• Journal of Chest Surgery
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• 제16권3호
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• pp.281-288
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• 1983

Heparinization is an essential step in extracorporeal circulation for open heart surgery. But wide individual variation to heparin effect sometimes makes it difficult to anticoagulate safely or neutralize appropriately. Because the conventional set protocol of heparinization did not consider this individual variation, a new method of control of heparinization was proposed by Dr. Brian Bull in 1974. We compared the group in which a conventional set protocol was used [Control group] with the other in which a new protocol modified from that of Bull was used [ACT group], on the aspects of the dosages of heparin and protamine administered and postoperative bleeding. Our conventional protocol [Control group] consisted of: 1. Initial heparin was given at dose of 350U/Kg into the right atrium prior to bypass. 2. Additional heparin was given every hour during E.C.C., as much as a half of the Initial dose. 3. 600U of heparin was mixed into every 100ml. of priming solution. 4. The protamine dose was calculated by totalling the units of heparin given to the patient and giving 1 .8mg. of protamine per 100 units of heparin. ACT protocol [ACT group] consisted of: 1. Initial heparinization was same as that of conventional protocol. 2. ACT`s were checked before [A point] and 10 minutes after initial heparinization [B point]. With these 2 points, a dose response curve was drawn. 3. Heparin for the priming solution was same as in control group. 4. Every 30 minutes during E.C.C., ACT`s were checked with Hemochron [International Technidyne Corp.]. ACT between 450 and 600 seconds was regarded as safety zone. If ACT checked at a time was below 450 seconds, heparin dose was calculated on the dose-response curve to lengthen ACT to 480 seconds and was given into the oxygenator. 5. About 10 minutes before the term of E.C.C., ACT was checked to estimate the blood heparin level at the time. Then, protamine dose was calculated at dose of 1.Stag per 100 units of heparin. The calculated dose of protamine was mixed into 50 to lO0ml of 5% Dextrose Water and dripped intravenously during the period of 15 minutes. Compared these two groups mentioned above, results were obtained as follows: 1. Mean value of normal ACT checked with Hemochron on 30 preoperative patients was 124 seconds [range 95-145 sec.]. 2. Doses of heparin and protamine given to the patient were decreased in ACT group as much as 32.2% and 62.2% respectively. 3. Postoperative bleeding and transfusion were also decreased in ACT group in 60.5% and 67.1% respectively. 4. Our modified dose-response curve did not cause any problems in the control of heparinization. 5. Initial heparinization [Heparin 350U/Kg] was sufficient for the most patients until 60 minutes under extracorporeal circulation. 6. We used 1.5mg of protamine to neutralize 100 units of heparin. But smaller dose of protamine may be sufficient for appropriate neutralization.

• Journal of Chest Surgery
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• 제53권5호
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• pp.258-262
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• 2020

Background: The aim of this study was to evaluate risk factors associated with difficult heparin reversal by protamine after cardiopulmonary bypass. Methods: Data from 120 consecutive patients who underwent open heart surgery from 2009 to 2017 were retrospectively reviewed. Patients were divided into 2 groups: (1) those in whom complete heparin reversal was achieved after a single infusion of protamine (group A, n=89); and (2) those who required more protamine for heparin reversal (group B, n=31). Results: Female sex, prolonged bypass time (>200 min), long aortic cross-clamping time (>120 min), and a lowest rectal temperature <26℃ were significant predictors of difficult heparin reversal. Larger amounts of fresh frozen plasma and platelet concentrate were transfused in group B than in group A. Conclusion: Surgeons' efforts to reduce operative time and avoid deep hypothermia may be helpful for increasing the likelihood of easy heparin reversal, especially in female patients.

• Clinical and Experimental Reproductive Medicine
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• 제47권1호
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• pp.68-76
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• 2020

Objective: Since sperm abnormalities are known to be a major reason for recurrent pregnancy loss (RPL), any defects in DNA structure and chromatin condensation can place embryos at risk in the early stage of development and implantation. As antioxidants such as vitamin C may play a protective role against the destruction of protamine genes in sperm chromatin, this study was conducted to evaluate the effects of vitamin C on chromatin and the expression of protamine genes in the male partners of couples with RPL. Methods: Twenty male partners of couples with RPL were selected as the intervention group and received vitamin C supplementation (250 mg daily for 3 months). Healthy fertile men (n = 20) were included as controls. Sperm chromatin, DNA integrity, and the expression levels of protamine genes were evaluated before and after treatment. Results: Significant differences were found in sperm morphology, protamine deficiency, and apoptosis between the two groups and before and after vitamin C administration. A significant change was found in mRNA levels of PRM1, PRM2, and the PRM1/PRM2 ratio after treatment. Conclusion: Daily oral administration of vitamin C may improve human sperm parameters and DNA integrity by increasing protamine gene expression levels in the male partners of couples with RPL. The beneficial effects of vitamin C supplementation as an antioxidant for the male partners of couples with RPL could lead to improved pregnancy outcomes in these cases.

• Clinical and Experimental Reproductive Medicine
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• 제44권2호
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• pp.73-78
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• 2017

Objective: Sperm morphology plays an important role in infertility, especially in cases of defects in the heads of spermatozoa. Tapered-head or elongated-head spermatozoa are examples of morphological abnormalities. The aim of this study was to compare the semen parameters, levels of protamine deficiency, and frequency of apoptosis between patients with normozoospermia and those with teratozoospermia with tapered-head spermatozoa. Methods: Fifty-two semen samples (27 patients with tapered-head sperm and 25 fertile men) were collected and semen analysis was performed according to the World Health Organization criteria for each sample. Protamine deficiency and the percentage of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3) staining and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays, respectively. Results: Sperm concentration, motility, and normal morphology in the tapered-head spermatozoa (cases) were significantly lower than in the normozoospermic samples (controls). CMA3-reactive spermatozoa (CMA3+) in the case group were more common than in the controls. Apoptotic spermatozoa (TUNEL-positive) were significantly more common in the cases than in the controls. Conclusion: This analysis showed that tapered-head spermatozoa contained abnormal chromatin packaging and exhibited a high rate of apoptosis, which can be considered to be an important reason for the impaired fertility potential in teratozoospermic patients with tapered-head spermatozoa.

• Journal of Chest Surgery
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• 제31권9호
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• pp.867-872
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• 1998

배경: 심폐바이패스에 의한 개심술시 적절한 항응고요법은 수술 관련 혈액응고장애를 최소화한다 는 점에서 중요하다. 심폐바이패스시 헤파린 및 프로타민 투여량을 결정하는데에는 활성화응고시간을 이 용한 고정용량법이 용량반응 곡선을 이용한 방법과 함께 보편적으로 사용되고 있다. 대상 및 방법: 저자들은 고정용량법에 대한 전향적 연구를 통해 고정용량 투여후 헤파린 및 프로 타민의 추가 투여 빈도, 헤파린 저항및 헤파린 유발성 혈소판감소증 빈도, 심폐바이패스시 활성화응고시 간 변화 양상, 그리고 아프로티닌 사용시 활성화응고시간 변화등을 분석하였다. 심폐바이패스 시작전 헤 파린 300 unit/kg를 투여하고 5분후 그리고 심폐바이패스 시작후 10분, 30분, 60분(이후 30분 간격)의 활성화응고시간을 측정하여 400초 이하인 경우 100 unit/kg 헤파린을 추가 투여하였다. 프로타민 중화는 사용 헤파린 100 unit당 1 mg을 투여한후 30분에 측정한 활성화응고시간이 130초 이상 또는 130초 미만이 라도 명백한 응고장애가 있을 경우 0.5mg/kg 추가 투여하였다. 결과: 서울대병원에서 개심술을 받은 환자중 80명(성인 50명, 소아 30명)을 대상으로 하였다. 수 술전 활성화응고시간은 성인에서 114.3$\pm$19.3초, 소아에서 119.5$\pm$18.2초로 나이, 체중, 체표면적 및 성 별에 따른 유의한 차이는 발견되지 않았다. 과거 개심술 병력도 수술전 활성화응고시간에 영향이 없었다. 고정용량법 투여후 헤린 추가 투여가 필요하였던 경우는 성인 환자 10례(20%), 소아 환자 3례(10%)였다. 프로타민 추가투여가 필요하였던 경우는 성인에서 9례(18%), 소아에서 10례(33%)였다. 심폐바이패스 시간 과 프로타민 추가 투여 사이에는 상관관계를 찾을수 없었다. 헤파린저항을 보였던 경우는 성인에서만 2명 (4%)이었고 소아는 없었다. 헤파린 유발성 혈소판감소증은 성인에서 2례(4%), 소아에서 1례(3.3%) 관찰되 었다. 심폐바이패스 운용중 활성화응고시간은 시간경과에 따라 길어지는 양상을 보였다. 성인환자에서 저 용량 아프로티닌이 12례(24%)에서 사용되었는데, 이들에서는 심폐바이패스중 활성화응고시간(celite를 활 성물질로 사용)이 비사용군에 비해 높았고, 활성 물질로 kaolin을 사용했을 때의 활성화응고시간은 celite ACT에 비하여 낮게 나타났다. 결론: 결론적으로 헤파린 및 프로타민 고정 용량 투여법은 큰 문제 없이 운용될 수있으나 추가 용량 투여 빈도의 측면에서는 만족할만한 수준은 되지 못하였다.

• Clinical and Experimental Reproductive Medicine
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• 제43권4호
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• pp.199-206
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• 2016

Objective: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. Methods: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. Results: The sperm DFI showed a significant correlation (r=-0.347, p< 0.001) with sperm motility and morphology (r=-0.114, p< 0.05) but not with other semen parameters. The DFI ($11.5%{\pm}2.0%$) of semen samples was significantly reduced by DGC ($8.1%{\pm}4.1%$) or MACS alone ($7.4%{\pm}3.9%$) (p< 0.05). The DFI was significantly further reduced by a combination of DGC and MACS ($4.1%{\pm}1.3%$, p< 0.05). Moreover, the combination of DGC and MACS ($1.6%{\pm}1.1%$, p< 0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC ($4.4%{\pm}3.2%$) or MACS alone ($3.4%{\pm}2.2%$). Conclusion: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.

• 한국식품과학회지
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• 제32권4호
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• pp.902-907
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• 2000

폐기되는 연어 정자로부터 프로타민을 제조하여 프로타민이 가지는 항균성 및 항산화성을 시험하였다. 그 결과 제조시 전처리 방법에 따라 추출 수율이 차이가 있었는데, 초음파와 균질기 전처리에 의한 추출 방법은 수율에 큰 차이를 보이지 않았지만, $80^{\circ}C$로 가열되는 마이크로파 전처리는 다른 전처리 시료에 비해 5%이상 수율이 낮았다. 각 조건별로 제조된 프로타민 시료들의 아미노산 분석 결과, 시판 프로타민의 경우 arginine 함량이 전체 61.1% 정도를 차지하였고, proline 함량은 11% 정도였다. $H_{2}SO_{4}$ 용액으로 균질화하여 추출한 M의 경우 arginine 함량이 19.3% 정도였고, 반면 methionine과 proline이 각각 34.3%, 28.1% 정도를 차지하였으며, 균질화시킨 다음 $H_{2}SO_{4}$ 용액으로 추출한 MC도 시판 프로타민과 유사한 조성을 나타내었다. 마이크로파 전처리 P60 및 초음파 전처리 UU6는 arginine이 각각 60.8%, 52.4%를 차지하였고, P60은 proline 함량이, UU6는 tryptophan 함량이 다소 많은 것이 차이였다. 제조된 프로타민은 곰팡이 이외에 그람양성 및 음성 세균에 대해 항균활성이 있었고, UU6나 P60이 다른 시료에 비해 활성이 높았고, 일부 세균에 대해서는 농도 의존성이 있으며, 시판 제품보다 높은 항균활성을 나타내었다. 프로타민의 항산화성은 전처리 방법에 관계없이 모든 시료에서 확인되었으며, 전처리 방법에 따라 어느정도 차이를 보였다. $EDA_{50}$을 측정한 결과, P60이 $101\;{\mu}g/mL$, UU6가 $410\;{\mu}g/mL$, M이 $520\;{\mu}g/mL$, MC가 $490\;{\mu}g/mL$였으며, 시판 프로타민은 $470\;{\mu}g/mL$였다.

• Molecules and Cells
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• 제37권8호
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• pp.613-619
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• 2014

The optic nerve often suffers regenerative failure after injury, leading to serious visual impairment such as glaucoma. The main inhibitory factors, including Nogo-A, oligodendrocyte myelin glycoprotein, and myelin-associated glycoprotein, exert their inhibitory effects on axonal growth through the same receptor, the Nogo-66 receptor (NgR). Oncomodulin (OM), a calcium-binding protein with a molecular weight of an ~12 kDa, which is secreted from activated macrophages, has been demonstrated to have high and specific affinity for retinal ganglion cells (RGC) and promote greater axonal regeneration than other known polypeptide growth factors. Protamine has been reported to effectively deliver small interference RNA (siRNA) into cells. Accordingly, a fusion protein of OM and truncated protamine (tp) may be used as a vehicle for the delivery of NgR siRNA into RGC for gene therapy. To test this hypothesis, we constructed OM and tp fusion protein (OM/tp) expression vectors. Using the indirect immunofluorescence labeling method, OM/tp fusion proteins were found to have a high affinity for RGC. The gel shift assay showed that the OM/tp fusion proteins retained the capacity to bind to DNA. Using OM/tp fusion proteins as a delivery tool, the siRNA of NgR was effectively transfected into cells and significantly down-regulated NgR expression levels. More importantly, OM/tp-NgR siRNA dramatically promoted axonal growth of RGC compared with the application of OM/tp recombinant protein or NgR siRNA alone in vitro. In addition, OM/tp-NgR siRNA highly elevated intracellular cyclic adenosine monophosphate (cAMP) levels and inhibited activation of the Ras homolog gene family, member A (RhoA). Taken together, our data demonstrated that the recombinant OM/tp fusion proteins retained the functions of both OM and tp, and that OM/tp-NgR siRNA might potentially be used for the treatment of optic nerve injury.

• Animal Bioscience
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• 제36권12호
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• pp.1796-1805
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• 2023

Objective: This study aims to identify heat shock protein70-2 (HSP70-2) and protamine-1 (PRM1) mRNA and protein in Madura bull sperm and demonstrate their relation as bull fertility biomarkers. Methods: The Madura bull fertility rates were grouped based on the percentage of first service conception rate (%FSCR) as high fertility (HF) (79.04%; n = 4), and low fertility (LF) (65.84%; n = 4). mRNA of HSP70-2 and PRM1 with peptidylprolyl isomerase A (PPIA) as a housekeeping gene were determined by quantitative real-time polymerase chain reaction, while enzyme-linked immunoassay was used to measure protein abundance. In the post-thawed semen samples, sperm motility, viability, acrosome integrity, and sperm DNA fragmentation index were analyzed. Data analysis was performed on the measured parameters of semen quality, relative mRNA expression, and protein abundance of HSP70-2 and PRM1, among the bulls with various fertility levels (HF and LF) in a one-way analysis of variance analysis. The Pearson correlation was used to analyze the relationship between semen quality, mRNA, proteins, and fertility rate. Results: Relative mRNA expression and protein abundance of HSP70-2 and PRM1 were detected and were found to be highly expressed in bulls with HF (p<0.05) and were associated with several parameters of semen quality. Conclusion: HSP70-2 and PRM1 mRNA and protein molecules have great potential to serve as molecular markers for determining bull fertility.

• 대한수의학회지
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• 제12권1호
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• pp.77-84
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• 1972

이 실험(實驗)은 뇌염(腦炎)바이러스(MVE virus)들 acetone, Tween-ether 그리고 Tween-ether-protamine-sulphate 로 처리한 후 그 바이러스가 지니고 있는 적혈구응집력가(赤血球凝集力價), 보체결합력가(補體結合力價) 그리고 감염력가(感染力價)의 변화 여부를 관찰함과 아울러 초원심분리법(超遠心分離法)에 의하여 이 바이러스가 지니고 있는 위의 세 가지 활성물질(活性物質)의 분획(分劃) 가능성(可能性)을 검토한 결과 다음과 같은 결과를 얻었다. 1) Acetone, Tween-ether 그리고 Tween-ether protamine-sulphate 처리에 의하여 MVE virus가 지니는 적혈구응집력가(赤血球凝集力價)는 8~16 배(倍)로 증가 하였다. 2) Acetone 및 Tween-ether 처리로 보체결합력가(補體結合力價)는 4 배(倍)로 증가 되었으나 Tween-ether protamine-sulphate 처리로는 저하되는 경향이 있었다. 3) Tween-ether로 처리하면 감염력가(感染力價)는 완전히 상실 되나 acetone으로 처리하면 $TCID50/log_{10}$ 3이 감퇴 되었다. 4) 위의 화학제의 처리와 관계없이 바이러스의 적혈구응집력가(赤血球凝集力價)는 $37^{\circ}C$ 에서 10 분간 가열(加熱)됨으로써 완전 소실 되었으나 보체결합력가(補體結合力價)는 상승 하였으며 이 활성도(活性度)는 $65^{\circ}C$ 에서 20 분간 까지도 계속 유지되었다. 5) 처리되지 않은 바이러스와 acetone 처리된 바이러스의 보체결합력가(補體結合力價)나 적혈구응집력가(赤血球凝集力價)는 Tween-ether 나 Tween-ether-protamine-sulphate 로 처리한 바이러스의 그것보다 열(熱)에 대해서 더 안정(安定)하였다. 6) 적혈구응집억제반응용(赤血球凝集抑制反應用) 항원제조(抗原製造)에 있어서 Tween-ether 로 처리하거나 acetone 으로 처리하여 만든 두가지 항원(抗原)은 모두 동일(同一)한 역가(力價)를 보였다. 7) 10~60%의 sucrose gradient centrifugation 에서 처리되지 않은 바이러스는 두개의 보체결합(補體結合)피크가 저농도(低濃渡)와 고농도(高濃渡)에서 각각 나타났으며 저농도(低濃渡)에서 얻은 재료는 적혈구응집력(赤血球凝集力)과 감염력(感染力)을 동반하고 있었다. 8) Acetone 으로 처리된 바이러스의 초원심분리(超遠心分離)에서는 위에 말한 두 보체결합(補體結合)피크 중 고농도부(高濃渡部)의 것은 감소 되었으나 다른 하나는 영향을 받지 않았다. 그리고 적혈구응집력가(赤血球凝集力價)가 증가된 반면 감염최고력가(感染最高力價)는 감퇴 되었으며 이 세 가지 활성(活性)은 acetone 처리 후에는 명확히 분획(分劃)되지 않았다. Tween-ether 로 처리된 바이러스는 초원심분리(超遠心分離)에 의하여 고농도부(高濃渡部)에 있는 보체결합(補體結合)피크는 완전 소실된 반면 저농도부(低濃渡部)에 있는 보체결합최고력가(補體結合最高力價)는 영향을 받지 아니 하였으며 이 피크는 감염력(感染力)을 동반하지 않는 적혈구응집소(赤血球凝集素)와 분획(分劃)이 가능 하였다.