• Asian-Australasian Journal of Animal Sciences
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• 제13권1호
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• pp.10-18
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• 2000

This paper describes the magnetic orientation of the intact and demembranated bull sperm treated by DTT or heparin in a 5,400 G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4{^{\circ}C}$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the sperm head area at 1, 3, and 6 days. After measuring the area, each sperm sample was exposed to a 5,400 G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the decondensation of bull sperm nuclei was not induced by the heparin treatment, however, incomplete decondensation was induced by the DTT treatment. During the magnetic orientation, bull sperms treated by DTT or heparin had low percentages of long axis perpendicular to the magnetic lines of force. However, different aspects were obtained for long axis perpendicular orientations following treatment of DTT or heparin. Through the DTT treatment, the decline of long axis perpendicularly oriented percentages was due to the increase of long axis parallel orientation with the head of the flat plane perpendicular to the magnetic lines of force, whereas, using the heparin treatment, the decline of long axis perpendicular orientation was due to the increment of long axis parallel orientation with the head of the flat plane parallel to the magnetic lines of force. Also, percentages of the head of the flat plane perpendicular were decreased by the heparin treatment. These findings suggest that maintaining the structure of protamine in the chromatin is necessary for the sperm head to orient with its flat plane perpendicular, and maintaining the disulfide bond in the chromatin is necessary for the long axis of sperm to orient perpendicularly.

• 한국미생물·생명공학회지
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• 제17권1호
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• pp.35-45
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• 1989

Streptococcus thermophilus 510으로부터 $\beta$-galactosidase의 생성조건은 탄소원으로 0.5% lactose를 첨가한 배지에서 초기 pH7.0, 배양온도 37$^{\circ}C$, 배양기간 18시간이었다. 배양여액으로부터 $\beta$-galactosidase를 ammonium sulfate 분획, 핵산의 제거, Sephadex G-200 gel filtration 및 DEAE-Sephadex A-50 ion exchange chromatography 등의 4단계 정제과정을 거쳐 정제한 결과 18배 정제되어 단일 단백질로 분리되었다. 정제효소의 활성 최적온도는 5$0^{\circ}C$, 최적 pH는 7.0이었고, 효소활성이 Mn$^{2+}$, $K^+$과 같은 금속이온과 dithiothreitol, 2-mercaptoethanol에 의해 촉진되었고, Hg$^{2+}$, $Zn^{2+}$, Co$^{2+}$, $Ca^{2+}$, EDTA, 8-hydroxyquinoline, galactose 등에 의해 저해되었다. 효소의 분자량이 520,000, 합성기질인 ONPG에 대한 $K_{m}$ 은 1,25mM, V$_{max}$는 88.50 $\mu$mole/min.mg protein이었고, 주종 아미노산은 glutainic acid, aspartic acid, leucine 및 valine이었다.

• Journal of Plant Biotechnology
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• 제47권3호
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• pp.209-217
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• 2020

The presence of high amounts of seed storage proteins (SSPs) improves the overall quality of soybean seeds. However, these SSPs pose a major limitation due to their high abundance in soybean seeds. Although various technical advancements including mass-spectrometry and bioinformatics resources were reported, only limited information has been derived to date on soybean seeds at proteome level. Here, we applied a tandem mass tags (TMT)-based quantitative proteomic analysis to identify the significantly modulated proteins in the seeds of two soybean cultivars showing varying protein contents. This approach led to the identification of 5,678 proteins of which 13 and 1,133 proteins showed significant changes in Daewon (low-protein content cultivar) and Saedanbaek (high-protein content cultivar) respectively. Functional annotation revealed that proteins with increased abundance in Saedanbaek were mainly associated with the amino acid and protein metabolism involved in protein synthesis, folding, targeting, and degradation. Taken together, the results presented here provide a pipeline for soybean seed proteome analysis and contribute a better understanding of proteomic changes that may lead to alteration in the protein contents in soybean seeds.

• Clinical and Experimental Reproductive Medicine
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• 제45권1호
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• pp.17-24
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• 2018

Objective: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. Methods: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1, DNMT3A, and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction. Global DNA methylation in sperm was determined by an enzyme-linked immunosorbent assay. Protamine deficiency and the proportion of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3), aniline blue (AB), and toluidine blue (TB) staining, as well as the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The p-values < 0.05 were considered to indicate statistical significance. Results: Significantly higher proportions of AB+, TB+, CMA3+, and TUNEL+ spermatozoa, as well as DNMT3A and DNMT3B transcription, were found in the OAT group. Positive correlations were detected between sperm parameters, DNA/chromatin damage, and DNMT3A and DNMT3B transcripts. Global DNA methylation was significantly higher in the OAT patients and had a significant correlation with abnormal results of all sperm chromatin integrity tests, but was not associated with DNMT1, DNMT3A, or DNMT3B expression. Conclusion: Oligoasthenoteratozoospermic men showed abnormal sperm parameters, abnormal chromatin/DNA integrity, and a higher global DNA methylation rate, as well as overexpression of DNMT mRNA.

• 한국임상수의학회지
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• 제36권1호
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• pp.74-77
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• 2019

A 10-year-old, 8.28 kg, intact female Cocker Spaniel was presented with continuous polyuria and polydipsia. The dog had proestrus bleeding 5 weeks earlier, and hyperglycemia, glucosuria, ketouria, and high level of serum fructosamine in laboratory findings. Based on hyperglycemia, glucosuria, ketouria, and ketosis, the patient was tentatively diagnosed as diabetes mellitus (DM) with ketouria. After diagnosis, 5 to 7 U/body porcine lente insulin was administered during 11 days from initial presentation as the remission of DM for the dog. But, blood glucose was still high level. Because there was no reaction to porcine lente insulin, it was replaced by 4-10 U/body neutral protamine Hagedorn (NPH) during 3 days. But, NPH also did not regulate blood glucose level. Because insulin therapy failed to regulate blood glucose level, the dog was considering insulin-resistant diabetes. The dog was tentatively diagnosed with progesterone-resistant DM on the basis of the history that had revealed proestrus bleeding 5 weeks earlier. Progesterone level was moderate high (43.7 ng/ml; reference range, 15.0-90.0 ng/ml). Ovariohysterectomy (OHE) was performed to remove the cause of the dog's diabetes. After OHE 11 days, blood glucose was gradually declined by insulin treatment. Consequently, blood glucose was well controlled in reference range without insulin treatment after 2 months. This case is a report on progesterone-induced DM treated with OHE and insulin treatment during the diestrus.

• Clinical and Experimental Reproductive Medicine
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• 제48권1호
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• pp.34-42
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• 2021

Objective: Studies of the effects of estrogens on the male reproductive system have emphasized the role of these hormones in male fertility. Sesame oil has many phytoestrogenic compounds and may improve male fertility. This study investigated the effects of sesame oil and different concentrations of estrogen on sperm parameters and DNA integrity in male mice. Methods: Twenty old NMRI (The Naval Medical Research Institute) male mice (40 weeks; weight, 30-35 g) were treated with sesame oil or different concentrations of estrogen (estradiol, 1 and 10 μL/kg/day) or received no treatment (controls). After 35 days, sperm parameters and DNA integrity were assessed and analyzed. Results: Sperm count, progressive motility, and morphology were decreased in the group that received 10 μL/kg of estradiol. A remarkably lower percentage of DNA fragmentation and protamine deficiency were detected in the group that received 1 μL/kg of estradiol. In the groups that received sesame oil and 1 μL/kg of estradiol, the numbers of spermatogonia and Leydig cells were higher than in controls. The combination of sesame oil and 1 μL/kg of estradiol led to improved sperm parameters and chromatin and testicular structure. Conclusion: Based on this study, consumption of sesame oil and a low concentration of estradiol may improve testicular function in older mice.

• 한국식품영양과학회지
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• 제14권4호
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• pp.345-352
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• 1985

칡잎단백질(蛋白質) 식량자원화(食糧資源化)를 위한 기초연구(基礎硏究)를 목적(目的)으로 칡잎으로부터 여러 가지 용매(溶媒)를 이용하여 단백질(蛋白質)을 추출(抽出), 분리(分離)한 후 그의 아미노산(酸) 조성(組成) 및 기능적(機能的) 특성(特性)을 검토한 결과는 다음과 같다. 용해도(溶解度)에 따른 칡잎단백질(蛋白質)은 수용성(水溶性) 단백질(蛋白質)이 18.5%, 염용해(鹽溶解) 단백질(蛋白質)이 33.5%, 알칼리 용해성(溶解性) 단백질(蛋白質)이 34.0%, protamine이 6.2%, 불용해성(不溶解性) 단백질(蛋白質)이 7.8%이었다. 증류수(蒸溜水), 1M NaCl, 0.015N NaOH의 용매(溶媒)에 의한 추출액(抽出液)의 단백질(蛋白質) 침전률(沈澱率)은 pH 3.0, 2.5, 4.0에서 가장 높다. 84.7, 76.4, 86.4% 이었으며 80%의 ethanol과 acetone의 첨가에 의해서는 약 90%가 침전(沈澱)되었다. 단백질농축물(蛋白質濃縮物)의 일반성분(一般成分)은 수분(水分) $1{\sim}2%$, 단백질(蛋白質)이 $59{\sim}67%$, 회분(灰分)이 $4{\sim}8$이었으며 지질함량에 있어서는 투석(透析)한 것은 $5{\sim}6%$, acetone 처리한 것은 $1{\sim}2%$이었다. 농축물(濃縮物)의 아미노산함량(酸含量)은 aspartic acid, glutamic acid, glycine의 순(順)으로 높았다. 농축물(濃縮物)의 용해도(溶解度)는 pH5이하에서 acetone 처리구(區)가 무처리구(區)에 비하여 낮았으며 그 이상의 pH에서는 높았다. 또 농축물(濃縮物)은 acetone 처리에 의하여 용적밀도(容積密度), 수분(水分) 및 지방흡착도(脂肪吸着度)에 상관관계를 나타내었으며 용적밀도(容積密度)에 대한 수분(水分) 및 지방흡착도(脂肪吸着度)는 반비례적이었다. 유화성(乳化性) 포립성(泡粒性)은 acetone 처리에 따라 별 차이가 없었다.

• Applied Biological Chemistry
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• 제22권4호
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• pp.191-197
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• 1979

Km값 및 분광학적성질(分光學的性質)를 조사하고 아울러 이 효소(酵素)의 결정화법(結晶化法)을 검토하여 다음과 같은 결과를 얻었다. 1. 효소(酵素)의 정제(精製)과정에서 유안분별침전(硫安分別沈澱)을 한 조효소(粗酵素)를 pH6.0 및 7.0의 인산염(燐酸鹽)완충액으로 48시간 이상의 충분한 투석(透析)을 시키고 DEAE-Sephadex column chromatography에서는 11mg protein/ml DEAE-Sephadex 정도의 DEAE-Sephadex를 사용하는 것이 효과적이였다. 2. 효소(酵素)의 정제과정(精製過程)에서 이미 알려진 protamin 황산(黃酸) 처리와 Sephadex G-150의 gel여과는 생략하여도 무방하였다. 3. 효소(酵素)의 결정화(結晶化)에서는 단백질을 20mg/ml의 농도가 되게끔 2-mercaptoethanol를 함유하는 0.01M 인산(燐酸)카리 완충액에 녹여 고체유안(固體硫安) 첨가(添加)법으로 결정(結晶)시키는 것이 가장 큰 육각봉상(六角棒狀)의 결정(結晶)이 얻어졌다. 4. Pyridoxal phosphate와 결합(結合)한 holo형 ${\beta}-tyrosinase$는 340nm와 430nm의 파장(波長)에서 각각 최대흡수(最大吸收)를 나타내었다. 5. ${\beta}-tyrosinase$의 L-tyrosine에 대한 Km값은 $2.31{\times}10^{-4}M$이였으며 SDS-polyacrylamide 전기영동법(電氣泳動法)에 의한 이 효소(酵素)의 subunit의 분자량(分子量)은 약 5,000이였다.

• Journal of Chest Surgery
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• 제3권2호
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• pp.73-90
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• 1970

Clinical perfusion data on 16 cases of cardiopulmonary bypass using Sigmamotor pump and RyggKyvsgaard Oxygenator which performed at Seoul National University Hospital during the period of Aug. 1968 to Aug. 1970 was analized. AIl cases were hemodiluted and the perfusion was carried out under the normothermic condition. The age of the patients ranged between 6 and 43 years. The b:dy weight varied between 18.3 and 54.0 kg and the body surface area between 0.78 and 1. 59$M^2$. The priming solution was consiste:I with fresh ACD blood. Hartmann solution and Mannitol. The average amount of priming was approximately 2242 ml. The average hemodilution rate was 17%. The flow rate ranged from 1.7L to 3.5L/Min/$M^2$ and averaged 2.4L/Min/$M^2$ or 78mI/Min/kg. The duration of perfusion varied from 22 to 110 min with average of 56.9 minutes. Some hemodynamic responses were observed. The arterial pressure dropped immediately after the initiation of partial perfusion and was more marked after the total perfusion foIlowed by gradual increase to the safety level. The central venous pressure reflected the reduced blood volume especially in the cases of prolonged perfusion which lasted over 60 min. In most of the cases, red blood cell count decreased and white blood ceIl count increased after the perfusion. Hemoglobin level was decreased, averaging of 12.5mg%, Hct 3.3% and platelets count of 18% postoperatively. Plasma hemoglobin increased mildly, from pre-perfusion average value of 4. 06mg% to postperfusion value of 22.5mg%. Serum potassium was 4.4mEq/L pre-operatively and was decreased to 3.7mEq/L postoperatively. Five cases showed definite hypopotassemia immediately after the operation. Sodium and chloride decreased mildly. These electrolyte changes are thought to be related with hemodilution. diuretics and reduced blood volume during and after the perfusion. Arterial blood pH value revealed minimal to moderate elevation from preperfusion average value of 7.376 to 7.461 during perfusion and then 7.365 after perfusion. The pC02 and hicarbonate showed minimal to moderately lowered values. The total CO2 was decreased. Buffer base decreased during perfusion (Av. 42.6mEq/L) and further decreased after the perfusion (Av. 40.8mEq/L). These arterial blood acid base changes suggested that the metabolic acidosis was accompanied by respiratory alkalosis during and immediately after the perfusion. Authors belived that the acidosis could more effectively be corrected with the more additional dose of bicarbonate than we used by this study. The chest tune drainage during the first 24 hours following operation was 1158 ml in average. One case (Case No. 15) showd definite bleeding tendency and it was believed that the cause might be due to the defect of heparin and protamine titration. The average urinary out put during 24 hours post-perfusion was 1291ml. One case (Case No. ]) showed definite post perfusion oliguria. As conclusion hemodilution using fresh ACD blood. Hartmann and Mannitol solution added with Bivon and high flow rate unler normothermia. was thought to amelioratc the severity of mctabolic acidosis during and after perfusion with relatively satisfactory effect on the diuresis and bleeding tendency.

• 대한약리학회지
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• 제7권1호
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• pp.53-65
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• 1971

It is well known that diabetes mellitus is associated with metabolic derangements, such as hyper-glycemia, ketosis, glycosuria, and also widespread alterations in the blood vessels, kidneys, eyes, peripheral nerves and heart. It is also recognized that healing of skin wound is delayed in diabetics. In bone, according to Aegerter, osteopenia develops in diabetes mellitus and it is chiefly ascribed to overutilization of protein. Shim claims that total blood flow to the entire skeletal system is approximately 4 to 8 percent of resting cardiac output and blood supply to the skeletal system would be decreased on account of secondary arteriosclerotic changes in the diabetics. An adequate blood supply is an essential factor in the healing process of fracture, and disturbed blood flow, either local or systemic, will invariably delay union of the fragments or the fragments from being fused. As the author has encountered several cases of diabetics in whom healing of fracture was delayed or incomplete, this experimental study was undertaken to elucidate the effects of hyperglycemia and diabetes mellitus on the healing process of fracture. In this experiment adult albino rabbits, weighing about 2 kg. were used and divided into 6 groups. The femur of each animal was fractured surgically, and then the healing process of fracture was periodically checked by radiography at an interval of one week for a period of 6 weeks. Thereafter, all the rabbits were killed to obtain tissue preparation of the femur. The experimental groups were as follows; 1) Control group: Six rabbits sustained a surgical fracture to the femur, without being given any other treatment or drug. 2) Alloxan-treated group: For inducing diabetes, alloxan was given intravenously to 17 rabbits in various dose as follows; to 7 of them 40 mg/kg, to 6 rabbits 80 mg/kg and to 4 rabbits 120 mg/kg of body weight, respectively. 3) Insulin-treated group: Protamine-zinc insulin was injected subcutaneously to each of 6 rabbits in a daily dose of 1 unit per kilogram of body weight. 4) Group treated with insulin after alloxan: Four rabbits were given 80 mg of alloxan once and than 1 unit of insulin per kilogram of body weight daily. Another 5 rabbits were injected 1 unit of insulin per kg of body weight daily following administration of alloxan in a dose of 120 mg/kg. 5) Homotransplantation group: Following intravenous injection of alloxan in a dose of 120 mg/kg, 10 rabbits underwent homotransplantation of a short bone segment to the femur. Five of them were subsequently given 1 unit/kg of insulin daily. 6) Sugar-treated group: six rabbits were fed $15{\sim}20$ gm of sugar daily throughout the period of experiment. The results obtained are summarized as follows; 1. Blood sugar level and damage to the pancreatic islet increased proportionately when alloxan was given to the rabbits in various doses. No appreciable change could be observed in the islets when the blood sugar level was altered by either oral administration of sugar or subcutaneous injection of insulin. 2. Comparing with the control group, healing of fracture was delayed in the alloxan-treated group, while callus formation and periosteal reaction were shown to be more prominent in this group and subsequently, the ultimate osseous tissue formed at the fracture site was significantly smaller in amount and less compact. These findings were more marked as the amount of alloxan increased. 3. Administration of insulin prevented the delay in healing process of fracture in the rabbits with alloxan-induced hyperglycemia. In this case, the course and progression of fracture healing were almost similar to those of control group. 4. Union between the host bone and the fragment transplanted from other rabbit of the same species was more delayed in the group treated with alloxan alone than in the group to which insulin was administered after development of alloxan-induced diabetes. In both groups periosteal new bone developed from the ends of the host bone, above and below the transplanted fragment, and directly fused with failure of periosteal callus to bridge the adjacent ends of the host bone and the transplanted fragment. 5. The healing process of fracture was not inhibited by alteration in blood sugar level when the blood sugar was abnormally increased by excessive sugar intake or lowered by administration of insulin alone. The healing of fracture in these groups progressed similarly as in the control group. In brief summary, it appears that the healing process of fracture would be definitely disturbed in diabetic state brought about by damage to the pancreatic islet. As such an inhibition could be overcome with insulin, it seems that insulin plays an important role in healing of fracture, but alteration in blood sugar level alone does not modify healing process of fracture to significant degree.