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Proteinase activity in the isolates of Trichomonas vaginalis according to their pathogenicity (질트리코모나스의 병원성과 단백 분해 효소와의 상관성)

  • 심영기;박경희
    • Parasites, Hosts and Diseases
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    • v.31 no.2
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    • pp.117-128
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    • 1993
  • Ten axonic isolates of Trichomonns uaginolis were subcutaneously injected to the BALB/c mice in order to assess their pathogenicity by means of so-called "mouse assay" method. All the isolates revealed neutral and acid proteinase activities both in their Iysates and in culture media, but the specific activities of both proteinases in the severely pathogenic group were significantly higher than the mildly pathogenic group (p < 0.05). In the SDS-PAGE system in which the electrophoretic gels contained 0.4% gelatin as the substrate, five different handing patterns of trichomonal proteinases were detected, and the patterns were closely related with the pathogenicity of the isolates of T. vosinalis. All five bands might be regarded as cysteine proteinases group in the inhibitor assays. The cytotoxicity of the Iysates of T. vaginalis to the target Chinese hamster ovarian (CHO) cell line was also significantly different according to the pathogenicity of the isolates, and generally lower in the Iysates treated with cysteine proteinase inhibitors than in the control Iysates. In summarizing the results, it might be considered that the proteinases of T.vaginalis showing five electrophoretic banding patterns are closely related with the pathogenicity and cytotoxicity of the isolates of T. voginolis.

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Ultrastructural Changes on the Secreting Cells of the Prothoracic Gland During the Larva-pupal Molt of Bombyx mori L. (누에나방(Bombyx mori L.) 종령유충일용 전환기(轉換期)의 전흉선(前胸腺) 분필세포(分泌細胞)의 미세구조적 변화)

  • Oh, S.J.;Kim, J.H.;Kim, C.W.;Kim, W.K.
    • Applied Microscopy
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    • v.12 no.2
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    • pp.69-79
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    • 1982
  • In order to define the morphological changes of the secreting cells of prothoracic gland during larva-pupal molt, ultrastructural observations were carried out using Bombyx mori L. as the experimental material. At first stage of present experiment, 4 day old 5th instar larva, the polyhedral secreting cells were centrally located in the prothoracic gland surrounded by the connective sheath. The secreting cells were attached to the neighboring cells by the prominent desmosomes, and the plasma membrane contacted with connective sheath were highly infolded. In cytoplasm, the most of the cell organelles, such as rod-like mitochondria, rough surfaced endoplasmic reticulum, ribosome were developed. As the stages advance from larva to pupa, general feature of the secreting cells were retained, but structural changes of the various cytoplasmic organelles-ribosome, rough surfaced endoplasmic reticulum, mitochondria, Golgi apparatus, lamellar body, and vesicle-were noted. In the perinuclear cytoplasm of the secreting cells at the stage of 6 day old 5th instar larva, it is peculiar that only a large amount of ribosomes were distributed and the other organelles were retreated from the juxtanuclear region. Just before and after spining cocoon, these features were more remarkable. Rough surfaced endoplasmic reticulum were gradually increased from the stage just before spining cocoon to the pharate pupa. Rod-like mitochondria with irregular cristae and the matrix showing low density were distributed throughout the cytoplasm in the secreting cells of the 4 day old 5th instar larva. Sometimes, longitudinally distended and curved mitochondria were observed. At the stage of pharate pupa, most of mitochondria were deformed. The rod-like mitochondria of the secreting cells of pupal prothoracic gland were narrower than those of 4 day old 5th instar larva, and the electron density of the mitochondrial matrix is increased in pupa. Golgi apparatus were a few in number in both stages, last instar larva and spining cocoon. In stages of the pharate pupa, the Golgi apparatus were frequently observed. Cytoplasmic vesicles were observed for the first time in the secreting cells of one day after spining cocoon, and the number and the size of cytoplasmic vesicles were distinctly increased inpharate pupa and just after pupation. In the secretory cells of the PG, it in concluded that the RER was closely related to syntheting the enzymes seem to produce the ecdysone.

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The Effects of the Epithelial Cells of Genital Tract on the Development of Mouse Early Embryos and Human Fertilized Oocytes (생쥐 초기배아와 사람의 수정란의 발생에 미치는 생식수관 상피세포의 영향에 관한 연구)

  • Lee, H.J.;Byun, H.K.;Kim, J.W.;Hwang, J.H.;Jun, J.Y.;Kim, M.K.
    • Clinical and Experimental Reproductive Medicine
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    • v.21 no.3
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    • pp.315-323
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    • 1994
  • Mammalian oviductal epithelial cells have been known to improve in vitro fertilization and embryonic development. Recently, co-cultured human embryos with the epithelial cells in human genital tract has been reported to improve the pregnancy rate. The purpose of the study was to investigate the effects of the epithelial cells of human genital tract on the development of mouse early embryos and human fertilized oocytes. The epithelial cells of human genital tract were collected from the fallopian tubes which were obtained during hysterectomy in fertile women and from the endometrium during endometrium biopsy. Collected human ampullary cells(HACs) and endometrial cells(HECs) were cultured for 10 days to establish primary monolayer. Second passaged HACs and HECs were obtained by trypsinization were cryopreserved in PBS with 1.5 M DMSO for later use. To investigate the effect when co-cultured with HACs and HECs, we tried to apply strict quality control on mouse embryo, from two cell to blastocyst prior to human trial. The results of quality control were as follows; In Group I (Ham's F10 with 10% FCS), Group IT (co-cultured with HACs) and Group ill (co-cultured with HECs), developmental rates to blastocyst were 63.3%(253/400), 76.0%(304/ 400),74.0%(296/400), respectively. Hatching rates were 36.8%(147/400), 41.80/0(167/400), 38.0%(152/400), respectively(p<0.05). To perform the human IVF, cryopreserved HACs were thawed at 37$^{\circ}C$ waterbath, seeded on the well dish and cultured for 48 hI'S. The pronuclear stage embryos were transferred to the seeded well dish. After 24 hRS, co-cultured embryos were examined and transferred to patient's uterus. The results of human IVF when co-cultured with HACs were that fertilization and developmental rates were 61.8% (256/414), 95.3% (244/256) as compared with 57.2% (279/488) and 94.6%(264/279) in Ham's F10 supplemented with 10% FCS(control). However, 62.9% (161/256) of co-cultured human embryos showed good embryos(no or slight fragmentation) as compared with 53.8 % (150/279) in control(p < 0.05). Pregnancy rate was 40.0% (12/30) when co-cultured with HACs whereas 30.6%(11/36) in control. In conclusions, co-culture system using HACs and HECs improved the developmental and hatching rates of mouse embryo. Also, in human IVF system when co-cultured with HACs, it improved both the quality of human embryos and the pregnancy rate.

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Pharmacological Action of Ginseng (인삼의 약리작용)

  • 홍사악;임정규;박찬웅;차인준
    • Journal of Ginseng Research
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    • v.3 no.1
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    • pp.66-93
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    • 1979
  • Panax ginseng C. A. Meyer, which has been known for more than EWO years. occupies a Particular prince in folk medicine as so called tonic remedy. The pharmacolgical investigations of ginseng, based on the scientific concepts and methodology, have been performed by many researchers through the past 50∼60 years at different parts of the world. The pharmacological action of Panax ginseng compiled from the numerous reports can be summarized as follows: 1. On central nervous system, the effect of Panax ginseng is timulatory in smaller doses and somewhat depressive in larger doses. From the psychopharmacological aspect, ginseng seems to increase the mental efficiency of man. 2. Ginseng has the effect tending to Protect organism from various physical and chemical stresses. 3. The growth and basal metabolic rates of experimental animals are stimulated by ginseng. Ginseng also prolongs the survival time of animals under adverse influences. 4. Increasing the physical and mental efficiency, ginseng postpones the onset of fatigue and increases the working capacities. 5. In the case of the intravenous administration of ginseng, a transitory and slight hypotensive effect is observed. These hypotensive effects seems to include that of a direct action and actions related to the release of histamine and/or serotonin by ginseng. 6. It is Presumed that ginseng lowers the elevated bleed ingar and cholesterol level. 7. Ginseng tends to increase the gastrointestinal motizity and tone 8. It is presumed that ginseng Promotes the iron metabolism and activates the hematopoietic factors. 9. Ginseng tends to stimulate the biosynthesis of nucleic acid and release of histamine and serotonin. 10. The toxicity end adverse reactions of ginseng appear to be nothing that warrants apprehension. 11. Anticancer erects of ginseng seem to be due to indirect action rather than direct action on cancer cell, by improving the host condition 12. Recent clinical trials of ginseng harts obtained sent good results, but Present trial is still limited in its range, so it is necessary to broaden the scope of trial covering many kinds of organs and diseases. From the above, although it appears that substantial advancements have been achieved in the studies on the Pharmacological actions of Panax ginseng there are many discrepancies noticed in the reported data. Furthermore the precise mechanisms of actions of ginseng are sometimes obscure, even unknown in other actions as the students stand now. The main reasons for this are considered to be that even though saponin has been identified at one of the active substances of ginseng, other components have not fully been identified and that the experimental approaches of the investigations varied with different researchers. Thus a thorough analysis of the chemical components and newer standardized concepts and metohds appear to be the pre-requisites for further study of the pharmacolgical effects and mechaisms of Panax ginseng.

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Structural Characteristics of Expression Module of Unidentified Genes from Metagenome (메타게놈 유래 미규명 유전자의 발현에 관련된 특성분석)

  • Park, Seung-Hye;Jeong, Young-Su;Kim, Won-Ho;Kim, Geun-Joong;Hur, Byung-Ki
    • KSBB Journal
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    • v.21 no.2
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    • pp.144-150
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    • 2006
  • The exploitation of metagenome, the access to the natural extant of enormous potential resources, is the way for elucidating the functions of organism in environmental communities, for genomic analyses of uncultured microorganism, and also for the recovery of entirely novel natural products from microbial communities. The major breakthrough in metagenomics is opened by the construction of libraries with total DNAs directly isolated from environmental samples and screening of these libraries by activity and sequence-based approaches. Screening with activity-based approach is presumed as a plausible route for finding new catabolic genes under designed conditions without any prior sequence information. The main limitation of these approaches, however, is the very low positive hits in a single round of screening because transcription, translation and appropriate folding are not always possible in E. coli, a typical surrogate host. Thus, to obtain information about these obstacles, we studied the genetic organization of individual URF's(unidentified open reading frame from metagenome sequenced and deposited in GenBank), especially on the expression factors such as codon usage, promoter region and ribosome binding site(rbs), based on DNA sequence analyses using bioinformatics tools. And then we also investigated the above-mentioned properties for 4100 ORFs(Open Reading Frames) of E. coli K-12 generally used as a host cell for the screening of noble genes from metagenome. Finally, we analyzed the differences between the properties of URFs of metagenome and ORFs of E. coli. Information derived from these comparative metagenomic analyses can provide some specific features or environmental blueprint available to screen a novel biocatalyst efficiently.

Effect of a Freshwater Bivalve (Unio douglasiae) and a Submerged Plant (Potamogeton crispus) on the Growth Inhibition of a Cyanobacterium Oscillatoria sp. (담수패류(Unio douglasiae)와 침수식물(Potamogeton crispus)의 유해 남조 Oscillatoria sp. 성장억제 효과)

  • Kim, Keun-Hee;Kim, Baik-Ho;Park, Myung-Hwan;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.41 no.spc
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    • pp.68-76
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    • 2008
  • This study examined the inhibition effects of a freshwater bivalve (Unio douglasiae) and a submerged plant (Potamogeton crispus) on the cyanobacterial bloom (Oscillatoria sp.). The experiment were conducted in aquarium $(50cm{\times}65cm{\times}120cm)$ with lake sediments in the bottom of the aquarium in 10 cm thick. Before the experiments, artificial cyanobacterial bloom was induced with the addition of lake sediment and CB medium. Total 12 transparent acrylic cylinders (${\Phi}19cm$, height 40 cm) were placed in the aquarium, and within which bivalves and plants were placed in various conditions such as the control (C), plant addition (P:5 stems), mussel addition (U:2 individuals), and both mussel and plant addition (PU: the same quantity as used in each treatment). The experiment was conducted in triplicate during 7 days. pH, dissolved oxygen (DO), electric conductivity (EC), salinity, cyanobacterial cell density, chlorophyll-${\alpha}$ concentration, and mussel filtering rate were monitored daily. At the end of the experiment, total phosphorus (TP), total nitrogen (TN), and plant height and weight were measured. Overall, a large degree of cyanobacterial growth inhibition appeared in both P and U treatments, and the effect was highest in the U treatment, followed by P and PU. The combined treatment of both U and P did not show any synergic effects compared to the effect in separated treatment. In all enclosures of the treatments chlorophyll-${alpha}$ (Chl-${alpha}$) concentration decreased until 36 hours after the additions of the plants and mussels. In contrast, Chl-${alpha}$ concentrations increased in PU enclosures after 36 hours. The same trend was shown in the cell density of Oscillatoria. pH and DO gradually decreased until 120 and 144 hours, respectively, in the P and PU enclosures. TP concentration increased in the mussel enclosures (U and PU), while TN concentration largely decreased in the plant enclosures (P and PU). Our results suggest that applied bivalve (Unio) and submerged plant (Potamogeton) seemed to have a potential effect on the growth inhibition of cyanobacteria, but their combined application may have an antagonistic effect to diminish the degree of the inhibition.

Role of Postoperative Radiation Therapy in the Management of Cervical Cancer (자궁경부암에서 수술 후 방사선치료의 역할)

  • Chun, Ha-Chung;Lee, Myung-Za
    • Radiation Oncology Journal
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    • v.22 no.4
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    • pp.265-270
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    • 2004
  • Purpose: To evaluate the effectiveness of postoperative radiation therapy in cervical cancer patients and define the prognostic factors to affect survival rates. Materials and Methods: Eighty one patients with cervical cancer who were treated with postoperative radiation therapy following surgery at our institution between May 1992 and April 2000 were retrospectivelv analyzed. Forty two patients had stage IB disease, 17 had stage IIA disease, and remaining 22 had stage IIB disease, respectively. Histological examination revealed 76 squamous cell carcinoma and 5 adenocarclnoma. Sixty one patients were noted to have stromal invasion greater than 8 mm and 20 patients were noted to have stromal invasion 7 mm or less. Sixteen patients had parametrial invasion and 65 patients did not. Positive vaginal resection margin was documented in only eight patients and positive lymphovascular invasion was in twelve patients. All of the patients were treated with external beam radiation therapy alone. Majority of the patients were treated with 4 field brick technique to encompass whole pelvis. Total of 5,500 cGy was delivered to the primary surgical tumor bed. Minimum follow up period was four years. Results: Actuarial disease free survival rates for entire group of the patients were 95% and 89% at 2 and 5 years, respectively Five year disease free survival rates for patients with stage IB, IIA, and IIB disease were 97%, 87% and 70%, respectivelv. Local recurrences were documented in 5 patients. Cumulative local failure rate at 3 years was 6% Five year disease free survival rates for patients with stromal invasion greater than 8 mm and 7 mm or less were 88% and 92%, respectively (p>0.05). Five year disease free survival rate for patients with parametrial invasion was significantly lower than those with no invasion (72% vs 92%, p<0.05). Also there was significantly lower survival in patients with positive vaginal resection margin, compared with patients with negative resection margin (64% vs 94%, p<0.05). However, lymphovascular invasion was not a statistically significant prognostic factor Parametrial invasion and positive surgical resection margins were noted to be significant prognostic factors. Conclusions: Postoperative radiation therapy appears to be beneficial in controlling local disease in cervical cancer patients with high pathologic risk factors. Parametrial invasion and positive resection margins were noted to be significant prognostic factors to affect survival and more effective treatment should be investigated in these patients.

A Clinical Study of Aseptic Meningitis in the Northern Area of Seoul in 2002 (2002년 서울 북부지역에서 유행한 무균성 수막염의 임상적 고찰)

  • Song, Myoung Hak;Chung, Ju Young;Kwon, Ji Won;Kim, Sang Woo
    • Pediatric Infection and Vaccine
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    • v.11 no.2
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    • pp.176-182
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    • 2004
  • Purpose : The etiologic agents of aseptic meningitis remain mostly unknown due to difficulty of viral culture and identification. There was an outbreak of aseptic meningitis in northern area of Seoul from June to August, 2002. We report the clinical features, laboratory data and causative viruses on 196 children with aseptic meningitis during this period. Methods : We retrospectively studied about clinical manifestations and laboratory findings 196 patients diagnosed as aseptic meningitis at Sanggye-Paik hospital. Virus isolation and serotype identification were performed by cell culture and reverse transcription polymerase chain reaction(RT-PCR) of the cerebrospinal fluid. Results : The male to female ratio was 1.39 : 1 and the mean age was 5.8+3 years. The clinical manifestations were fever, headache and vomiting. It occurred mostly in June, July and August. The numbers of peripheral blood leukocytes were $4,800{\sim}24,360/mm^3$. On cerebrospinal fluid examinations, leukocytes were in range of 10~2,000(mean 105)/$mm^3$, protein level in range of 15~171(mean 41.4) mg/dL and glucose level from 16~97(mean 57.9) mg/dL. Viral culture of cerebrospinal fluid showed 3 cases of Echovirus 9, 1 case of 25 and 30. In stool culture, 2 cases of Echovirus 6, 2 cases of Echovirus 13 and 1 case of Echovirus 30 were isolated. Conclusion : The etiologic viruses of the aseptic meningitis in northern area of Seoul in 2002 are presumed to be Echovirus 6, 9, 13, 25, 30.

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Effects of Non-ionic Surfactant Tween 80 on the in vitro Gas Production, Dry Matter Digestibility, Enzyme Activity and Microbial Growth Rate by Rumen Mixed Microorganisms (비이온성 계면활성제 Tween 80의 첨가가 반추위 혼합 미생물에 의한 in vitro 가스발생량, 건물소화율, 효소활력 및 미생물 성장율에 미치는 영향)

  • Lee, Shin-Ja;Kim, Wan-Young;Moon, Yea-Hwang;Kim, Hyeon-Shup;Kim, Kyoung-Hoon;Ha, Jong-Kyu;Lee, Sung-Sil
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1660-1668
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    • 2007
  • The non-ionic surfactant (NIS) Tween 80 was evaluated for its ability to influence invitro cumulative gas production, dry matter digestibility, cellulolytic enzyme activities, anaerobic microbial growth rates, and adhesion to substrates by mixed rumen microorganisms on rice straw, alfalfa hay, cellulose filter paper and tall fescue hay. The addition of NIS Tween 80 at a level of 0.05% increased significantly (P<0.05) in vitro DM digestibility, cumulative gas production, microbial growth rate and cellulolytic enzyme activity from all of substrates used in this study. In vitro cumulative gas production from the NIS-treated substrates; rice straw, alfalfa hay, filter paper and tall fescue hay was significantly (P<0.05) improved by 274.8, 235.2, 231.1 and 719.5% compared with the control, when substrates were incubated for 48 hr in vitro. The addition of 0.05% NIS Tween 80 to cultures growing on alfalfa hay resulted in a significant increase in CMCase (38.1%), xylanase (121.4%), Avicelase (not changed) and amylase (38.2%) activities after 36 h incubation. These results indicated that the addition of 0.05% Tween 80 could greatly stimulate the release of some kinds of cellulolytic enzymes without decreasing cell growth rate in contrast to trends reported with aerobic microorganism. Our SEM observation showed that NIS Tween. 80 did not influence the microbial adhesion to substrates used in the study. Present data clearly show that improved gas production, DM digestibility and cellulolytic enzyme activity by Tween 80 is not due to increased bacterial adhesion on the substrates.

Quality Characteristics and Inhibition Activity against Helicobacter pylori KCCM 40449 of Liquorice Yogurts Manufactured by Exopolysaccharide Producing Lactic Acid Bacteria (Exopolysaccharide 생성 유산균을 이용한 감초 추출물 첨가 Yoghurt의 품질특성 및 Helicobacter pylori KCCM 40449 억제활성)

  • Jung, Seung-Won;Kim, Cheol Woo;Lee, Su Han
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.346-354
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    • 2011
  • This study was carried out to fortify the antimicrobial activity of yoghurt by adding liquorice extract to it. The liquorice extracts (1 mg/mL) showed relatively high antibacterial activity against H. pylori KCCM 40449 (p < 0.05). The solvent liquorice extracts of minimal inhibitory concentrations (MIC) against H. pylori KCCM 40449 were 25- 100 ${\mu}g$/mL. Lactobacillus amylovorus DU-21 with high EPS production ability were inoulated to milk after the addition of different amounts of liquorice extracts (0.0%, 0.05%, 0.1% and 0.2%). The physico-chemical characteristics of yoghurts added with liquorice extracts were examined. The initial pH, titratable acidity, viscosity and viable cell counts of the yoghurt added liquorice extracts were 3.41-3.51, 1.021-1.091%, 1,686-1,930 cp and 9.41-9.38 Log CFU/mL, respectively. The viscosity and syneresis of yoghurt were better than that of the control. Antimicrobial activity against H. pylori KCCM 40449 increased with increasing addition of liquorice extract. However, the sensory score of yoghurt added with different amounts of liquorice extracts was lower than that of the control (p < 0.05). As a result of the sensory evaluations, the flavor, taste, texture, color and overall acceptability of the yoghurt with 0.05% liquorice extract were found to be much better than those of the other groups (p < 0.05). Overall, the optimal amount of liquorice extract added in the manufacture of yoghurt was 0.05% of the total weight. Further studies on increment of antimicrobial activity and palatability of liquorice extract added yoghurt are necessary.