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Maturational Changes in Binding Capacity of Fowl Sperm to the Epithelium of the Sperm Storage Tubules during Their Passage through the Male Reproductive Tract

  • Ahammad, Muslah U.;Okamoto, S.;Kawamoto, Yasuhiro;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1199-1203
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    • 2011
  • The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).

Porcine somatic cell nuclear transfer using telomerase reverse transcriptase-transfected mesenchymal stem cells reduces apoptosis induced by replicative senescence

  • Jeon, Ryounghoon;Rho, Gyu-Jin
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.3
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    • pp.215-222
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    • 2020
  • Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

A Study of Ice-Formation Phenomena on Freezing of Flowing Water in a Stenotic Tube

  • Suh, Jeong-Se;Kim, Moo-Geun;Ro, Sung-Tack;Yim, Chang-Soon
    • International Journal of Air-Conditioning and Refrigeration
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    • v.7
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    • pp.1-10
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    • 1999
  • In this study, a numerical analysis is made on the ice-formation for laminar water flow inside a stenotic tube. The study takes into account the interaction between the laminar flow and the stenotic port in the circular tube. The purpose of the present numerical investigation is to assess the effect of a stenotic shape on the instantaneous shape of the flow passage during freezing upstream/downstream of the stenotic channel. In the solution strategy, the present study is substantially distinguished from the existing works in that the complete set of governing equations in both the solid and liquid regions are resolved. In a channel flow between parallel plates, the agreement between the of predictions and the available experimental data is very good. Numerical analyses are performed for parametric variations of the position and heights of stenotic shape and flow rate. The results show that the stenotic shape has the great effect on the thickness of the solidification layer inside the tube. As the height of a stenosis grows and the length of a stenosis decreases, the ice layer thickness near the stenotic port is thinner, due to backward flow caused by the sudden expansion of a water tunnel. It is found that the flow passage has a slight uniform taper up to the stenotic channel, at which a sudden expansion is observed. It is also shown that the ice layer becomes more fat in accordance with its Reynolds number.

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Eating Behaviour, VFA Production, Passage Rate and Nutrient Digestibilities in Cattle Fed on Wheat Straw Supplemented with Different Levels of Berseem

  • Das, A.;Singh, G.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.7
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    • pp.1040-1048
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    • 1999
  • Four ruminally fistulated crossbred $(Sahiwal{\times}Holstein\;Friesan)$ adult (~6 yr.) cattle of about $318{\pm}16kg$ body weight were randomly assigned in an experiment based on $4{\times}4$ latin square design to study the effect of different level of barseem (Trifolium alexendrinum) supplementation to wheat straw based diet on intake, digestion, VFA production, eating behaviour and passage rate. Four dietary treatments were wheat straw ad lib. (I), supplementation to wheat either 15(II), 30(III) or 45(IV)% of berseem. Mean total volatile fatty acids (TVFA) concentration in rumen liquor was 58.45, 66.14, 77.92 and 78.64 mmol/l. TVFA concentration in rumen liquor increased significantly (p<0.01) with increased level of berseem supplementation upon 30% level of breseem. Two peaks of TVFA concentration was observed at 4 and 8 h post feeding. Brseem supplementation showed no significant effect on daily time spent for eating, ruminating or idling, chews per minute or number of rumination boli ingested per minute. Time spent for eating and ruminating one kg neutral detergent fibre (NDF) was significantly (p<0.05) less in breseem supplemented groups. Animals in groups I, II, III and IV consumed 26, 34, 47 and 57% of DM within 1st 4 h and 64, 70, 70 and 77% of total DM within 1st 8 h of offering. All the animals consumed more than 90% of their DMI within 12 h. Active period of rumination was observed 8h post feeding reached the peak at 16 h post feeding, then declined and animals spend considerable time idling in last 4 h of 24 h feeding cycle. Berseem supplementation showed no significant effect on eating and rumination time. It is concluded that berseem supplementation upon 30% increases the efficiency of chewing during eating and rumination, which results in increase intake and TVFA production and nutrient digestibility.

Establishment of an Efficient System for the Production of Transgenic Somatic Cell Nuclear Transfer Embryos

  • Cho, J.K.;Bhuiyan, M.M.U.;Jang, G.;Park, E.S.;Chang, K.H.;Park, H.J.;Lim, J.M.;Kang, S.K.;Lee, B.C.;Hwang, W.S.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.75-75
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    • 2002
  • The present study was conducted for the production of transgenic cloned cows by somatic cell nuclear transfer (SCNT) that secrete human prourokinase into milk. To establish an efficient production system for bovine transgenic SCNT embryos, the offset was examined of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos. An expression plasmid far human prourokinase (pbeta-ProU) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected using a lipid-meidated method. In Experiment 1, developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In Experiment 2, the effect of cellular senescence in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed. In Experient 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 ${\mu}{\textrm}{m}$) or small cell (<30 ${\mu}{\textrm}{m}$)] were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.

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Steroid Effects on Cell Proliferation, Differentiation and Steroid Receptor Gene Expression in Adult Bovine Satellite Cells

  • Lee, Eun Ju;Choi, Jinho;Hyun, Jin Hee;Cho, Kyung-Hyun;Hwang, Inho;Lee, Hyun-Jeong;Chang, Jongsoo;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.501-510
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    • 2007
  • The present study was conducted to establish primary bovine muscle satellite cell (MSC) culture conditions and to investigate the effects of various steroid hormones on transcription of the genes involved in muscle cell proliferation and differentiation. Of three different types of proteases (type II collagenase, pronase and trypsin-EDTA) used to hydrolyze the myogenic satellite cells from muscle tissues, trypsin-EDTA treatment yielded the highest number of cells. The cells separated by hydrolysis with type II collagenase and incubated on gelatin-coated plates showed an enhanced cell attachment onto the culture plate and cell proliferation at an initial stage of cell growth. In this study, the bovine MSCs were maintained in vitro up to passage 16 without revealing any significant morphological change, and even to when the cells died at passage 21 with decreased or almost no cell growth or deformities. When the cells were incubated in a steroid-depleted environment (DMEM(-)/10% CDFBS (charcoal-dextran stripped FBS)), they grew slowly initially, and were widened and deformed. In addition, when the cells were transferred to an incubation medium containing steroid (DMEM(+)/10% FBS), the deformed cells resumed their growth and returned to a normal morphology, suggesting that steroid hormones are crucial in maintaining normal MSC morphology and growth. The results demonstrated that treatments with 19-nortestosterone and testosterone significantly increased AR gene expression (p<0.05), implying that both testosterone and 19-nortestosterone bind with AR and that the hormone bound-AR complex up-regulates the genes of its own receptor (AR) plus other genes involved in satellite cell growth and differentiation in bovine muscle.

Effects of several suspending media on behavior of Toxoplasma gondii (보존액이 Toxoplasma gondii의 생존성에 미치는 영향)

  • Jin, Joo-Eun;Chung, Kyung-Tae;Lee, Woo-Won;Yang, Ju;Lee, Gang-Rog;Kim, Geun-Kyu
    • Korean Journal of Veterinary Service
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    • v.22 no.2
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    • pp.129-134
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    • 1999
  • A preservation test of Toxoplasma gondii tachyzoites for considerable time was tried to obtain simple and economical methods using various suspending media at $4^{\circ}C$ rather than serial passage of the parasite in mice. The preservation period was a term that the tachyzoites were detected from the peritoneal fluid of mice after inoculation of 2$\times$$10^5$ organisms preserved according to the lapse of time. The numbers of tachyzoite per 1\textrm{mm}^3$ of the peritoneal fluid with 2$m\ell$ of the saline solution taken in 4days after inoculation were presented as percentage in proportion to the control. The numbers. of tachyzoite per 1\textrm{mm}^3$ of the peritoneal fluid of the control were consisted of the average number of the tachyzoites of 10 mice inoculated with 2$\times$$10^5$ organisms by serial mouse passage. The tachyzoites could be preserved for 26 days in the suspending medium of saline solution at $4^{\circ}C$ Ringer's solution for 18 days, Hank's solution for 28 days, and egg-glycerine solution for 50 days.

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Effects of Bleeding on Heat/Mass Transfer in a Rotating Channel with Transverse Ribs (90도 요철이 설치된 회전덕트에서 유출이 열/물질전달에 미치는 영향)

  • Park, Suk-Hwan;Jeon, Yun-Heung;Kim, Kyung-Min;Lee, Dong-Hyun;Cho, Hyung-Hee
    • The KSFM Journal of Fluid Machinery
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    • v.9 no.1 s.34
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    • pp.25-31
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    • 2006
  • The present study investigates the effects of bleed flow on heat/mass transfer and pressure drop in a rotating channel with transverse rib turbulators. The hydraulic diameter $(D_h)$ of the square channel is 40.0mm. The bleed holes are located between the rib turbulators on leading surface and the hole diameter (d) is 4.5 mm. The square rib turbulators are installed on both leading and trailing surfaces. The rib-to-rib pitch is 10.0 times of the rib height(e) and the rib height-to-hydraulic diameter ratio $(e/D_h)$ is 0.055. The tests were conducted at various rotation numbers (0, 0.2, 0.4), while the Reynolds number and the rate of bleed flow to main flow (BR) were fixed at 10,000 and $10\%$, respectively. The results suggest that the heat/mass transfer characteristics in the internal cooling passage are influenced by rib turbulators, bleed flow and the Coriolis force induced by rotation. For the rotating ribbed passage with bleed flow, the heat/mass transfer on the leading surface is hardly affected by bleed flow, but that on the trailing surface decreases due to the diminution of main flow. The results also show that the friction factor decreases with the bleed flow.

Falling Accident Case Analysis on Construction Working Platform and Working Passage (건설현장 작업발판 및 안전통로 관련 추락 및 전도재해 사고사례 분석)

  • Kim, Hyunsoo;Lee, Yong-Soo;Oh, Inhwan;Ahn, Hongseob
    • Journal of the Korea Institute of Construction Safety
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    • v.2 no.1
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    • pp.9-15
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    • 2019
  • Despite the efforts for enhancing the safety record, construction industry has been suffered from higher fatalities than other industries. The poor record of safety in construction industry means that there is a clear need for an effective countermeasure. As mentioned in previous studies, it is important to identify the type of activities or risks that are likely to cause accidents and to develop appropriate safety measures. Considering the large number of accident cases on the temporary installations including work platforms and work passages, the temporary installations should be managed first. To support it, this study aims to analyze falling accident cases on construction working platforms and passages which can lead to develop proper safety measures. Through the analysis of 1663 accident cases in the perspective of cost, progress, activity, and type of workers, this study identifies how the recent accidents occur and what is the cause of the accident occurrence. The identified causes of accident occurrence will help us to improve current construction safety.

Factors which contribute to time of first stool and first urine passage in Newborns (신생아에서 첫 태변 배출과 초뇨 배출 시간 결정인자)

  • Lee, Hye Jin;Jae, Hyun Gon;Son, Sang Hee
    • Clinical and Experimental Pediatrics
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    • v.49 no.5
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    • pp.482-488
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    • 2006
  • Purpose : To evaluate the factors which contribute to the time of the first stool and the first urine passage. Methods : We retrospectively reviewed a chart of 1,221 infants ${\geq}34$ weeks of gestational age admitted to the normal newborn nursery of Il Sin Christian Hospital, Busan, from November 2004 to April 2005. We compared the time to first stool and urine according to maternal factors(maternal age, parity, mode of delivery, meconium-stained amniotic fluid, and maternal diabetes) and infant factors (gender, Apgar score, gestational age, type of feeding during first 24 hours, age at the first feeding, number of feeds during the first 12 hours and age at discharge). Results : In total, 95.3 percent of our infants had passed their first stool by 24 hours and 99.8 percent of them had a stool by 36 hours. A total of 95.8 percent of our infants had passed urine by 24 hours of age and 98.3 percent of them by 36 hours. Comparing preterm and term infants, the time to first urine is $6.5{\pm}5.8$ hours and $12.1{\pm}6.6$ hours, respectively(P=0.000). The time to first stool is $20.7{\pm}13.5$ hours and $10.0{\pm}6.3$ hours, respectively(P=0.000) Early-fed infants were significantly earlier in time to first urine(P=0.023) and first stool(P=0.012). There was no statistically significant relationship between the number of feeds in 0-12 hours, mode of delivery, Apgar score, parity, gender, type of feeding, maternal diabetes and the time of the first urine and first stool. Conclusion : Gestational age, birth weight and age at first feeding were significantly related to the time of the first urine and first stool passage. When there is delayed passage of the first urine and first stool, we should consider close observation of other associated symptoms and other factors previously mentioned, to avoid extensive evaluation and intervention.