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Detection of Human Taurine Transporter and Production of Monoclonal Antibody

  • An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.102-102
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    • 2001
  • Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

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A Survey on the Immune Status and Productivity of Vaccinated Poultry Flocks against Newcastle Disease in the Epizootic Area (뉴캣슬병백신 접종계군에 있어서 면역상태와 ND 발생시 닭의 생산성에 미치는 영향)

  • 박근식;김선중
    • Korean Journal of Poultry Science
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    • v.11 no.1
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    • pp.49-64
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    • 1984
  • During the 1978-79 Newcastle disease (ND) epizootic period, a detailed survey was conducted on the five representative farms which had been following one of the recommended vaccination programs. When the disease broke out during laying period, clinical symptoms were mild to moderate respiratory distress and greenish diarrhea. Affected flocks experienced weekly mortality from less than 1% to 17%. Egg production returned to normal 18 to 36 days after the initial signs appeared although some flocks never returned to normal. On postmortem examination,, most affected chickens showed severe hemorrhagic lesions in the duodenum, hematoma on ova, and heavy fat accmulation on various visceral organs. Most of the NO affected flocks had geometric mean hemagglutination inhibition antibody(HIA) titers of 7 log$_2$ or higher two to three weeks after the appearance of clinical signs. These HIA titers were at least 16-fold higher than those before infection. Flock mean HIA titers before infection were usually lower than 3 log$_2$. Severity of clinical signs and anamnestic antibody response were maximum in the flocks whose vaccination immunity was insufficient or waned considerably. Observations showed that even young birds, if properly vaccinated, could get effective protection from field ND exposure.

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Clinical Analysis of C.N.S. Cysticercosis (뇌 낭미충증에 관한 임상적 연구)

  • Cho, Soo-Ho;Moon, Choong-Bae;Choi, Byung-Yearn
    • Journal of Yeungnam Medical Science
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    • v.1 no.1
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    • pp.25-34
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    • 1984
  • During last 10 years we experienced 25 cases of C.N.S. cysticercosis. Now clinical analysis and evaluation of our cases were made and the results are followings: 1. Prevalence in man and women are nearly same and about 70% of cases are distributed between 20-50 years old. 2. According to Nieto's classification, mostly are ventricular (44%) and parenchymal type (36%). 3. Clinical manifestations were IICP (92%), focal neurological deficits (68%), seizure (48%), altered mental status (36%) and others listed on table 7. 4. In ventricular type, IICP and cerebellar dysfuction signs were predominated but seizure and focal neurological deficits were commonly seen in parenchymal type. 5. Subcutaneous cysticercus nodules were palpated in 32 % of cases. 6. Positive stool ova was observed in 29% of cases. 7. Radiologic studies revealed as followings: 16 % of cases showed abnormal findings on plain film, 84% on angiography, 94% on ventriculography and 100% on computed tomography and myelography. Computed tomography looks like most helpful diagnostic method for C.N.S. cysticercosis, they ususally revealed lucent cystic lesion, hydrocephalic findings and contrast enhancement. 8. Suboccipital craniectomy, craniotomy with removal of parenchymal cyst or laminectomy were done according to location and types of lesion. 72 % of operated cases revealed good results and motarlity was 4 % of cases.

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Parasitic Infection Among Inhabitants in Urban Area of Seoul (서울지역(地域)에 있어서 기생충(寄生虫) 감염(感染) 조사성적(調査成績))

  • Lee, Joon-Sang;Joo, Kyung-Hwan;Chung, Young-Suk;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.10 no.1
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    • pp.36-41
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    • 1985
  • To evaluate the parasitic infection rates among inhabitants in urban area of Seoul, Korea, a total of 5,275 stool and anal swab specimens was obtained from 2,527 male and Z,748 female living in 24 Dongs of 17 Kus. Cases were sampled randomely to represent 1,000 inhabitants refereed to the census in 1980. The methods employed were formalin ether technique to detect helminth ova and protozoan cysts and scotch tape anal swab technique to detect eggs of Enterobius vermicularis. The results obtained are as follows ; 1) Total positive rate of helminthes was 23.5% among 5,275 (male 22.5% and female 24.5%) specimens. Nine kinds of the helminthes were detected and infection rates of each helminth were; Ascaris lumbricoides 4.1%, Hookworm 0.1%, Trichuris trichiura 11.1%, Trichostrongylus orientalis 0.1%. Clonorchis sinensis 1.2%, Metagonimus yokogawai 0.1%, Taenia spp. 0.2%, Hymenolepis nana 0.3 and Enterobius vermicularis 9.6%. 2) Total positive rate of intestinal protozoa was 1.4%. Four kinds of the protozoan cysts were detected and the infection rates of each protozoa were; Entamoeba histolytica 0.15%, E. coli 0.4%, Endolimax nana 0.04% and Giardia lamblia 0.89%. 3) No significant differences in the parasitic infection rate by sex was noticed although male group showed lower infection rate than female group. However the incidence of C. sinensis, M. yokogawai and Taenia spp. was twice as much in female group as in male group. 4) No difference in the infection rate by age was found although E. vermicularis positive rate was highest in "0~9 years group" by 2.1% and C. sinensis infection rate was higher in over "30~39 years group". The parasitic infection rate of the present study was significantly lower than those of previous reports in Seoul area and other provinces.

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Production of In-Vivo Embryos by Superovulation and Result of Transfer with Fresh or Frozen Embryos for Hanwoo and Holstein Cattle (한우 및 젖소에서 과배란 처리를 이용한 체내수정란 생산과 신선 및 동결 수정란 이식 결과)

  • Kim Y. J.;Song J. W.;Seo S. H.;Jeong K. N.;Kim Y. S.;Lee H. R.;Shin D. S.;Jo S. W.;Kim S. H.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.209-218
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    • 2004
  • To investigate the feasibility of embryo transfer technology to promote productivity of cattle, 36 cows(18 Holstein, 18 Hanwoo) were superovulated. Fresh embryos were transferred to 25 recipients(14 Holstein, 11 Hanwoo), whereas frozen embryos were transferred to 17 recipients(10 Holstein, 7 Hanwoo). Two embryos were transferred at a time to 13 recipients(9 Holstein, 4 Hanwoo) to produce twin calves. 1. 75.0% of donor cattle were reacted to hormonal treatment far superovulation. 2. The rate of embryo recovery by non-surgical method for Holstein and Hanwoo was 90.4 and 95.8% in comparison with numbers of corpus luteum. 3. Of all the ova collected non-surgically, the rate of viable blastocyst was 66.4% and the rate of transferrable blastocysts was 48.6%. 4. The rate of embryo collection by one-catheter method was 75.0%. 5. The rate of pregnancy/delivery following embryo transfer with fresh embryos was 60.0%. 6. The rate of pregnancy/delivery following embryo transfer with frozen embryos was 35.3%. 7. In embryo transfer to produce twin calves, the rate of pregnancy/delivery was 28.6% with fresh embryos and 16.7% with frozen embryos.

Inhibitory effects of Paridis Rhizoma in the activation and proliferation of eosinophils: implications on its regulatory roles for asthma (rmIL-5로 유도된 호산구의 활성화 및 성장에서 중루의 천식반응 억제효과)

  • Sin, Mi-Kyung;Kil, Ki-Jeong;Lee, Young-Chul;Kim, Jin-Suk;Seo, Young-Bae;Roh, Seong-Soo
    • The Korea Journal of Herbology
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    • v.20 no.2
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    • pp.159-169
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    • 2005
  • Objectives : This study was carried out for the purpose of knowing the effect from anti-arthma action of the abstraction from a extract of Paridis Rhizoma(EPR). In order to know what the effect of controlling an abstraction from Paridis Rhizoma. and about the expression of B cells and Ig E cells, mast cells it was necessary for it to be activated by ovalbumin. Methods : In order to know what the effect was on the organization of cytokine gene expression from The increase and divorce of the B cells and allergic acting by EPR, we found it necessary to examine the BALF. At the same time, as we examined the histamine release by ELISA method, we also examined the effect of EPR. Results : EPR at $100\;{\mu}g/ml$, the highest concentration examined did not have any cytotoxic effects on mLFCs. In FACS analysis, number of granulocyte/lymphocyte, $CD3e^+/CCR3^+,\;CD4^+\;and\;CD23^+/B220^+$ in asthma-induced lung cells were significantly decreased by EPR treatment compared to the control group. In RT-PCR analysis, mRNA expression for CCR3, eotaxin and histamine in asthma-induced lung cells, which was induced by rIL-3 plus rmIL-5 treatments, was significantly decreased by EPR treatment. In ELISA analysis, production levels of IL-4, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by EPR treatment. EPR treatments significantly inhibited the proliferation of eosinohils prepared from asthma-induced mouse lung tissues compared to the non-EPR treated control cells. Immunohistochemical analysis revealed that EPR treatment significantly decreased the levels of eosipnphil activation compared to non-treated cells. Conclusion : The present data suggested that Paridis Rhizoma may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Paridis Rhizoma.

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A Study on the Use and Extention of Native Species of Trees through the Field Survey of Present Status of Landscape -The Case Study on 38 Elementary, Middle and Schools in Iri city- (학교 조경수목 직재현황 조사를 통한 향토수종 이용 확대에 관한 연구 -이리시 초.中고등학교 38개교를 대상으로-)

  • 박영수;심경구
    • Journal of the Korean Institute of Landscape Architecture
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    • v.21 no.1
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    • pp.31-50
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    • 1993
  • This study was conducted to investigate the plants status of the woody landscape plants species of 38 elementary, middle, and high school in Iri city, and to obtain some fundamental information for development and use of native species of trees. The results are summarized as follows: 1. Landscape plants species of 38 elementary, middle and high school in Iri city was 55 families, 164 species, and 85, 045 trees and the average of landscape plants species of 17 elementary schools was 50.5 families, 3, 010 trees, on the other hand, middle and high school was 42.4 families, 1, 609 trees. 2. Plants species by type was Deciduous Broadleaved Tree 32.9%, Deciduous Broadleaved Shrub 25.6%, Evergreen Coniferous Tree 14.6%, Evergreen Broadleaved Shrub 9.7%, Vine 4.8%. Evergreen Trees on the other hand, according to the utilization rate 'others' was 32.2%. 3. The rate of plants species of the Evergreen Trees to the Deciduous (except Vine and 'others') was 34:66. on the other hand, rate of quantity was 50.5:49.5 and the rate of the trees to the shrubs(except Vine and 'others') was 58:42. to the plants species and to the quantity of the number of trees was 25.9:74.1. 4. Plants species of fruit plants of 38 elementary, middle and high school in Iri city was 31 species to the 164 species. The rate of frequency was Poncirus trifoliata 27.8%, Lycium chinense 25.1%, Ginkgo biloba 16.7%, to the quantity of the number of trees, fruit plants was 2, 605 trees to the 85, 045 trees and the frequency was 3%. 5. School trees and flowers of 38 elementary, middle and high school in Iri city were appointed 17 elementary school, on the other hand, middle and high schools were only 4 middle and high schools. Plants species of the best of frequency school tree was Zelkova serrata and school flower was Rosa spp.. 6. The rate of the native species to the introduced species was 73.8:26.2. on the other hand, the rate of quantity by the number of trees was 84.7:15.3 to the type, the rate of frequency of plants of native species Evergreen trees was Thuja orientalis, deciduous was Ginkgo biloba, Vine was Wisteria floribunda and 'others' was Sasa borealis. on the other hand, to the introduced species, Evergreen Trees was Juniperus chinensis. Deciduous Trees was Metasequoia glyptostroboides, and Vine was Vitis labrusca and 'others' was Phyllostachys pubescens. 7. As the fundament of the results of above the optimum selecting of woody landscape plants according to native species of trees was 29 families, 58 species. on the basis of 30 points, 28 points was 8 species Ginkgo biloba, Ulmus davidiana var. japonica, Parthenocissus tricuspidata, Elaeagnus umbellata, Fraxinus rhynchophylla, Weigela subsessilis, Koelreuteria paniculata, Lonicera japonica, 26 points was 27 species Thuja orientalis, Zelk ova serrata, Euonymus japonica etc., and 24 points was 23 species Populus tomen tiglandulosa, Juglans sinensis, Sophora japonica etc..

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Studies on In Vitro Fertilization and Development of Bovine Follicular Oocytes Matured In Vitro III. Effect of Anti-Cumulus Cell Antibody on In Vitro Fertilization and Development of Bovine Follicular Oocytes Matured In Vitro (체외성숙 우난포란의 체외수정과 발달에 관한 연구 III. 항난구세포 항체가 체외성숙 우난포란의 체외수정과 발달에 미치는 영향)

  • 박세필;김은영;정형민;고대환;김종배;정길생
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.101-106
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    • 1990
  • These mxperiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro fertilization and following development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2~6mm of diameter. Bovine oocytes were matured in vitro for 24~26hrs in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$ and subsequently cultured in medium containing cumulus cell antibody for 1 hour. The medium used for maturation was TCM-199 supplemented with hormones, pyruvate, FCS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2~3 hrs in BO solution 10~15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after insemination the eggs were transferred to TCM-199 supplemented with FCS(10%) and then cultured for 7 days. The results obtained in these experiments were summarized as follows : 1. When the follicular oocytes matured in vitro were treated with antibody to intact cumulus cells, the fertilization rate of cumulus intact and removed oocytes was ranged to 45.0 to 53.7%. These value is slightly lower than that(64.3%) of follicular oocytes not treated with the antibody, and increased frequency of both male and female pronuclear formation was found in cumulus intact oocytes cultred in medium without the antibody(p<0.05). 2. The fertilization rate of cumulus intact and removed oocytes treated with antibody to solubilized cumulus cells was ranged 45.0 to 52.5%, significantly lowre than that(62.8%) of oocytes cultured in antibody free medium, and increased frequency of ova with male and female pronuclei was found when cumulus cells were present(p<0.05). 3. The rates of cumulus cell intact and removed oocytes developed to 8-, 16-cell and morula or blastocyst after treatment of intact and solubilized cumulus cell antibody were ranged 7.1 to 14.5, 2.9 to 5.9 and 1.5 to 2.9%, respectively, slightly lower than 18.6, 10.0 and 8.6% of cumulus intact oocytes cultured in medium without the antibody. The results of this stduy indicate that cumulus cells promote not only normal fertilization with proper pronuclear formation, but embryo development and that the beneficial effect of cumulus cell to the pronuclear formation and embryo development is blocked by the action of antibody to cumulus cell.

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The effect of Jackyakgamcho-tang on Follicular Maturation and Estrogen Production in the Immature Rat (작약감초탕이 미성숙 흰쥐에서 난포성숙 및 Estrogen생성에 미치는 효과)

  • Jung, Ki-Kyung;Kang, Seog-Youn;Kim, Tae-Gyun;Kim, Chang-Ok;Moon, A-Ree;Ryu, Kyung-Za;Lee, Song-Deuk;Ryeu, Hang-Mook;Kim, Seung-Hee
    • Korean Journal of Pharmacognosy
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    • v.28 no.3
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    • pp.104-111
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    • 1997
  • The traditional herbal medicine, Jackyakgamcho-tang(JGT), was reported to decrease serum testosterone levels and make pregnancy possibel in anovulatory woman and rat. JGT contains Paeoniae Radix(PR) and Glycyrrhizae Radix(GR) in equal amount. This study was designed to investigate the effect of JGT and its components(PR, GR, paeoniflorin and glycyrrhizin) on uterine and ovarian responses, follicular development, and estrogen secretion in the immature rat. The samples(water extracts of JGT, PR, GR; pure compound of paeoniflorin and glycyrrhizin) were administered orally to rats from the 21th day of age to the 28th or 30th days of age for 7 or 9 days. JGT(400mg/kg) and PR(100mg/kg, 200mg/kg) treatments significantly increased serum estradiol above levels in control rats, but both GR and glycyrrhizin had no effect on this parameter. Gross observation and histological analysis revealed that an increased number of growing follicules was observed in the ovaries of JGT and PR treated rat. However the lutenized follicles and ova present in the oviducts were not observed in all rats except one treated with estrogen as a positive control. These results indicate that JGT stimulates the estrogen production and follicular maturation in the immature rat and PR is the main component to induce such reaction.

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Effect of the Addition of "Royal Jelly" on Post-thaw Viability and Longevity of Canine Spermatozoa (Royal Jelly 첨가가 동결융해 후 개 정자의 활력도 및 생존성에 미치는 영향)

  • 공일근;조성균
    • Journal of Embryo Transfer
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    • v.16 no.1
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    • pp.53-60
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    • 2001
  • This study was conducted to evaluate whether \"Royal jelly\" (RJ) added to Tris-buffer dilute contributed to supporting post-thaw viability and longevity of frozen canine spermatozoa. Two Japanese spitzs (2 to 4 years of age) were used as a semen donor. Semen was collected by manual masturbation and separated into 3 fractions. Only the sperm-rich fraction having sperm motility of more than 70%, containing sperm concentration of 2~4$\times$10$^{8}$ cells/ml and having dead or abnormal spermatozoa of less than 15% was used for the experiment. Each ejaculated semen was centrifuged at 400 $\times$ g for 5 min and then diluted in a Tris-buffer supplemented with 20 ml egg yolk (Ext I), 4% glycero1 and 1% Equex STM Paste (Ext II) or g1ycero1, Equex STM paste and RJ of various concentrations (Ext II-RJ). After freezing and thawing, viability of spermatozoa in Ext II -RJ containing 1% RJ immediately after thawing (67.5$\pm$9.6) was significantly lower than that of Ext II , Ext II -RJ containing 0.01 or 0.1% RJ (77.5$\pm$12.5, 78.7$\pm$8.2 and 80.0$\pm$6.3). However, Ext II-RJ containing 0.1% RJ yielded higher viability than Ext II, Ext II-RJ containing 0.01% at or 1% 1 h after thawing (69.5$\pm$8.1 vs. 55.0$\pm$12.9, 57.5$\pm$9.6 and 41.5$\pm$12.6; P<0.05). At 1 h after thawing, the viability of spermatozoa thawed in 7$0^{\circ}C$ (68.8$\pm$12.5) was significantly higher than that of spermatozoa thawed in 38$^{\circ}C$ (48.8$\pm$16.3), although there was no difference in the viability between both groups immediately after thawing (77.5$\pm$9.6 and 81.3$\pm$8.1). Post-thaw viability and longevity of post-thaw spermatozoa in Ext II-RJ containing 0.1% RJ was higher in those in Ext II at 1 h (65.0$\pm$12.9 vs. 42.5$\pm$12.6), 2 h (52.5$\pm$12.6 vs. 27.5$\pm$17.1) and 3 h (40.0$\pm$14.1 vs. 20.0$\pm$12.1) after thawing. These results indicated that addition of 0.1% af to Tris-buffer enhanced post-thaw viability and longevity of canine spermatozoa and this additive can be used for increasing the possibility of collision between spermatozoa and ova during insemination.emination.

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