• 대한치과보철학회지
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• 제30권2호
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• pp.247-257
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• 1992

The present study quantitates the in vitro cytotoxicity of a variety of denture base acrylic resins using cell culture techniques combined with image analysis to measure nuclear area and DNA contents. In this study, a comparison was made among direct curing, heat curing and microwave curing resins. The results obtained from this study were as follows : 1. Morphologically, cell process and nucleus became prominent but macroscopic difference according to the resins were nit observed. In addition, increased cellular density around the specimen were observed. 2. In DNA contents measurements, $S-G_2M$ phase cell was 15.47%, 14.58% in control and heat curing resin on 1st day and the others group $21.39\sim33.36%$ were measured. 3. Nuclear area and DNA contents were increased on 3rd day except DNA content of the microwave curing resin group. These results suggest that denture base acrylic resins stimulate gingival fibroblasts in vitro, especially stimulation of direct curing resin is larger and longer than the others.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.812-817
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• 2012

Nitrate can serve as a terminal electron acceptor in place of carbon dioxide and inhibit methane emission in the rumen and nitrate reducing bacteria might help enhance the reduction of nitrate/nitrite, which depends on the type of feed offered to animals. In this study the effects of three levels of sodium nitrate (0, 5, 10 mM) on fermentation of three diets varying in their wheat straw to concentrate ratio (700:300, low concentrate, LC; 500:500, medium concentrate, MC and 300:700, high concentrate, HC diet) were investigated in vitro using buffalo rumen liquor as inoculum. Nitrate reducing bacteria, isolated from the rumen of buffalo were tested as a probiotic to study if it could help in enhancing methane inhibition in vitro. Inclusion of sodium nitrate at 5 or 10 mM reduced (p<0.01) methane production (9.56, 7.93 vs. 21.76 ml/g DM; 12.20, 10.42 vs. 25.76 ml/g DM; 15.49, 12.33 vs. 26.86 ml/g DM) in LC, MC and HC diets, respectively. Inclusion of nitrate at both 5 and 10 mM also reduced (p<0.01) gas production in all the diets, but in vitro true digestibility (IVTD) of feed reduced (p<0.05) only in LC and MC diets. In the medium at 10 mM sodium nitrate level, there was 0.76 to 1.18 mM of residual nitrate and nitrite (p<0.01) also accumulated. In an attempt to eliminate residual nitrate and nitrite in the medium, the nitrate reducing bacteria were isolated from buffalo adapted to nitrate feeding and introduced individually (3 ml containing 1.2 to $2.3{\times}10^6$ cfu/ml) into in vitro incubations containing the MC diet with 10 mM sodium nitrate. Addition of live culture of NRBB 57 resulted in complete removal of nitrate and nitrite from the medium with a further reduction in methane and no effect on IVTD compared to the control treatments containing nitrate with autoclaved cultures or nitrate without any culture. The data revealed that nitrate reducing bacteria can be used as probiotic to prevent the accumulation of nitrite when sodium nitrate is used to reduce in vitro methane emissions.

• Parasites, Hosts and Diseases
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• 제48권1호
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• pp.49-55
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• 2010

In vitro cultivation of trematodes would assist studies on the basic biology of the parasites and their hosts. This is the first study to use the yolk of unfertilized chicken eggs as a simple and successful method of ovocultivation and the first time to obtain the adult-stage of the trematode Cymatocarpus solearis Braun, 1899 (Digenea: Brachycoeliidae). Chicken eggs were inoculated with metacercariae from the muscle of the spiny lobster, Panulirus argus (Latreille, 1804). The metacercariae were excysted and incubated for 576 hr (24 days) at $38^{\circ}C$ to obtain the adult stage. Eggs in utero were normal in shape and light brown color. The metacercariae developed into mature parasites that have been identified as the adultstage found in marine turtles. The adult lobsters collected in Quintana Roo State, Mexico, showed the prevalence of 49.4% and the mean intensity of 26.0 per host (n=87). A statistical study was performed to determine that no parasitic preference was detected for male versus female parasitized lobsters. Morphometric measurements of the adult-stage of C. solearis obtained in our study have been deposited in the National Helminths Collection of the Institute of Biology of the National Autonomous University of Mexico. This study is significant because it is the first time that a digenean of the family Brachycoeliidae has been demonstrated to develop in vitro from metacercariae into adults capable of producing eggs using the yolk of unfertilized chicken eggs. Secondly, this technique allows to obtain the adult stage of C. solearis without the presence of its marine turtle host, allows us to describe the mature parasites, and thus contribute to our understanding of the biology of C. solearis.

• 한국초지조사료학회지
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• 제27권3호
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• pp.219-228
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• 2007

본 연구는 호밀 건초를 급여하는 조건하에 알코올 발효사료에 대한 면실, 아마 및 대두종실의 첨가가 in vitro 반추위 발효특성에 미치는 영향을 구명하고자 실시하였다. 시험구 처리는 전지종실을 첨가하지 않고 발효한 알코올 발효사료구(대조구: T1), 면실종실을 첨가하여 발효한 면실 알코올 발효사료구(T2), 아마종실을 첨가하여 발효한 알코올 발효사료구(T3) 및 대두종실을 첨가하여 발효한 알코올 발효사료구(T4)의 4처리로 하였다. 배양시간에 따른 pH는 면실 종실을 첨가한 T2구는 대조구에 비해 배양 6시간에서 12시간까지 감소하는 결과를 보였으며(p<0.05), 대두종실을 첨가하는 T4구의 pH는 대조구보다 배양 3시간에서 12시간까지 감소하는 것으로 나타났다(p<0.05). 그러나 아마종실을 첨가하는 T3구의 pH는 배양 초기부터 배양 9시간까지 대조구에 비해 차이가 없는 결과를 보였다(p>0.05). 배양액의 ammonia 농도는 T2구가 대조구에 비해 배양 6시간경에 현저하게 증가하는 것으로 나타났으며(p<0.05), T4구의 ammonia 농도는 대조구에 비해 배양 6 및 9시간에서 유의적으로 높은 결과를 보였다(p<0.05). 배양시간에 따른 반추위액 배양액의 휘발성 지방산 농도는 대조구에 비해 전지종실 처리구들이 전반적으로 증가하는 경향을 보였으며, 배양 3시간경에서 면실, 아마 및 대두종실을 첨가하는 T2, T3 및 T4구의 acetate, butyrate 및 propionate 농도가 대조구에 비해 유의적으로 증가하는 결과를 보였다(p<0.05). 특히, 대두 종실을 첨가한 T4구의 휘발성지방산은 배양초기부터 12시간까지 전체 배양시간 동안 대조구에 비해 현저하게 증가하는 결과를 보였다(p<0.05). 이상의 결과에서 면실, 아마 및 대두의 전지종실을 첨가하여 발효한 알코올 발효사료는 반추위 배양액의 pH를 다소 감소시키며, ammonia 및 휘발성지방산 농도를 증가시키는 것으로 밝혀졌다. 따라서 호밀 건초를 급여하는 조건에서 면실, 아마 및 대두 종실 처리하는 알코올 발효사료는 반추위내 발효양상을 개선할 수 있는 처리임을 알 수 있었다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권12호
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• pp.1864-1872
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• 2019

Objective: This study was conducted to evaluate the effects of Ecklonia stolonifera (E. stolonifera) extract addition on in vitro ruminal fermentation characteristics, methanogenesis and microbial populations. Methods: One cannulated Holstein cow ($450{\pm}30kg$) consuming timothy hay and a commercial concentrate (60:40, w/w) twice daily (09:00 and 17:00) at 2% of body weight with free access to water and mineral block were used as rumen fluid donors. In vitro fermentation experiment, with timothy hay as substrate, was conducted for up to 72 h, with E. stolonifera extract added to achieve final concentration 1%, 3%, and 5% on timothy hay basis. Results: Administration of E. stolonifera extract to a ruminant fluid-artificial saliva mixture in vitro increased the total gas production. Unexpectedly, E. stolonifera extracts appeared to increase both methane emissions and hydrogen production, which is contrasts with previous observations with brown algae extracts used under in vitro fermentation conditions. Interestingly, real-time polymerase chain reaction indicated that as compared with the untreated control the ciliate-associated methanogen and Fibrobacter succinogenes populations decreased, whereas the Ruminococcus flavefaciens population increased as a result of E. stolonifera extract supplementation. Conclusion: E. stolonifera showed no detrimental effect on rumen fermentation characteristics and microbial population. Through these results E. stolonifera has potential as a viable feed supplement to ruminants.

• Advances in Traditional Medicine
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• 제5권1호
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• pp.29-36
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• 2005

Polysaccharide hydrogels constitute a structurally diverse class of biological macromolecules with a wide range of physicochemical properties. They also constitute important members of the family of industrial water-soluble polymers. They find application in Pharmacy as binders, disintegrants, suspending, emulsifying and sustaining agents. According to the International Pharmaceutical Excipients Council (IPEC), an excipient must have an established safety profile. Hence, in the present study, in vitro cytotoxicity on Vero and HEp-2 cell lines, and in vivo acute toxicity in rats were carried out to establish the safety of polysaccharide hydrogels from the seeds of Plantago ovata and Ocimum basilicum. The in vitro cytotoxicity was determined by MTT and SRB assays. In the in vivo acute toxicity, the effects of three different doses of hydrogels (100, 200 and 400 mg/kg body weight) on food and water intake, body weight, biochemical and hematological parameters were studied. The results of in vitro did not show any cytotoxicity on both the cell lines used. In the in vivo acute toxicity, the hydrogels did not show any toxic symptoms in all three dose levels. This establishes the safety of the selected hydrogels. Hence, they can be used as excipients in pharmaceutical dosage forms.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1728-1733
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• 2015

This study was done to establish a mouse model for catheter-related biofilm infection suitable to bioluminescence imaging (BLI). Biofilm formation of Pseudomonas aeruginosa (P. aeruginosa) Xen5 grown on catheter disks in vitro and in an implanted mouse model was real-time monitored during a 7-day study period using BLI. The numbers of integrated brightness (IB) and viable bacterial count (VBC) in the biofilm disks in vitro were highest at 24 h after inoculation; the IB of biofilm in vivo was increased until 24 h after implantation. A statistical correlation was observed between IB and VBC in vitro by linear regression analysis. The actual VBC value in vivo can be estimated accurately by IB without sacrifice. In addition, we monitored the change in white blood cells (WBCs) during infection. The number of WBCs on day 7 was significantly higher in the infection group than in the control group. This study indicates that BLI is a simple, fast, and sensitive method to measure catheter biofilm infection in mice.

• 한국가축번식학회지
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• 제21권3호
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• pp.293-302
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• 1997

Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

• Parasites, Hosts and Diseases
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• 제38권2호
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• pp.59-64
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• 2000

To determine a suitable condition for in vitro infection model of cryptosporidium parvum, four different cell lines, AGS, MDCK, HCT-8 and Caco-2, were used as host cell lines which were cultured at various concentrations of added supplements. These supplement include fetal bovine serum (FBS), sodium choleate, ascorbic acid, folic acid, calcium pantothenate, para-aminobenzoic acid and pyruvate and their effects on the cell lines which were infected with C. parvum were evaluated. The results of this study showed that the AGS cell line was most susceptible to C. parvum whereas the Caco-2 cells appeared to be least susceptible to C. parvum. In regards to the serum condition, 10% FBS was suitable for the growth of AGS and HCT-8 cells, and 1% FBS was good for the growth of the MDCK cells when they were inoculated with C. parvum. Vitamines had a positive effect on the AGS cells, and pyruvate also showed positive effects on all of the cell lines except for Caco-2. Modified medium for each cell line was prepared by adding appropriate amounts of each supplement which resulted in the highest parasite infection number. Modified media increased the number of parasites infected on AGS cells to 2.3-fold higher when compared to the control media. In this study, we found that the AGS cell line was a suitable host model for evaluating C. parvum in vitro study and the media contents for the optimal infection conditions were suggested.

• 생명과학회지
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• 제15권5호
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• pp.755-762
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• 2005

세포 내에서 매우 적은 양으로 존재하는 RNA 전사체와 기능적으로 동일하거나 비슷한 RNA를 RNA 중합효소를 써서 in vitro에서 생화학적으로 의미 있는 양 만큼을 합성할 수 있다. T7 RNA중합효소를 발현하는 재조합 유전자를 지닌 대장균주 BL21/pAR1219로부터 순수한 T7 RNA중합효소를 손쉽게 얻는 방법을 본 논문에서 소개한다. 황산암모늄 분획화와 sephadex SP 컬럼 크로마토그래피법으로써 여타의 방법과 비교하여 더 간단하고 빠르게, 그리고 경제적으로 T7 RNA 중합효소를 분리할 수 있었다. 정제된 T7 RNA중합효소를 이용하여 보통의 화학적 합성법으로 불가능한 긴 길이(1.54 kb)의 RNA전사체를 합성 하였다. 한편,정제된 T7 RNA중합효소에 의해 생성된 망치머리 리보자임은 표적 RNA를 in vitro에서 절단함으로써, 생성된 RNA가 생화학적 기능성을 유지한다는 것을 입증하였다 따라서 본 연구에서 소개되는 절차들은 다양한 길이의 RNA를 목적에 따라 간단하고 경제적으로 합성하는데 유용하게 이용될 수 있다.