• Journal of Chest Surgery
• /
• v.31 no.9
• /
• pp.867-872
• /
• 1998

Background: The fixed dose regimen with activated coagulation time(ACT) is the most commonly employed method for determining the required dosage of heparin and protamine during cardiopulmonary bypass(CPB). Material and Method: We performed a prospective study on a fixed dose regimen for analyzing adequate dosages of heparin and protamine, the incidence of heparin resistance and heparin-induced thrombocyt openia, factors affecting ACT during CPB, and changes of ACT during aprotinin usage. 300 units/kg of heparin were administered to patients, and ACTs were measured after 5 mins. ACTs were checked at 10 mins and 30 mins after the onset of CPB, and then at 30 min intervals thereafter. If the measured ACT was under 400 secs, we added 100 units/kg of heparin. The heparin was reversed with 1 mg of protamine for each 100 units administered. If the measured ACT was longer than 130 secs 30 mins after protamine administration or if there was definitive evidence of a coagulation defect, we administered a further 0.5 mg/kg of protamine. Result: We studied 80 patients(50 adults and 30 children) who underwent open heart surgery(OHS) at Seoul National University Hospital. Preoperative ACT was 114.3${\pm}$19.3 secs in adults, and 119.5${\pm}$18.2 secs in children. There were no differences in preoperative ACT due to age, body weight, body surface area, or sex. The preoperative ACT was not influenced by a positive past history of OHS. Ten adults(20%) and 3 pediatric patients(10%) needed additional doses of heparin to maintain the ACT above 400 secs. Additional protamine administration was needed in 9 adults(18%) and 10 children(33%). Heparin resistance was found in only two adults. Heparin-induced thrombocytopenia was detected in 2 adults and 1 child. During CPB, ACT was prolonged. 12 adult patients received a low dose of aprotinin and showed longer celite activated ACT compared to the control group.The kaolin activated ACT showed a lower tendency than the celite activated ACT in aprotinin users. Conclusion: In conclusion, fixed dose regimen of heparin and protamine can be used without significant problems, but the incidence of need of additional dosage remains unsatisfactory.

• Animal Bioscience
• /
• v.36 no.12
• /
• pp.1796-1805
• /
• 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.

• Korean Journal of Food Science and Technology
• /
• v.32 no.4
• /
• pp.902-907
• /
• 2000

Protamine-strong basic protein was prepared from salmon(chum salmon, Oncorhynchus keta) sperm by several pretreatment method. And there were determined yield, amino acid composition, antimicrobial and antioxidant activity of protamine on each pretreatment condition. The yield of protamine was different according to pretreatment, and ultrasonicating, homogenizing and microwaving pretreatment were about 16.0%, 15.5% and 10%, respectively. The main amino acid of P60(microwaving pretreatment for 10 min at $80^{\circ}C$) and UU6(ultrasonicating pretreatment for 60 min at $20^{\circ}C$) were arginine, proline and tryptophan, and arginine content of P60 and UU6 were 61%, 53%, respectively. On the other hand, main amino acid of M(homogenizing pretreatment by mixer) were methionine, proline and arginine, the content were 34%, 28% and 11%, respectively. Also MC(homogenizing pretreatment with $H_{2}SO_{4}$ soln. by mixer) was very different with P60, UU6 and M, the content of MC were proline 44.8% and arginine 39.7%. Prepared protamines showed antimicrobial activity to several gram(+) and gram(-) strain. In particular, the UU6 and P60 protamine has strong antimicrobial activity to Bacillus subtilis and Escherichia coli, and the activity was increased with concentration increasing. Regardless of pretreatment method, all protamine showed antioxidant activity and the $EDA_{50}$ of P60, UU6, M and MC were $101\;{\mu}g/mL$, $410\;{\mu}g/mL$, $523\;{\mu}g/mL$ and $490\;{\mu}g/mL$, respectively.

• Korean Journal of Veterinary Research
• /
• v.12 no.1
• /
• pp.77-84
• /
• 1972

Throughout the studies the following experimental results were obtained and summarized. 1. Treatment of MVE virus with acetone, Tween-ether and Tween-ether-protamine sulphate caused an eight to 16 fold increase in HA activity. 2. Treatment with acetone and Tween-ether resulted in a four fold increase in CF activity. Treatment with Tween-ether-protamine sulphate decreased the activity. 3. The crude virus showed a complete loss of infectivity after treatment with Tween-ether, but three log unit was, decreased with acetone treatment. 4. The HA activity of treated and crude virus was disappeared after heating at $37^{\circ}C$ for 60 minutes but CF activity was increased. 5. Tween-ether or acetone treatment equally applicable to the preparation of haemagglutinin for HI test. 6. Zonal centrifugation of crude virus in a linear ten to 60 percent sucrose gradient showed two peaks of CF activity, and one of high buoy ant density part accompanied by HA activity and infectivity and the other of lower density part. Acetone treatment brought a decrease of the high density CF activity but not affected the second peak of low density found with crude virus, and resulted in increased HA activity and decreased infectivity. The peaks of HA, CF and infectivity after acetone treatment were not clearly separated. Tween-ether treatment caused a loss of the peak of CF activity found in the area of high density with crude virus, but the peak in the area of low density was not affected. This peak of CF activity was separated from noninfectious HA activity. The HA and CF activities were considered to be contributed by different parts of the varion.

• Archives of Pharmacal Research
• /
• v.27 no.7
• /
• pp.797-805
• /
• 2004

LPD vectors are non-viral vehicles for gene delivery comprised of polycation-condensed plasmid DNA and liposomes. Here, we described a novel anionic LPD formulation containing protamine-DNA complexes and pH sensitive liposomes composed of DOPE and cholesteryl hemisuccinate (Chems). Central composite design (CCD) was employed to optimize stable LPD formulation with small particle size. A three factor, five-level CCD design was used for the optimization procedure, with the weight ratio of protamine/DNA ($X_1$), the weight ratio of Chems/DNA ($X_2$) and the molar ratio of Chems/DOPE in the anionic liposomes ($X_3$) as the independent variables. LPD size ($Y_1$) and LPD protection efficiency against nuclease ($Y_2$) were response variables. Zeta potential determination was utilized to define the experimental design region. Based on experimental design, responses for the 15 formulations were obtained. Mathematical equations and response surface plots were used to relate the dependent and independent variables. The mathematical model predicted optimized $X_1-X_3$ levels that achieve the desired particle size and the protection efficiency against nuclease. According to these levels, an optimized LPD formulation was prepared, resulting in a particle size of 185.3 nm and protection efficiency of 80.22％.

• Korean Journal of Veterinary Research
• /
• v.25 no.2
• /
• pp.187-195
• /
• 1985

The Present study has been carried out to elucidate the antiestrogenic effects of tamoxifen in uteri of immature rats. Immature female Sprague-Dawley rats were allocated into 4, groups and injected with $5{\mu}g$ of estradiol-$17{\beta}$, $50{\mu}g$ of tamoxifen, a combination of both or vehicle only subcutaneously three times after an interval of 24 hours respectively. The concentrations, of cytosol estradiol receptor in uterus were measured by DCC method before and 1, 3, 6, 12, 24, 48 and 72 hours after the above treatments and those of nuclear estradiol were measured by protamine exchange method 72 hours and those of nuclear estradiol were measured by protamine exchange method 72 hours after the above treatments. The results obtained were summarized as follows: 1. The binding affinity of tamoxifen to estradiol receptor in uterine cytosol was lower than that of estradiol-$17{\beta}$, accordingly the translocation of estradiol receptor into the nucleus was found to be delayed. 2. Tamoxifen caused the retention of estradiol receptor in nucleus over 24 hours and inhibited the replenishment of the receptor from nucleus to cytosol in uterus.

• Clinical and Experimental Reproductive Medicine
• /
• v.32 no.3
• /
• pp.279-286
• /
• 2005

Objective: Although several genetic factors have been associated with defects in human spermatogenesis, the unambiguous causative genes have not been elucidated. The male infertility by haploinsufficiency of PRM1 or PRM2 has been reported in mouse model. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) of PRM1 and PRM2, related to the genotype of Korean infertile men. Methods: Genomic DNAs were extracted from peripheral bloods of infertile men with oligozoospermia or azoospermia, and analyzed using polymerase chain reaction (PCR) and direct sequencing. We carried out the direct sequencing analysis of amplified fragments in PRM1 (557 nucleotides from -42 to 515) and PRM2 (599 nucleotides from 49 to 648) genes, respectively. Results: Three SNPs of coding region in the PRM1 gene was found in the analysis of 130 infertile men. However, the SNPs at a133g (aa 96.9%, ag 3.1% and gg 0.0%), c160a (cc 99.2%, ca 0.8% and aa 0.0%) and c321a (cc 56.9%, ca 35.4% and aa 7.7%) coded the same amino acids, in terms of silence phenotypes. On the other hand, as results of the PRM2 gene sequencing in 164 infertile men, only two SNPs, g398c (gg 62.2%, gc 31.1% and ga 6.7%) and a473c (aa 63.4%, ac 29.9% and cc 6.7%), were identified in the intron of the PRM2 gene. Conclusions: There was no mutation and significant SNPs on PRM1 and PRM2 gene in Korean infertile men. These results suggest that the PRM1 and PRM2 genes are highly conserved and essential for normal fertility of men.

• Korean Journal of Microbiology
• /
• v.9 no.4
• /
• pp.175-178
• /
• 1971

Because of the cases of Japanese Encephalitis(J.E.) were reported every year in Korea. We, Dong-A Pharmaceutical Co., Ltd., produced J.E. virus vaccine, with lower price, since 1970 in order to prevent ourselves from being infected by the disease. And inoculated the J.E. virus vaccine for the children with a great success. We are going to report several questions which brought about in producing the J.E. virus vaccine by alcohol precipitation, protamine sulfate treatment method. The results obtained were as folows ; 1) In process treated with 40% alcohol, we used to ethanol made in Germany, but it was too expensive to use it. As the result which we had studied about it, we were satisfied with J.E. virus vaccine which produced with alcohol made in Korea, and then, we treated with accurate specific gravity of 40% ethanol for the precipitation of the virus. And also, we knew that it was the best method to be treated it for 3hrs, $13^{\circ}C$. 2) When we treated with protamine sulfate (0.025mg/ml), we acquired the highest potent titer, and suited into purpose for the nitrogen concentration. 3) The filtration of the purified J.E. virus vaccine, in case of millipore filter paper of large pore size was not suitable for the sterility. Therefore the pore size less than 0.8.$\mu$ (AA filter paper) in millipore filter paper was very suitable. But it seemed to be important subhects that the smaller was the pore size, the lower was the potent titer.

• Microbiology and Biotechnology Letters
• /
• v.19 no.5
• /
• pp.456-463
• /
• 1991

$\beta$-Galactosidase of Bifidobacterium longum KCTC 3215 was studied on the production, purification, and characterization. Optimum conditions for the enzyme production were in the medium of 1.0% lactose as carbon source, initial pH 7.0 and in 17 hours of cultivation at $37^{\circ}C$. The enzyme was purified 9.25 folds by protamine sulfate precipitation, ammonium sulfate fractionation, DEAE-Sephadex A-50 ion exchange chromatography and Sephadex G-150 gel filtration. The maximal P-galactosidase activity was observed at pH 6.5 and at the temperature of $40^{\circ}C$ This enzyme was stable at pH 6.0-8.5. Metal ions such as $Ca^{2+} \;and \; Co^{2+}$, 2-mercaptoethanol, cysteine, and glutathione stimulated B-galactosidase activity. The enzyme activity was inhibited by addition of $Mg^{2+}, Fe^{2+}, Cs^{1+}, Li^{1+}$, DETA, galactose, and $\rho$-chloromercuribenzoic acid. The kinetics of o-nitrophenyl-$\beta$-D-galactopyranoside and lactose were $K_m$ = 1.66 mM, $V_{max}= 0.30 mM/min\cdot mg\cdot protein$ and $KK_m = 3.18 mM, \; V_{max}= 0.42 mM/min \cdot mg\cdot$ protein, respectively. The molecular weight of native enzyme was about 360, 000 dalton and the enzyme consisted of 2 identical subunits with a molecular weight of 180, 000.

• Journal of Chest Surgery
• /
• v.33 no.7
• /
• pp.560-564
• /
• 2000

Background; Aprotinin, which is a nonspecific serine protease inhibitor, has an antiinflammatory and thrombogenic effect. However, it has an antithrombogenic effect during the cardiopulmonary bypass. This study was performed to evaluated the effects of aprotinin on the activated clotting time(ACT) and the total amount of the heparin used during the cardiopulmonary bypass. Marterial and Method; From December 1998 to November 1999, 82 consecutive patients electively underwent open heart surgery at Gachon medical school. The patients were older than 18 years. Eighty two patients were classified into a control group(group C, n=36) and a aprotinin-treated group(group A, n=46). Body weight, height, body surface area(BSA), pump time(PT), aortic cross clamping time(ACCT), and body temperature(BT) were determined. Total amount of heparin and protamine during the CPB were also measured. ACT was determined before heparin administration, at 20, 40 and 60 minutes after heparin administration, and after protamine administration. Result; No significant differences were noted in either group in body weight, height, BSA, BT, and the total amoun of heparin and protamine. Group A demonstrated a significant(p <0.05) increase in age, PT, ACCT, and ACT at 20, 40, and 60 minutes after heparin administration. Conclusion; In summary, the use of aprotinin prime resulted in an increase in ACT. The total amount of heparin in aproinin-treated patient was similar to that of the control group in spite of having the prolonged pump time. Therefore aprotinin may reduce the requirement of heparin.