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Dynamics of Rouleaux Patterns of Red Blood Cells under Pulse Magnetic Field (강한 펄스자기장 자극에 의한 적혈구 연전현상의 활동성 조사)

  • Hwang, Do Guwn
    • Journal of the Korean Magnetics Society
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    • v.27 no.3
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    • pp.92-97
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    • 2017
  • It is widely known that pulsed magnetic field (PMF) is very useful tool to manipulate chemical and physiological processes in human body. The purpose of our study is to observe dynamics of rouleaux patterns of red blood cells (RBC) under PMF. The aggregation of RBCs or rouleaux formation is caused by fibrinogen in blood plasma. The maximum magnetic field intensity is 0.27 T and pulse time of 0.102 msec and pulse repetition rate was 1 Hz. PMF stimulus was applied to the palm of left hand for 5, 10, 15 and 20 min. Live blood analysis was used in vitro in order to quantitatively estimate the velocity of RBC exposed to PMF stimulus. The velocity of stacked-RBC of 10 minute PMF stimulus was increased up to $8{\times}10^{-4}m/sec$, but it decreased rapidly as the time passed. The results of present study have adduced that PMF stimulus on hand provide the improvement of RBC rouleaux formation, increase of RBC's moving velocity as well as low blood viscosity.

Embryonic Developmental Capacity and Pregnancy Rates of Fertilized Oocytes in IVF, ICSI and TESE-ICSI Cycles (IVF, ICSI 또는 TESE-ICSI에서 수정을 유도한 난자의 배아 발생능력 및 임신율)

  • Park, Kee-Sang;Park, Yoon-Kyu;Song, Hai-Bum;Lee, Taek-Hoo;Chun, Sang-Sik
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.3
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    • pp.169-176
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    • 2004
  • Objective: This study was performed to evaluate and compare the embryonic developmental capacity and pregnancy rates in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with ejaculated sperm or testicular sperm cycles. Materials and Methods: Fertilization was examined in the following morning after IVF (group I), ICSI (group II) or TESE-ICSI cycles (group III). Fertilized oocytes were co-cultured with Vero cells until embryo transfer (ET). On day 2 and $5{\sim}7$, grades of embryos (<4- or $\geq$4-cell) and blastocysts (BG1, 2, 3 or early) were evaluated. Clinical pregnancy rate was determined by detecting G-sac with transvaginal ultrasonogram. We analyzed the results by $X^2$ and Student's t-test and considered statistically significant when P value was less than 0.05. Results: Fertilization rate was significantly higher (p<0.05) in group I ($79.0{\pm}21.2%$) than in group II and III ($56.8{\pm}21.6%$ and $36.7{\pm}25.3%$). Cleavage and blastulation rate of group I ($95.8{\pm}13.8%$ and $59.5{\pm}25.3%$) were significantly higher (p<0.05) than those of group III ($83.4{\pm}18.6%$ and $40.4{\pm}36.5%$). Clinical pregnancy rate was significantly higher (p<0.05) in group I and II (40.7% and 41.7%) than that in group III (12.5%). No differences were found in the rates of multiple pregnancy and abortion among three groups. Embryonic implantation rate was higher in group I ($15.1{\pm}20.2%$, p<0.05) and II ($14.7{\pm}20.6%$, NS) than that in group III ($5.1{\pm}15.6%$). However, embryonic implantation rate was increased in ET with blastocyst(s) among three groups. Conclusions: Fertilized oocytes obtained from TESE-ICSI were harder to be successfully cultured to blastocyst stage for 5$\sim$7 days than that from IVF cycles. However, all blastocyst(s) ET increased the embryonic implantation rate equally in IVF, ICSI and TESE-ICSI cycles.

Nuclear Transfer using Human CD59 and IL-18BP Double Transgenic Fetal Fibroblasts in Miniature Pigs

  • Ryu, Junghyun;Kim, Minjeong;Ahn, Jin Seop;Ahn, Kwang Sung;Shim, Hosup
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.1-7
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    • 2016
  • Xenotransplantation involves multiple steps of immune rejection. The present study was designed to produce nuclear transfer embryos, prior to the production of transgenic pigs, using fibroblasts carrying transgenes human complement regulatory protein hCD59 and interleukin-18 binding protein (hIL-18BP) to reduce hyperacute rejection (HAR) and cellular rejection in pig-to-human xenotransplantation. In addition to the hCD59-mediated reduction of HAR, hIL-18BP may prevent cellular rejection by inhibiting the activation of natural killer cells, activated T-cell proliferation, and induction of $IFN-{\gamma}$. Transgene construct including hCD59 and ILI-18BP was introduced into miniature pig fetal fibroblasts. After antibiotic selection of double transgenic fibroblasts, integration of the transgene was screened by PCR, and the transgene expression was confirmed by RT-PCR. Treatment of human serum did not affect the survival of double-transgenic fibroblasts, whereas the treatment significantly reduced the survival of non-transgenic fibroblasts (p<0.01), suggesting alleviation of HAR. Among 337 reconstituted oocytes produced by nuclear transfer using the double transgenic fibroblasts, 28 (15.3%) developed to the blastocyst stage. Analysis of individual embryos indicated that 53.6% (15/28) of embryos contained the transgene. The result of the present study demonstrates the resistance of hCD59 and IL-18BP double-transgenic fibroblasts against HAR, and the usefulness of the transgenic approach may be predicted by RT-PCR and cytolytic assessment prior to actual production of transgenic pigs. Further study on the transfer of these embryos to surrogates may produce transgenic clone miniature pigs expressing hCD59 and hIL-18BP for xenotransplantation.

In Vitro Maturation, Fertilization and Development of Mouse Oocytes Derived from In Vitro Grown Preantyal Follicles (체외성장된 Preantral Follicle에서 유래된 생쥐난자의 체외성숙, 수정 및 발달)

  • Kim, D.H.;Lee, H.J.;K.S. Chung;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.133-142
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    • 2002
  • The objective of this study was to examine maturation, fertilization and developmental rate of the in vitro-grown mouse oocytes, and to compare these results with those of oocytes grown and matured in vivo. The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After in vitro growth and maturation, 72.5 % of oocytes grown in vitro produced polar body which can be comparable to in vivo growth (70.5 %). However, the mean oocyte diameter of the in vitro group (69.6$\pm$2.1$\mu$m) was smaller than that of the in vivo group (73.3$\pm$3.0$\mu$m). The fertilization rate was significantly lower (p<0.05) in the in vitro group (76.5%) than in the in vivo group (90.2%), however, there was no difference in the percentage of monospermic and polyspermic oocytes between two groups. The capacities of in vitro grown ova to cleave and develop to blastocyst were (57.8 and 14.4%, respectively) significantly lower (p<0.001) than those of the in vivo counterpart (84.4 and 56.6%, respectively). Moreover, the mean number of cells per blastocyst was significantly lower (p<0.05) in the in vitro group (39.0$\pm$10.8) than in the in vivo group (60.5$\pm$12.5). Live young were produced from transferred 2-cell embryos derived from in vitro-grown and matured oocytes. In conclusion, the results show that in vitro-grown oocytes did not achieve the developmental capacity of in vitro-grown oocytes.

Sex Determination in Somatic and Embryonic Cells of the Pig by FISH and PCR (FISH와 PCR에 의한 돼지 체세포 및 배아세포의 성 판정)

  • Chung, Y.;Jeon, J.T.;Kim, K.D.;Lee, S.H.;Hong, K.C.
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.323-331
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    • 1996
  • Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.

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Studies on the Glucose Isomerizing Enzyme -Part II. Some Properties of the Glucose Isomerizing Enzyme of K-17 strain- (포도당(葡萄糖) 이성화(異性化) 효소(酵素)에 관(關)한 연구(硏究) -제2보(第二報); 분리선정(分離選定)된 균주효소(菌株酵素)의 성질(性質)에 대(對)해서-)

  • Seu, J.H.;Kim, C.K.;Ki, W.K.;Rhee, I.K.;Kwon, T.J.;Woo, D.L.
    • Applied Biological Chemistry
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    • v.11
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    • pp.49-53
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    • 1969
  • Some properties of glucose isomerizing enzyme which produced by the strain K-17 in xylose containing nutrient broth medium were investigated. The optimal pH for enzyme reaction was indicated about 7.2 and optimal temperature was about $75^{\circ}C$. The same optimal temprature was indicated by both cell free extract and acetone dried cells using as enzyme. The glucose isomerizing enzyme from strain K-17 was not inhibited by the high concentration of substrate even in a suturated glucose solution, but most enzyme was inactivated by the heat treatment at $80^{\circ}C$. The maximum fructose forming ratio from glucose was about 50 percents.

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The Changes of Quality and Microflora during the Preservation of Korean Takju (탁주(濁酒) 보존중(保存中) 품질변화(品質變化)와 미생물군(微生物群) 소장(消長))

  • Jung, Ji-Heun;Jung, Soon-Teck
    • Applied Biological Chemistry
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    • v.28 no.4
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    • pp.252-260
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    • 1985
  • The changes of the qualities and the components of 5 different Takju (Korean wine) in various stroage temperatures were evaluated. Commercial Takju didn't change in acidity and alcoholic degree at $5{\sim}10^{\circ}C$ for 36 hours. But at $30{\sim}35^{\circ}C$ the alcohollic degree has inversed to $5.0{\sim}7.0$. As time went by, pH nearly went down to pH 3.7 and went up back over 4.0. Amino acidity was risen continuously during the stroage terms and the change of reducing sugar was irregular. The number of vegetable cell of yeast in commercial Takju at $30{\sim}35^{\circ}C$ after 18 hours was $2.4{\times}10^8$, at $5{\sim}10^{\circ}C$ after 36 hours $2.4{\times}10^8$, and each of them was reduced below $1{\times}10^8$ after exceeding the limit of shelf-life. Number of general bacteria was increased suddenly in a high temperature at the first and it was decreased afterwards, but the tendency of increase and decrease was slow in a low temperature. It was possible to keep the quality for 5 days at $5{\sim}10^{\circ}C$ in case of commercial Takju and the shelf-life of pasteurized Takju was extended over 7 days.

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Differential Gene Expression in the Bovine Transgenic Nuclear Trasnsfer Embryos (소 형질전환 복제란의 유전자 이상발현 규명)

  • Cho, Jong-Ki;Song, Bong-Seok;Yong, Hwan-Yul;Lee, Doo-Soo;Koo, Deok-Bon;Lee, Kyung-Kwang;Shin, Sang-Tae
    • Journal of Veterinary Clinics
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    • v.24 no.3
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    • pp.295-299
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    • 2007
  • The detrimental effects of gene transfection on embryo development and the molecular mechanism behind the differential expression of genes related to early embryo development were assessed in the production of transgenic cow embryos through somatic cell nuclear transfer (NT). Parthenogenetic, IVF, and transgenic NT embryos derived from ${\alpha}_1$-antitrypsin transfected ear fibroblast cells was produced. To investigate the molecular mechanism behind lower developmental competence of transgenic NT embryos, the differential mRNA expression of three genes ($IFN-{\tau}$, Oct4, Fgf4) in the 3 types of embryo (Parthenogenetic, IVF, transgenic NT) was examined. RNA was extracted from ten blastocysts derived from 3 types of embryos and reverse-transcripted for synthesis of the first cDNA. The quantification of 3 gene transcripts ($IFN-{\tau}$, Oct4, and Fgf4) was carried out in three replicate by quantitative real-time reverse transcriptase PCR. Expression level of $IFN-{\tau}$ mRNA was significantly higher in transgenic NT embryos than parthenogenetic and IVF embryos (P<0.05). However, expression level of Oct4 and Fgf4 of transgenic NT embryos was significantly lower than IVF embryos (P<0.05). Altered levels of these three mRNA transcripts may explain some of the embryonic/fetal/neonatal abnormalities observed in offspring from transgenic NT embryos.

Assessment of Pubertal Development to Parabens-induced Estrogenic Effect in Male Mice (파라벤류가 수컷 성 성숙에 미치는 시험연구)

  • Kim, Sun-Jung;Hwang, Jae-Woong;Park, Jung-Ran;Lee, Seong-Hun;Lee, Yeong-Geon;Chung, Ji-Hye;Jeong, Yun-Hyeok;Lee, Soo-Jin;Jung, Ji-Won;Jung, Ji-Youn;Lee, Yong-Soon;Kang, Kyung-Sun
    • Journal of Food Hygiene and Safety
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    • v.21 no.4
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    • pp.197-203
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    • 2006
  • Parabens are most wildly used in food, cosmetics and pharmaceutic products as preservatives caused of safety and cheap. we had examined that paraben had estrogenic activity through the in vivo and in vitro experiments in last year. We demonstrated that most of parabens(ethyl, butyl, propyl, isobutyl, isopropyl) increased significantly uterus weight as well as induced proliferation of MCF-7 cell and binding of estrogen receptor as endocrine disrupter compounds. In this study, we evaluated that whether parabens have effect on male reproductive system or not. the male rats were administrated parabens by oral injection then examined separation of preputial day for $PND23\simPND52$. As the results, most parabens delayed pubertal development compare to control group. The separation of preputial day of Butyl and Propyl parabens at high concentration were PND 44 days and PND 45days compared to control group as PND 40 days. Even though, parabens as endocrine disrupter wildly spread in food, cosmetics and pharmaceutic products, we didn't have the safe guideline. In abroad, they are re-evaluating safety assessment for parabens. In conclusion, parabens delayed pubertal development in juvenile parabens are consider as endocrine disrupter chemicals.

Effect of Irrigation Frequency on Growth of Tomato Plug Seedlings in Media Containing Cellular Glass Foam and Peatmoss (Cellular Glass Foam과 Peatmoss를 함유한 혼합배지에서 관수빈도가 토마토 플러그묘의 생육에 미치는 영향)

  • 박정호;정병룡
    • Journal of Bio-Environment Control
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    • v.12 no.1
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    • pp.7-11
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    • 2003
  • Cellular glass foam (CGF), the reprocessed glass, has a possibility to be used as a medium component in plug culture of horticultural crops due to the its excellent air and water permeability as comparable to perlite. An experiment was conducted to investigate growth of plug seedlings of Lycopersicum esculentum 'Segye' as influenced by irrigation frequency in various medium combinations of CGF (2.0-4.0 mm particle size) and peatmoss. Seeds were sown in 200-cell plug trays, filled with mixtures of CGF and peatmoss either at 33:67 or 25:75 (%. v/v) and were germinated on a fogged propagation bed. The irrigation frequencies used were one, two or three times per every two days. A commercial plug medium (Tosilee medium) was used as the control, and the irrigation frequency in the control was one time per day. Growth of seedlings, and medium pH and EC were measured at 33 days after sowing. The medium composition had little influence on overall growth of seedlings. Irrigation frequency very significant affected number of leaves, leaf area, chlorophyll concentration, fresh and dry weights of shoots and roots, and dry matter. Growth of seedlings was the greatest with the highest irrigation frequency in the 25% CGF+75% peatmoss mixture.