• Title/Summary/Keyword: and biotin

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Studies on the Production of Lysine by Fermentation Process (2) -Lysine Production by Auxotrophs- (발효에 의한 라이신(L-Lysine) 생산에 관한 연구 (2) -영양요구성 변이주에 의한 Lysine 생산-)

  • Min, Tae-Ick;Kim, Hang-Mook;Kwon, Tai-Wan
    • Korean Journal of Food Science and Technology
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    • v.4 no.2
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    • pp.123-133
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    • 1972
  • Over 90 of lysine producing auxotrophs were obtained from Corynebacterium sp. S-27-12, Brevibacterium flavum ATCC 15168 and Micrococcus glutamicus ATCC 13032 by UV light, $Co^{60}$ irradiation and N-methyl-N'-nitro-N-nitrosoguanidine treatment. One of the mutant, Brev. flavum U46-N59, was identified as a leucine auxotroph and accumulated lysine during flask (500 ml) cultivation (180 strokes/min.) up to 21.6 mg per ml of broth at pH 7.5 and $28^{\circ}C$ after 4 days. The medium consisted of glucose, 100; urea, 10; corn steep liquor, 40; $KH_2PO_4,\;2;\;K_2HPO_4,\;0.5;\; MgSO_4.\;7H_2O,\;0.4;\;antifoam\;S-57,\;1g;\;Fe_2(SO_4)_3.XH-2O,\;10;\; MnCl_2,\;4H_2O,\;10mg;\;biotin,\;30;\;thiamine-HCl,\;100{\mu}g$in 1l of distilled water, and 40 U/ml of penicillin was added after 36 hrs fermentation.

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Correlation between Cyclin D1, p53, Bcl-2 Protein Expression and Prognosis in Primary, Resected Stage IIIA Non-Small Cell Lung Cancer (NSCLC) (완전 절제된 제IIIA기 비소세포폐암에서 Cyclin D1, p53, Bcl-2 단백질 발현의 의의)

  • 정경영;양우익
    • Journal of Chest Surgery
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    • v.31 no.12
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    • pp.1200-1205
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    • 1998
  • Background: Lung cancer formation is a multistage process involving activation of protooncogene and inactivation of tumor suppressor genes. We evaluate the significance of cyclin D1, p53, bcl-2 gene mutations in patients with curatively resected stage IIIA non-small cell lung cancer(NSCLC). Material and Method: One hundred consecutive cases of stage IIIA lung cancers from patients operated on curatvely between 1990 and 1995 for which adequate paraffin blocks and clinical history were available. Immunohistochemical studies were performed on the representative tissue sections from each case by the labelled streptovidin- biotin method. Sections for cyclin D1, p53, Bcl-2 immunostaining were pretreated in a microwave oven for 10 to 20 minutes in citrate buffer before immunostaining. The overnight incubation with NCL-cyclin D1-GM for cyclin D1, with clone DO-7 for p53, with clone 124 for bcl-2 was done. Mean follow-up was 24.1 months (range 2-84 months) after operation. Result: One hundred cases of lung cancers were composed of 56 cases of squamous cell carcinoma, 37 cases of adenocarcinoma, 5 cases of adenosquamous cell carcinoma, and 2 cases of large cell carcinoma. The 5-year survival was 32.1%. The positive expression rate of cyclin D1 was 35%, p53 was 56%, and bcl-2 was 17%. But there were no correlation between cyclin D1, p53, Bcl-2 protein expression and survival. Conclusion: These observation indicate that cyclin D1, p53, bcl-2 protein overexpression might be implicated in the oncogenesis of non-small cell lung carcinomas but they have no usefulness as a prognostic marker.

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Serum IgE levels in rats infected with Parugonimus westermani (폐흡충(Paragonimus westermani) 감염백서에서의 혈청내 IgE 항체가의 변동)

  • Sin, Myeong-Heon;Ryu, Jae-Suk;Min, Deuk-Yeong
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.397-402
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    • 1991
  • Paragonimus westermani is a common fluke in Korea. The present study aimed to determine serum total IgE and specific IgG levels in experimental paragonimiasis of rats. Each Wistar rat was inoculated orally with 20~25 metacercariae of P. westermani from Cambaroides similis. Before and after infection(1,2,3,4,6,8 weeks) of P. westermani, the blood was collected from the retro- orbital venous plexus of rats and kept serum at $-70^{\circ}C$. Serum total IgE and specific IgG levels were determined by the capture and conventional enzyme-linked immunosorbent assay, respectively. The results were as follows; 1. Serum IgE values were increased to 0. 18${\pm}$0.042 at 2 weeks, $0.28{\pm}0.151$ at 4 weeks and 0.43${\pm}0.055$ at 8 weeks after infection. The absorbances of non-infected rats ranged $0.07{\pm}0.021~0.12{\pm}0.025$. 2. Specific IgG values were slightly increased at 3 weeks ($0.20{\pm}$0.032) and gradually increased up to 8 weeks($0.31{\pm}0.067$) after infection. The absorbances of non-infected rats ranged $0.11{\pm}0.035~0.18{\pm}0.019$. The present results suggested that p. westermani could elevate serum IgE and specific IgG antibodies in Wistar rats which were not a good definitive host.

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Studies on the Effect of Stress-Induced Gastric Ulcer on Gastric Carcinogenesis in Rats treated with N-methyl-N'-nitro-N-nitrosoguanidine (랫드에서 스트레스에 의한 위궤양이 위발암화과정에 미치는 영향에 관한 연구)

  • 이종권;김형진;이영순
    • Journal of Food Hygiene and Safety
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    • v.6 no.1
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    • pp.27-40
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    • 1991
  • ;ABSTRACT-The effects of gastric ulcer induced by restraint and water-immersion stress on gastric carcinogenesis in Wistar male rats treated with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were examined. Rats of group 1 were 2iven stress for 8 hours before they were received MNNG (100 mg/l) in drinking water for 20 weeks. Rats of group 2 were received MNNG first for 2 weeks and then were given stress once a week from 3rd to 12th weeks, with simultaneous MNNG adminitration and followed by MNNG only until 20th weeks. Rats of group 3 were received MNNG only as a positive control and rats of group 4 were not treated with carcinogen. All groups were sacrificed in 20 weeks. Sections of the pyloric mucosa were stained by avidin-biotin peroxidase complex (ABC) immuno-histochemical method. PAPG (pepsinogen isozyme 1 altered pyloric gland), body weight change, gross lesions and histopathological changes were examined. The results obtained from these studies were summarized as follows: 1. The mean body weight gains of the rats fed with carcinogens (group 1, 2, 3) were significantly lower than that of group 4 (control group, without carcinogen. p<0.05). However, the differences of the mean body weight of rats treated with carcinogen were not significant. 2. Stress treatment (group 1 and 2) increased the appearance of the numbers of PAPG (Pepsinogen 1 Altered Pyloric Gland) induced by carcinogen significantly compared with that of group 3 (carcinogen only, p<0.01). 3. The incidence rate of mucosal hyperplasia in pylorus was significantly increased in group 2 compared with group 3 (p<0.05).0.05).

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Detection of Pneumocvstis carinii by in situ hybridization in the lungs of immunosuppressed rats (면역억제 흰쥐에서 조직내교잡법을 이용한 페포자충의 검출)

  • Jin KIM;Jae-Ran YU;Sung-Tae HONG;Chang-Soo PARK
    • Parasites, Hosts and Diseases
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    • v.34 no.3
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    • pp.177-184
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    • 1996
  • In situ hybridization was performed to detect rat heumocwstis ca4nii in the lung sections. Rats were immunosuppressed by weekly subcutaneous injection of 10 mg/kg methylprednisolone. On the 6th, 8th and 9th week of immunosuppression, the lungs were removed and fled in 10% neutral formalin. A 22 base oligonucleotide probe complementary to p. carinii 5S ribosomal RMh was commercially synhesized and its 3' terminal was labeled wiH biotin. In situ hybridization was performed utilizing manual capillary action technolog)r on the Microprobe system. p. cnrinii were detected along the luminal surface of alveolar pneumocytes, in exudate of alveolar cavities, and also in secretory material of bronchioles. In the 6th week group, positive reaction was observed focally in the peripheral region of the lung sections, but the reaction was observed diffusely in the 8th or 9th week groups. In comparison with Grocott's methenamine silver stain, in situ hybridization technique can detect the organism rapidly, and can detect trophic forms very well. Furthermore, no nonspecific reaction with other pathogenic fungi and protozoa was recognized. Therefore, in situ hybridization can be a good technique to detect p. carinii in the lungs of infected rats.

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Studies on Scintillation Proximity Assay for the mesurement of alpha-hCG (Alpha-hCG 측정을 위한 섬광 근접 측정법 (Scintillation Proximity Assay)에 관한 연구)

  • Choi, Tae-Hyun;Lim, Sang-Moo;Choi, Chang-Woon;Chung, Wee-Sup;Lim, Soo-Jeong;Lee, Su-Jin;Lee, Tae-Sup;Awh, Ok-Doo
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.2
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    • pp.133-139
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    • 2002
  • Purpose: Scintillation Proximity Assay (SPA) does not require the physical separation of receptor bound form from free form. SPA was applied to the study of interaction of human chorionic gonadotropin (hCG) and $anti-{\alpha}$ hCG in serum. Materials and methods: $Anti-{\alpha}$ hCG was biotinylated for the binding to streptavidin. The assay was based on the simple competitive binding method between $[^{125}I]hCG$ and the hCG in sample serum, with $anti-{\alpha}$ hCG-coated beads. Aliquots of biotinylated $anti-{\alpha}$ hCG were dispensed into scintillation vials containing $100{\mu}{\ell}\;[^125}I]hCG\;and\;200{\mu}{\ell}$ of either a standard concentration of hCG for preparation of standard curve or unknown sample, and incubated for 20 min. at room temperature. Then $20{\mu}{\ell}$ streptavidin-coated beads were added to vials, and finally incubated for 10 min at room temperature. Values for unknown samples were then calculated from the standard curve. Results: Optimal background counts were certificated using varied radioactivity of radionuclides. Appropriate standard curve was obtained from SPA method successively, and the concentration of hCG from unknown serum was determined by standard curve. The result from SPA assay was similar to that of RIA. Conclusion: This observation confirms that SPA method could be useful for clinical diagnosis.

Studies on the Main Factors Affecting the Mycelial Growth of Phellinus linteus (목질진흙버섯균 Phellinus linteus의 균사체 생육에 미치는 주요 인자에 관한 연구)

  • Chi, Jeong-Hyun;Ha, Tae-Moon;Kim, Young-Ho;Rho, Yeong-Dock
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.214-222
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    • 1996
  • This study was carried out to obtain the basic data on artificial culture of Phellinus linteus. The optimum condition for the mycelial growth was $25{\sim}30^{\circ}C$ and pH $6.0{\sim}7.0$. The carbon sources such as D-glucose, D-mannose and Dextrose were favorable to mycelial growth. As nitrogen sources, peptone, cassamino acid and glutamic acid appeared to be favorable. The optimum C/N ratio was about 20:1, when 2% of glucose was provided as a carbon source. The better organic acids and vitamin among tested ones were gallic, silicic acids and biotin. The mineral nutrients of $KH_2PO_4,\;FeSO_4{\cdot}7H_2O,\; MgSO_4{\cdot}7H_2O,\;ZnSO_4{\cdot}7H_2O$ were effective and the optimum concentrations were 0.05, 0.001, 0.02 and 0.003%, respectively.

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Detection of Mycobacterium Tuberculosis by In Situ Hybridization (조직내교잡법을 이용한 결핵균의 검출)

  • Park, Chang-Soo;Kim, Young-Chul;Lee, Jee-Shin;Jung, Jong-Jae;Kim, Doo-Hong;Kim, Jin
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.5
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    • pp.699-708
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    • 2000
  • Background : A presumptive histopathologic diagnosis of tuberculosis is commonly based on the finding of acid-fast bacilli upon microscopic examination of a diagnostic specimens. Although this traditional histochemical staining method is satisfactory, it is time-consuming and not species-specific. For more specific assessment, in situ hybridization assay with oligonucleotide probes is introduced. Methods : The human surgical specimens were obtained from tuberculosis patients, and experimental specimens were made by injecting cultured M. tuberculosis organisms into fresh rat liver. Oligonucleotide probes complementary to ribosomal RNA portion were synthesized and labeled with multiple biotin molecules. For a rapid detection, all procedures were carried out using manual capillary action technology on the Microprobe staining system. Results : The in situ hybridization assay produced a positive reaction in experimental specimens (80-90% sensitivity) after pepsin-HCl pre-treatment for a good permeabilization of probes, but reliable result was not obtained from human surgical specimens. Conclusion : It is, therefore, suggested that biotin-labeled oligonucleotide probes have considerable potential for identification and in situ detection of M. tuberculosis but, there are some barriers to overcome for the diagnostic use of this method.

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Study on the Vitamin Contents of Commercial Powdered Infant Formula (국내 시판 유아용 고형 조제분유의 비타민 함량에 관한 조사)

  • Bae, Hae-Jin;Jun, Mi-Ra;Kim, Young-Gil
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1689-1694
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    • 2007
  • This study was conducted to investigate both fat-soluble and water-soluble vitamin contents of commercial powdered infant formula for obtaining basic data on infant nutrition. Ten commercial infant formula based on cow's milk were collected and the contents of fat-soluble vitamins (vitamin A, D, E, K) and water-soluble vitamins (vitamin C, thiamin, riboflavin, niacin, $B_6$, folate, $B_{12}$, pantothenic acid, biotin) were compared with Dietary Reference Intakes for Koreans (KDRIs). The overall vitamin contents in 100 g and in 100 kcal of infant formula satisfied the recommended formula regulation (KDRIs) and Codex. In infant formula during 0-5 monthly age, fat-soluble vitamin A, D, E, K could supply 178.6%, 205.3%, 208.4%, 976.3% of adequate daily vitamin intakes, respectively. Water soluble vitamins, vitamin C, thiamin, riboflavin, niacin, $B_6$, folate, $B_{12}$, pantothenic acid, biotin could supply 173.2%, 237.2%, 269.8%, 295.9%, 431.6%, 165.8%, 1186.3%, 203.8%, 408.3% of adequate daily vitamin intakes, respectively. In infant formula during 6-11 monthly age, all vitamins satisfied their adequate daily intakes as well. Vitamin A, D, E, K supplied 199.2%, 262.3%, 220.5%, 626.46% of adequate daily vitamin intakes. Vitamin C, thiamin, riboflavin, niacin, $B_6$, folate, $B_{12}$, pantothenic acid, biotin could supply 179.5%, 210.2%, 264.7%, 241.5%, 206.0%, 166.9%, 699.5%, 247.0%, 475.0% of adequate intake of KDRIs. From this study, evaluation of vitamin contents of commercial infant formula was established, which could strengthen the basic information on infant nutrition.

Qualitative Enzyme-Linked Immunosorbent Assay (ELISA) for the Diagnosis of Edwardsiellosis (Edwardsiellosis의 진단을 위한 정성적 ELISA법)

  • Kim, Myoung-Sug;Hwang, Eun-A;Huh, Min-Do;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.12 no.1
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    • pp.24-31
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    • 1999
  • Optimization and standardization of solid phase enzyme immunoassay were done for the diagnosis of edwardsiellosis in fish. The analyzed degree of immobilized antibody on surface of solid phase with peroxidase saturation method showed the optimized result by using partially purified $50{\mu}g/ml$ of rabbit anti-E. tarda Edk-2 antibody in sodium bicarbonate buffer for overnight incubation to cover the surface of polystyrene beads. Optimized immunoreaction was observed in the treatment of $50{\mu}g/ml$ of biotin conjugated antibody followed extravidin-peroxidase diluted 1 : 2,000 in PBS. The detectable concentrations of the this method were $1{\times}10^5$ cells/ml and $1{\times}10^5$ cells/ml expressed as the source of antigen amount for EDTA extraction and heat extraction, respectively. High cross-reaction of solid phase ELISA with the prepared rabbit and-E. tarda Edk-2 was observed against E. tarda strains isolated from flounder suffering from edwardsiellosis in aquatic farms of Korea. It suggested that the potential of this solid phase of ELISA technique is very powerful for the application to different strains of E. tarda isolated in farms of many different areas.

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