• Food Science and Biotechnology
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• v.18 no.3
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• pp.641-648
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• 2009

A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.12
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• pp.7675-7679
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• 2013

Background: A missense mutation in exon 7 (R249S) of the p53 tumor suppressor gene is characteristic of aflatoxin B1 (AFB1) exposure. AFB1 is believed to have a synergistic effect on hepatitis virus B (HBV) carcinogenesis. However, results of studies comparing R249S prevalence among patients are conflicting. The aim of this study was to determine the prevalence of the R249S mutation in hepatocellular carcinoma (HCC) patients with or without positive HBsAg. Materials and Methods: Paraffin embedded liver tissues were obtained from 124 HCC patients who underwent liver resection and liver biopsy in King Chulalongkorn Memorial Hospital. Restriction fragment length polymorphism (RFLP) was utilized to detect the R249S mutation. Positive results were confirmed by direct sequencing. Results: Sixty four (52%) patients were positive for HBsAg and 18 (15%) were anti-HCV positive. 12 specimens tested positive by RFLP. Ten HCC patients (8.1%) were confirmed to be R249S positive by Sanger sequencing (AGG to AGT). Out of these 10, six were HBsAg positive, and out of the remaining 4, two were anti-HCV positive. The R249S prevalence among HCC patients with positive HBsAg was 9.4% compared to 6.7% for HBsAg negative samples. Patients with the R249S mutation were younger ($55{\pm}10$ vs $60{\pm}13$ year-old) and tended to have a more advanced Edmonson-Steiner grade of HCC, although differences did not reach statistical significance. Conclusions: Our study shows moderate prevalence of aflatoxin B1-related p53 mutation (R249S) in HCC with or without HBsAg. HBsAg positive status was not associated with R249S prevalence.

• Current Research on Agriculture and Life Sciences
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• v.2
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• pp.68-76
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• 1984

A $2{\times}4$ factorial study was carried out to investigate the interaction of aflatoxin and vitamin $D_3$ in broiler chicks. The day-old 336 chicks were allocated to triplicate 8 treatments. The 0 or 0.5 ppm of aflatoxin $B_1$ (AFB) and 0, 500, 1,000 or 1,500 IU/Kg of vitamin $D_3$ (VD) were supplemented to the basal diet. There were no significant differences among treatments in respect to the body weight gain, feed intake, feed conversion, shank color, mortality and incidence of weak legs. The utilization efficiencies of dry matter, crude protein, ether extract, N-free extract and crude ash showed also no significant differences among treatments, respectively. The mean utilization efficiency of crude fiber in AFB group was lower than that in normal groups (P<.01). However, no significant difference was found among groups fed different levels of VD, and no interaction between AFB and VD was found. The utilization efficiency of Ca in AFB group was somewhat higher than that in normal group without statistical significance, and the similar values were found among groups fed different VD. The utilization efficiencies of P and Na were not significantly different among treatments, respectively. The tibia ash appeared to be similar among treatments fed different levels of AFB and VD. However, the Ca content in tibia of birds fed 0.5 ppm of AFB was higher than that of normal chicks (P<.05). The slightly increasing trend was shown in Ca contents when fed increasing revel of VD, and the interaction between AFB and VD was recognized(P<.01). The P content of tibia was increased by feeding AFB(P<.05). However, there was no significant difference among groups fed different level of VD and no interaction between AFB and VD in respect to the P content of tibia. Feeding AFB did not affect the Na content in tibia. However, there was a highly significant difference among groups fed different levels of VD(P<.01), the highest values were at 1,000 IU/Kg group, and the interaction between AFB and VD was not significant. The Ca content in serum of birds fed AFB was higher than in control group (P<.01). The Ca of serum increased when fed more VD, although no significance was found among groups, and there was an interaction between AFB and VD(P<.05). The P content of serum showed no significant difference among treatments. The alkaline phosphatase activity in serum of chicks fed AFB was higher than that of control group (P<.01). The enzyme activity increased slightly with increasing level of VD, however, there was no interaction between AFB and VD.

• Korean Journal of Poultry Science
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• v.21 no.2
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• pp.113-117
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• 1994

Of the 200, 000 known species of molds, only 50 or 60 are known to be harmful to humans or livestock. Certain fungi that grow on grains and grasses can produce chemical substances called mycotoxins that adversely affect performance in poultry. There are several methods of preserving feed ingredients. The list includes: drying, antioxidants, mold inhibitors, organic acids, phosphates, cooking or toasting, fat extraction, blending and fermenting. Mold inhibitors are manufactured to inhibit mold growth and prevent the production of toxic substances. They are fungistats and not fungicides, that is, they only stop the growth of molds. Practical and cost-effective methods to detoxify mycotoxin containing feedstuffs are in great demand. 0.5% hydrated sodium calcium aluminosilicate added to the diet protected chicks from the deleterious effects of aflatoxin-contaminated feed. The dietary addition of antioxidants and methionine also significantly diminished the negative effects on body weight in chicks toxicated with 3.0 ppm aflatoxin B1.

• Journal of Environmental Health Sciences
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• v.32 no.5 s.92
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• pp.515-521
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• 2006

아플라톡신(aflatoxin)은 곰팡이의 2차 대사산물이며 인간발암원으로 보건상 위해를 준다. 쌀은 아플라톡신을 생성하는 곰팡이인 Aspergillus flavus와 A. parasitidcus에 좋은 기질이다. 이 연구에서는 쌀과 쌀제품 및 쌀부산물에서 전세계적으로 나타나고 있는 아플라톡신 오염 정도와 오염 수준에 대하여 고찰하였다. 아플라톡신 오염 정도와 오염 수준은 시료의 유형과 시료채취 지역에 따라 다르게 나타나는 것으로 보인다. 쌀과 쌀제품 2,511 시료 중 36.6%에서, 그리고 쌀부산물 374 시료 중 57.8%에서 아플라톡신이 검출되었다. 이들 시료에서 aflatoxin $B_{1}$ 및 total aflatoxin오염도는 $0-185{\mu}g/kg$였다. 일부 백미 시료에서는 $20{\mu}g/kg$을 초과하는 시료가 있었다. 쌀의 아플라톡신 관련 위생관리지표를 고안할 수 있는 기초자료를 제공하기 위하여 쌀의 아플라톡신 오염에 대한 모니터링이 필요하다. 또 쌀과 쌀제품 생산의 모든 단계와 저장에서 아플라톡신 관리를 위한 통합적 접근법이 필요하다. 가축을 아플라톡신으로부터 보호하는 전략은 인간집단을 보호할 수 있는 효과적인 접근법이 될 수 있다.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.873-886
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• 1996

For better understanding the interrelationship of hemorrhage and aggregation mechanism, cyclopiazonic acid(CPA) known as promoting the aggregation of platelet, aflatoxin $B_1(AFB_1)$ inhibiting platelet aggregation were used as toxic mycotoxins in these studies. In order to investigate the potential role of prostaglandin metabolism on the platelet aggregation, a variety of prostaglandin metabolites such as $PGF_{2{\alpha}}$, $PGE_2$ and $TXB_2$ were measured in homogenized rabbit platelets by TLC and LSC. And the role of $Ca^{{+}{+}}$ on the platelet aggregation was investigated by flow cytometer. Finally, the morphological effects of mycotoxins on platelet were determined by transmission electron microscope. The results and conclusions obtained from these studies are: 1) CPA induced no changes but $AFB_1$ increased $PGE_2$ and $TXB_2$. 2) CPA promoted ADP, collagen, thrombin, A.A., and PAF-induced $Ca^{{+}{+}}$ release. $AFB_1$, however, decreased $Ca^{{+}{+}}$ level except collagen-induced $Ca^{{+}{+}}$ release. When the calcium blocker, verapamil, was used, CPA decreased thrombin-induced $Ca^{{+}{+}}$ release and increased collagen, ADP, PAF and A.A.-induced $Ca^{{+}{+}}$ release. $AFB_1$ in contrast decreased the all factors induced $Ca^{{+}{+}}$ release. 3) $AFB_1$ did not induce any ultrastructural changes except large vacuole formation in a few platelets. And CPA also did not induce any changes except moderate shape change, indicator of platelet activation. In conclusion, CPA promoted platelet aggregation by the increases of $Ca^{{+}{+}}$ release but had no changes in A.A. metabolites. Antiaggregating effects of $AFB_1$ may be due to decreases of $Ca^{{+}{+}}$ release and increases of $PGE_2$ and $PGF_{2{\alpha}}$ formation. These data provide the basis for the future study of mobilization and function of $Ca^{{+}{+}}$ in platelet aggregation.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.623-630
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• 2008

A non-instrumental immunochromatographic (ICG) strip-test and direct competitive enzyme-linked immunosorbent assay (DC-ELISA) for aflatoxin B1 (AFB1) determination were developed and optimized. The detection limits of ICG strip-test and DC-ELISA were 0.5 and 0.004 ng/mL, respectively, and these methods possessed a cross-reaction to aflatoxins. The results of spiked samples by both methods were coincided with the amount spiked AFB1 and the comparative analyses of 172 real samples by 2 immunoassays and high performance liquid chromatography (HPLC) showed a good agreement. Especially, the ICG strip-test is easier to perform and quicker, but less sensitivity than DC-ELISA. Both methods could analyze a high sample throughput with short time, but the sample throughput of ICG strip-test was better. Therefore, the ICG strip-test can be used as a simple, easy, non-instrumental, and fast screening technique for AFB1 determination.

• Food Science and Preservation
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• v.18 no.3
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• pp.374-382
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• 2011

Aflatoxin ($AFB_1$) is a potent hepatotoxic and hepatocarcinogenic mycotoxin in humans. It is also well-known to be accumulated in animal tissues via various metabolic pathways. This study was conducted to determine the effects of vitamin C on the residual $AFB_1$ in rat sera that were treated with radiation and $AFB_1$. Six week-old male Sprague-Dawley rats were randomly divided into five groups: a control group, $AFB_1$-treated group, the group treated with $AFB_1$ and vitamin C, the group treated with X-ray and AFB1, and the group treated with X-ray and $AFB_1$ with vitamin C. On the first day of the experiment, only one dose of X-rays was exposed to the entire liver at 1,500 cGy. Next, vitamin C was injected at 10 mg/kg body weight via intraperitoneal injection, followed 1 hr later by the administration of 0.4 mg/kg of $AFB_1$ via intraperitoneal injection. These treatments were then administered every three days over a period of 15 days. On the 16th day of treatments, the animals were sacrificed. The contents of $AFB_1$ in rat sera were determined via indirect competitive ELISA and HPLC method. In the quantitative analysis of $AFB_1$ in rat sera via ELISA, $5.17{\pm}0.34$ ng/mL of $AFB_1$ was detected in the $AFB_1$-treated groups, but the amount more significantly decreased to $3.23{\pm}0.76$ ng/mL in the groups treated with $AFB_1$ and vitamin C (p<0.01) than in the $AFB_1$-treated groups. The $AFB_1$ contents of the rat sera of the groups treated with X-ray and $AFB_1$ did not significantly decreased with the administration of vitamin C. The $AFB_1$ content of the rat sera that was analyzed via HPLC showed a tendency similar to that of the content that was analyzed via ELISA. With regard to these data, vitamin C was very effective in reducing $AFB_1$ residue in rat sera.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.172.2-173
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• 2003

3-Alkoxy-6-allylthiopyridazine derivatives showed the strongest protective effect against oxidative stress and their anticancer effect determined on the growth of SK-Hep-l hepatocellular carcinomar cells. The allythio group as a pharmacologically active group was introduced into pyridazine nucleus and a substituent such as halogen or alkoxy was also introduced into paraposition of allylthio group. Five kinds of 3-alkoxy-6-allylthiopyridazine derivatives were synthesized and their chemoprotective activities examined in rats exposed to aflatoxin $B_1$-toxicant. (omitted)

• Toxicological Research
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• v.3 no.1
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• pp.1-8
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• 1987

In vitro induction of cytochrome 450 associated monooxygenase activities by phenobarbital (PB) and 3-methylcholanthrene (MC) was investigated in primary cultures of adult rat hepatocytes. PB and MC were added to the culture 24 hr after the initial plating of hepatocytes. A signiftcant increase of the activities of 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase were observed in MC and PB treated culture. MC caused about 500% induction of the initial oxidation rates of both enzymes in 48 hr. However the PB maintained both enzyme activities close to the level of freshly isolated hepatocytes. Biphenyl 4-hydroxylase and aminopyrine N-demethylase activities were also induced by MC and PB. But the level of induction was less than that occuring with 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase. When aflatoxin $B_1$ was added to the hepatocyte cultures which have been treated with MC or PB, it caused a significant increase of the unscheduled DNA synthesis at higher dose of aflatoxin $B_1$ as compared to those of untreated control hepatocyte cultures. The results suggest that microsomal enzyme activities can be selectively controlled preferably in hepatocyte cultures by the in vitro induction method. This principle may be useful for studying the metabolism and other toxicological studies.