In Vitro 고삼투압이 정자 원형질막의 Protein Tyrosine Phosphorylation에 미치는 영향

오영근,장재호,최인호,정노팔,신형철,곽병주 대한의생명과학회 2000 Journal of biomedical laboratory sciences Vol.6 No.4

정자의 원형질막은 삼투압에 의해서 영향을 받는다고 보고되고 있다. 이중 세포막내 분자구조의 변화 특히 막지질 구조의 변화와 동반되는 이온채널의 변화 그리고 Ca2+과 HCO-3의 유동성과도 밀접한 관련이 있는 것으로 보고되고 있다. 지금까지의 연구보고에 의하면, 정자의 첨체반응 (acrosome reaction)이 일어날 경우 protein tyrosine phosphorylation이 증가되는데 이것은 cAMP, protein kinase A 등을 통하여 작용되는 것으로 보고되고 있다. 막의 지질변화를 유도하는 물질로 일종의 sterol acceptor인 BSA가 알려져 있는 바, 실제로 BSA가 막지질 성분에 미치는 영향을 관찰한 결과 cholesterol이 유출되고 이온 등의 유동성 변화가 일어나, 이 유동성 변화가 정자의 adenylyl cyclase를 활성화시켜 cAMP를 증가시키고, PKA가 활성화되어 결과적으로 Protein tyosine phosphorycation이 유도된다고 보는 것이다. 침체반응과 protein tyrosine phosphorylation과는 밀접한 관계가 있는 것으로 사료되고 있다. 본 연구는 정자 원형질막에서 cholesterol이 유출되어 protein tyrosine phosphorylation이 유도될 때, BSA와 같은 sterol acceptor가 작용할 것이라는 전제하에, 고삼투압 하에서 탈수로 인해 원형질막이 위축되더라도 sterol acceptor가 존재한다면 막지질 성분의 구조적 변화가 억제 될 수 있을 것이라는 가설을 설정하였다. 실험결과, 저온 및 고삼투압 하에서 정자운동은 감소되지만 원형질막의 구조적 변화는 없고, 삼투압에 대한 반응정도는 원형질막을 통한 수분이동과 세포용적 변화에 따라 비례적으로 일어난다고 하는 사실을 발견하였다. 이 결과는 정자보존에 있어서 저온변화에 영향을 미치는 여러 인자들 특히 protein fgrosine phosphorylation의 증가와 밀접한 관계가 있음을 시시해 준다. 또한 sterol acceptor로 알려진 BSA는 삼투압이 변화되더라도 역시 중요한 인자로 작용할 수 있음을 알 수 있었으며, 특히 고삼투압으로의 변화는 cAMP와 protein kiname A를 거치는 신호전달과정에 있어서 중요한 요인이라는 사실을 확인할 수 있었다. It has been reported that plasma membrane activity of the spermatozoa may be susceptible to be influenced by extracellular osmolality and such membranous changes involve intracellular molecular changes, special regard to the structure of membranous lipids, and the accompanying ion-channel of which are closely related with their fluidity of Ca2+ and HCO-3. It is of common recognition that a certain kind of sterol acceptor player an important to induce lipid fluctuation of the sperm plasma membrane which have been influenced by BSA administration and came in effect to outflow of cholesterol from the spermatozoa and resulted in changes of ionic fluidity to facilitate adenylyl cyclase, and to induce protein tyrosine phosphorylation by increase of cAMP and activation of PKA. Thus it seems likely that an augmentation of the acrosomal reaction is closely related with protein tyrosine phosphorylation. The following experimental results were obtained in the present study; Under the high osmolality conditions, the spermatozoa motility declined significantly and the structural change of the plasma membrane diminished to confirm that the response degrees to the osmolality depended upon the water transfer volume through the plasma membrane and the changes of cellular volume. Those experimental results suggest that a physiological parameter such as low temperature condition played an important role for presentation of spermatozoa and that inducement of spermatozoa activation for reinforcement of protein tyrosine phosphorylation. On the other hand, it seemed likely that the BSA administration as one of sterol acceptors might represent a key role also under the high osmolality condition and their result also suggests that osmolality change, special regard to high osmolality condition may play an important role also in the processes of signal transmission

Rat의 복강내에 투여한 5-fluorouracil-polyglycolic acid 제형이 장기에 미치는 영향에 관한 연구

강대영,송규상,최정목,노승무,정경수,오정연,김진향,양준묵,최선웅,이진호,조준식,민병무,김용백,김창식,박근성,김승영,김학용,인현빈 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

For more effective intraperitoneal chemotherapy, the authers made 5-fluorouracil-polyglycolic acid composite disks(FU-PGA disks) with 5-fluorouracil and polyglycolic acid, a biodegradable polymer. We inserted the FU-PGA disk into the peritoneal cavites of Sprague-Dawley male rats, the control and three experimental groups; one FU-PGA disk insert group(300mg/kg), two FU-PGA disk insert group(600mg/kg), and three FU-PGA disk insert group(900mg/kg). The control group received a similar number of PGA disk inserts. A pharmacokinetic study was performed to measure the 5-fluorouracil concentration in the peritoneal fluid, blood, and tissues(liver, kidney and heart) at 24 hours, 72hours and 168 hours after insertion of the FU-PGA disk. Light and electron microscopic studies were done. The results were as follows: 1) The number of white blood cells and platelets decreased after FU-PGA insertion. The degree was proportional to the duration and amounts inserted. The change in the number of red blood cells varied slightly. 2) Light microscopically, slight changes were noted at 168 hours in the 3 disk insert group (900mg/kg). Mild fatty change and hepatocyte degeneration around the central veins of the liver were noted, with vacuolar degeneration in the proximal convoluted tubules of the kidney. 3) Electron microscopically,the liver showed focal increases of lipid droplets in the hepatocytes, and irregular nuclear membrane with focal nucleolar segregation of the fibrillar and granular elements. Also the double membranous structure of the hepatocytes and Kupffer cells was widened with bleb formation. The kidney showed wide separation of the nuclear double membrane in the proximal convoluted tubular cells and mesangial cells, with vacuolation and myelin figure formation in the mitochondria of the proximal convoluted tubular cells, at 72 hours and 168 hours in the 2 to 3 FU-PGA insert groups. The heart showed focal loss of cristae,vacuolation and myelin figure formation in the mitochondria of the 2 to 3 FU-PGA insert groups. In conclusion, despite the large amont of FU-PGA inserted, the histopathological changes in the liver, kidney and heart were slight, and consonant with the very low amount of 5-fluorouracil concentrations detected in the liver, kidney and heart. The above results suggest that the FU-PGA composite can serve as a new device for releasing drugs in a controlled manner and easily targeted to intraperitoneal organs. This device can improve the efficacy of intraperitoneal chemotherapy for gastric cancer.

Rat의 복강에 삽입한 Filorouracil-Polyglycolic acid제형의 Fluorouracil용출에 관한 연구

노승무,정경수,오정연,김진향,양준묵,강대영,송규상,최정목,최선웅,이진호,조준식,민병무,김용백,김창식,박근성,김승영,김학용,인현빈 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

A common form of relapse in adenocarcinoma of the stomach is intraperitioneal dissemination, in fact, among gastric adenocarcinoma patients who have undergone surgery intended to cure, approximately 50% of the patients develope initial recurrence in the peritoneal cavity regardless of the anatomic site of the primary tumor within the stomach. The efficacy of systemic postoperative chemotherapy to prevent peritoneal recurrence of gastrric adenocarcinoma is not satisfactory. There is still a great need for improved therapeutic strategies on the disseminated microscopic disease and small miliary nodules remaining on the peritoneal surface or lymphatics after operation. The authers have made fluorouracil-polyglycolic acid composite disks(Fu-PGA disks) with fluorouracil and biodegradable polymer: polyglycolic acid for more effective intraperitoneal chemotherapy. We inserted the Fu-PGA disk(s) in the peritoneal cavity of rat and pharmacokinetic study was performed to measure fluorouracil concentration in the peritoneal fluid, plasma, liver, kidney and heart tissue at 24 hour, 72 hour and 168 hour after insertion of Fu-PGA disk(s). Myelosuppressive action of this composite also was determined following its administration. The data of this study suggested that Fu-PGA composite will be a new device releasing drugs in a controlled manner and having targetability to peritoneum, and this device will be improving the efficacy of intraperitoneal chemotherapy for gastric adenocarcinoma.

Rat의 복강에 삽입한 Fluorouracil-Polyglycolic acid 제형의 Fluorouracil 용출에 관한 연구

노승무,정경수,오정연,김진향,양준묵,강대영,송규상,최정목,최선웅,이진호,조준식,민병무,김용백,김창식,박근성,김승영,김학용,인현빈 忠南大學校 癌共同硏究所 1998 癌共同硏究所 硏究誌 Vol.2 No.1

A common form of relapse in adenocarcinoma of the stomach is intraperitoneal dissemination, in fact, among gastric adenocarcinoma patients who have undergone surgery intended to cure, approximately 50% of the patients develope initial recurrence in the peritoneal cavity regardless of the anatomic site of the primary tumor within the stomach. The efficacy of systemic postoperative chemotherapy to prevent peritoneal recurrence of gastrric adcnocarcinoma is not satisfactory. There is still a great need for improved therapeutic strategies on the disseminated microscopic disease and small miliary nodules remaining on the peritoneal surface or lymphatics after operation. The authers have made fluorouracil-polyglycolic acid composite disks(Fu-PGA disks) with fluorouracil and biodegradable polymer: polyglycolic acid for more effective intraperitoneal chemotherapy. We inserted the Fu-PGA disk(s) in the peritoneal cavity of rat and pharmacokinetic study was performed to measure fluorouracil concentration in the peritoneal fluid, plasma, liver, kidney and heart tissue at 24 hour, 72 hour and 168 hour after insertion of Fu-PGA disk(s). Myelosuppressive action of this composite also was determined following its administration. The data of this study suggested that Fu-PGA composite will be a new device releasing drugs in a controlled manner and having targetability to peritoneum, and this device will be improving the efficacy of intraperitoneal chemotherapy for gastric adenocarcinoma.

위암환자의 복강내에 투여한 Mitomycin C-Carbon Particle의 Mitomycin 용출에 관한 연구

노승무,조영훈,정경수,오정연,김진향,양준묵,강대영,송규상,조준식,최선웅,이진호,민병무,김용백,김창식,박근성,인현빈,정현용,김학용 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

Locoregional recurrence is the most common type of recurrence in surgical operation of gastric adenocarcinoma, and peritoneal dissemination is one of the most difficult problems in advanced gastric adenocarcinoma treatment. Because the peritoneal cavity is the most common site of the first recurrence after gastric cancer resection, intraperitoneal chemotherpy seems a logical choice for cancer chemotherapy. The Mitomycin C(MMC) adsorbed by the activated charcoal particles(CH) is relatively released when the drug concentration surrounding the carbon particles becomes low in the peritoneum of the peritoneal cavity. For the intraperitoneal chemotherapy on the advanced gastric adenocarcnoma, mitomycin C adsorbed on activated carbon particles was administered in the peritoneal cavity just before abdominal wall closure. The closed drainage tubes were inserted in the peritoneal cavity and clamped for tuo hours after completion of operation. MMC concentrations were serially measured in peritoneal fluid, plasma and urine at 2hour, 48 hour, 72 hour and 168 hour following its administration in order to study the efficacy of the MMC-CH as a drug delivery system. There were minimal toxicities in born marrow, liver, and gastrointestinal system after intraperitoneal MMC-CH administration. The data of this study suggested that MMC-CH may have a somewhat more beneficial effect than surgery alone when administered in optimal dose and schedules, but the MMC concentration of the peritoneal fluid was not sufficient to eradicate remnant cancer cells, and effective duration of maintenance was only below 24 hours in the peritoneal fluid and plasma.

반월, 시화공단 악취물질의 안산지역 확산 연구

송동웅,송창근,김영노 漢陽大學校 環境科學硏究所 2003 環境科學論文集 Vol.24 No.-

안산지역은 인근 반월, 시화공단에서 발생하는 악취 및 부유물질로 인한 민원이 그치지 않고 있으며 그 동안 지속적인 악취저감대책에도 불구하고 근원적 해결이 안 되고 있다. 특히, 안산지역은 구지풍인 해륙풍의 영향을 강하게 받는 연안도시로 해풍의 유입과 관련한 오염물질의 이동과 확산과정을 시뮬레이션함으로써 악취민원 발생지역의 범위 예측과 고농도 발생일의 원인규명들을 위한 과학적이고도 정확한 분석이 필요하다. 따라서 본 연구에서는 반월, 시화공단에서 배출되는 악취물질이 해륙풍등의 국지순환에 의하여 이동, 확산되는 과정을 규명하기 위해 대기확산모델링을 실시하였으며 모니터링자료를 이용하여 모델의 재현성을 평가하였다. There have been persistent civil appeals in Ansan area against the odor and aerosols emitted from nearby Banwol/Sinhwa industrial complex. A fundamental solution for the good air quality has not been addressed yet in spite of the continuous counterplan to reduce odor emission. A systematic and scientific study is needed to examine the reason for the odor episode and to predict the impact coverage of odor pollution. An approach by computational simulation is considered to be adequate to investigate the transportation and the dispersion processes of air pollutants blown by sea breeze toward the coastal city, Ansan. This study has employed various dispersion models to simulate the transportation and the dispersion processes of odor pollutants by a local circulation between land/sea breeze using the data set of emission rates of odorous species from the Banwol/Sihwa industrial complex.

레이저를 이용한 얇은 원통형 튜브에서의 유도초음파 발생

김현묵,정경일,장경영,안형근,곽노권,이창목 한국비파괴검사학회 2003 학술대회 논문집 Vol.- No.1

레이저를 이용하여 두께가 얇은 원통형 튜브에 유도초음파를 발생하고, 공기-결합 트랜스듀서를 이용하여 비접촉식으로 수신하였다. 원통형 튜브는 발전설비의 열교환기 및 증기발생기 튜브에 사용되는 인코넬(Inconnel)튜브이다. 레이저의 조사 경로에 선 배열 슬릿을 삽입하여 유도 초음파의 파장을 조절하였다. 파장이 일정할 때 발생되는 모드를 이론적인 분산선도로부터 예측하고, 예측된 위상속도로부터 공기-결합 트랜스듀서의 경사각을 결정하였다. 원통형 튜브에는 축대칭 모드가 지배적으로 발생하는 것으로 나타났다. 이러한 결과는 웨이블렛 변환에 의한 시간-주파수 선도와 이론적인 분산선도를 비교한 결과 증명되었다.

백서의 복강내에 투여한 Mitomycin-C-Polyglycolic acid 제형이 장기에 미치는 영향에 관한 연구

송규상,강대영,최정목,노승무,정경수,오정연,김진향,양준목,최선웅,이진호,조준식,민병무,김용백,김창식,박근성,김승영,김학용,인현빈 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

To evaluate the in-vivo effects of mitomycin-C-polyglycolic acid composite(MMC-PGA) the disk: shaped composite were implanted into the peritoneal cavity of the Sprague-Dawley rats. The pathologic changes were examined at various time points up to 12 weeks. Initially the soft tissue around the inserted disks showed capillary congestion at 3 days. Inflammatory infiltrates with foreign body giant cells appeared from the 2nd week and reached peak response at 6-8 weeks. These reaction diminished prominently at 12 weeks. No specific pathologic change was found in the liver, the kidneys, and the heart. The above results suggest that the MMC-PGA composite can serve as a new device for intraperitoneal chemotherapy of various types of cancers.

생체분해성 고분자를 이용한 사시수술용 제형개발 및 효용성에 대한 연구

민병무,김용백,김승영,김창식,박근성,길숙종,조항진,이성복,노승무,송규상,강대영,조준식,양준묵,정경수,최선웅,이진호,김학용,인현빈 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

A new device, muscle clamping system was developed to facilitate exact quatifying technique A strabismus surgery, and reduce the risk of complications. The device is composed of a lower fixing body with three jaws and an upper supporting body. They are used to clip an extraocular muscle and fix it to the sclera with a single bite. Superior rectus recession on 16 rabbit eyes were performed with this new device. Conjunctival injection, muscle adhesion strength, and light microscopic findings were examined at 1, 2, 4 and 8 weeks postoperatively. The Conjunctival injection were minimal, adhesion power ranged from 420 to 600 gram gravity, which is sufficient in withstand the normal pull of human extraocular muscle. In microscopic exam, some Inflammatory cells and fibrosis were found. The new device was technically easy, fast, and accurate, so it may be useful in stabismus surgery.

생체분해성 망막압정을 이용한 망막고정에 대한 실험적 연구

김용백,민병무,김창식,박근성,김승영,길숙종,조항진,이성복,노승무,송규상,강대영,조준식,양준묵,정경수,최선웅,이진호,김학용,인현빈 충남대학교 의과대학 지역사회의학연구소 1998 충남의대잡지 Vol.25 No.1

Biodegradable retinal fixation devices obtain mechnical fixation of the retina with desirable chorioretinal scarring and with the potential for local, sustained release of antimetabolites and steroids to inhibit proliferative vitreoretinopathy. We manufactured a biodegradable retinal tack with barb that was designed in order to prevent intrusion from implantation of retinal tacks. This study was carried to evaluate the efficacy for retinal fixation and the capability for sustained release of drugs with a newly designed biodegradable retinal tack Biodegradable retinal tacks were made of polymers of glycolic acids and were designed with barbs in a shape to prevent the disinsertion. Biodegradale retinal tacks are divided into 3 parts, a conical portion that is inserted into the sclera, a cylinder portion that remains in the vitreous, and a neck portion between the pin and the cylinder. The tapered conical end was manufactured to allow easy insertion through the retina and choroid into the sclera. A cylinder portion was manufactured with a tapered angle that fixes firmly into the orifice of 19 gauge spinal needle. A neck portion, 0.4 mm in diameter, was designed to prevent disinsertion from following implantation of retinal tack. The applicator was a 19 gauge spinal needle and its orifice was prepared to 15°angle to accept the tapered cylinder portion of the retinal tack. The retinal tacks, secured in the needles, were passed through the formed vitreous and inserted into the retina, choroid, and sclera and were released by pushing the internal needle, usually within 2-3mm of the medullary ray of the posterior rabbit retina A retinal tack was placed in each of 8 pigmented rabbit eyes. Slit-lamp biomicroscopy, indirect ophthalmoscopy and fundus photography were performed periodically from 1 day to 8 weeks after surgery. Eight eyes were enucleated and studied by light microscopy at 8 weeks. Biomicroscopic evaluation of the animals revealed edemas adjacent to the retinal surfaces immediately after insertion of the biodegradable retinal tacks in all the animals. These edemas disappeared after 1 week. The first noticeable change in the size of retinal tacks was shown after 2weeks. The size of the retinal tacks gradually got smaller, decreasing to about one-half at 4 weeks and about one-third at 8 weeks. All retinal tacks remained in inserted places without any movement for an 8 week period. On light microscopy, epiretinal proliferations were seen to extend into the vitreous cavity. Cellular capsules that lined the inner aspect of the scleral defect caused by tack insertion were found. However the adjacent retina had a normal cytologic appearance and architecture in all specimens. We manufactured a biodegradable retinal tack that is designed to prevent intrusion from implantation of retinal tacks. All biodegradable retinal tacks reduce in size with time, but no retinal tacks extruded from the inserted place. The newly designed biodegradable retinal tack can be used for retinal fixation and may be used as a vehicle for the introduction of pharmacologic agents to prevent the cellular events that promote proliferative vitreoretinopathy.