• Clinical and Experimental Reproductive Medicine
• /
• 제11권2호
• /
• pp.59-67
• /
• 1984

In vitro fertilization have been performed to know whether the frozen semen has fertilizing ability and can be used clinically. The results of cultured and developed embryos obtained are as follows: 1. The semen was frozen in three media for the good viability. The viability was more than 50% and the motility was also moderate (grade III), 2. As the 33 oocytes were collected from 45 follicles, the oocyte recovery rate was 73.3%. Among them, mature and immature ova were 5% each, and premature ova were 69.7%, When the first polar body was appeared, above ova were inseminated after adequate incubation with activated sperms. 3. The main components of three freezing medium containing egg yolk, glycerol and pyruvate respectively were the best for sperm viability, and Ham's F-10 medium was used for the fertilization and culture of eggs. 4. The results of in vitro fertilization of 33 ova, showed the second polar body developed in 12%, polyspermia in 24%, 1-cell embryo in 21% and 2-cell embryo in 9%. One mature ova developed to blastocyst via 16-cell to 32-cell embryo. The fertilization rate was 66%. 5. Above mentioned results represent that the frozen semen has fertilizing ability and can be used practically in the clinic.

• BMB Reports
• /
• 제51권11호
• /
• pp.596-601
• /
• 2018

Colon cancer is one of the most lethal and common malignancies worldwide. STK899704, a novel synthetic agent, has been reported to exhibit anticancer effects towards numerous cancer cells. However, the effect of STK899704 on the biological properties of colon cancer, including cancer cell migration and cancer stem cells (CSCs), remains unknown. Here, we examined the inhibitory effect of STK899704 on cell migration and CSC stemness. In the wound healing assay, STK899704 significantly inhibited the motility of colon cancer cells. Furthermore, STK899704 downregulated the mRNA expression levels of the cell migration mediator focal adhesion kinase (FAK). STK899704 also suppressed mitogen-activated protein kinase kinase and extracellular signal-regulated kinase, which are downstream signaling molecules of FAK. Additionally, STK899704 inhibited stemness gene expression and sphere formation in colon cancer stem cells. These results suggest that STK899704 can be used to treat human colon cancer.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권2호
• /
• pp.831-836
• /
• 2015

Tumor fluid accumulation occurs in both human cancer and experimental tumor models. Solid tumors show a tendency to tumor fluid accumulation because of their anatomical and physiological features and this may be influenced by molecular factors. Fluid accumulation in the peri-tumor area also occurs in the Dunning model of rat prostate cancer as the tumor grows. In this study, the effects of tumor fluids that were obtained from Dunning prostate tumor-bearing Copenhagen rats on the strongly metastatic MAT-LyLu cell line were investigatedby examining the cell's migration and tumor fluid's toxicity and the kinetic parameters such as cell proliferation, mitotic index, and labelling index. In this research, tumor fluids were obtained from rats injected with $2{\times}10^5$ MAT-LyLu cells and treated with saline solution, and 200 nM tetrodotoxin (TTX), highly specific sodium channel blocker was used. Sterilized tumor fluids were added to medium of MAT-LyLu cells with the proportion of 20% in vitro. Consequently, it was demonstrated that Dunning rat prostate tumor fluid significantly inhibited proliferation (up to 50%), mitotic index, and labeling index of MAT-LyLu cells (up to 75%) (p<0.05) but stimulated the motility of the cells in vitro.

• Molecules and Cells
• /
• 제26권4호
• /
• pp.329-337
• /
• 2008

Src-family kinases are critically involved in the control of cytoskeleton organization and in the generation of integrin-dependent signaling responses, inducing tyrosine phosphorylation of many signaling and cytoskeletal proteins. Activity of the Src family of tyrosine kinases is tightly controlled by inhibitory phosphorylation of a carboxy-terminal tyrosine residue, inducing an inactive conformation through binding with its SH2 domain. Dephosphorylation of C-ter tyrosine, as well as its deletion of substitution with phenylalanine in oncogenic Src kinases, leads to autophosphorylation at a tyrosine in the activation loop, thereby leading to enhanced Src activity. Beside this phophorylation/dephosphorylation circuitry, cysteine oxidation has been recently reported as a further mechanism of enzyme activation. Mounting evidence describes Src activation via its redox regulation as a key outcome in several circumstances, including growth factor and cytokines signaling, integrin-mediated cell adhesion and motility, membrane receptor cross-talk as well in cell transformation and tumor progression. Among the plethora of data involving Src kinase in physiological and pathophysiological processes, this review will give emphasis to the redox component of the regulation of this master kinase.

• 환경생물
• /
• 제33권1호
• /
• pp.45-52
• /
• 2015

본 연구에서는 새로운 국내 생태독성평가 시험 생물종으로서 유글레나의 적용가능성을 평가하기 위하여 국내 서식종인 E. agilis를 이용하여 5종의 휘발성 유기용 매(Acetone, Dimethyl sulfoxide (DMSO), Ethanol, Methanol)에 의한 독성영향을 평가하였다. E. agilis의 식물적 특성을 이용한 급성독성영향 평가 결과, D. magna에 비해 DMSO에 대한 독성민감도가 상대적으로 낮았으나 전반적으로 비교생물 종과 유사하거나 민감하였다. E. agilis의 동물적 특성을 이용한 세포반응 시험 결과, Chloroform의 경우 반수영향을 야기하는 시험물질농도(96시간 $EC_{50}$)에 상당히 신속하고 민감한 독성반응을 나타내었다. 환경시료에 대한 모니터링 목적으로 다양한 생물 종을 이용한 생물검정 기법 적용이 증가하고 있는 추세이다. 수 생태 환경관리를 목적으로 새로운 생태독성시험 개발에 적용할 생물 종은 국내 수질환경에 적합하고 배양이 용이하며 다양한 유해화학물질에 대한 독성이 민감해야 한다. 또한 반응시간이 짧아 실시간으로 모니터링이 가능할 뿐만 아니라 경제성도 고려되어야한다. 본 연구의 결과 E. agilis는 배양이 용이하고 다루기 쉬우며 생장율(growth rate), 이동속도(velocity), 활동도(motility), 세포형태(com-pactness) 등 식물적 특성과 더불어 동물적 특성과 연계된 인자들을 종말점으로 독성시험을 수행할 수 있다. 따라서 E. agilis는 다양한 유해화학 물질 및 환경시료에 대하여 신속하고 민감한 독성반응 도출이 가능할 것으로 예상되며 향후 국내 수질관리를 위한 새로운 생태독성 영향평가 생물 종으로 적용될 수 있을 것으로 판단된다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권3호
• /
• pp.1099-1104
• /
• 2014

Aims and Background: Ginsenoside Rh2, which exerts the potent anticancer action both in vitro and in vivo, is one of the most well characterized ginsenosides extracted from ginseng. Although its effects on cancer are significant, the underlying mechanisms remain unknown. In this study, we sought to elucidate possible links between ginsenoside Rh2 and phosphoglucose isomerase/autocrine motility factor (PGI/AMF). Methods: $KG1{\alpha}$, a leukemia cell line highly expressing PGI/AMF was assessed by western blot analysis and reverse transcription- PCR (RT-PCR) assay after transfection of a small interfering (si)-RNA to silence PGI/AMF. The effect of PGI/AMF on proliferation was measured by typan blue assay and antibody array. A cell counting kit (CCK)-8 and flow cytometry (FCM) were adopted to investigate the effects of Rh2 on PGI/AMF. The relationships between PGI/AMF and Rh2 associated with Akt, mTOR, Raptor, Rag were detected by western blot analysis. Results: KG1${\alpha}$ cells expressed PGI/AMF and its down-regulation significantly inhibited proliferation. The antibody array indicated that the probable mechanism was reduced expression of PARP, State1, SAPK/JNK and Erk1/2, while those of PRAS40 and p38 were up-regulated. Silencing of PGI/AMF enhanced the sensibility of $KG1{\alpha}$ to Rh2 by suppressing the expression of mTOR, Raptor and Akt. Conclusion: These results suggested that ginsenoside Rh2 suppressed the proliferation of $KG1{\alpha}$, the same as down-regulation of PGI/AMF. Down-regulation of PGI/AMF enhanced the pharmacological effects of ginsenoside Rh2 on KG1${\alpha}$ by reducing Akt/mTOR signaling.

• 한국미생물·생명공학회지
• /
• 제43권4호
• /
• pp.357-366
• /
• 2015

쿼럼센싱은 세포 간의 의사소통 방법이며 박테리아의 병원성과 관련된 유전자들의 조절메커니즘이다. 박테리아는 다양한 생리학적 과정들을 제어하기 위해 이 쿼럼센싱 시스템을 활용한다. 본 연구에서 석류(Punica granatum L.) 껍질 추출물이 바이오 리포터 균주인 Chromobacterium violaceum 과 C. violaceum CV026에서 쿼럼센싱 억제능을 갖는 것으로 1차 선별되어 식중독균인 Y. enterocolitica에서 편모에 의한 운동능과 바이오필름형성 억제능에 대한 석류껍질 추출물의 효과에 대한 다음 실험을 수행하였다. 추가로 N-acylhomoserine lactones (AHLs)의 합성(yenI and yenR)과 편모 레귤론(fliA, fleB and flhDC) 에 관련된 특정유전자의 발현변화를 역전사 중합효소연쇄반응법으로 평가하였다. 결과는 석류껍질 추출물이 C. violaceum CV026에서 쿼럼센싱으로 제어되는 바이오레신 생산을 78.5%까지 억제하였으며, Y. enterocolitica에서는 세포의 성장에 영향을 주지 않고 바이오필름 형성과 편모 운동성을 현저히 감소시키는 것을 확인할 수 있었다. 이러한 억제 효과는 AHLs의 합성과 운동성에 관여하는 유전자 발현을 down-regulation 하는 결과와도 일치하였다. 본 연구의 결과는 석류껍질 추출물의 임상 적용을 위하여 생체 내 특성에 대한 추가적인 연구가 필요하다는 것뿐 아니라 석류껍질 추출물이 사람의 위장관염을 방지하기 위한 잠재적인 치료제가 될 수 있다는 것을 보여준다.

• 한국동물생명공학회지
• /
• 제36권2호
• /
• pp.106-115
• /
• 2021

Sperm cryopreservation is an important method of assisted reproductive techniques and storing genetic resources. It plays a vital role in genetic improvement, livestock industrial preservation of endangered species, and clinical practice. Consequently, the cryopreservation technique is well organized through various studies, especially on Korean native cattle (Hanwoo). However, the cryopreservation technique of Korean native brindled cattle, which is one of the native cattle species in Korea, is not well organized. Therefore, it is necessary to develop a Supplementary Table technique for the cryopreservation of Korean native brindled cattle. For this purpose, it is important to first evaluate the quality of the currently produced cryopreserved sperm of Korean native brindled cattle. In this study, we randomly selected 72 individual Korean native brindled cattle semen samples collected from 8 different region research centers and used them to evaluate sperm functions. We focused on the quality evaluation of cryopreserved Korean native brindled cattle semen following the measurement of motion kinematics, capacitation status, intracellular ATP level, sperm motility, and cell viability. Then, the values of each of the eight groups were derived from various sperm parameters of nine individual samples, including sperm motility, kinematics, cellular motility, and intracellular ATP levels, which were used to compare and evaluate sperm function. Overall, differences in various sperm parameters were observed between most of the research centers. Particularly, the deviations of motility and motion kinematics were high according to the sample. Therefore, we suggest that it is necessary to develop a standard method for the cryopreservation of Korean native brindled cattle semen. We also suggest the need for sperm quality evaluation of the cryopreserved semen of Korean native brindled cattle before using artificial insemination to attain a high fertility rate.

• 한국가축번식학회지
• /
• 제23권1호
• /
• pp.29-36
• /
• 1999

본 연구는 Hamster 정소상체 정자의 운동성에 영향을 미치는 methylxanthine derivatives 의 효과를 알아보고자 수행하였다 . Pentoxifylline 을 첨가하였을 때 운동 형태에 대한 첨가효과는 정소상체 체부에서 차이를 보였으며, 농도에 대한 변화는 1 mM 농도군에서 현저한 차이를 나타냈다. 2-Deoxya-denosine 첨가군에서도 pentoxifylline 첨가군과 같이 특히 VCL 이 정소상체 체부 정자부터 증가하였으며 다른 농도군에 비해 1 mM 과 2 mM 에서 차이가 났다. Hypoxanthine 첨가군은 1 mM 농도에서 다른 첨가군에 비해 운동 형태가 증가하였다. 그러나 pentoxifylline 과 adenosine 첨가군과는 달리 농도를 달리하여도 뚜렷한 운동형태의 변화는 관찰되지 않았다. VCL 과 VAP 는 pentoxifylline 1 mM 과 2-Deoxyadenosine 1 mM 첨가군에서 정소상체 체부 정자의 운동성이 증가하고 hypoxanthine 1 mM 첨가군도 유의하게 증가하였다. 결론적으로 hamster 정소상체 정자의 운동성은 적정한 농도의 methylxanthine derivatives 의 첨가로 증가됨을 알 수 있었다.

• Journal of Microbiology and Biotechnology
• /
• 제24권8호
• /
• pp.1034-1043
• /
• 2014

This study determined the effect of light intensity and photoperiod on the dry cell weight and total amount of carotenoids in four isolates of purple non-sulfur bacteria obtained from shaded and exposed microhabitats of a mangrove ecosystem in Kota Kinabalu, Sabah, Malaysia. The initial isolation of the bacteria was carried out using synthetic 112 medium under anaerobic conditions (2.5 klx) at $30{\pm}2^{\circ}C$. On the basis of colony appearance, cell morphology, gram staining, motility test, and 16S rRNA gene sequencing analyses, all four bacteria were identified as Afifella marina. One of the bacterial isolates, designated as Af. marina strain ME, which was extracted from an exposed mud habitat within the mangrove ecosystem, showed the highest yield in dry cell weight ($4.32{\pm}0.03g/l$) as well as total carotenoids ($0.783{\pm}0.002mg/g$ dry cell weight). These values were significantly higher than those for dry cell weight ($3.77{\pm}0.02g/l$) and total carotenoid content ($0.706{\pm}0.008mg/g$) produced by the isolates from shaded habitats. Further analysis of the effect of 10 levels of light intensity on the growth characteristics of Af. marina strain ME showed that the optimum production of dry cell weight and total carotenoids was achieved at different light intensities and incubation periods. The bacterium produced the highest dry cell weight of 4.98 g/l at 3 klx in 72 h incubation, but the carotenoid production of 0.783 mg/g was achieved at 2.5 klx in 48 h incubation. Subsequent analysis of the effect of photoperiod on the production of dry cell weight and total carotenoids at optimum light intensities (3 and 2.5 klx, respectively) revealed that 18 and 24 h were the optimum photoperiods for the production of dry cell weight and total carotenoids, respectively. The unique growth characteristics of the Af. marina strain ME can be exploited for biotechnology applications.