• Journal of Chest Surgery
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• v.13 no.1
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• pp.13-20
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• 1980

Heparin would have been used for preventing clotting of blood during extracorporeal circulation and subsequent use of protamine sulfate and made possible the neutralization of heparin. This procedure has been adopted for eliminating one of the great causes of bleeding, especially in cardiac surgery. In this experiment, the hypocoagulability of blood induced by heparin followed by neutralization with treatment of protamine sulfate were estimated by the Lee-White clotting time [CT], partial thromboplastin time [PTT] and protamine titration test. The results were as follows: 1] Comparison of clotting time between the heparinized [2.0 mg/kg] and non-heparinized dogs was done using CT and PT`I` of the blood. In heparinized group [Group I], the CT lasted infinitively and prolongation of PTT [4 times than normal] until 60 minutes. The CT [2 times] and PTT [3 times] has been shortened after 90 minutes, however they returned to normal limit level within 180 minutes. 2] The determination of appropriate ratio of heparin and protamine In vivo were performed. The group II [heparin 2.0 mg/kg, protamine 1.0 mg/kg] revealed rapid decrease of CT and PTT, but returned to normal after 120 minutes. The group III [heparin 2.0 mg/kg, protamine 2.0 mg/kg] returned rapidly to normal within 15 minutes. The group IV [heparin 2.0 mg/kg, protamine 3.0 mg/kg] recovered its normal level after 60 minutes. The group V [heparin 2.0 mg/kg, protamine 4.0 mg/kg] recovered its normal level after 90 minutes. 3] In the combined experimental study In vivo and vitro, the protamine titration test was done using the dog which were given 2.0 mg/kg and 3.0 mg/kg of heparin, respectively and coagulation time were checked after 15, 30, 60 and 120 minutes. The complete neutralization was showed to be heparin-protamine ratio of 1:1 to 1.5. 4] In vitro study, fresh blood was drawn into known amount of heparin content [20, 40, 60 and 100/ug per 1 ml of blood] syringe, thereafter protamine titration test was done. In all cases, the complete neutralization was found in heparin-protamine ratio of 1:0.85 to 1.5. 5] It was found by the present experiment that the ideal heparin-protamine ratio was 1:1 within 60 minutes and 1:0.5 after 60 minutes for avoiding the serious side effect due to overadministration of protamine sulfate.

• Journal of Chest Surgery
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• v.24 no.3
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• pp.253-260
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• 1991

The adequacy of anticoagulation with heparin during cardiopulmonary bypass, and precise neutralization with protamine at the conclusion of cardiopulmonary bypass, were important. In sixty children undergoing cardiopulmonary bypass, ACT and heparin dose-response curve were studied. Total dose of heparin before bypass were 2.80$\pm$0.74 mg/kg and the amount of protamine administered after bypass were 3.0$\pm$1.23 mg/kg. So protamine: heparin ratio was 1.07: l.c After administration of protamine which dose is calculated with heparin dose-response curve, ACTs were returned to normal range[mean 114.8 $\pm$13 second]. The heparin sensitivity and its half-life do not have relationship with age, weight, height, surface area and urine amount during operation. And there are too much individual variations in heparin sensitivity and its half-life. So conventional heparin protocols can overestimate or underestimate the amount of heparin and protamine. Heparin dose-response curve makes it possible to maintain anticoagulation in a safe range during bypass with adequate amount of heparin individually. At the conclusion of bypass, this curve can be used to predict the precise amount of protamine amount of protamine needed for neutralization of the heparin. But heparin dose-response curve to be used clinically, further studies will be needed about relationship between ACT and heparin level in the high range, influence of hemodilution and hypothermia to ACT and discrepancy between true adequate amount of protamine and calculated amount by heparin dose-response curve.

• Journal of Chest Surgery
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• v.13 no.4
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• pp.346-355
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• 1980

It has been proposed that wide individual variation in response to heparin be not considered in the conventional set protocol for the control of heparin and protamine during extracorporeal circulation. In this paper, two protocol of heparin and protamine therapy were compared to assess the role of the Activated Clotting Time [ACT] in relation to heparin, protamine, and postoperative blood loss and transfusion. The study groups consisted of the 31 patients [adults 15 and children 16] anticoagulated with the conventional heparin protocol and the 31 patients [adults 15 and children 16] anticoagulated with ACT protocol during extracorporeal circulation. In the conventional heparin protocol, two mg of heparin per kg was administered initially with an additional 0.75 mg of heparin per kg every 30 minutes of extracorporeal circulation, and reversal was accomplished with protamine in a dose of 1.5 times the total milligram of heparin. In the ACT protocol, two mg of heparin per kg was administered initially with an additional dose of heparin enough to reach an ACT of 480 seconds [within safe zone 300 to 600 seconds] from the patient`s dose response curve every 1 hour of extracorporeal circulation, and reversal was done with protamine in a dose of 1.3 times the milligram of the residual heparin. The results were summarized as follows. After a dose of 2 mg per kg of heparin, the patient`s ACT varied from 240 to 600 seconds in adults and from 240 t~ 660 seconds in children. In the ACT group the total amount of heparin administered was markedly reduced when compared to the conventional group, and less protamine was required to neutralize heparin. The dose of heparin administered decreased from 7.07 [SE 0.42] mg/kg of the conventional group to 4.92 [SE 0.32] mg/k8 of the ACT group in adults and from 10.17 [SE 1.15] mg/kg to 5.23 [SE 0.24] mg/kg in children, which represent 30.4% and 48.6% decrease respectively. The dose of protamine administered for reversal decreased from 10.6 [SE 0.63] mg/kg of the conventional group to 3.35 [SE 0.35] mg/kg of the ACT group in adults and from 15.7 [SE 1.70] mg/kg to 3.26 [SE 0.27] mg/kg in children, which represent 68.4% and 79.2% respectively. The ratio of protamine to heparin administered in the conventional group was 1.50:1 in adults and 1.54:1 in children, but in the ACT group 0.68:1 in adults and 0.62:1 in children. Postoperative blood loss and transfusion revealed no statistically significant difference between the two groups. Although six patients in the conventional group and one in the ACT group needed re-exploration for continuous hemorrhage, no case of generalized oozing was encountered, and in each case a definite bleeding site was identified. Author would like emphasizing the value of the ACT protocol in controlling heparin and protamine administration during extracorporeal circulation.

• Journal of Veterinary Clinics
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• v.34 no.3
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• pp.197-199
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• 2017

A 14-year-old castrated male ShihTzu diagnosed with chronic kidney disease (CKD) 6 months prior was referred to our clinic. The patient had been experiencing symptoms such as vomiting, poor appetite and hind limbs weakness. Hematology tests showed that he had a non-regenerative anemia. With aggressive treatment, the patient's state had gotten worse. He showed ragged breath, vomiting blood and loss of consciousness temporarily. Hematocrit maintained low level. Gastric hemorrhage was strongly suspected by hematemesis. Whole blood transfusion was performed and heparin was used as an anticoagulant. Prior to transfusion, the blood cross matching between donor and patient was performed and the result was compatible. After the transfusion was stabilized, 1 mg of protamine sulfate for each 100 units of heparin was prepared and given intravenously over 3 minutes to reverse the effects of heparin. Immediately after protamine injection, the patient conducted severe anaphylactic shock. Protamine sulfate is used to reverse the anticoagulant action of heparin in dogs and humans. The adverse reaction of protamine sulfate range from mild reaction to fetal cardiac arrest. When using protamine sulfate as heparin neutralization, it can lead to the death of a patient cause of anaphylactic shock. For this reason, the protamine sulfate should be injected slowly with antihistamine and the clinician should carefully monitor patients.

• Journal of Chest Surgery
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• v.37 no.7
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• pp.606-608
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• 2004

Anaphylactic reaction to protamine sulfate, which is used widely to reverse the anticoagulative effect of heparin after cardiopulmonary bypass, is very rare. But the result of anaphylactic reaction can be very fatal and the mechanism of it is still not clear. We report. a. case of severe anaphylactic reaction to protamine sulfate following the replacement of the mitral valve and .Maze procedure using microwave in a non-diabetic 57-year-old female patient.

• Journal of Chest Surgery
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• v.23 no.2
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• pp.222-230
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• 1990

Protamine, a polycationic peptide extracted from fish, has been widely used for the reversal of anticoagulant action of heparin. However it may cause untoward circulatory side effects including hypotension and bradyarrhythmia. Nowadays, histamine and prostacyclin are regarded as one of the causative agents in the underlying mechanism of hemodynamic changes. To certify the possible role of histamine and prostacyclin, we observed simultaneous changes of the hemodynamic status, plasma concentration of thromboxane B, and circulating platelet count before and after intravenous injection of protamine. Experimental dogs, weighing 12-14kg, were divided into 2 groups; group A animals [n=10], were pretreated with indomethacin[2.5mg/kg] and group B animals[n=10] were pretreated with chlorpheniramine[0.5mg/kg] Heparin[3mg/kg] and protamine [3mg/kg] were administered sequentially in both groups. The results were as follows ; 1. The mean systemic arterial pressure was maintained well in groups A, whereas in group B it decreased from 165\ulcorner18mmHg to 138\ulcorner30mmHg[p<0.01] and 151\ulcorner21 mmHg[p<0.05] at 1 minute and 2 minutes after protamine injection. The mean pulmonary arterial pressure was not changed significantly in group A, whereas in group B it increased from 852 mmHg to 11\ulcorner3 mmHg[p<0.05], 11\ulcorner3 mmHg[p<0.05] and 10\ulcorner3 mmHg[p<0.05] at 1 minute, 3 minutes and 5 minutes after protamine injection. 2 The thromboxane B2 was not changed significantly in group A, whereas in group B it increased from 399\ulcorner401 \ulcornerg/ml to 744\ulcorner615 \ulcornerg/ml[p<0.05] and 814\ulcorner1070 \ulcornerg/ml [p<0.0 5] at 1 minute and 3 minutes after protamine injection without concomitant changes of pulmonary vascular resistance and pulmonary capillary wedge pressure. 3. The number of circulating platelet was not changed in group A, whereas in group B it decreased from 207100\ulcorner103600/\ulcornerl to 159700\ulcorner90900/\ulcornerl [p<0.05] at 1 minute after protamine injection, Although thromboxane B2 and platelet count were changed significantly after protamine injection, they did not cause the remarkable hemodynamic changes. Considering the above results, hemodynamic changes may be caused mainly by prostacyclin rather than thromboxane or platelet. Therefore, the pretreatment with cyclooxygenase inhibitor would be beneficial to prevent circulatory adverse effects of protamine for the patients undergoing cardiac surgery.

• Journal of Chest Surgery
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• v.16 no.3
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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.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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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.

• Journal of Chest Surgery
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• v.53 no.5
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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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• v.43 no.4
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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.