• Title/Summary/Keyword: P19 cells

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Early Mouse and Human Embryonic Development in vitro by Co-culture with Human Oviduct Epithelial Cells (인간 난관 상피세포와의 공동배양이 생쥐와 인간수정란의 체외발달에 미치는 영향에 관한 연구)

  • Ko, J.J.;Chung, M.K.;Do, B.R.;Oum, K.B.;Yoon, T.K.;Cha, K.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.2
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    • pp.133-141
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    • 1992
  • We examined effects of co-culture with human oviduct epithelial cells (HOEC) on the development of mouse and human embryos from early embryonic· stage to late morula or blastocyst stage (LM or B). In human, embryos were transferred and pregnancy rate was investigated. The HOEC, collected from surgically removed fallopian tube, were cultured in medium-199 supplemented with 20 % fetal cord serum (FCS). The HOEC were characterized by using immunocytochemical staining with anticytokeratin antibody and then used for cultures of mouse and human embryos. Results obtained from co-culture system were as follows. Development rate of mouse embryos was improved by co-culture system at late developmental stage (p<0.025). Human supernumerary embryos remained after transfer, unsuitable for freezing because of their poor quality, were co-cultured for 72hrs. Co-culture (78.79%) or conditioned medium (78.26%) system improved the developmemt rate, significantly, in comparision with control (11.11%)(p<0.00l). Co-cultured (85.71%) human zygotes for 24hrs showed the better development rate in comparision with control (50.00%) (p<0.01). When we transferred embryos cultured with the HOEC to patients, we obtained one pregnancy. Co-cultured human zygotes for 24hrs showed the better quality and viability for the replacement in comparision with control (p<0.01). In addition, improved pregnancy rate was obtained. Our results suggest that the co-culture system can rescue early degenerating embryos by improving early development and yield a resonable number of blastocyst for the appropriate replacement. The effect provided by cultured HOEC is not species specific for the development of embryos and it can be used to overcome in vitro blocks for the development. And also the co-culture system offers the possibility to freeze embryos at blastocyst stage which is more sucessful stage for the freezing. The HOEC monolayer may provide some stimulus via specific factor, which is unknown, to the development of embryos. Our results showed that the co-culture system with HOEC can be an alternative to conventional culture system.

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Effects of Herba Taraxaci Herbal-Acupuncture on Adjuvant Arthritis in Rats (포공영(蒲公英) 약침(藥鍼)이 Rat의 Adjuvant 관절염(關節炎)에 미치는 영향(影響))

  • Ha, Ji-Yong
    • Journal of Pharmacopuncture
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    • v.2 no.1 s.2
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    • pp.135-152
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    • 1999
  • To investigate effects of Taraxaci herbal-acupuncture on Adjuvant Arthritis in rats, the edema rate, the number of WBC, the quantity of total protein, albumin and globulin in the blood serum and histological test of the muscular tissue were measured in the arthritis part. 1. After elicitating arthritis of Sprague Dawely(SD) rats by injection of Freund's complets adjuvant for 2 weeks, saline was injected for the Exp. I group and Taraxaci herbal-acupuncture was injected for the Exp. II group during 30days. Selected point was $ST_{35}$ in both the groups. And then the volume of the paw were checked. The volume of paw was $0.84{\pm}0.14mm$ in the Exp. I group and $0.38{\pm}0.17mm$ in the Exp. II group, the swelling of the paw was restricted significantly in the Exp. II group(p<0.05) 2. The number of WBC was $10.34{\pm}0.14(10^3/ml)$ in the normal group and $37.47{\pm}5.46(10^3/ml)$ in the Exp. I group. It was $20.39{\pm}4.23(10^3/ml)$ in the Exp. II group. This fact showed that the group Exp. II was more effective than the Exp. I group in the treatment of arthritis(p<0.05) 3. The content of the total protein in the blood serum was $6.14{\pm}0.43g/dl$ in the normal group, $7.95{\pm}0.94g/dl$ in the Exp. I group, and $6.38{\pm}1.75g/dl$ in the Exp. II group. This fact showed that the group Exp. II was more effective than the Exp. I group in the treatment of arthritis(p<0.05) 4. The contents of albumin in the blood serum was $2.94{\pm}0.13g/dl$ in the normal group, $2.01{\pm}0.48g/dl$ in the Exp. I group. and $2.71{\pm}0.34g/dl$ in the Exp. II group. This fact showed that the group Exp. II was more effective than the Exp. I group in the treatment of arthritis(p<0.05) 5. The contents of globulin in the blood serum was $3.19{\pm}0.48g/dl$ in the normal group, $4.70{\pm}1.26g/dl$ in the Exp. I group. and $3.67{\pm}0.56g/dl$ in the Exp. II group. There was no significance in the serum globulin between Exp. II group and Exp. I group from the stastical analysis 6. In histological finding, because of severe inflammatory reaction, remarkably irregular tissue and large amount of inflammatory cells were found in the Exp. I group. But the Exp. II group showed small amount of inflammatory cells, the refrained inflammatory state and even recovering state. From these results, it is showed Taraxaci herbal-acupuncture refrain inflammatory reaction and muscular tissue necrosis in SD rats paw were induced by Freund's complete adjuvant

Effects of Nutritional Interventions based on Enteral Nutrition Guidelines for Neurosurgical Intensive Care Unit Patients (신경외과 중환자실 환자의 경장영양 지침서 적용에 따른 영양적 중재 효과)

  • Han, Jung-Hee;Kim, Young Ran;Lee, Young Min;Kim, Ae Lee;Kim, Ho Jin;Jeong, Young-Gyun;Ryu, Jeong Ha
    • Quality Improvement in Health Care
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    • v.17 no.1
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    • pp.79-88
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    • 2011
  • Background : Evidence-based guidelines are now used for enteral nutrition(EN) in neurosurgical intensive care unit patients who mostly depend on EN. This study compared and analyzed the nutritive conditions of patients before and after they underwent guideline based nutritional interventions in order to determine whether using these guidelines improved their calorie supply. Methods : Data on the patients' nutritional requirements, maximum calorie supply through EN, serum albumin level, and total lymphocyte count were collected and analyzed using SAS version 9.1.3. All the statistical analyses were performed at a significance level of P<0.05. Result : The maximum calorie supply through EN was $923.1{\pm}359.7$ kcal before the intervention and $1254.4{\pm}196.3$ kcal after the intervention; this difference was statistically significant(P<0.05). The ratio of nutritional requirements to maximum calorie supply through EN was $55.5{\pm}22.4%$ and $74.2{\pm}13.9%$ before and after the intervention, respectively; this difference was statistically significant(P<0.05). This indicates a 19% increase in the ratio after the nutritional intervention. The serum albumin level also significantly increased from $2.7{\pm}0.6g/dL$ before the intervention to $3.2{\pm}0.4g/dL$ after the intervention(P<0.05). The total lymphocyte count slightly increased from $1267.7{\pm}728.2cells/mm^3$ before the intervention to $1801.9{\pm}1211.5cells/mm^3$ after the intervention; this difference was not statistically significant. Conclusion : The results showed that using the evidence-based feeding guidelines for interventions increased the calorie supply and improved the patients' nutritive conditions from moderate malnutrition to mild malnutrition.

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Effect of Mori Folium. on the anti-allergic inflammatory response (상엽(桑葉)이 항알러지 염증반응에 미치는 영향)

  • Lee, Jin-Yong;Kim, Deog-Gon;Cho, Kyu-Seok
    • The Journal of Pediatrics of Korean Medicine
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    • v.19 no.1
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    • pp.185-201
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    • 2005
  • Objective : This experimental study was performed to examine the anti-allergic infiammatory effects of Mori Folium. Method : Macrophage 264.7 cells were pretreated for 1hour with Sangyup. After pretreatment, macrophage were incubated with lipopolysaccharide(LPS) $100ng/m{\ell}$ 12h and media collected and $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, IL-10 concentrations in supernatants were measured each by Enzyme linked immuno-sorbent assay. Sangyup were used $1mg/m{\ell}$, $500{\mu}g/m{\ell}$, $250{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $50{\mu}g/m{\ell}$. Hydrocortisones were used $10^{-4}M,\;10^{-5}M,\;10^{-6}M,\;10^{-6}M,\;10^{-6}M$. Results : Macrophage 264.7 cells were pretreated with Hydrocortisones and Sangyup. After pretreatment, macrophage were incubated with lipopolysaccharide(LPS). To investigate the anti-inflammatory effect of Sangyup, we measured the amount of cytokines, and the results are as follows; 1. Mori Folium. showed statistically significant inhibitory effect on antiinflammation in $TNF-{\alpha}(p<0.02)$ in all five concentrations compared with the (-)controls treated with LPS. Mori Folium showed inhibitory effect on antiinflammation in $TNF-{\alpha}$ in similarpattern in all five concentrations compared with the (+)control pretreated with hydrocortisone. 2. Mori Folium. showed statistically significant inhibitory effect on antiinflammation in IL-6(p<0.01) in all five concentrations compared the (-)controls treated with LPS. Mori Folium. showed inhibitory effect on antiinflammation IL-6 similar pattern in all five concentrations compared with the (+)control pretreated with hydrocortisone. 3. Experimental Group pretreated with Mori Folium. showed statistically significant difference of antiinflammation in $IL-1{\beta}$ in concentrations of $50{\mu}g/m{\ell}(p<0.01)$, $250{\mu}g/m{\ell}(p<0.05)$ compared with the (-)control treated with LPS. Experimental Group pretreated with Mori Folium. showed increased level of $IL-1{\beta}$ in all concentrations compared the (+)control treated with hydrocortisone. 4. Experimental Group pretreated with Mori Folium. did not show statistically significant effect on IL-10 compared with the (-)control. Conclusion : By the findings of this experiment, Mori Folium is observed to have antiallergic and antiinflammation effect.

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Quality Characteristics of Yellow Layer Cake Prepared with Different Levels of Adenophora remotiflora Powder (모시대 분말 첨가 수준에 따른 옐로우 레이어 케이크의 품질 특성)

  • Choi, Yoon-Hee;Bae, Song-Hwan;Park, Jeong-Woo;Cho, Nam-Ji;Han, Myung-Ryun;Kim, Young-Ho;Yoon, Seong-Jun;Kim, Myung-Hwan;Kim, Ae-Jung
    • Journal of the East Asian Society of Dietary Life
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    • v.19 no.5
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    • pp.736-742
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    • 2009
  • The purpose of this study was to improve the quality of yellow layer cake by adding the Adenophora remotiflora powder. The effects of the Adenophora remotiflora powder in on the final product quality of yellow layer cake and the optimum amount of the Adenophora remotiflora powder in the yellow layer cake formula were investigated. The more increased amounts of Adenophora remotiflora powder was added to the samples, the resulted in "L", and Hunter's "a" and "b" values of the crust color of yellow layer cake were significantly decreased (p<0.05). In the case of crumb color, the more increased levels of Adenophora remotiflora powder led to the samples, the "L" and "a" values were significantly decreased (p<0.05), but "b" value increased significantly (p<0.05). As more Adenophora remotiflora powder was added to the cake samples, the Hardness, Gumminess and Chewiness were significantly increased (p<0.05), but Springiness and Cohesiveness were did not. The results of sensory evaluation showed revealed that the addition of 2% of Adenophora remotiflora powder of overall acceptability were most preferred. Larger scanning electron microscopy revealed the presence of numbers of greater concentrations of cells with different sizes were observed for in samples from products that received 1 % Adenophora remotiflora powder addition. Collapsed cells and cell coalescence with big large and irregular shapes were shown observed in samples that were amended with at 2, 4, and 8% addition Adenophora remotiflora power. The addition of Adenophora remotiflora powder was shown to improved the functionality and quality characteristics such as color, taste and flavor of yellow layer cake. Therefore, it was expected that the 2% addition of Adenophora remotiflora powder will improve the preference to the yellow layer cake.

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HDAC6 siRNA Inhibits Proliferation and Induces Apoptosis of HeLa Cells and its Related Molecular Mechanism

  • Qin, Hai-Xia;Cui, Hong-Kai;Pan, Ying;Yang, Jun;Ren, Yan-Fang;Hua, Cai-Hong;Hua, Fang-Fang;Qiao, Yu-Huan
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3367-3371
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    • 2012
  • Objective: To investigate the effects of histone deacetylase 6 (HDAC6) siRNA on cell proliferation and cell apoptosis of the HeLa cervical carcinoma cell line and the molecular mechanisms involved. Methods: Division was into three groups: A, the untreated group; B, the control siRNA group; and C, the HDAC6 siRNA group. Lipofectamine 2000 was used for siRNA transfection, and Western blot analysis was used to determine the protein levels. Cell proliferation and apoptosis were characterized using a CCK-8 assay and flow cytometry, respectively. Results: HDAC6 protein expression in the HDAC6 siRNA-transfection group was significantly lower (P < 0.05) than in the untreated and control siRNA groups. The CCK-8 kit results demonstrated that the proliferation of HeLa cells was clearly inhibited in the HDAC6 siRNA transfection group (P < 0.05). In addition, flow cytometry revealed that the early apoptotic rate ($26.0%{\pm}0.87%$) was significantly elevated (P < 0.05) as compared with the untreated group ($10.6%{\pm}1.19%$) and control siRNA group ($8.61%{\pm}0.98%$). Furthermore, Western blot analysis indicated that bcl-2 protein expression in the HDAC6 siRNA-transfection group was down-regulated, whereas the expression of p21 and bax was up-regulated. Conclusion: HDAC6 plays an essential role in the occurrence and development of cervical carcinoma, and the down-regulation of HDAC6 expression may be useful molecular therapeutic method.

Effects of Culture Systems on In Vitro Fertilization and Development of In Vitro Matured Bovine Follicular Oocytes (배양체계가 체외성숙 소 난포란의 체외수정 및 배 발달에 미치는 효과)

  • 조성근;송상현;정기화;강대진;박충생
    • Journal of Embryo Transfer
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    • v.11 no.1
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    • pp.15-26
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    • 1996
  • This study was conducted to improve the in vitro maturation(JVM), in vitro fertilization (IVF) and in vitro developmental capacity of oocytes derived from slaughtered Korean native cattle. The recoverd oocytes, obtained from a local slaughter house, were used completely surrounded by at least 3 layers of cumulus cells in combination with a homogeneous cytoplasmic pigmentation. In vitro maturation was induced in TCM-199 or Ham's F-10 supplemented with LH(1O $\mu$g/rnl), FSH(35 $\mu$g/ml), estradiol-17$\beta$(1 $\mu$g/ml) at 39$^{\circ}C$ under 5% $CO_2$ in air for 24 hours. Sperm from caudal epididyrnis and previously matured cumulus-oocytes complexes were cultured for 24 hours in 100 $\mu$l droplets of fertilization media under paraffin oil. The zygotes were cultured with media(TCM-199 with bovine oviductal epithelial cells or CRlaa) for 7 to 10 days. The cleavage rate of IVM-IVF oocytes was significantly (P<0.05) higher following maturation using Ham's F-10 (59.9%) than TCM-199 (51.6%). Development to the blastocysts among cleaved embryos was not signficantly different between maturation media: Ham's F-10 (16.0%) and TCM-199(11.9%). However, the hatching rate was affected significantly (P<0.05) on rnaturation media as 62.9% in Ham's F-10, compared with 41.2% in TCM-199. The cleavage rate of IVM-IVF oocytes was significantly (P<0.05) higher following IVF using m-TALP medium (80.1%) than BO medium (51.6%). The percentage of in vitro developed blastocysts among cleaved embryos was not signficantly different between fertimization media: BO (11.7%) and m-TALP (17.6%). The cleavage and the developmental rate to the blastocysts after IVF in m-TALP or condition medium(CM) with or without oviduct epithelial cell monolayer(OECM) was similar(80.1% and 17.6% in m-TALP, 83.8% and 19.4% in M-TALP with OECM. 82.9% and 18.9% in CM, 87.6% and 16.0% in CM with OECM, respectively). The percentage of in vitro developed blastocysts among cleaved embryos was significantly (P<0.05) higher in TCM-199 medium co-cul tured with bovine oviduatal epithelial cell monolayers(35.2%) than CRlaa medium(1.9%). These results stggest that the most transferable IVF embryos could be produced from Ham's F-10, m-TALP and TCM-199 medium with bovine oviductal epithelial cell monolayers for IVM, IVF and IVC, respectively.

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Heterologous Expression of Interferon α-2b in Lactococcus lactis and its Biological Activity against Colorectal Cancer Cells

  • Meilina, Lita;Budiarti, Sri;Mustopa, Apon Zaenal;Darusman, Huda Shalahudin;Triratna, Lita;Nugraha, Muhammad Ajietuta;Bilhaq, Muhammad Sabiq;Ningrum, Ratih Asmana
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.75-87
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    • 2021
  • Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp45 signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SPUsp45-IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC50 values of 33.22 ㎍/ml and 127.2 ㎍/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 ㎍/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 ㎍/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codon-optimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

Impact of Absorber Thickness on Bifacial Performance Characteristics of Semitransparent Amorphous Silicon Thin-Film Solar Cells (광흡수층 두께에 따른 투광형 비정질 실리콘 박막 태양전지의 양면발전 성능특성)

  • Seo, Yeong Hun;Lee, Ahruem;Shin, Min Jeong;Cho, Ara;Ahn, Seungkyu;Park, Joo Hyung;Yoo, Jinsu;Choi, Bo-Hun;Cho, Jun-Sik
    • Current Photovoltaic Research
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    • v.7 no.4
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    • pp.97-102
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    • 2019
  • Bifacial and semitransparent hydrogenated amorphous silicon (a-Si:H) thin-film solar cells in p-i-n configuration were prepared with front and rear transparent conducting oxide (TCO) electrodes using plasma-enhanced chemical vapor deposition method. Fluorine-doped tin oxide and tin-doped indium oxide films were used as front and rear TCO contacts, respectively. Film thickness of intrinsic a-Si:H absorber layers were controlled from 150 nm to 450 nm by changing deposition time. The dependence of performance characteristics of solar cells on the front and rear illumination direction were investigated. For front illumination, gradual increase in the short-circuit current density (JSC) from 10.59 mA/㎠ to 14.19 mA/㎠ was obtained, whereas slight decreases from 0.83 V to 0.81 V for the open-circuit voltage (VOC) and from 68.43% to 65.75% for fill factor (FF) were observed. The average optical transmittance in the wavelength region of 380 ~ 780 nm of the solar cells decreased gradually from 22.76% to 15.67% as the absorber thickness was changed from 150 nm to 450 nm. In case of the solar cells under rear illumination condition, the JSC increased from 10.81 to 12.64 mA/㎠ and the FF deceased from 66.63% to 61.85%, while the VOC values were maintained at 0.80 V with increasing the absorber thickness from 150 nm to 450 nm. By optimizing the deposition parameters, a high-quality bifacial and semitransparent a-Si:H solar cell with 350 nm-thick i-a-Si:H absorber layer exhibited the conversion efficiencies of 7.69% for front illumination and 6.40% for rear illumination, and average visible optical transmittance of 17.20%.

The Usefulness of Vaginal Cytology during the Estrous Cycle in Miniature Schnauzer Dogs (Miniatur Schnauzer견에서 발정 주기 동안 질세포 검사의 유용성)

  • Kim, Jung-Bae;Kim, Bang-Sil;Park, Sang-Guk;Kim, Jae-Pung;Moon, Jin-San;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
    • Journal of Embryo Transfer
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    • v.23 no.1
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    • pp.13-18
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    • 2008
  • Vaginal cytology with behavioral observation was performed in 34 estrous cycles in 16 Miniature Schnauzer dogs to evaluate the its usefulness. The mean duration of proestrus and estrus in Miniature Schnauzer based on behavioral observation and vaginal cytology was $7.58{\pm}1.50$ (Mean ${\pm}$S.D., range: $5{\sim}10$) and $8.46{\pm}2.82\;(6{\sim}17)$ days for proestrus, and $6.97{\pm}1.66\;(4{\sim}10)$ and $10.29{\pm}2.61\;(6{\sim}19)$ days for estrus, respectively. The duration of each phase of the estrous cycle was not significantly different based on between behavioral observation and vaginal cytology. The gestational length from the first day of male acceptance was $66.55{\pm}2.91\;(64{\sim}76)$ days, $57.70{\pm}1.92\;(54{\sim}62)$ days from the first male refusal, and $56.90{\pm}1.62\;(54{\sim}60)$ days from the onset of cytologic diestrus, respectively. Vaginal cytology during the estrous cycle were significantly characteristic of large intermediate cells in proestrus, anuclear cells in estrus, small intermediate cells in diestrus, and parabasal cells and small intermediate cells in anestrus (p<0.001), respectively. Cornification index (CI) by vaginal cytology was higher in proestrus ($69.90{\pm}1.44%$) and estrus ($91.35{\pm}1.09%$), then it was decreased in diestrus ($13.53{\pm}1.28%$) and anestrus ($1.16{\pm}1.26%$).