• Title/Summary/Keyword: microbial hormone

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A Continuous Spectrophotometric Assay for NADPH-cytochrome P450 Reductase Activity Using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide

  • Yim, Sung-Kun;Yun, Chul-Ho;Ahn, Tae-Ho;Jung, Heung-Chae;Pan, Jae-Gu
    • BMB Reports
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    • v.38 no.3
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    • pp.366-369
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    • 2005
  • NADPH-cytochrome P450 reductase (CPR) transfers electrons from NADPH to cytochrome P450 and also catalyzes the one-electron reduction of many drugs and foreign compounds. Various spectrophotometric assays have been performed to examine electron-accepting properties of CPR and its ability to reduce cytochrome $b_5$, cytochrome c, and ferricyanide. In this report, reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) by CPR has been assessed as a method for monitoring CPR activity. The principle advantage of this substance is that the reduction of MTT can be assayed directly in the reaction medium by a continuous spectrophotometric method. The electrons released from NADPH by CPR were transferred to MTT. MTT reduction activity was then assessed spectrophotometrically by measuring the increase of $A_{610}$. MTT reduction followed classical Michaelis-Menten kinetics ($K_m\;=\;20\;{\mu}M$, $k_{cat}\;=\;1,910\;min^{-1}$). This method offers the advantages of a commercially available substrate and short analysis time by a simple measurement of enzymatic activity of CPR.

The Effect of Collagen Supplementation from Pork Skin on Serum Collagen, Serum Sex Steroid Hormone, Serum Lipid and Skin Crack in Korean Middle-aged Women (돈피 추출 콜라겐 보충 식이가 중년 여성의 혈중 콜라겐, 성호르몬, 지질대사 및 피부 갈라짐에 미치는 영향)

  • Han, Chae-Jeong;Kang, Sang-Mo
    • Korean Journal of Community Nutrition
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    • v.13 no.6
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    • pp.912-921
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    • 2008
  • This study was performed to examine if the effects of collagen supplementation from pork skin could improve the sex steroid hormone, serum lipid and skin crack in Korean middle-aged women. Middle-aged women (40-55 years) who were not diagnosed with any type of disease were included in this study and thirty subjects were randomly assigned to a control group (n = 15) or a collagen supplemented group (n = 15). The collagen supplemented group ingested collagen flour 2 g, 3 times a day for 12 weeks. We measured serum collagen, estrogen, estradiol, estriol, progesterone, total cholesterol, triglyceride, HDL-cholesterol and LDL-cholesterol concentration. The collagen supplementation group had significantly increased serum collagen (p < 0.05) compared with the control group. In addition, skin crack was improved. But, there were no differences for sex steroid hormone and lipid profile in control and collagen supplemented groups. The result of the present study demonstrated that supplementation of 6 g collagen per day for 12 weeks can give beneficial effects on skin crack reduction and serum collagen concentration.

Temperature Effect on the Functional Expression of Human Cytochromes P450 2A6 and 2E1 in Escherichia coli

  • Yim Sung-Kun;Ahn Taeho;Jung Heung-Chae;Pan Jae-Gu;Yun Chul-Ho
    • Archives of Pharmacal Research
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    • v.28 no.4
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    • pp.433-437
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    • 2005
  • Human cytochromes P450 (GYP) 2A6 and 2E1 are of great interest because of their important roles in the oxidation of numerous drugs and carcinogens. Bacterial expression systems, especially Escherichia coli cells, have been widely used for the production of various GYP enzymes in order to obtain high yield of proteins. The expression methods usually employ longer culture time (30-72 h) at lower temperature (usually under $30^{\circ}C$). Expression levels of GYPs 2A6 and 2E1 at $37^{\circ}C$ were compared to those at $28^{\circ}C$, which is a usual temperature used in most bacterial expression systems for human GYP expression. Within 18 h the expression levels of GYPs 2A6 and 2E1 reached up to 360 and 560 nmol per liter culture at $37^{\circ}C$, respectively, which are compatible with those of 36 h culture at $28^{\circ}C$. The activities of GYPs expressed at $37^{\circ}C$ were also comparable to those expressed at $28^{\circ}C$. The present over-expression system can be useful for rapid production of large amounts of active human GYPs 2A6 and 2E1 in E. coli.

Comparison of Salivary Stress Hormone Levels between Periodontitis Patients and Healthy Subjects in Korea

  • Lee, Sol;Heo, Narae;Heo, Seok-Mo
    • The Journal of the Korean dental association
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    • v.54 no.11
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    • pp.897-906
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    • 2016
  • Objectives: Periodontitis is multifactorial disease mainly caused by microbial community. Recently, some research has been conducted to find other possible risk factors including stress hormones related to periodontitis. Psychological stress can affect the periodontal health by a variety of biological mechanisms. This study compared the stress hormone levels in healthy subjects and patients with periodontal disease using saliva in order to investigate the association between periodontitis and stress. Methods: The human saliva was collected from 38 periodontally healthy individuals and 34 patients with chronic periodontitis under Institutional Review Board. Their age was 20-60 years ($40.3{\pm}10.45$). From these samples, determination of salivary levels of cortisol and Dehydroepiandrosterone (DHEA) performed by enzyme immunoassay kit (Salimetrics Europe, Suffolk, UK). The independent t-test and Mann-Whitney test for trend was applied using IBM SPSS statistics version 12.0 Program to analyze statistically significant differences. Results: Salivary cortisol levels of periodontitis patients were higher than those levels of healthy subjects (P < 0.001), while salivary DHEA levels of periodontitis patients were not significantly different (P = 0.431). Salivary cortisol/DHEA ratio of periodontitis patients was higher than those levels of healthy subjects (P < 0.001). Conclusions: Our study demonstrates the high levels of cortisol concentrations and cortisol/DHEA ratio in saliva of periodontitis patients than those of healthy subjects. Since cortisol levels and cortisol/DHEA ratio can be significant factors related to the severity of periodontal disease, our study would be helpful for early diagnosis and treatment of periodontal disease.

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Evaluation of immunocontraceptive vaccine composed of gonadotropin-releasing hormone conjugated with granulocyte-macrophage colony-stimulating factor in male rats

  • Park, Byung-Joo;Kim, Yong-Hyun;Ahn, Hee-Seop;Han, Sang-Hoon;Go, Hyeon-Jeong;Lee, Joong-Bok;Park, Seung-Yong;Song, Chang-Seon;Lee, Sang-Won;Choi, In-Soo
    • Korean Journal of Veterinary Research
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    • v.57 no.3
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    • pp.155-158
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    • 2017
  • Surgical castration performed to reduce male-associated problems results in pain and microbial infections in male animals. Therefore, immunocontraception, which is mediated by the animal's own antibodies against reproductive hormones, has been recommended as an alternative to surgical castration when considering the animal's welfare. In this study, a new immunocontraceptive vaccine composed of six tandem copies of gonadotropin-releasing hormone (GnRH) fused to rat granulocyte-macrophage colony-stimulating factor (GM-CSF) was developed, and its efficacy was evaluated in male rats. Three different doses (10, 50, and $100{\mu}g$) of recombinant GM-CSF-GnRH protein were injected three times at intervals of two weeks into male rats. The rats vaccinated with three doses of GM-CSF-GnRH produced a significantly higher level of antibodies against GnRH than that in the negative control rats. Severe atrophy of gonads was observed in rats vaccinated with three doses of GM-CSF-GnRH but not in the negative control rats. The results reveal that the new GnRH vaccine conjugated with rat GM-CSF induces efficient immunocontraception in male rats. This formulation of the immunocontraceptive vaccine would be applicable to both domestic and pet male animals.

Recent Advances of Therapeutic Targets for the Treatment of Periodontal Disease

  • Kim, Woo Jin;Soh, Yunjo;Heo, Seok-Mo
    • Biomolecules & Therapeutics
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    • v.29 no.3
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    • pp.263-267
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    • 2021
  • Periodontal disease is primarily associated with bacterial infection such as dental plaque. Dental plaque, an oral biofilm harboring a complex microbial community, can cause various inflammatory reactions in periodontal tissue. In many cases, the local bacterial invasion and host-mediated immune responses lead to severe alveolar bone destruction. To date, plaque control, non-surgical, and surgical interventions have been the conventional periodontal treatment modalities. Although adjuvant therapies including antibiotics or supplements have accompanied these procedures, their usage has been limited by antibiotic resistance, as well as their partial effectiveness. Therefore, new strategies are needed to control local inflammation in the periodontium and host immune responses. In recent years, target molecules that modulate microbial signaling mechanisms, host inflammatory substances, and bone immune responses have received considerable attention by researchers. In this review, we introduce three approaches that suggest a way forward for the development of new treatments for periodontal disease; (1) quorum quenching using quorum sensing inhibitors, (2) inflammasome targeting, and (3) use of FDA-approved anabolic agents, including Teriparatide and sclerostin antibody.

Aging effects on the diurnal patterns of gut microbial composition in male and female mice

  • Kim, Hyun-Jung;Moon, Chang Mo;Kang, Jihee Lee;Park, Eun-Mi
    • The Korean Journal of Physiology and Pharmacology
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    • v.25 no.6
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    • pp.575-583
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    • 2021
  • Composition of the gut microbiota changes with aging and plays an important role in age-associated disease such as metabolic syndrome, cancer, and neurodegeneration. The gut microbiota composition oscillates through the day, and the disruption of their diurnal rhythm results in gut dysbiosis leading to metabolic and immune dysfunctions. It is well documented that circadian rhythm changes with age in several biological functions such as sleep, body temperature, and hormone secretion. However, it is not defined whether the diurnal pattern of gut microbial composition is affected by aging. To evaluate aging effects on the diurnal pattern of the gut microbiome, we evaluated the taxa profiles of cecal contents obtained from young and aged mice of both sexes at daytime and nighttime points by 16S rRNA gene sequencing. At the phylum level, the ratio of Firmicutes to Bacteroidetes and the relative abundances of Verrucomicrobia and Cyanobacteria were increased in aged male mice at night compared with that of young male mice. Meanwhile, the relative abundances of Sutterellaceae, Alloprevotella, Lachnospiraceae UCG-001, and Parasutterella increased in aged female mice at night compared with that of young female mice. The Lachnospiraceae NK4A136 group relative abundance increased in aged mice of both sexes but at opposite time points. These results showed the changes in diurnal patterns of gut microbial composition with aging, which varied depending on the sex of the host. We suggest that disturbed diurnal patterns of the gut microbiome can be a factor for the underlying mechanism of age-associated gut dysbiosis.

Studies of Tarak, a Korean Traditional Fermented Milk Product (한국 전통 발효유 타락에 관한 연구 고찰)

  • Yoon, Jin A;Shin, Kyung-Ok
    • Journal of Dairy Science and Biotechnology
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    • v.36 no.1
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    • pp.26-31
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    • 2018
  • Tarak is a traditional Korean fermented milk product, which is prepared by the addition of rice wine to milk. The major microbial strains found in Tarak are Leuconostoc citreum, Lactobacillus plantarum, Lactococcus lactis, Saccharomyces cerevisiae, and Pichia kudriavzevii. The activity of lactic acid bacteria isolated from traditional Korean foods of Taraki against the carcinogenic bacteria Helicobacter pylori, Escherichia coli O157:H7, and Cronobacter sakazakii was characterized. Tarak extract significantly increased the proliferation of T-lymphocyte Jurkat (clone E6-1) cells. Tarak also inhibited the tyrosinase activity and melanin biosynthesis induced by an ${\alpha}$-melanocyte-stimulating hormone in pituitary intermediate lobe.

Evaluation of Freshness Prolonging Ability of Antibacterial Packaging Bags and Corrugated Boxes (항균포장지와 포장상자에 저장된 과채류의 선도평가)

  • Kim, Chul-Hwan;Kim, Jae-Ok;Park, Woo-Po;Jung, Jun-Ho;Cho, Sung-Hwan
    • Korean Journal of Food Science and Technology
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    • v.37 no.3
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    • pp.364-371
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    • 2005
  • Packaging bags and corrugated boxes prepared from newly developed antimicrobial packaging paper had greater ability to prolong freshness of packed fruits, and lower weight loss of fruits and microbial growth than non-treated bags and boxes. Addition of zeolite to paper greatly increased absorbance of aging hormone, $C_{2}H_{4}$, emitted from stored fruits, and contributed to decrease in putrefied fruits during storage. Water repellent agent such as alkyl ketone dimer played significant role in preventing functional paper from absorbing water from stored fruits, leading to preservation of fresh weight in stored fruits. Because rise in production cost of functional packaging paper could be offset by reduction of fruit and vegetable wastes, storing fruits and vegetables in functional paper containers was more economical than storing in non-treated containers.

Continuous Production of Natural Colorant, Betacyanin, by Beta vulgaris L. Hairy Root

  • Kim, Sun-Hee;Ahn, Sang-Wook;Bai, Dong-Kyu;Kim, Kwang-Soo;Hwang, Baik;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.716-721
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    • 1999
  • It has been known that continuous cultivation of hairy root is difficult to maintain for a long period of time compared to the microbial and callus cultures. Chemostat cultivation was successfully carried out in order to economically produce a plant-based colorant, betacyanin, from red beet hairy root for more than 85 days in a 14-1 fermentor. The result from the chemostat cultivation was compared to those of the batch and fed-batch cultivations of red beet hairy roots. It was shown that hairy root reached its steady state within 50 days of the cultivation, and then maintained for about 25-30 days in a wide range of dilution rates. Total betacyanin production from the continuous process was also calculated to be 2.65g at 0.28(l/d) of dilution rate, compared to 0.196g from fed-batch cultivation. It was found that betacyanin production was a partially growth related process, yielding 0.376 mg/g-fresh wt. cell and $1.89{\times}10^{-5}$ mg/g-fresh wt. cell/d, with 0.92 of correlation factor in a partial growth-product model. It was also shown that the cell growth required was relatively large for maintenance amount of energy at a low dilution rate. The growth of hairy root was inhibited by high light intensity in following a photo-inhibition model. The growth parameters were estimated to be 0.3(l/d), $10.56kcal/\textrm{m}^2/h$,{\;}and{\;}35.81kcal/\textrm{m}^2/h$ for the maximum specific growth rate, half saturation light intensity, and inhibition light intensity, respectively.

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