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Viabilities of Biopsied Mouse Embryos after Ultrarapid Refreezing and Thawing (미세조작된 생쥐수정란의 초급속 재동결융해 후 생존성)

  • 신상태;임준호;강만종;한용만;이경광
    • Korean Journal of Animal Reproduction
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    • v.20 no.2
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    • pp.207-214
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    • 1996
  • To examine the developmental capacity of manipulated embryos after ultrarapid refreezing and thawing, mouse embryos were biopsied at 4-cell stage, frozen twice at 4-cell and morula stages, respectively, and then transferred to rec-ipients. Single blastomeres were biopsied from 4-cell embryos by a modified aspiration method. Biopsied 4-cell embryos were equilibrated into freezing medium at room temperature for 2.5 min, loaded into 40 $\mu$I of freezing medium in 0.25 ml plastic straw and then directly immersed into liqiud nitrogen. Freezing medium for 4-cell embryos consisted of 4.0 M ethylene glycol and O.25 M sucrose in dPBS supplemented with 6 mg/lm BSA. Morulae were frozen into freezing medium containing 5.0 M glycerol instead of ethylene glycol. Thawing was conducted by agitating each straw in 3TC water for 20 sec. The c content of each straw was expelled into 0.5 ml of dilution medium, which consisted of 0.25 M sucrose and 3 mg/ml BSA in dPBS. The thawed embryos were rehydrated in dilution medium for 10 min, washed 3 times with dPBS and then cultured in M16 medium at 37$^{\circ}C$, 5% CO$_2$ in air. Blastocysts that developed from frozen or refrozne biopsied embryos were transferred to recipients on Day 3 of pseudopregnancy, respectively. In vitro and in vivo developmental rates of the biopsied and intact 4~cell embryos after freezing and thawing were 78 (10l/130) and 25% (10/40), and 91 (114/125) and 30% (12/40), respectively. Although the rates of in vitro development of biopsied and intact embryos to blastocyst stage were significantly different after freezing and thawing (P

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T-cell Mediated Immunity in Pulmonary and Extrapulmonary Tuberculosis (폐 및 폐외결핵환자에서의 T 림프구 매개성 면역기능의 변화에 관한 연구)

  • Choi, Dong-Chull;Shim, Tae-Sun;Cho, Sang-Heon;Jung, Ki-Ho;Hyun, In-Gyu;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.1
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    • pp.62-72
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    • 1992
  • Background: T-cell mediated cellular immunity has been suggested as an important mechanism in mycobacterial infection and imbalance between helper/inducer and suppressor/cytotoxic T-cell has been suggested as an important immunological abnormality in the pathogenesis of tuberculosis in human. Method: To determine whether there is any difference in T-cell mediated immunity in the pathogenesis of pulmonary and extra pulmonary tuberculosis, total numbers of WBC&lymphocytes were counted and helper/inducer and suppressor/cytotoxic cells were calculated by flow cytometry. Blastogenesis after stimulation with Concanavalin-A, Phytohemagglutinin and PPD were measured by $^3H$-thymidine uptake. PPD skin test was performed as an in vivo test. Results: 1)There was no significant difference in the size of PPD skin test between pulmonary and extrapulmonary tuberculosis groups. 2)Number of total lymphocytes significantly decreased in tuberculosis patients compared with healthy control group. But there was no significant difference between pulmonary and extrapulmonary tuberculosis groups. 3) Number of HLA-DR and Interleukin-2 receptor (+) cells were significantly increased in tuberculosis patients. But there was no significant difference between pulmonary and extra pulmonary tuberculosis groups. 4) There was no significant difference in the numbers of WBC, $T_3$, $T_4$ and $T_8$ lymphocytes and $T_4/T_8$ ratio between tuberculosis patients and healthy controls. 5) There was no significant difference in the blastogenesis after stimulation with specific and non-specific blastogens between tuberculosis patients and healthy controls. 6) The percentage and absolute number of $T_4$ lymphocyte were significantly correlated with the size of PPD skin test. (r=0.689 and 0.598). Conclusion: From these results, it is concluded that there was no difference in T-cell mediated immunity between pulmonary and extra pulmonary tuberculosis group. But, because it is suspected that there might be some difference in the role of T-cell mediated immunity in the pathogenesis of pulmonary and extra pulmonary tuberculosis or even among the extrapulmonary tuberculosis patients, further studies would be required.

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Factors Influencing the Preference for German farm Tourism: A Path Model Approach

  • Sidali, Katia Laura;Spiller, A.
    • Journal of Global Scholars of Marketing Science
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    • v.18 no.4
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    • pp.33-59
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    • 2008
  • This paper aims to analyse the preference for German farm tourism among the German population. For this reason, we conducted an empirical study in Germany during summer 2007 and we applieda structural equation model based on partial leasts quares(PLS) to analyse the data. In the following chapters we will introduce the literature review and our conceptual frame work. We will then outline the procedures we adopted and the results of the empirical analysis. In the final part so me conclusions will be presented and a discussion will follow in order to draw the future directions of our research. According to our hypotheses, the possibility that agri-tourism enters in the evoked set of an individual is higher: H1: The higher the information degree about it. H2: The lower the influence of the social stimuli. H3: The higher the physical exposure to it (experience). H4: The higher the wellness image of agri-tourism. H5: The higher the traditional image of agri-tourism. H6: The higher the exciting image of agri-tourism. H7: The higher the perceived value for money. Our further hypotheses affirm that the possibility that agri-tourism enters in the evoked set of an individual is higher: H8: The lower the perceived risk. H9: The higher the motive to enjoy a holiday in the nature. H10: The higher the motive to enjoy a sport holiday. H11: The lower the motive to have an organized holiday. H12: The lower the motive to have a holiday abroad. H13: The lower the motive of action and night life. H14: The higher the motive to spend a holiday with the family. H15: The lower the motive to spend a city holiday. Finally, our model has some socio-demographics data. As we mentioned before, German agri-tourism has traditionally been the travel destination of large-size families, with low-to-middle income. For that reason, our final hypothesises are the following: the possibility that agri-tourism enters in the evoked-set of an individual is higher: H16: The higher the number of family members. H17: The lower the family income. Since in this study we use a path model with a PLS approach, we are able to state some interrelations among the exogenous latent variables: H18: The motive of sport holiday has a positive influence towards nature motives. H19: The physical exposition to agri-tourism has a positive influence toward information. H20: The motive of family holiday has a negative influence toward the motive of action and night life. H21: Social stimuli have a positive influence towards individuals risk perceptions. H22: Social stimuli have negative influence towards experience. Data for this study were gathered via administrated questionnaires during the summer 2007 within the frame of an academic "marketing research" course. The corresponding t-values are assessed using the bootstrapping method with 500 re-samples. In our model 61% of the degree of appreciation of German agri-tourism (evoked set) is explained by five independent variables: value for money ($0.335^{{\ast}{\ast}{\ast}}$) (H7) experience ($0.267^{{\ast}{\ast}}$) (H3), exciting image ($0.204^{\ast}$) (H6) organisation ($-0.162^{\ast}$) (H11) and holiday abroad ($-0.156^{\ast}$) (H12). The variance explained ($R^2$) for the other endogenous variables are the following: nature 24.3%, information 14.1%, action holiday 13.8%, risk perception 5.8% and experience 2.4%. An overview can be inferred from table 5. The results also allow us to test each of the proposed hypotheses. With exception of organization and abroad, none of the others travel style factors (H9 to H15) seem to have any significant impact towards evoked set which leads to the rejection of the respective hypotheses. As expected, social stimuli have a significant influence on individuals' risk perception (H21 accepted), however neither the former nor the latter have a valuable impact on evoked set (rejection of H2 and H8). Besides, since the influence of social stimuli towards experience is not significant, also H22 has to be rejected. Experience influences information (H19 accepted) but the latter does not affect significantly the evoked set (H1 rejected). Both H4 as well as H5, referring respectively to the perceived images of German agri-tourism as a wellness destination and the traditional image of the German farm tourism have to be rejected. Finally, none of the demographic data included in the model explains significantly the variance of the factor evoked set. Therefore neither H16 nor H17 has been accepted. As far as the interrelation between sport and nature (H18) and family and action (H20) are concerned, the stated relationship among these variables has been statistically confirmed. Our path model based on partial least squares shows the factors influencing the preference for farm tourism in Germany. Among others value for money and experience are the most significant ones. Practical implications are discussed.

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Adequate Standard Pot and Number of Plants Per Tree of Raising Seeding Pot on the Foxtail Millet Transplanting Culture in the Southern Province (남부지방 조 이식재배시 육묘폿트의 적정규격 및 주당본수)

  • Kim, Yong-Soon;Kim, Dong-Kwan;Choi, Jin-Gyung;Park, Heung-Gyu;Kim, Myeong-Seok;Shin, Hae-Ryoung;Choi, Gyung-Ju;Yun, Jong-Tag
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.1
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    • pp.23-28
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    • 2015
  • This study was performed to investigate the adequate standard pot and number of plants per tree of raising seeding pot on the foxtail millet transplanting culture in the southern province. Due to the various application of wellbeing-health food recently, for upbringing of the foxtail millet, millet and sorghum in minor cereals, R & D and policy support is being promoted actively. The foxtail millet growing season is so short from 90 to 130 days, and it is large variations for a growth temperature. The main results are as follows. When it comes to foxtail millet transplantation, seedling quality of 406 holes, 200 holes and 162 holes of raising seeding pot type were not all significant, and field rooting percentage is accounted for all 94 to 95%. Yield of a foxtail millet was exposed in 406holes 305 kg/10a>162holes 303 kg> 200holes 302 kg order, and it was no significance between test processing. When it's the raising seeding transplanting culture, in case of pot culture, 406holes pot culture were reduced the bed soil cost 63%, pot 50%, working hours 18% for 200holes pot. Transplanting seedling quality per a foxtail millet transplanting culture method, dry weight was high inclination as transplanting number of plant is less, and field rooting percentage displayed more than all 95%. Yield appeared to 2 plants seedling transplanting 315kg/10a> 3 plants seedling transplanting 304kg>1 plant seedling transplanting 256kg order. The projected cost per the pot-sort on the raising seeding transplanting culture of foxtail millet, the seedling transplanting culture of 406holes was reduced 40% percentages compared to 200holes as 76,230won/10a. As a result, 406holes pot and 2plants seedling transplanting culture, labor-saving culture was possible.

Structural analysis of Precipitates in a Nickel based Cast Single Crystal of CMSX 6 (니켈계 초합금 CMSX 6 단결정 주조조직의 석출물구조 분석)

  • An, Seong-Uk;Larionov, V.;Grafas, I.;Kim, Su-Cheol;Im, Ok-Dong;Kim, Seung-Ho;Jin, Yeong-Hun;Choe, Jong-Su;Lee, Jae-Hun;Lee, Sang-Jun;Seo, Dong-Lee;Lee, Tae-Hun;Heo, Mu-Yeong
    • Korean Journal of Materials Research
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    • v.8 no.12
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    • pp.1165-1169
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    • 1998
  • A single crystal cast blade was manufactured by CMSX 6, one of the first generarion nickel based single crystal superalloys by the selector method in a vacuum furnace. The single crystal has been grown with cooling rate of 2.5 mm/min, after pouring the molten alloy of 163$0^{\circ}C$ to the mold heated to 150$0^{\circ}C$. The cast structure could be classified into matrix (dendrite) and eutectic regions in ${\gamma}$'shape and size. The eutectic region showed higher Ti content. As the additional results of ${\gamma}$'precipitates by EPMA and CBED analysis the ${\gamma}$'size was less than 0.5~0.7$\mu\textrm{m}$, showing the chemical composition close to Ni$_3$Al of Ll$_2$ lattice structure. But ${\gamma}$'size has increased to bigger than 1.0$\mu\textrm{m}$, being near to eutectic region, changing its shape to bar or huge block types. These showed the chemical structure near to Ni$_3$Ti of D $O_{24}$ lattice structure. Therefore, ${\gamma}$'morphology of dendrite and eutectic regions depends absolutely on its chemical composition and lattice structure.

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RNA Interference of Chitinase Gene in Spodoptera litura (담배거세미나방(Spodoptera litura) Chitinase gene의 RNA interference)

  • Jeon, Mi Jin;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.202-209
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    • 2014
  • RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

COMPARATIVE STUDY OF BEHAVIOR AND COGNITIVE FUNCTION BY ADMINISTRATION OF METHYLPHENIDATE AND IMIPRAMINE IN ATTENTION DEFICIT-HYPERACTIVITY DISORDER (Methylphenidate와 Imipramine투여에 따른 주의력 결핍${\cdot}$과잉운동장애 환아의 행동 및 인지기능 변화에 대한 연구)

  • Ahn, D.H;Hong, K.E;Oh, K.J;Shin, M.S;Yoo, B.C;Chung, K.M
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.3 no.1
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    • pp.26-45
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    • 1992
  • This study presents the behavioral and cognitive changes by administration of methylphenidate(MPH) and imipramine(IMI) for the treatment of attention-deficit hyperactivity disorder(ADHD) in $5_{1/2}{\sim}12$ years old children referred to child psychiatric clinics. Behavioral changes are assessed with parent's and teacher's ratings. Drug effects on attention. short-term memory, and impulsivity are evaluated with psychological tests in laboratory. The changes were assessed twice in a 8-week periods. The data were analyzed seperately for 15 subjects each drug using repeated measured analysis of variance(ANOVA). The findings indicates that behavioral and cognitive impairments are improved by both drugs, but impulsivity is not. And MPH is superior to IMI on the improvement of attentional problem ; especially the findings indicates important differences between simple task and complex. perceptual-search task. These data confirm the effectiveness of MPH for treatment of ADHD, also raise questions regarding assessment method of attention and impulsivity as fell as importance of impulsivity in ADHD.

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A STUDY ON FRACTURAL BEHAVIOR OF DENTIN-RESIN INTERFACE (상아질-복합레진 접착계면의 파괴거동에 대한 연구)

  • Ryu, Gil-Joo;Choi, Gi-Woon;Park, Sang-Jin;Choi, Kyung-Kyu
    • Restorative Dentistry and Endodontics
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    • v.32 no.3
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    • pp.208-221
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    • 2007
  • The fracture toughness test is believed as a clinically relevant method for assessing the fracture resistance of the dentinal restoratives. The objectives of this study were to measure the fracture toughness $(K_{1C})$ and microtensile bond strength of dentin-resin composite interface and compare their relationship for their use in evaluation of the integrity of the dentin-resin bond. A minimum of six short-rod specimens for fracture toughness test and fifteen specimens for microtensile bond strength test was fabricated for each group of materials used. After all specimens storing for 24 hours in distilled water at $37^{\circ}C$, they were tensile-loaded with an EZ tester universal testing machin. Statistical analysis was performed using ANOVA and Tukey's test at the 95% confidence level, Pearson's coefficient was used to verify the correlation between the mean of fracture toughness and microtensile bond strength. FE-SEM was employed on fractured surface to describe the crack propagation. Fracture toughness value of Clearfil SE Bond (SE) was the highest, followed by Adper Single Bond 2 (SB), OptiBond Solo (OB), ONE-STEP PLUS (0S), ScotchBond Multi-purpose (SM) and there was significant difference between SE and other 4 groups (p < 0.05). There were, however, no significant difference among SB, OB, OS, SM (p > 0.05). Microtensile bond strength of SE was the highest, followed by SB, OB, SM, OS and OS only showed significant lower value (p < 0.05). There was no correlation between fracture toughness and microtensile bond strength values. FE-SEM examination revealed that dentin bonding agent showed different film thickness and different failure pattern according to the film thickness. From the limited results of this study, it was noted that there was statistically no correlation between K1C and ${\mu}TBS$. We can conclude that for obtaining the reliability of bond strength test of dentin bonding agent, we must pay more attention to the test procedure and its profound scrutiny.

Influences of Rate of Artificial Insemination Following Estrus Induction in Dog (개에서 발정유도가 인공수정효율에 미치는 영향)

  • 이영락;강태영;최상용
    • Journal of Embryo Transfer
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    • v.18 no.1
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    • pp.61-68
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    • 2003
  • Considerable attention has been focused on the cryopreservation of semen and estrus induction in dog, as consequence of poor productivity caused by long anestrus period, in order to enhance the productivity of youngs and to preserve the breeds. The objectives of this study were to improve reproductive efficiency of artificial insemination with fresh- and frozen-semen following estrus induction in dog. Fifty infertilie dogs (age 2~3 years) were selected fur the study and divided into three different estrus induction treatment groups. Group 1 : dogs (n=15) were given clomifene (0.1 mg/kg) orally f3r five days at 12 hr intervals. Croup 2: dogs (n=15) were given bromocriptine (50 $\mu$g/kg) orally for five days at 12 hr intervals, followed by single injection intravenously of 500 IU GnRH (Croup 3, n=20) when pro-estrus occurred. After being treated, the dogs were evaluated fur the rates of estrus induction and time interval lapses from treatment to beginning of the pro-estrus. The rates of pregnancy in estrus inducted dogs mated naturally compared to those inseminated artificially with ejaculated fresh semen and frozen-thawed semen. Estrus detection was performed using the method of vaginal smear and confirmed by the plasma progesterone assay. Pregnancy was confirmed by ultrasonograpy on day 25, 35 and 55 post insemination. The ejaculated semen was exposed to a mixture of Tris extender with cryoprotectant (Trisma, 81 mM; TES, 209 mM; citric acid, 6 mM; glucose, 5 mM; glycerol, 8%) and cryopreserved gradually by slow-cooling at 17 co above the surface of liquid nitrogen (L$N_2$) for 23 min. The use of fresh semen, the pregnancy rates were observed 66.6, 66.6, 75.0 and 83.3% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. The use of frozen-thawed semen, the pregnancy rates were observed 66.6, 33.3, 50.0 and 60.0% in natural estrus, clomifene induced, bromocriptine induced and a combination of GnRH and bromocriptine, respectively. No difference was observed in the number of offspring produced among natural estrus and treated groups inseminated with fresh or frozen-thawed semen. In conclusion, there was no significant differences in the pregnancy rate of dogs between group treated with a combination of GnRH and bromocriptine and group treated clomifene or bromocriptine only. However, frozen-thawed semen can be used successfully fur artificial insemination in dog.

Effect of Proteinase Activity on the Cheddar Cheese Quality (단백분해 효소 활성(蛋白分解 酵素 活性)이 Cheddar Cheese의 품질(品質)에 미치는 영향(影響))

  • Kim, Min-Bae
    • Journal of Dairy Science and Biotechnology
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    • v.14 no.2
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    • pp.157-164
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    • 1996
  • This study aimed increase the quality during ripening of Cheddar cheese made with proteinase-negative mutant of Streptococcus lactis KCTC 1913 selected by curing. The degradation of protein during cheese ripening were investigated by electrophoresis and chromatography. The results were summarized as follow ; 1. The number of lactic acid bacteria decreased with the ripening stage, and that of the control cheese decreased faster than that of the cheese made with mutant. 2. Polyacrylamide gel electrophoretic analysis of cheese caseins revealed no difference between the cheese made with mutant and the control cheese, but differences along with the ripening stage were evident. 3. On Sephadex G-25 column chromatography, the extracts of bitter components from the green cheese and 3 month ripended cheese were fractionated into 3 fractions. With the progress of ripening, bitter peptides were degraded to rather small peptides or free amino acids. 4. Sensory evaluation of the 3 month ripended Cheddar cheese found no significant differences in color but the cheese made with mutant evidenced higher palatability in flavor and better texture than the control cheese. 5. The yields of the cheddar cheese made with mutant was 0.14% higher than that of the control cheese.

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