• Journal of Microbiology
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• 제44권6호
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• pp.622-631
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• 2006

In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and $37^{\circ}C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$-$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $Cu^{2+}$ and $Co^{2+}$, but enhanced by the additions of $Ca^{2+}$ and $Mg^{2+}$ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it's a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

• Asian-Australasian Journal of Animal Sciences
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• 제26권11호
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• pp.1553-1561
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• 2013

Forty-eight crossbred heifers ($378.1{\pm}18$ kg) were used in a 56-d feeding trial (four pens per treatment in a randomised complete block design) to evaluate the influence of ionophore supplementation on growth performance, dietary energetics and carcass characteristics in finishing cattle during a period of heat stress. Heifers were fed a diet based on steam-flaked corn (2.22 Mcal $NE_m/kg$) with and without an ionophore. Treatments were: i) control, no ionophore; ii) 30 mg/kg monensin sodium (RUM30); iii) 20 mg/kg lasalocid sodium (BOV20), and iv) 30 mg/kg lasalocid sodium (BOV30). Both dry matter intake (DMI) and climatic variables were measured daily and the temperature humidity index (THI) was estimated. The maximum THI during the study averaged 93, while the minimum was 70 (THI average = $79.2{\pm}2.3$). Compared to controls, monensin supplementation did not influence average daily gain, the estimated NE value of the diet, or observed-to-expected DMI, but tended (p = 0.07) to increase (4.8%) gain to feed. Compared to controls, the group fed BOV30 increased ($p{\leq}0.03$) daily gain (11.8%), gain to feed (8.3%), net energy of the diet (5%), and observed-to-expected DMI (5.2%). Daily weight gain was greater (7.6%, p = 0.05) for heifers fed BOV30 than for heifers fed MON30. Otherwise, differences between the two treatments in DMI, gain to feed, and dietary NE were not statistically significant (p>0.11). Plotting weekly intakes versus THI, observed intake of controls was greater (p<0.05) at THI values ${\leq}77$ than ionophore groups. When THI values were greater than 79, DMI of control and MON30 were not different (p = 0.42), although less than that of groups fed lasalocid (p = 0.04). Variation in energy intake was lower (p>0.05) in the ionophores group (CV = 1.7%) than in the control group (CV = 4.5%). Inclusion of ionophores in the diet resulted in relatively minor changes in carcass characteristics. It is concluded that ionophore supplementation did not exacerbate the decline of DM intake in heat-stressed cattle fed a high-energy finishing diet; on the contrary, it stabilised feed intake and favoured feed efficiency. Ionophore supplementation reduced estimated maintenance coefficients around 10% in finishing cattle during a period of heat stress. This effect was greatest for heifers supplemented with 30 mg lasalocid/kg of diet.

• Toxicological Research
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• 제24권1호
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• pp.51-58
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• 2008

The present study was designed to explore the immunotoxic effects of orally administered aluminum (AI) on pregnant rats (n = 60) and their growing fetuses and consequently on the animal wealth. The animals were randomly allocated into three equal groups of 20 rats each. The first group has no treatment and kept as a control (G1). The second and third groups of pregnant rats were treated orally with aluminum chloride at 345 mg/Kg b.wt. The second group (G2) received the tested compound from the $6^{th}$ day of gestation to the end of weaning, whereas the third group (G3) received the tested compound from the $15^{th}$h day of gestation to the end of weaning. Control and treated animals (dams and offspring) were immunized ip with (0.5 ml) 20% sheep red blood cell (SRBC) suspension seven days before the end of experiments. At the end of exposure, ten dams and ten offspring from each group were used for assessment of cell-mediated immunity and a similar number of animals were sacrificed for evaluating the humoral immune response and serum protein profile. Aluminum chloride exposure of dams ($G_2&G_3$) caused significant suppression of both cell mediated and humoral immune responses in the obtained offsprings compared to the control group ($G_1$) without any significant effect on the immune responses of these dams. Moreover, the serum total globulins, albumin/ globulin (A/G) ratio and gamma globulin fraction were significantly decreased in the treated dam's offsprings compared to the corresponding controls while the serum total protein and all serum protein fractions showed non significant difference between the control and treated dams and between the two treated dam groups themselves. There were no histopathological changes observed in thymus, spleen and liver of the control and treated dams. Thymus of treated dam's offsprings (G2) showed lymphoid depletion in both cortex and medulla. Their spleens showed lymphoid depletion in the white pulps and congestion with hemosiderosis in the red pulps. Liver of treated dam's offsprings showed dilation and congestion of its central vein with degenerative changes in the hepatocytes. These histopathological changes were more severe in G2 than in G3 offsprings. It can be concluded that gestational and/ or lactation exposure of pregnant dams to AI chloride caused suppression of both cellular and humoral immune responses of their offsprings.

• 한국동물생명공학회지
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• 제34권2호
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• pp.117-122
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• 2019

The objective of this study was to establish an in vitro culture system for ovarian preantral follicles of B6D2F1. First, we optimized the in vitro preantral-follicle culture by culture duration, follicle stimulating hormone (FSH) type, and activin A concentration. Duration of in vitro culture for 9, 11, and 13 days was sufficient for the normal development of preantral follicles to antral follicles. Formation of cumulus cell-oocyte complex (COC) was induced by treatment with human chorionic gonadotropin (hCG; 2.5 IU/mL) and epidermal growth factor (EGF; 5 ng/mL). In addition, metaphase II (MII) oocytes formed during this in vitro culture of preantral follicles. In vitro preantralfollicle culture for 9 days showed higher rates of growth and maturation, thus yielding a greater number of antral follicles, and there were significant differences (p < 0.05) in the number of MII oocytes (that formed from these preantral follicles via differentiation) between the 9-day culture and 11-day or 13-day culture. The follicles cultured for 9 days contained a tightly packed well-defined COC, whereas in follicles cultured for 11 days, the COC was not well defined (spreading was observed in the culture dish); the follicles cultured for 13 days disintegrated and released the oocyte. Second, we compared the growth of the preantral follicles in vitro in the presence of various FSH types. There were no significant differences in the growth and maturation rates and in differentiation into MII oocytes during in vitro culture between preantral follicles supplemented with FSH from Merck and those supplemented with FSH from Sigma. To increase the efficiency of MII oocyte formation, the preantral follicles were cultured at different activin A concentrations (0 to 200 ng/mL). The control follicles, which were not treated with activin A, showed the highest rate of differentiation into antral follicles and into MII oocytes among all the groups (0 to 200 ng/mL). Therefore, activin A (50 to 200 ng/mL) had a negative effect on oocyte maturation. Thus, in this study, we propose an in vitro system of preantral-follicle culture that can serve as a therapeutic strategy for fertility preservation of human oocytes for assisted reproductive medicine, for conservation of endangered species, and for creation of superior breeds.

• 한국식품영양학회지
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• 제29권4호
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• pp.547-556
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• 2016

The purpose of this study was to investigate the relationship between frequency of coffee consumption, metabolic biomarkers, and nutrition intake in adult participants in the combined 2007~2009 Korean National Health and Nutrition Examination Survey (KNHANES). Subjects (2,095 males and 3,297 females) were classified according to sex and frequency of coffee consumption (${\leq}1$ time/month, ${\geq}2$ times/month and ${\leq}6$ times/week, 1 time/day, 2 times/day, 3 times/day) using food frequency questionnaires. Nutrition intake was analyzed using 24 h recall data. The 3 times/day coffee consumption group had a significantly higher age, and frequency of smokers and drinkers compared to the ${\leq}1$ time/month coffee consumption group in both male and female participants. Males in the 3 times/day coffee consumption group had a significantly lower HDL-cholesterol level, but females had a higher waist circumference compared with the ${\leq}1$ time/month coffee consumption group. Males in the 3 times/day coffee consumption group had a significantly lower nutrient density of fiber, vitamin B2, vitamin C, calcium and phosphorus compared with the ${\leq}1$ time/month coffee intake group. Females in the 3 times/day coffee consumption group had a significantly higher nutrient density of fat and niacin, but lower nutrient density of carbohydrate, calcium, phosphorus, and iron compared with the ${\leq}1$ time/month coffee intake group. In males, the frequency of coffee consumption was not associated with the levels of metabolic biomarkers. In females, the frequency of coffee consumption was positively associated with diastolic blood pressure after adjustments for multiple confounding factors, including age, BMI, smoking status, alcohol consumption, physical activity and energy intake. Coffee consumption was associated with decreased diastolic blood pressure in females. These findings suggest the importance of an awareness of the association between coffee consumption and metabolic risk.

• 한국식품영양학회지
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• 제30권3호
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• pp.609-617
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• 2017

This experiment was carried out to enhance the availability of blackberry. Since it is difficult to use blackberry as a fresh fruit, we investigated the quality characteristics of blackberry powder obtained by various drying methods (freeze drying and hot-air drying at $40{\sim}80^{\circ}C$). The L- and b-values of freeze-dried powder was higher than hot-air dried powder. The pH (3.2) was lowest and the acidity (14.4%) was highest in freeze-dried powder. In freeze drying, the brix degree was $65.7^{\circ}Bx$, but it increased from $54.7^{\circ}Bx$ to $68.5^{\circ}Bx$ with increasing temperature during hot air drying. The total polyphenol and flavonoids contents were the highest in freeze-dried powder, at 9.3 and 6.2 mg/g, respectively. The levels increased as temperature increased in hot air drying. Anthocyanin content in freeze-dried powder was 8.51 mg/g, while it sharply decreased to 1.17~2.45 mg/g in hot-air drying. Vitamin C content in freeze drying ($979.4{\mu}g/g$) was higher than that in hot-air drying ($48.3{\sim}303.2{\mu}g/g$). The sample concentration required for 50% reduction of DPPH free radical scavenging ($RC_{50}$) was $79.7{\mu}g/mL$ in freeze drying, and showed high antioxidant activity. Also it decreased from $122.4{\mu}g/mL$ to $87.7{\mu}g/mL$ with temperature increase during hot air drying. We therefore conclude from the above results that freeze drying is more suitable for the production of blackberry powder, because this method showed high value of chromaticity, total polyphenol, flavonoid, anthocyanin content, vitamin C and antioxidant activity.

• 한국축산식품학회지
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• 제32권5호
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• pp.604-611
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• 2012

The purpose of this study was to explore the effect of fermented persimmon diet on the meat quality of fattening pigs. Pigs (75 kg) were grouped and housed as 15 animals per pen and 3 replications per treatment. The basal diet (C) was substituted with 3% (T1), 5% (T2) and 10% (T3) of fermented diet. The 180 pigs were fed experimental diet for 42 d and 10 longissimus dorsi (LD) per treatment were collected when each swine reached 105.6 kg of body weight. The crude fat concentration of LD was lower (p<0.05) in treatments, but the pH value of T3 and the shear force values of T2 and T3 were higher (p<0.05) than those of C. The CIE $L^*$ value (lightness) was significantly (p<0.05) higher in T2 and T3 than C, but the CIE $b^*$ (yellowness) of T2 and T3 was significantly (p<0.05) lower than C. The composition of myristic acid and oleic acid was significantly (p<0.05) higher in treatments than in C, while the composition of palmitic acid and stearic acid of treatments were significantly (p<0.05) lower than C in LD. Sensory evaluation of cooked meat as scores of aroma, taste, juiciness and overall acceptability showed higher (p<0.05) in treatments than in C. In conclusion, the dietary of fermented persimmon diet decreased the crude fat concentration, improved the fatty acids composition (increased composition of unsaturated fatty acid and decreased composition of saturated fatty acid) and improved the sensory evaluation of pork meat from fattening pigs.

• 한국식품과학회지
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• 제39권6호
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• pp.614-618
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• 2007

미숙성김치와 숙성김치를 동결건조한 후 방사선량 별로 나누어 이에 대한 이화학적 분석, 잔존 미생물수 분석 및 관능평가를 통하여 김치의 양념소재화 가능성을 평가하였다. 미숙성김치와 숙성김치는 pH 및 적정산도는 차이가 있었으나 이의 동결건조물의 방사선 조사 처리구들 간에는 큰 차이가 없는 것으로 나타났다. 방사선을 조사하기 전 숙성김치의 색도는 미숙성김치에 비해 L, a 및 b값에서 다소 높은 값을 보였으며, 조사처리 후에는 조사선량의 강도에 의한 영향은 거의 받지 않았고 미숙성김치는 숙성김치보다 색상에서 다소 변화를 보였다. 미숙성김치분말보다 숙성김치분말은 미생물에 대한 방사선 조사에 의한 살균 효과가 전반적으로 높게 나타났으며, 30 kGy 조사 시에는 특히 총균수와 젖산균이 검출되지 않았다. 숙성김치분말에는 ethanol, 1-propene-3-methyl, disulfide dimethyl 및 disulfide methyl propyl 등의 휘발성 성분이 다소 많았고 특히 disulfide dipropyl 성분은 숙성김치분말에만 존재하는 특성을 보인 반면 그 외의 성분에서는 오히려 미숙성김치분말에 많은 것으로 나타났다. 방사선을 조사함에 따라 숙성김치분말에서는 2-butanone과 acetic acid, 핵산 및 butanal 2-methyl 성분 등이, 미숙성김치분말에서는 hexane 및 butanal 2-methyl 성분이 새롭게 생성되거나 증가되는 경향을 보였으며 전반적으로 황화물의 함량이 감소하는 경향을 보였다. 조사한 김치분말에 대하여 이미, 이취 및 매운맛에 대한 관능평가결과 모두 조사량이 증가함에 따라 이미와 이취는 증가하였으나 조사랑 30 kGy 까지는 수용가능한 것으로 평가되었다.

• 한국응용곤충학회지
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• 제7권
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• pp.39-51
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• 1969

The study involved determination of resistance levels of spider mites ta argano-phosphates using topical application and slide dip techniques; laboratory serening tests of alternative acaricides using an O/P resistant strain and a field trial of the screened materials. 1. Strains of Tetranychus were from Timaru(TR), Havelock Narth (HNR), Lincaln (LN). Germany (GR, GN). Comparisons of the resistant strains and normal strains at the LD50 and LC50 levels were as follows : (a) Using the topical application tochnique; with Parathian. resistant levels of the GR. TR and HNR strains of T. urticae were respeativuly, 1035. 484 and 452 times as resistant' as the LN strain. (b) Using the slide dip technique; with Phosdrin, resistant of GR, TR and HNR strains of T. urticae were 635, 274 and 266 times greater respeativuly, than the GN strain. 2. The laboratory sereaning tests were carried out far their contact plus stomach and residual effect to assess the toxicities of eleven alternative materials which would be used far control of O/P resistant strain of T. urticae. The acaricide groups represented were 3 organo-chlorines (Spidex, Kelthane and C 8514), 2 nitrophenyls (UC 19786 and Morocide), 2 cyclic carbonates(Eradex and Morestan). I carbamate (UC2004 7A), 1 mixture of carbamate and orano-chlorine and 2 other chemicals (C 8677 and M2527). From all acaricide tested. Kelthane and Morocide were the most effective, folowed by Spidex and M2527. Morestan, C8514. C8677 and RS 143 were intermediate, but Eradex, UC 19786 and UC 20046A were poor. 3, The number of sapmles required for estimation of the population in the field evaluation of acaricidal effects was one giving the highest practical precision. It was decided, after preliminary sampling trials. to use samples of 30 leaves per replicate which gave a $5.7\%$ standard error. 4. In the field trials, Morocide applied at the $0.05\%\;and\; 0.04\%$ a. i. conc. to black currant trees gave excellent control of O/P resistant population of T. urticae for about 12 days, but Morocide 0.025 and Kel thane $0.02\%$ a. i. cone. gave efficient control for about 6 days. In other words. first applications of Kel thane ane Moroeide gave very high degrees of control of O/P resistant population of the two-spotted spider mite. However, the results indicate that secondary application would sometimes be necessary. There was no foliage damage of black Currants and strawberries by either acaricides at the concentrations used. Acknowledgment ... The authors are grateful to: Dr. R. P. pottinger, Senior Lecturer in Agricultural Zoology. Lincoln college. New Zealand. for his helpful assistance in aiding with the organization of thd field work. Department of agriculture officers for mite colonies. Mr. D. A. Slade, Technical Advisor. Fruitgrowers' Federation (now at Massey University) for his assistance and provision of mites for testing. Mr T. McRae of Timaru for permission to use his crops for field tests. The following chemical companies and I or their New Zealand agents for so readily supplying samples of acarides; Ivan Watkins-Dow Limited. Fruitgrowers Chemical Company Limited. Henry H. York & company (New Zealand). Shell Oil (New Zealand) Limited.

• 한국식품저장유통학회지
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• 제12권1호
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• pp.80-85
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• 2005

일상생활에서 식용 또는 약용으로 이용되고 있는 12종의 버섯 추출물을 제조하여 이들 추출물의 항산화능 검색과 함께 인간유래의 혈액암 세포주 및 정상 임파구세포의 성장에 미치는 영향에 대하여 조사하였다. 그 결과, 향버섯, 번데기동충하초, 만가닥버섯, 아가리쿠스, 영지버섯, 표고버섯 메탄올 추출물에서 $30\~60\%$정도의 DPPH radical 소거 활성을 보였고, 목이버섯, 그물버섯, 새송이버섯에서 $10\~30\%$의 활성을 보였다. 또한 chinese hamster V79 cells에서 만가닥 버섯, 번데기동충하초, 향버섯 메탄올추출물의 경우 H2O2로 유도된 세포 독성에 대한 $39\~53\%$ 정도의 유의적인 생존율을 나타내었다. 그리고 인간유래 혈액암세포인 HL6O과 U937에 대해 향버섯, 만가닥버섯, 번데기동충하초, 아가리쿠스, 영지버섯 추출물들이 높은 증식 억제 활성이 보였다. 특히 버섯 메탄올추출물 1.0 mg/mL 처리시 HL 60에 대해 향버섯이 $70.5\%$로 가장 높은 저해율을, U937에 대해서는 번데기동충하초가 $81.5\%$의 가장 높은 저해율을 나타내었다. 그리고 버섯 열수추출물들은 메탄올추출물보다 낮은 저해 활성을 보였다. 그러나 모든 버섯추출물들은 인간의 정상 임파구세포에 대해 $95\%$ 이상의 높은 생존율을 나타내어, 정상 세포에 대한 성장 저해 효과가 없음을 보여주었다. 번데기동충하초, 아가리쿠스, 만가닥버섯, 영지버섯, 향버섯 추출물은 항산화 할성 및 혈액암 세포주에 대한 증식 억제 활성이 높음을 확인할 수 있었다.