• Journal of Microbiology and Biotechnology
• /
• 제25권7호
• /
• pp.1015-1025
• /
• 2015

The development of diverse polymyxin derivatives is needed to solve the toxicity and resistance problems of polymyxins. However, no platform has generated polymyxin derivatives by genetically engineering a polymyxin synthetase, which is a nonribosomal peptide synthetase. In this study, we present a two-step approach for the construction of engineered polymyxin synthetases by substituting the adenylation (A) domains of polymyxin A synthetase, which is encoded by the pmxABCDE gene cluster of Paenibacillus polymyxa E681. First, the seventh L-threonine-specific A-domain region in pmxA was substituted with the L-leucine-specific A-domain region obtained from P. polymyxa ATCC21830 to make polymyxin E synthetase, and then the sixth D-leucine-specific A-domain region (A_6-D-Leu-domain) was substituted with the D-phenylalanine-specific A-domain region (A_6-D-Phe-domain) obtained from P. polymyxa F4 to make polymyxin B synthetase. This step was performed in Escherichia coli on a pmxA-containing fosmid, using the lambda Red recombination system and the sacB gene as a counter-selectable marker. Next, the modified pmxA gene was fused to pmxBCDE on the chromosome of Bacillus subtilis BSK4dA, and the resulting recombinant strains BSK4-PB and BSK4-PE were confirmed to produce polymyxins B and E, respectively. We also succeeded in constructing the B. subtilis BSK4-PP strain, which produces polymyxin P, by singly substituting the A_6-D-Leu-domain with the A_6-D-Phe-domain. This is the first report in which polymyxin derivatives were generated by genetically engineering polymyxin synthetases. The two recombinant B. subtilis strains will be useful for improving the commercial production of polymyxins B and E, and they will facilitate the generation of novel polymyxin derivatives.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권3호
• /
• pp.1023-1035
• /
• 2016

The purpose of this study was to investigate the role of colon cancer stem cells (CSCs) during chemically-induced rat multi-step colon carcinogenesis with or without the treatment with a specific cyclooxygenase-2 inhibitor drug (celecoxib). Two experiments were performed, the first, a short term 12 week colon carcinogenesis bioassay in which only surrogate markers for colon cancer, aberrant crypt foci (ACF) lesions, were formed. The other experiment was a medium term colon cancer rat assay in which tumors had developed after 32 weeks. Treatment with celecoxib lowered the numbers of ACF, as well as the tumor volumes and multiplicities after 32 weeks. Immunohistochemical proliferating cell nuclear antigen (PCNA) labeling indexes LI (%) were downregulated after treatment by celecoxib. Also different cell surface antigens known to associate with CSCs such as the epithelial cell adhesion molecule (EpCAM), CD44 and CD133 were compared between the two experiments and showed differential expression patterns depending on the stage of carcinogenesis and treatment with celecoxib. Flow cytometric analysis demonstrated that the numbers of CD133 cells were increased in the colonic epithelium after 12 weeks while those of CD44 but not CD133 cells were increased after 32 weeks. Moreover, aldehyde dehydrogenase-1 activity levels in the colonic epithelium (a known CSC marker) detected by ELISA assay were found down-regulated after 12 weeks, but were up-regulated after 32 weeks. The data have also shown that the protective effect of celecoxib on these specific markers and populations of CSCs and on other molecular processes such as apoptosis targeted by this drug may vary depending on the genetic and phenotypic stages of carcinogenesis. Therefore, uncovering these distinction roles of CSCs during different phases of carcinogenesis and during specific treatment could be useful for targeted therapy.

• Preventive Nutrition and Food Science
• /
• 제19권3호
• /
• pp.145-155
• /
• 2014

Vascular dementia is characterized by white matter lesions involving the demyelination and activation of astrocytes and microglia. In a previous study, we showed that the supernatant of a laboratory-scale, hot water extract of ground whole wheat (TALE) attenuated white matter injury and astrocytic activation in a rat model of bilateral common carotid artery occlusion (BCCAO). In the present study, we made several modifications to the hot water extraction process to remove starch and enable large-scale production. We used wheat bran (WB), which contains less starch, instead of ground whole wheat. In addition, we removed starch granules with a decanter before hot water extraction. The final product, wheat bran extract (WBE), contained 2.42% arabinose, a surrogate marker of arabinoxylan, which is an active constituent of WBE. Supplementation of the rat model of BCCAO with WBE (400 mg/kg/day) for 33 days attenuated white matter injury, which was assessed by Luxol Fast Blue staining, in the corpus callosum (cc) and optic tract (opt) regions. Attenuation of white matter injury in the opt region was accompanied by improvement of the pupillary light reflex. Immunochemical staining revealed that supplementation with WBE reduced astrocytic activation in the cc and opt regions and reduced microglial activation in the opt region. These findings indicate that supplementation with WBE is effective at attenuating white matter injury accompanied by the inhibition of astrocytic and microglial activation. Therefore, extracts from WB, a cheap by-product of wheat milling, can be developed as a nutraceutical to prevent vascular dementia, a disease for which there is no approved pharmaceutical treatment.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권5호
• /
• pp.2399-2403
• /
• 2012

Objective: To compare expression level of serum tumor associated materials (TAM) with several conventional serum tumor biomarkers, eg., carcinoembryonic antigen (CEA), carbohydrate antigen19-9 (CA19-9), carbohydrate antigen 15-3 (CA15-3), alpha-fetoprotein(AFP), in selected solid tumors. Methods: Patients diagnosed histologically or cytologically with liver, breast, esophageal, gastric, colorectal or pancreatic cancers were enrolled into this study. After diagnosis, the level of TAM was determined by chemical colorimetry, and levels of conventional tumor markers was measured by chemiluminescence methods. Results: A total of 560 patients were enrolled into this study. No statistically significant difference was detected in TAM and the above mentioned tumor biomarkers in terms of their positivity and negativity ( P>0. 05). Conclusions: Detection of TAM in liver, breast, esophageal, gastric, colorectal, and pancreatic cancer patients demonstrates a good accordance with CEA, CA199, CA153, and AFP, thus suggesting that further study is warranted to verify whether TAM could be a surrogate for these conventional biomarkers.

• 대한한방부인과학회지
• /
• 제31권3호
• /
• pp.152-163
• /
• 2018

Objectives: In recent studies, increased AMH level has been suggested as objective surrogate marker for diagnosis PCOS, one of the major causes of oligomenorrhea. The purpose of this study is to report the clinical effect of Korean medicine treatment on oligomenorrhea patient with increased Anti-mullerian hormone level, who can be diagnosed who can be ruled out PCOS. Methods: A 27 year old woman with oligomenorrhea was enrolled in this study. We measured serum hormone levels and ruled out PCOS. The patient received Korean medicine treatment for 3 months, we assessed the result of treatment through observation of the menstrual cycle and follow-up measurements of serum hormone levels. Results: 1. The patient had menstrual cycle regularly. 2. Increased serum AMH level of the patient decreased from 12.16 ng/ml to 8.51 ng/ml. 3. The other serum hormone levels such as testosterone, LH/FSH ratio decreased Conclusion: This case shows that Korean medicine treatment could have a beneficial effect on menstrual cycle and decrease the increased serum hormone levels of ruled out PCOS patient.

• Journal of Breast Cancer
• /
• 제21권4호
• /
• pp.399-405
• /
• 2018

Purpose: Vesicle-associated membrane protein 8 (VAMP8) is a soluble N-ethylmaleimide-sensitive factor receptor protein that participates in autophagy by directly regulating autophagosome membrane fusion and has been reported to be involved in tumor progression. Nevertheless, the expression and prognostic value of VAMP8 in breast cancer (BC) remain unknown. This study aimed to evaluate the clinical significance and biological function of VAMP8 in BC. Methods: A total of 112 BC samples and 30 normal mammary gland samples were collected. The expression of VAMP8 was assessed in both BC tissues and normal mammary gland tissues via a two-step immunohistochemical detection method. Results: The expression of VAMP8 in BC tissues was significantly higher than that in normal breast tissues. Furthermore, increased VAMP8 expression was significantly correlated with tumor size (p=0.007), lymph node metastasis (p=0.024) and recurrence (p=0.001). Patients with high VAMP8 expression had significantly lower cumulative recurrence-free survival and overall survival (p<0.001 for both) than patients with low VAMP8 expression. In multivariate logistic regression and Cox regression analyses, lymph node metastasis and VAMP8 expression were independent prognostic factors for BC. Conclusion: VAMP8 is significantly upregulated in human BC tissues and can thus be a practical and potentially effective surrogate marker for survival in BC patients.

• Journal of Preventive Medicine and Public Health
• /
• 제33권3호
• /
• pp.306-312
• /
• 2000

Background : Polycyclic aromatic hydrocarbons (PAH) are well known environmental pollutants. The measurement of PAH in ambient air is not commonly used, because it is quite difficult to perform and is unreliable. Using biomarkers of PAH can be an alternative approach to this problem. The PAH in ambient air is absorbed in particulate matter. Total suspended particulate(TSP) or particulate matter of less than $10{\mu}m$ in diameter (PM10) can be easily measured. Therefore, TSP or PM10 can be used as a surrogate measurements of ambient air PAH. Objectives : We investigated whether the urinary concentration of two biomarkers of PAH, 1-hydroxypyrene (1-OHP) and 2-naphthol, could reflect the total suspended particulate in the general population. Methods : In order to exclude the effects of occupational exposure and smoking, first grade middle school students were included in this study. Four middle schools within a one kilometer boundary of ambient air monitoring stations were selected. Total suspended particulate was regarded as the marker of airborne PAH. Diet and smoking data were collected by self administered questionnaires, and spot urine samples were collected. Urinary 1-OHP and 2-naphthol were analyzed by high performance liquid chromatography. Results : The correlation between urinary 1-OHP, 2-naphthol and passive smoking was not statistically significant. The correlation between urinary 1-OHP and TSP indices was not statistically significant. The correlations between urinary 2-naphthol and TSP of two lag days, one lag day, and zero lag days were statistically significant. The statistical significance of two lag days was the strongest (p=0.001), one lag day was the next (p=0.0275), and zero lag days was the weakest (p=0.0349). Conclusion : Our results imply that the urinary concentration of 2-naphthol can be applied as a PAH exposure marker for the general population with low PAH exposure.

• Tuberculosis and Respiratory Diseases
• /
• 제66권6호
• /
• pp.467-470
• /
• 2009

CA 125의 상승은 난소의 악성종양에 나타날 수 있지만 비특이적이다. CA 125의 상승은 다양한 양성 질환 혹은 악성 종양에서 나타날 수 있다. 저자들은 최근 지속적으로 혈중 CA 125 수치가 증가되어 있지만 경질 초음파, 흉부 CT, PET-CT 등에서 악성 종양이 발견되지 않은 기관지확장증 1예를 경험하였다. 14개월의 추적 관찰 기간 중에도 난소암과 같은 악성 종양은 발견되지 않고 기관지확장증에 병발된 폐렴 이외에는 특이 소견이 없어, 혈중 CA 125의 상승은 기관지확장증에 의한 급성 폐 손상에 의한 것으로 추정된다. 혈중 CA 125 수치의 상승은 환자의 임상 양상과 연관하여 조심스럽게 판단하여야 될 것으로 생각된다.

• Tuberculosis and Respiratory Diseases
• /
• 제71권6호
• /
• pp.417-424
• /
• 2011

Background: Recent evidences have revealed metabolic functions of p53 in cancer cells; adaptation or survival to metabolic stress and metabolic shift toward oxidative phosphorylation. However, further studies in clinical setting are needed. We investigated whether p53 protein expression, as a surrogate marker for loss of p53 function, is associated with metabolic features of stage I non-small cell lung cancer (NSCLC), focusing on tumor necrosis and maximal standardized uptake value (SUVmax) on $^{18}F$-fluorodeoxyglucose positron emission tomography. Methods: Clinical information was obtained from retrospective review of medical records. p53 expression was assessed by immunohistochemical staining. Results: p53 protein expression was detected in 112 (46%) of 241 NSCLC cases included in this study. p53 expression was independently associated with the presence of necrosis (odds ratio [OR], 2.316; 95% confidence interval [CI], 1.215~4.416; p=0.011). Non-adenocarcinoma histology (OR, 8.049; 95% CI, 4.072~15.911; p<0.001) and poorly differentiation (OR, 6.474; 95% CI, 2.998~13.979; p<0.001) were also independently associated with the presence of necrosis. However, p53 expression was not a significant factor for SUVmax. Conclusion: p53 protein expression is independently associated with the presence of necrosis, but not SUVmax.

• 한국콘텐츠학회논문지
• /
• 제10권7호
• /
• pp.268-275
• /
• 2010

관상동맥 석회수치(CACS)는 조직학적으로 동맥경화반의 총량과 높은 상관관계를 보이며, 관상동맥의 협착을 예측하는 지표와 향후 허혈성 심장질환 발생의 독립적인 위험인자가 된다. 본 연구는 2006년 1월부터 2008년 12월까지 화순전남대학교병원 건강증진센터에서 건강검진을 목적으로 CACS검사를 시행한 1042명을 대상자로 심혈관질환 위험요소 및 관상동맥 석회수치와 Framingham Risk Scores(FRS)와의 상관관계를 알아보고자 하였다. CACS와 FRS는 남성(OR, 2.38; 95% CI, 1.83-3.11), 여성(OR, 2.12; 95% CI, 1.03-4.35) 모두 관상동맥 석회수치가 없는 군과 비교하여 통계적으로 유의하게 높았으며, CACS와 심혈관질환 위험요인과의 관계에서는 연령과 성별을 통제한 다변량분석에서 여자는 연령(OR, 1.10; 95% CI, 1.06-1.15), 고밀도지단백콜레스테롤(OR, 2.38; 95% CI, 1.04-5.44), 공복혈당(OR, 2.89; 95% CI, 1.16-7.21)이 남자에서는 연령(OR, 1.11; 95% CI, 1.08-1.14), 저밀도지단백콜레스테롤(OR, 2.12; 95% CI, 1.28-3.50), 감마-글루타밀전이효소(OR, 1.73; 95% CI, 1.17-2.55), 당뇨병(OR, 3.92; 95% CI, 1.73-8.89)이 관상동맥 석회수치의 유의한 위험인자로 나타났다.