significantly improved the silage in view of both dry matter content and nutritive value.

EFFECTS OF ACUTE AND SUBACUTE ADMINISTRATION OF COCAINE ON DOPAMINERGIC SYSTEMS IN THE RAT STRIATUM

  • Lim, D.K.;Ho, I.K.
    • Toxicological Research
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    • v.6 no.1
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    • pp.75-88
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    • 1990
  • The characteristics of dopamine uptake, D-1 and D-2 receptors after acute and subacute cocaine administration were determind in striatum from WKY and SHR. Cocaine was administered either acutely (40 mg/kg, s.c.) or twice daily (20 mg/kg, s.c.) for 3 and 7 days in 9-wk old WKY and SHR. Rats were sacrificed 30 min, 2 or 24 h after the single injection and 18 h after the last administration to the subacutely treated group. The changes in dopamine uptake, dopamine uptake sites, D-1 and D-2 receptors were determined using $(^3H)$dopamine, $(^3H)$-GBR-12935, $(^3H)$SCH-23390 and $(^3H)$sulpiride, respectively. In acutely treated rats, significant increases in $V_{max}$of dopamine uptake were observed 30 min after the cocanine injection in both strains without changes in $K_m$ values. The in vitro $IC_{50}$for cocaine was significantly decreased 30 min in WKY and 2 h in SHR. However, that for in vitro GBR-12909 was significantly increased 30 min and 2 h in both strains. Also densities of $(^3H)$-GBR-12935 binding sites were significantly increased 30 min and 2 h without changes in their $K_d$. Significant increases in D-2 receptor density were observed 30 min, 2 or 24 h after acute injection in both strains without changes in their affinities. The density of D-1 receptor was significantly decreased 30 min after the injection in WKY, but not in SHR. In subacutely treated rats, a significant increase in $K_m$ of dopamine uptake was observed in 7-day treated SHR. The in vitro $IC_{50}$fot GBR-12909 was significantly increased in 3-day treated WKY. The density of D-1 receptors was significantly increased in 3- and 7-day treated WKY, but not in SHR. The affinity of both binding sites remained unchanged. The results suggest that cocanine administration alters dopamine uptake, characteristics of dopamine uptake sites and dopamine receptor binding characteristics in rat brain. Furthermore, D-1 and D-2 dopamine receptors appear to be differently regulated.

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국내 연근해 및 환자로부터 분리된 vibrio vulnificus의 세균학적 특징

  • 신광훈;신영학;이종삼
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.15-29
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    • 1992
  • Vibrio vuln$cus has been recognized as a pathogen of septicemia and wound infection, when the organism attacks high-risk persons with a history of hepatic disease. alcohol abuse. diabetes or any debilitative disease. Forty six strains of K vulnzjicus. isolated from 1025 marine specimens from May to Novemver for three years. from 1985 to 1987. were studied for their biochemical properties. growth requirements, serotype and drug susceptibilities. The isolates were different in their various biochemical reactions. Ninety-five percent of isolated strains were able to ferment lactose, while most strains didn't utilize sucrose in their biochemical test, for example ornithine, gelatin and mannitol were quite dit'ferent composition than those described in other reports. It was found that the biochemical test wasn't useful for identifying strain. The type of somatic 0 antiserum was determined in isolates from marine sources and in patients with Vibrio septicemia. In patient isolates. 1-2 group were 24% and 1-4 group were 42%. However. 02 group(33%) were more abundant in isolates from marine sources. Minimal inhibitory concentrations(M1Cs) of chloramphenicol, tetracycline. erythromycin and ampicillin were determinef for V vuln~ficus by broth dilution method. MIC90 was I , 0.25, :! and 4,ug/ml in patient isolates. 1, 0.25, 2 and 2 ,ug/ml in marine isolates. The divalent chelating agent, IDTA. inhibited the growth of V. vuln!'ficus at 6.25 mMlml of MIC90.

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