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Improving Corsican pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology and germination

  • Wtpsk, Senarath;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.61-62
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    • 2003
  • Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

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Establishment of rhizosphere microbes for plant protection on soil-borne diseases -Benificial antagonist and its mode of action toward ginseng root rot pathogen- (근권미생물과 토양병방제 -유용길항균이 인삼근부병원에 미치는 영향-)

  • Kim, S.I.;Lee, M.W.
    • The Korean Journal of Mycology
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    • v.22 no.1
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    • pp.50-61
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    • 1994
  • From soil samples, 380 antagonistic microorgnisms were isolated. Among the isolates, 42 strains had mycelia growing inhibition ability against Fusariun solani, ginseng root rot causing pathogen. Isolates CHA 1 and S-PFHR 6 were proposed as antagonists for this study and they were identified as Promicromonospora sp. and Pseudomonas pseudoalcaligenes respectively. As an antagonism against hyphae of F. solani in dual culture test, CHA 1 and S-PFHR 6 inhibited linear growing, caused abnormal branching, and the membrane projection which formed by cell wall destruction. The secondary metabolites contained in the culture filtrates which prepared from PD broth and Nutrient broth inhibited the spore germination to 14.3%. The culture filtrate of S-PFHR 6 which prepared by a little amount of soil extract addition to nutrient rich medium had more strongly. inhibited the spore germination and spore germination decreased to less than 4.0% in it. The soil used in this study had fungistasis and the germination rate of macroconidia and chlamydospore of F.solani was 19.4% and 17.7% respectively. The steam sterilized soil lost fungistasis and germination rate of conidia increased to more than 97.9%. The soils amended with the propagule of CHA 1 and S-PFHR 6 increased fungistasis and the germination rate of macroconidia decreased to 14.7% and 11.7% respectively in each treatments. But the soil ammended with glucose and asparagine annulled fungistatic ability and the germination rate of macroconidia increased to more than 48.0%. As an antagonistic activity of the secondary metabolites of two antagonistic isolates in soil, the germination rate of macroconidia of F. solani was 9.3% in the soil amended with the culture filtrate of CHA 1 but the culture filtrate of S-PFHR 6 had no such activity. In the soil which treated with antagonist propagule or culture filtrate, the chlamydospore germination rate was lower than that in natural soil. The addition of glucose and asparagine to antagonist propagule treated soil did not enhanced the chlamydospore germination.

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Factors Predicting the Development of Radiation Pneumonitis in the Patients Receiving Radiation Therapy for Lung Cancer (방사선 치료를 시행 받은 폐암 환자에서 방사선 폐렴의 발생에 관한 예측 인자)

  • An, Jin Yong;Lee, Yun Sun;Kwon, Sun Jung;Park, Hee Sun;Jung, Sung Soo;Kim, Jin whan;Kim, Ju Ock;Jo, Moon Jun;Kim, Sun Young
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.1
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    • pp.40-50
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    • 2004
  • Background : Radiation pneumonitis(RP) is the major serious complication of thoracic irradiation treatment. In this study, we attempted to retrospectively evaluate the long-term prognosis of patients who experienced acute RP and to identify factor that might allow prediction of RP. Methods : Of the 114 lung cancer patients who underwent thoracic radiotherapy between December 2000 and December 2002, We performed analysis using a database of 90 patients who were capable of being evaluated. Results : Of the 44 patients(48.9%) who experienced clinical RP in this study, the RP was mild in 33(36.6%) and severe in 11(12.3%). All of severe RP were treated with corticosteroids. The median starting corticosteroids dose was 34 mg(30~40) and median treatment duration was 68 days(8~97). The median survival time of the 11 patients who experienced severe RP was significantly poorer than the mild RP group. (p=0.046) The higher total radiation dose(${\geq}60Gy$) was significantly associated with developing in RP.(p=0.001) The incidence of RP did not correlate with any of the ECOG performance, pulmonary function test, age, cell type, history of smoking, radiotherapy combined with chemotherapy, once-daily radiotherapy dose fraction. Also, serum albumin level, uric acid level at onset of RP did not influence the risk of severe RP in our study. Conclusion : Only the higher total radiation dose(${\geq}60Gy$) was a significant risk factor predictive of RP. Also severe RP was an adverse prognostic factor.

The Physiological Efficacy of Aloe Gel (Aloe Gel의 생리 효과에 대한 고찰 - Gel의 다당류와 미량 성분을 중심으로 -)

  • 서화중
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.6
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    • pp.1026-1038
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    • 1995
  • Although aloe lost a lot of its previous popularity in modern clinical uses as medicine numerous scientific researches still have claimed the beneficial properties(curing and general tonic effect) of aloe gel. Whereas considerable contradictory reports have helped to confuse the aloe gel issue and continually aroused controversy about aloe gel efficacy. However health food, cosmetic and medicinal products made from aloe gel are widely available in the world market especially in U.S.A. so the growing of Aloe plant and the processing of A. vera gel have become big industries in some countries. In some previous papers the salicylic acid, one of the common trace gel components, was thought to have an analgetic and antinflammatory effect. Large amount of Mg ion in the gel was suggested to act as anesthetic, Mg-lactate as antihistamic, and Aloctin A(a glycoprotein) as wound healer by promoting the cell growth. The carboxypeptidase and bradykinase activity in the gel were proposed to have the pain relieving and antiinflammatory effect. But any of thes etheories concerining the physiological action of the trace gel components has not been demonstrated by modern pharmacology, and failed to be supported by clinical research. It was suggested by some research workers that trace amount of anthraquinone compounds in the gel play an important role to act as false substrate inhibitors for PG and TX production(antiprostanoid effect), by which, they believed, inflammation, burn and frostbite, and infected wound could be healed. This hypothesis has not been substantiated. Butthe suggested antimicrobial action, antidiabetic, and antidotic effect of aloe gel are likely to be attributed to the trace anthraquinone compounds. In a lot of recent experimental reports it has been claimed that aloe gel polysaccharides(acetylglucomannan, acetylmannan, and glycoprotein) have the antimicrobial, antinflammatory, antitumour, and infected wound healing effect by immunoenhancement. It is hoped that these effects will be soon documented in clinical studies, then the controversy on aloe gel beneficial effect will cease. In the 30 days subchronic toxicity test the lowest observed adverse effect level of acemannan(acetylmannan) on dog was 5.0 mg/kg, IP. But the aloe gel is generally agreed to be harmless and non toxic even for the internal use such as health food. In the case of idiosynrasy one must keep the delayed type hypersensitivity reaction of aloe gel in mind. In conclusion it seem to be impossible to simply refuse a lot of evidences made by research workers who have claimed aloe gel's beneficial effects and to deny the fact that there had been long therapeutic histories of Aloe plants.

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Adhesion Characteristics and the High Pressure Resistance of Biofilm Bacteria in Seawater Reverse Osmosis Desalination Process (역삼투 해수담수화 공정 내 바이오필름 형성 미생물의 부착 및 고압내성 특성)

  • Jung, Ji-Yeon;Lee, Jin-Wook;Kim, Sung-Youn;Kim, In-S.
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.51-57
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    • 2009
  • Biofouling in seawater reverse osmosis (SWRO) desalination process causes many problems such as flux decline, biodegradation of membrane, increased cleaning time, and increased energy consumption and operational cost. Therefore biofouling is considered as the most critical problem in system operation. To control biofouling in early stage, detection of the most problematic bacteria causing biofouling is required. In this study, six model bacteria were chosen; Bacillus sp., Flavobacterium sp., Mycobacterium sp., Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhodobacter sp. based on report in the literature and phylogenetic analysis of seawater intake and fouled RO membrane. The adhesion to RO membrane, the high pressure resistance, and the hydrophobicity of the six model bacteria were examined to find out their fouling potential. Rhodobacter sp. and Mycobacterium sp. were found to attach very well to RO membrane surface compared to others used in this study. The test of hydrophobicity revealed that the bacteria which have high hydrophobicity or similar contact angle with RO membrane ($63^{\circ}$ of contact angle) easily attached to RO membrane surface. P. aeruginosa which is highly hydrophilic ($23.07^{\circ}$ of contact angle) showed the least adhesion characteristic among six model bacteria. After applying a pressure of 800 psi to the sample, Rhodobacter sp. was found to show the highest reduction rate; with 59-73% of the cells removed from the membrane under pressure. P. fluorescens on the other hand analyzed as the most pressure resistant bacteria among six model bacteria. The difference between reduction rates using direct counting and plate counting indicates that the viability of each model bacteria was affected significantly from the high pressure. Most cells subjected to high pressure were unable to form colonies even thought they maintained their structural integrity.

Radiotherapy Results of Uterine Cervix Cancer Stape IIB : Overall Survival, Prognostic factors, Patterns of Failure and Late Complications (자궁경부암 병기 IIB에서의 방사선 치료 성적: 생존율 및 예후인자, 치료 후 실패양상, 만성 합병증)

  • Kim Eun-Seog;Choi Doo-Ho;Huh Seung-Jae
    • Radiation Oncology Journal
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    • v.16 no.1
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    • pp.51-61
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    • 1998
  • Purpose : Treatment of choice for uterine cervix cancer stage IIB is radiotherapy. We analyzed survivals, Prognostic factors, patterns of failure and complications. Materials and Methods : This is a retrospective analysis of 167 patients with stage IIB carcinoma of uterine cervix treated with curative external pelvic and high dose rate intracavitary radiotherapy at the Department of Therapeutic Radiology, Soonchunhyang University Hospital from August 1985 to August 1994. All the patients followed up from 3 to 141 months(mean 60 months) and age of patients ranged from 31 to 78 years at presentation(mean : 55 years). Results : Overall complete response rate was $84\%$. The response rate for squamous cell carcimoma and adenocarcinoma were $86\%$ and $60\%$, respectively. Overall 5-years survival rate and disease free survival rate was 62 and $59\%$, respectively Mass size and treatment response were significant Prognostic factors for survival Pathologic type and parametrial involvement were marginally significants Prognostic factors. Local failure was 43 cases, distant metastasis was 14 cases and local failure plus distant metastasis was 3 cases, and most of local failures occurred within 24 months, distant metastasis within 12 months after treatment Twenty eight($16.8\%$) patients developed late rectal and urinary complications There were tendency to increasing severity and frequency according to increased fractional dose and total(rectal and bladder) dose. Conclusions : Survival rate was significantly related to tumor size and radiotherapy response. Tumor size should be considered in the clinical s1aging. To increased survival and local control, clinical trials such as decreasing duration of radiotherapy or addition of chemotherapy is needed. To detect early recurrence, regular follow up after RT is important. Because total rectal and bladder dose affected late complications. meticulous vaginal packing is needed to optimize dose of normal tissues and to decrease late complications.

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Predictors of Meningitis in Febrile Infants Aged 3 Months or Younger (열이 있는 3개월 이하의 영아에서 수막염의 예측에 대한 연구)

  • Song, Hyang Soon;Kim, Eun Ok;Jang, Young Taek
    • Pediatric Infection and Vaccine
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    • v.16 no.1
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    • pp.40-46
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    • 2009
  • Purpose : The purpose of this study was to identify useful predictors for diagnosing bacterial meningitis and performing CSF studies in febrile infants three months or younger. Methods : Six hundred and fifty two febrile infants with a rectal temperature ${\geq}38.0^{\circ}C$ presented from January 2003 to April 2008 and were retrospectively studied. The total white blood cell count (WBC), band count, absolute neutrophil count (ANC), quantitative C-reactive protein (CRP) and blood cultures were performed on admission. The clinical variables associated with bacterial meningitis were analyzed. Results : In patients with bacterial meningitis, the clinical variables including CRP (P=0.036), band count (P=0.037), ANC (P=0.036) and age (P=0.001) were significantly different. The area under the receiver-operating characteristic curve was 0.969 for CRP, 0.946 for the band count, 0.765 for the ANC and 0.235 for age. A CRP cutoff point of 8 mg/dL was determined to maximize both the sensitivity and specificity (sensitivity 83%, specificity 95%, likelihood ratio 16.6). A CRP concentration of <7 mg/dL "ruled-out" bacterial meningitis, with a likelihood ratio of 0.17, a posttest probability of <0.1% and negative predictive value 91%. A CRP concentration greater than 9 mg/dL had a much higher likelihood ratio (20.1) than the band count (16.6) and ANC (2.2). Conclusion : The CRP concentration was a useful laboratory test for the differential diagnosis of bacterial meningitis among febrile infants three months of age or younger. A CRP concentration of <7 mg/dL effectively ruled out bacterial meningitis; a value ${\geq}9mg/dL$ increased the clinical suspicion of bacterial meningitis and the need for CSF evaluation.

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Differences in the Clinical Characteristics of Children with Urinary Tract Infections Based on the Results of $^{99m}Tc$-Dimercaptosuccinic Acid Renal Scanning (요로감염 소아에서 입원 초기 시행한 DMSA 신 스캔 결과에 따른 임상양상의 차이에 대한 연구: DMSA 신 스캔의 임상적 의미)

  • Kim, Dong Ouk;Lee, Sang Min;Lee, Jeong Bong;Ko, Young Bin;Kim, Su Jin
    • Childhood Kidney Diseases
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    • v.17 no.2
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    • pp.110-116
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    • 2013
  • Purpose: The $^{99m}Tc$-Dimercaptosuccinic acid (DMSA) renal scan is used primarily for the diagnosis of renal scarring and acute pyelonephritis in children with urinary tract infections (UTI). This study aimed to evaluate clinical differences based on the positive or negative results of DMSA scans and kidney ultrasonography (US) in pediatric UTI. Method: We retrospectively reviewed 142 pediatric patients with UTI who were admitted to Myongji Hospital from January 2004 to December 2012. We performed a comparative analysis of clinical parameters such as age, sex, white blood cell (WBC) count, neutrophil count, blood urea nitrogen (BUN) level, creatinine (Cr) level, C-reactive protein (CRP) level, and durations of hospitalization and fever, grouped by the results of the DMSA scans and kidney US. Results: The mean age of the patients was $33.8{\pm}48.3$ months, and 78 (55%) were male. Fifty-two patients had abnormal DMSA findings, and 71 patients had abormal kidney US findings (test positive groups). In the DMSA scan positive group, there were significant differences in age, WBC counts, neutrophil counts, CRP level, BUN level, Cr level, hospitalization duration, number of abnormal findings on kidney US, and incidence of vesicoureteral reflux (VUR) compared with the scan negative group. The kidney US positive group had significant differences in age, neutrophil count, CRP level, BUN level, Cr level, hospitalization duration, number of abnormal findings on the DMSA scans, and more frequent VUR compared with the US negative group. Conclusion: Our data suggest that there were no major differences in clinical parameters based on the results of the DMSA scans compared with kidney US in pediatric UTI. However, as kidney US and DMSA scan were performed to predict VUR, the sensitivity and negative predictive value was increased.

Comparative Evaluation of 25-OH-VitD and 25-OH-VitD3 by Radioimmunoassay (방사면역 측정법에 의한 25-OH-VitD와 25-OH-VitD3의 비교 평가)

  • Lee, Young-ji;Park, Ji-hye;Lim, Soo-yeon;Cheon, Jun-hong;Lee, Sun-ho
    • The Korean Journal of Nuclear Medicine Technology
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    • v.24 no.1
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    • pp.33-38
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    • 2020
  • Purpose Vitamin D is essential for maintaining bone health, controling cell proliferation or differentiation, strengthening immune function by controlling calcium metabolism in the body. Vitamin D deficiency can lead to increase the risk of rickets, osteoporosis, cardiovascular disease, diabetes and cancer. Especially, South Korea is one of the highest population proportion of vitamin D deficiency. Accurate determination of levels of 25-OH-VitD or 25-OH-VitD3 in blood serum is required for the diagnosis and treatment of vitamin D deficiency. In this study, radioimmunoassay of 25-OH-VitD and 25-OH-VitD3 was performed and compared to evaluate the effectiveness of Vitamin D radioimmunoassay. Materials and Methods Serum 25-OH-VitD and 25-OH-VitD3 levels were measured using radioimmunoassay. The interrelationship, reproducibility and population distribution rate were evaluated. In addition, the internal quality control was performed at Asan Medical Center from April 2017 to June 2019 and the result of external quality control (Interagency proficiency evaluation) of first and second half of 2018 hosted by the Korean Society of Nuclear Medicine Technology (KSNMT). Both tests were measured by same manufacturer's reagent. Results 25-OH-VitD showed a strong positive correlation on 97 samples, as 25-OH-VitD3 x 0.9 + 0.3 (R>0.9). In repeated measurement, the average Diff(%) value of the reproducibility evaluation of 25-OH-VitD and 25-OH-VitD3 were 7.7% and 7.4%, respectively. Population distribution results showed no statistically significant differences(p>0.05). The resultant value of internal quality control, which measured from April, 2017 to June 2019 in Blood test room of Nuclear Medicine at Asan Medical Center, showed average (CV%) 6.2% and 6.8%, respectively. As a result of the external quality control (interagency proficiency evaluation) Z value obtained under 2.0, as shown below; Conclusion The interrelationship, reproducibility, population distribution rate, internal quality control and external quality control between 25-OH-VitD and 25-OH-VitD3 radioimmunoassay shows superior outcome. Radioimmunoassay, which can be alone measured in the blood as 25-OH-VitD or 25-OH-VitD3, is considered suitable screening tests for the diagnosis of vitamin D deficiency.

Production and physicochemical properties of black bean yogurt made with lactic acid bacteria isolated from vinegar and kimchi (식초와 김치 유래 젖산균으로 제조한 서리태 발효우유 제조 및 물리화학적 품질 특성 연구)

  • Jeong, Seong-Yeop;Kang, Suna;Lee, Na Ra;Ryu, Myeong Seon;Wu, Xuangao;Kim, Da Sol;Park, Sunmin
    • Korean Journal of Food Science and Technology
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    • v.50 no.1
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    • pp.76-82
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    • 2018
  • This study evaluated the physicochemical properties of a novel black bean milk yogurt made by lactic acid producing bacteria isolated from vinegar (Bacillus acidiproducens, BA) and kimchi (Lactobacillus plantarum, LP). Commercially available Lactobacilli mixture for yogurt (YF) were regarded as control bacteria. To investigate optimal conditions to make black bean yogurt, different types of bacteria (BA and LP) and dilution ratios of black bean milk and water (1:1, 1:1.5, and 1:2) were used. Titratable acidity and pH were affected by these dilution ratios, but not by the different types of Lactobacilli used. Titratable acidity and pH were lowest in the 1:1 diluted sample. The visible cell count, sugar content, and viscosity were significantly higher in the sample containing the black bean milk: water ratio of 1:1 than the rest of the samples. In the sensory test, black bean yogurt made with BA showed a higher overall acceptability score, similar to yogurt made with YF. In conclusion, black bean yogurt made by fermentation with BA and with 1:1 black bean milk:water dilution ratio was better than ones made with LP and at other dilution ratios.