• Toxicological Research
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• v.32 no.1
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• pp.81-87
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• 2016

Aflatoxin B1 (AFB1) is produced by Aspergillus flavus growing in feedstuffs. Early detection of maize contamination by aflatoxigenic fungi is advantageous since aflatoxins exert adverse health effects. In this study, we report the development of an optimized conventional PCR for AFB1 detection and a rapid, sensitive and simple screening Real-time PCR (qPCR) with SYBR Green and two pairs of primers targeting the aflR genes which involved aflatoxin biosynthesis. AFB1 contaminated maize samples were divided into three groups by the toxin concentration. Genomic DNA was extracted from those samples. The target genes for A. flavus were tested by conventional PCR and the PCR products were analyzed by electrophoresis. A conventional PCR was carried out as nested PCR to verify the gene amplicon sizes. PCR-RFLP patterns, obtained with Hinc II and Pvu II enzyme analysis showed the differences to distinguish aflatoxin-producing fungi. However, they are not quantitative and need a separation of the products on gel and their visualization under UV light. On the other hand, qPCR facilitates the monitoring of the reaction as it progresses. It does not require post-PCR handling, which reduces the risk of cross-contamination and handling errors. It results in a much faster throughout. We found that the optimal primer annealing temperature was $65^{\circ}C$. The optimized template and primer concentration were $1.5{\mu}L\;(50ng/{\mu}L)$ and $3{\mu}L\;(10{\mu}M/{\mu}L)$ respectively. SYBR Green qPCR of four genes demonstrated amplification curves and melting peaks for tub1, afIM, afIR, and afID genes are at $88.0^{\circ}C$, $87.5^{\circ}C$, $83.5^{\circ}C$, and $89.5^{\circ}C$ respectively. Consequently, it was found that the four primers had elevated annealing temperatures, nevertheless it is desirable since it enhances the DNA binding specificity of the dye. New qPCR protocol could be employed for the determination of aflatoxin content in feedstuff samples.

• Tuberculosis and Respiratory Diseases
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• v.38 no.4
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• pp.340-346
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• 1991

Tuberculous tracheobronchitis is defined as a specific inflammation of the trachea or major bronchi caused by the tubercle bacillus and recognized as one of the most common and serious complication of pulmonary tuberculosis. It had been a diagnostic challenge in prebronchoscopic era and since 1968, fiberoptic bronchoscopy has been accepted as a safe and valuable diagnostic procedure of tuberculous tracheobronchitis. Now, it remains a troublesome therapeutic problem due to its sequelae such as bronchostenosis, bronchiectasis and bronchial deformity. The authors analyzed the clinical features, radiological findings and bronchoscopic findings with pathologic and bacteriologic study on 61 cases of tuberculous tracheobronchitis and following results were obtained. 1) The peak incidence was in the fourth decade and male to female ratio was 1:3.4. 2) The most common symptom was cough (86.9%) and followed by sputum (49.2%), dyspnea (27.9%), fever (19.8%), weight loss (11.5%), hemoptysis (6.6%), hoarseness (6.6%) and chest discomfort (3.3%) and localized wheezing was heard in 18%. 3) In chest X-ray, consolidation with collapse was observed in 70.5%, and followed by consolidation only (18.0%), mediastinal node enlargement (8.2%), cavitary lesion (6.6%), suspicious hilar mass (3.3%) and miliary lesion (1.6%) and there was no abnormal findings in 4.9%. 4) Bronchoscopy showed hyperplastic lesion in 67.2%, mucosal lesion (18.0%), ulcerative lesion (9.8%) and stenotic lesion (4.9%). The most common site of bronchial lesion was right upper bronchus (36.1%) and followed by right main bronchus (34.4%), left main bronchus (29.5%), left upper bronchus (16.4%), right middle bronchus (8.2%), right lower bronchus (6.6%) and left lower bronchus (3.3%). 5) Chronic granulomatous inflammation with or without caseation necrosis on microscopic examination was confirmed in 69.7%, bronchial washing AFB stain was positive in 34.1%, prebronchoscopic sputum AFB stain was positive in 88.1% and postbronchoscopic sputum AFB stain was positive in 30.1%.

• Journal of Yeungnam Medical Science
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• v.2 no.1
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• pp.161-166
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• 1985

Endobronchial tuberculosis is frequently associated with lymphadenopathy, bronchial stenosis, atelectasis which needs to be differentiated with other pulmonary disease. Authors are presenting 25 cases of endobronchial tuberculosis which were confirmed by bronchoscopic biopsy and AFB smear and/or culture. The study was done on patients who visited Yeungnam University Hospital from May 1983 to July 1985. The following results were obtained: 1. Of the 25 cases, 8 cases were male and 17 cases were female (male : female= 1 : 2). 32 percent of the cases was older than 60 years of age. 2. Abnormal findings on chest x-ray were seen on 22 cases of 25 cases (88%) and its most predilective site was right upper lobe.(24%). 3. Symptoms were coughing (56%), hemoptysis, hoarseness, chest pain, dyspnea and fever in orders. 4. The positive results were obtained in 73 percent of sputum AFB sme ar (11 cases of 15 cases), 60 percent of AFB culture (3 cases of 5 cases) and 58 percent of bronchoscopic biopsy (14 cases of 24 cases). 5. Complete pulmonary function test was done on 19 cases and showed normal result in 6 cases, restrictive pattern in 8 cases and small airway dysfunction in 5 cases. No case of obstructive airway disease was detected. 6. Associated disease were hypertension (2 cases), tuberculous meningitis (1 case), diabetes mellitus (1 case), and cataract (1 case). 7. The site of bronchoscopic lesion on bronchoscopic examination were as follows; 8. All 25 patients were treated with anti-tuberculosis theyapy and 6 patients were treated with additional steroid therapy because of severe respiratory symptom and showed dramatic improvement of symptom as well as lung function.

• Preventive Nutrition and Food Science
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• v.3 no.2
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• pp.169-174
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• 1998

Antimutagenic and anticarcinogenic effects of alginic acids extracted from sea mustard(SM) and sporophyII of sea mustard(SSM) were studied by Salmonella typhimurium assay system and cytotoxicity and transformation tests using C3H/10T1/2 cells, respectively. alginic acid-SM andalginic acid-SSM showed antimutagenic effects on aflatoxin B1(AFB1)and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in Salmonella typhimurium TA100 strain. The antimutagenic effect showed concentration dependent manner. At the 2.5mg/plate concentration , alginic acid-SSM exhibited 92% antimutagenicity against AFB1 ,while alginic acid-SM revealed 54% antimutagenictity ,s howing effectiveness of the alginic acid-SSM for the antimutagenicity. Alginic acidSSm also significantly decreased the cytotoxicity induced by 3-methylcholanthrene(MCA) and MNNG in C3H/10T1/2 cells (p<0.05). The type II and type IIItransformation foci formation by MCA and MNNG were also decreased when the alginic acid-SSM was treated, indicating that the alginic acid -SSM reduces the carcinogenesis induced by these carcinogens. The MCA-treated culture produced 10.5foci of type II +III in C3H/10T1/2 cells, however, MCA + 0.2mg/ml alginic acid-SSM treated culture formed only 1.8 foci of the types II + III in C3H/10T1/2 cells, however , MCA+0.2mg/ml alginic acid -SSM treated culture formed only 1.8 foci of the types II+ III(p<0.05). While MNNG-treated culture formed 13.0 foci, MNNG + 0.2mg/ml alginic acid -SSM treated one produced 3.0 foci of type II+III(p<0.05). These results suggest that alginic acid-SSM can effectively prevent the mutagenicities and also decrease cytotoxicity and transformation induced by some carcinogens.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.424-434
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• 2011

The increase in the consumption of herb medicines have made their use a public health problem due to the potential fungal contamination and the risk of the presence of my cot ox ins. 360 samples of herb medicines were evaluated for the aflatoxin contamination. The natural occurrence of aflatoxins in these samples were determined using immunoaffinity column clean up and high performance liquid chromatography (HPLC) with post-column derivatization. For samples analyzed, mean levels (incidence) of AFB1, AFB2, AFG1 and AFG2 in positive samples were $1.4\;{\mu}g/kg$(46.4%), $0.4\;{\mu}g/kg$(25.4%), $1.1\;{\mu}g/kg$(37.8%) and $0.9\;{\mu}g/kg$(24.3%), respectively. Recoveries of the full analytical procedure were 71.7~99.7% for AFB1, 88.1~99.2% for AFB2, 82.8~95.5% for AFG1 and 77.9~90.0% for AFG2. The excess cancer risk estimated using the cancer potency of aflatoxin B1 (7 $(mg/kg/day)^{-1}$ for $HBsAg^-$ and 230 $(mg/kg/day)^{-1}$ for $HBsAg^+$) were $1.30{\times}10^{-5}{\sim}1.22{\times}10^{-7}$ for hepatits B surface antigen negative ($HBsAg^-$) and $3.31{\times}10^{-4}{\sim}3.12{\times}10^{-6}$ for hepatits B surface antigen positive ($HBsAg^+$) respectively. In conclusion, although the contamination levels of samples used in the study were low, further actions are also required to undertake a program of herbal surveys in order to access mycotoxin contamination overall so that the safety of public will be protected.

• Microbiology and Biotechnology Letters
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• v.21 no.6
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• pp.628-634
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• 1993

The inhibitory effects of various extracts from Chinese pepper on the mutagenicity and the growth of MG-63 human osteosarcoma cells were studied. Chinese pepper was extracted with methanol and then the methanol extract was further fractionated by using hexane, chloroform, ethyl acetate and butanol. The methanol extract of Chinese pepper revealed the strong antimutagenic activity on the aflatoxin B1(AFB1) and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in Ames mutagenicity test and SOS chromotest.

• Investigative Magnetic Resonance Imaging
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• v.26 no.1
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• pp.71-75
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• 2022

Tuberculous epididymo-orchitis, a rare form of extrapulmonary tuberculosis, results from hematogenous dissemination or retrograde extension from the lower urinary tract. Herein, we studied the case of a 22-year-old male patient who presented with refractory left scrotal pain and inflammation. The patient also complained of multifocal pain involving the right buttock, posterior thigh, and right wrist, without trauma history. The patient was diagnosed with multifocal tuberculosis by sputum AFB study and right sacroiliac joint biopsy.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.2
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• pp.156-161
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• 1991

The mutagenic, comutagenic and antimutagenic effects of red pepper powder were studied by using Ames mutagenicity test. extracts(3 fractions) of the red pepper powder did not show any mutagenicity with or without S9 mix in Salmonella typhimurium strains of TA100 and TA98. These extracts did not show any comutagenicity on N-methyl-N'-nitro-N-nitrosoguanidine(MNNG). Capsaicin also did not exhibit any mutagenicity in the absence or presence of S9 mix prepared from rat or hamster livers. However, the red pepper powder showed antimutagenicity aganist aflatoxin $B_1(AFB_1)$ mediatdd mutagenicity. Especially first fraction of the pepper powder inhibited strongly the mutagenicity of $AFB_1$. There was no difference of these activities between hotter tasted pepper powder and plain hot tasted pepper powder.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.7
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• pp.3499-3503
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• 2016

Our recent studies conducted in South America have shown that mycotoxin contamination of red chili peppers (RCPs) may be associated with an increased risk of gallbladder cancer (GBC). Whether this relationship exists in India, a country with a high incidence of GBC and high consumption of RCPs, is unclear. We therefore measured concentrations of aflatoxins (AFs) and ochratoxin A (OTA) in RCPs from areas of low, medium, and high incidence of GBC in India, and compared these concentrations with GBC incidence in each area. Twenty-one RCP samples were collected from nine cities (eight from a low-incidence area, five from a medium-incidence area, and eight from a high-incidence area). Concentrations of AFs and OTA were measured using high-performance liquid chromatography. No significant differences in mean concentrations of AFs and OTA were found in the three areas. AFB1 levels in the low-incidence area ($10.81{\mu}g/kg$) and high-incidence area ($12.00{\mu}g/kg$) were more than 2.2 and 2.4 times higher compared with the maximum permitted level of AFB1 in spices ($5.0{\mu}g/kg$) set by the Commission of the European Communities, or that ($4.4{\mu}g/kg$) obtained in our previous study in Chile. Our results show that the mean concentrations of mycotoxins in RCPs are similar among the three areas in India with different incidences of GBC. Further studies with human subjects are needed to evaluate any association between AFB1 and GBC.

• Tuberculosis and Respiratory Diseases
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• v.72 no.1
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• pp.55-58
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• 2012

Mycobacterium szulgai is a rare nontuberculous mycobacterium found in Korea. It is an opportunistic pathogen and is usually isolated from patients with a history of alcoholism, chronic pulmonary disease, or an immunocompromising condition. We present here a case of M. szulgai isolated from a patient with a history of pulmonary tuberculosis. A 54-year-old man was admitted with dyspnea and febrile sensation. He had a history of pulmonary tuberculosis which occurred 30 years earlier and treatment with anti-tuberculosis medication. His chest computed tomography scan showed cavitary consolidation in both upper lungs. A sputum acid-fast bacilli (AFB) smear was positive and anti-tuberculous medication was started. However, a polymerase chain reaction for mycobacterium tuberculosis was negative and anti-tuberculous medication was stopped. M. szulgai was isolated on 3 separate sputum and bronchial wash fluid AFB cultures. He was treated with clarithromycin, rifampicin, and ethambutol. After 1 month, a sputum AFB smear and culture became negative and no additional M. szulgai were isolated during a 16-month treatment.