Kim, Tae-Yong;Kim, Ji-Young;Chang, Kyung-Soo;Kim, Myung-Cheol;Park, Chang-Sik;Han, Hong-Ryul;Jun, Moo-Hyung
Korean Journal of Veterinary Research
/
v.45
no.1
/
pp.45-53
/
2005
GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.
In our previous study, we observed that hydrosalpingeal fluid (HSF) adversely effect mouswe embryo development and hatching. The aim of this study was to evaluate the effect of HSF as assessed by the blastocyst development rate (BDR) and by cell counting in vitro. HSF was collected from ninie patients undergoing salpingoneostomy to correct hydrosalpinx. Two-cell embryos were obtained from superovulated ICR mice. T6 medium and $T6{\pm}0.4%$ bovine serum albumin were used as control media. T6 medium containing 10% or 50% HSF and 100% HSF from each patient were used as test media. Nine to 15 embryos were cultured in micro drops prepared from each of these media. To assess the total cell number within each blastocyst, the blastocysts were fixed and stained with Hoechst 33342 to facilitate cell counting. The mean BDR in two control media were 88.89% and 85.40%. The mean BDR in media containing 10%, 50%, 100% HSF were 85.87%, 89.58% and $75.57%^*$, respectively ($^*$: p<0.05). The overall mean cell count $({\pm}SEM)$ in control media were $87.6{\pm}9.65\;and\;90.12{\pm}11.38$. The BDR was affected adversely only by 100% HSF and not in media containing 10% or 50% HSF. Mean cell counts were decreased significantly only in blastocysts cultured 100% HSF ($63.8{\pm}13.66$; p<0.01) but not in blastocysts cultured in 10% or 50% HSF ($91.3{\pm}12.44\;and\;82.9{\pm}18.27$, respectively). Thus, it is concluded that HSF has no embyotoxic effect but has a mildly negatively effect on embryonic growth and development.
Park, Kee-Sang;Song, Hai-Bum;Lee, Taek-Hoo;Jeon, Sang-Sik
Clinical and Experimental Reproductive Medicine
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v.27
no.2
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pp.165-172
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2000
Objective: In vitro fertilization (IVF) and a prolonging the time of culture may be helpful in establishing a viable pregnancy through a selection effect. Some embryos do not develop beyond the 4-cell stage and some may not develop to the blastocyst stage. We have evaluated the safety of SET and the outcomes of pregnancy. Methods: Sperms were treated with Ham's F-10 supplemented with 10% human follicular fluid (hFF). oocytes or fertilized oocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% or 20% hFF respectively. Up to five oocytes were inseminated with approximately 200,000 sperm cells/2 ml in each well. Fertilization was examined in the following morning and fertilized oocytes were co-cultured until embryo transfer. Vero cells for co-culture were prepared in Tissue Culture Medium - 199 (TCM-199) with 10% fetal bovine serum. At the two to four cell and blastocyst on day 2 and day 5, embryo and blstocyst grading were evaluated. Pregnancy rate was determined after transfer of human embryos at the two to four cell stage on day 2 (Group I) or subsequent transfer of embryos on day 2 and at the blastocyst stage on day 5 (Group II). For statistical analysis, Student's t-test and Chi-square (${\chi}^2$_test) were used. Results were considered statistically significant when p value was less than 0.05. Results: No differences was found in the fertilization between Group I (81.0%, 98/121) and Group II (81.8%, 180/220). In case of cleavage rate, no difference was found in Group I (95.9%, 94/98) and Group II (97.8%, 174/178). However, the rate of-clinical pregnancy was significantly higher (p=0.014) in Group II (66.7%, 12/18) than in Group I (26.3%, 5/19). Conclusion: The results of this study showed that SET is safe and effective, and significantly increases the pregnancy rate.
Using differential scanning calorimetry (DSC), changes underwent by a mixture of $\alpha$-lactalbumin ($\alpha$-La) and $\beta$-lactoglobulin ($\beta$-Lg) during heat treatment were studied, yielding useful information for the dairy industry. Results of the DSC showed that the heat denaturation temperature of the hobo-$\alpha$-La was higher than that of apo-$\alpha$-La, suggesting hole-$\alpha$-La‘s greater stability. The denaturation temperature of a mixture of holo-$\alpha$-La and $\beta$-Lg was also slightly lower than that of holo-$\alpha$-La alone. The denaturation temperature of an apo-$\alpha$-La and $\beta$-Lg mixture was higher than that of holo-$\alpha$-La and $\beta$-Lg, suggesting that the heat stability of apo-$\alpha$-La was increased by $\beta$-Lg. Based on these results, it is possible to conclude that a mixture of holo-$\alpha$-La and $\beta$-Lg is more intensively affected by an increase in temperature than other samples, and that free sulphydryl groups seem to take part in this heat-induced denaturation.
Journal of the Korean Data and Information Science Society
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v.26
no.6
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pp.1249-1258
/
2015
DGAT1(diacylglycerol O-acyltransferase1) gene is well known as a major gene of milk production in dairy cattle. This study was conducted to investigate how the DGAT1 gene effect on milk yield was appeared from the genome wide association (GWA) using high density whole genome SNP chip. The data set used in this study consisted of 353 Korean Holstein sires with 50k SNP genotypes and deregressed estimated breeding values of milk yield. After quality control 41,051 SNPs were selected and locations on chromosome were mapped using UMD 3.1. Bayesian regression of BayesB method (pi=0.99) was used to estimate the SNP effects and genomic breeding values. Percentages of variance explained by 1 Mb non-overlapping windows were calculated to detect the QTL region. As the result of this study, top 1 and 3 of 2,516 windows were seen around DGAT1 gene region and 0.51% and 0.48% of genetic variance were explained by these two windows. Although SNPs on the DGAT1 gene region are excluded in commercial 50k SNP chip, the effect of DGAT1 gene seem to be reflected on GWA by the SNPs which are in linkage disequilibrium with DGAT1 gene.
The purpose of this study was to evaluate the effects of commercial home-tooth bleaching agents on the color of tooth. Twenty five sound extracted teeth were randomly divided into five groups. The color differences between before and after treatment with five types of tooth bleaching agents (7.5% hydrogen peroxide Nite White $Excel^{(R)}$, 10% carbamide peroxide Nite White $Excel^{(R)}$, 16% carbamide peroxide Nite White $Excel^{(R)}$, 10% carbamide peroxide Insta-BriteTM, 20% carbamide peroxide Insta-$Brite^{TM}$) were evaluated. The results were as follows: 1. By 2 week home tooth bleaching agent applications, the values ($L^*$) of bovine teeth increased as high as 4.38 $\sim$ 8.80 when comparing to those of the samples before treatment, and the color difference (${\Delta}E^*$) showed as high as 10.16 $\sim$ 15.04. 2. 16% carbamide peroxide Nite White Excel induced significantly greater ${\Delta}L^*$ than other test edgroups except for 7.5% hydrogen peroxide Day White Excel, and significantly greater ${\Delta}E^*$ than other tested groups by 2 week bleaching agent treatments (p<0.01). 3. 16% carbamide peroxide Nite White Excel(${\Delta}L^*$=8.80, ${\Delta}E^*$=15.04) induced significantly greater ${\Delta}L^*$ and ${\Delta}E^*$ than 10% carbamide peroxide Nite White Excel(${\Delta}L^*$=5.01, ${\Delta}E^*$=10.16)(p<0.01), but significant difference between 10% carbamide peroxide Insta-Brite(${\Delta}L^*$=4.38, ${\Delta}E^*$=10.51) and 20% carbamide peroxide Insta-Brite(${\Delta}L^*$=5.63, ${\Delta}E^*$=11.23) was not shown in ${\Delta}L^*$ and ${\Delta}E^*$(p>0.01). 4. 16% carbamide peroxide Nite White Excel(${\Delta}L^*$=8.80, ${\Delta}E^*$=15.04) which were applied in night time induced significantly greater ${\Delta}L^*$ and ${\Delta}E^*$ than 7.5% hydrogen peroxide Day White Excel(${\Delta}L^*$=8.47, ${\Delta}E^*$=12.75) which were applied in day time. Conclusions: These results demonstrate that all the commercial home-tooth bleaching agents have appreciable bleaching effect on teeth, and the effects of home-tooth bleaching agents which are used during night time are affected by content of carbamide peroxide. Especially the whitening effect of home tooth bleaching agents that are used through night time is greater than that of short time-applying tooth bleaching agent.
To provide a basis for studying the pharmacological actions of tetracaine HCl, we analyzed the membrane activities of this local anesthetic. The n-(9-anthroyloxy) stearic and palmitic acid (n-AS) probes (n = 2, 6, 9, 12 and 16) have been used previously to examine fluorescence polarization gradients. These probes can report the environment at a graded series of depths from the surface to the center of the membrane bilayer structure. In a dosedependent manner, tetracaine HCl decreased the anisotropies of 6-AS, 9-AS, 12-AS and 16-AP in the hydrocarbon interior of synaptosomal plasma membrane vesicles isolated from bovine cerebral cortex (SPMV), and liposomes derived from total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. However, this compound increased the anisotropy of 2-AS at the membrane interface. The magnitude of the membrane rotational mobility reflects the carbon atom numbers of the phospholipids comprising SPMV, SPMVTL and SPMVPL and was in the order of the 16, 12, 9, 6, and 2 positions of the aliphatic chains. The sensitivity of the effects of tetracaine HCl on the rotational mobility of the hydrocarbon interior or surface region was dependent on the carbon atom numbers in the descending order 16-AP, 12-AS, 9-AS, 6-AS and 2-AS and on whether neuronal or model membranes were involved in the descending order SPMV, SPMVPL and SPMVTL.
The aim of this study was to provide a basis for the molecular mechanism underlying the pharmacological action of ethanol. We studied the effects of 1-propanol on the location of n-(9-anthroyloxy)palmitic acid or stearic acid (n-AS) within the phospholipids of synaptosomal plasma membrane vesicles (SPMV). The SPMV were isolated from the bovine cerebral cortex and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL). 1-Propanol increased the rotational mobility of inner hydrocarbons, while decreasing the mobility of membrane interface, in native and model membranes. The degree of rotational mobility varied with the number of carbon atoms at positions 16, 12, 9, 6 and 2 in the aliphatic chain of phospholipids in the neuronal and model membranes. The sensitivity of increasing or decreasing rotational mobility of hydrocarbon interior or surface by 1-propanol varied with the neuronal and model membranes in the following order: SPMV, SPMVPL and SPMVTL.
Statement of problem. Cortical bone plays an important role in the primary implant stability, which is essential to immediate/early loading. However, immediate load-bearing capacity and primary implant stability according to the change of the cortical bone thickness have not been reported. Purpose. The objectives of this study were (1) to measure the immediate load-bearing capacity of implant and primary implant stability according to the change of cortical bone thickness, and (2) to evaluate the correlation between them. Material and methods.48, screw-shaped implants (3.75 mm$\times$7 mm) were placed into bovine rib bone blocks with different upper cortical bone thickness (0-2.5 mm) and resonance frequency (RF) values were measured subsequently. After fastening of healing abutment. implants were subjected to a compressive load until tolerated micromotion threshold known for the osseointegration and load values at threshold were recorded. Thereafter, RF measurement after loading, CT taking and image analysis were performed serially to evaluate the cortical bone quality and quantity. Immediate load-bearing capacity and RF values were analyzed statistically with ANOVA and post-hoc method at 95% confidence level (P<0.05). Regression analysis and correlation test were also performed. Results. Existence and increase of cortical bone thickness increased the immediate load-bearing capacity and RF value (P<0.05) With the result of regression analysis, all parameter's of cortical bone thickness to immediate load-bearing capacity and resonance frequency showed significant positive values (P<0.0001). A significant high correlation was observed between the cortical bone thickness and immediate load-beating capacity (r=0.706, P<0.0001), between the cortical bone thickness and resonance frequency (r=0.753, P<0.0001) and between the immediate load-bearing capacity and resonance frequency (r=0.755, P<0.0001). Conclusion. In summary, cortical bone thickness change affected the immediate load-baring capacity and the RF value. Although RF analysis (RFA) is based on the measurement of implant/bone interfacial stiffness, when the implant is inserted stably, RFA is also considered to reflect implant/bone interfacial strength of immediately after placement from high correlation with the immediate load-baring capacity. RFA and measuring the cortical bone thickness with X-ray before and during surgery could be an effective diagnosis tool for the success of immediate loading of implant.
This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.
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