Micetismos: Parte 4: Síndromes tempranos con síntomas complejos / Mushroom poisonings. Part 4: early-onset syndromes with complex symptoms / Intoxicações por cogumelos. Parte 4: Síndromes precoces com sintomas complexos

En esta Parte 4 de la serie de cuatro artículos sobre micetismos se analizan los síndromes que se caracterizan por presentar un período de latencia muy corto, con la aparición de síntomas complejos en menos de 6 horas después de la ingestión de los macromicetos. Se discuten los siguientes micetismos: 1) Toxíndrome muscarínico o colinérgico periférico por especies de Inocybe y Clitocybe. 2) Toxíndrome inmunohemolítico o hemolítico por Paxillus. 3) Toxíndrome neumónico alérgico por Lycoperdon perlatum y por Pholiota nameko. 4) Toxíndrome panterínico o neurotóxico glutaminérgico por compuestos isoxazólicos o síndrome pantherina/muscaria. 5) Toxíndrome coprínico o cardiovascular. 6) Toxíndrome neurotóxico alucinogénico por psilocibina y derivados indólicos. 7) Toxíndrome psicotrópico por estirilpironas y gimnopilinas de Gymnopilus spectabilis o G. junonius. 8) Toxíndrome agudo de rabdomiólisis por Russula subnigricans. 9) Toxíndrome cianogénico por Marasmius oreades. 10) Toxíndrome inmunosupresor por tricotecenos macrocíclicos de Podostroma cornu-damae. 11) Toxíndrome hemolítico debido a ostreolisina de Pleurotus ostreatus y especies relacionadas. Se analizan los síntomas, las toxinas involucradas, los mecanismos de acción, cuando se conocen, y las especies causantes de los micetismos.

This Part 4 of the series of four articles on mushroom poisonings refers to early-onset syndromes, which are characterized by a very short latency period, and the appearance of complex symptoms in less than 6 hours after mushroom ingestion. The following mycetisms are discussed, (1) Peripheral cholinergic, or muscarinic syndrome due to Inocybe and Clitocybe species. (2) Immunohaemolytic or haemolytic syndrome by Paxillus. (3) Allergic pneumonic syndrome due to Lycoperdon perlatum, and Pholiota nameko. (4) Glutaminergic neurotoxic, or pantherinic syndrome by isoxazole compounds or pantherina/muscaria syndrome. (5) Coprinic or cardiovascular syndrome. (6) Hallucinogenic neurotoxic syndrome due to psilocybin and indole derivatives. (7) Psychotropic syndrome by styrylpirones and gymnopilins of Gymnopilus spectabilis or G. junonius. (8) Rhabdomyolysis acute syndrome due to Russula subnigricans. (9) Cyanogenic syndrome by Marasmius oreades. (10) Immunosuppressive syndrome by macrocyclic trichothecenes of Podostroma cornu-damae. (11) Haemolytic syndrome due to ostreolisine of Pleurotus ostreatus, and related species. The symptoms, toxins involved, mechanisms of action, when known, and the species of mushrooms responsible for the mycetisms are analyzed.

Nesta parte 4 da série de quatro artigos sobre intoxicação por cogumelos são analisadas síndromes que se caracterizam por apresentar um período de latência muito breve, com aparecimento de sintomas complexos em menos de 6 horas após a ingestão dos macromicetos. As seguintes intoxicações com cogumelos são discutidas: (1) Toxíndrome muscarínico ou colinérgico periférico por espécies de Inocybe e Clitocybe. (2) Toxíndrome imuno-hemolítica ou hemolítica por Paxillus. (3) Toxíndrome pneumônica alérgica por Lycoperdon perlatum e por Pholiota nameko. (4) Toxíndrome panterínica ou neurotóxica glutaminérgica por compostos isoxazólicos ou síndrome pantherina/muscaria. (5) Toxíndrome coprínica ou cardiovascular (6) Toxíndrome neurotóxico-alucinogênica por psilocibina e derivados indólicos. (7) Toxíndrome psicotrópica por estirilpironas e gimnopilinas de Gymnopilus spectabilis ou G. junonius. (8) Toxíndrome aguda de rabdomiólise por Russula subnigricans. (9) Toxíndrome cianogênica por Marasmius oreades. (10) Toxíndrome imunossupressora por tricotecenos macrocíclicos de Podostroma cornu-damae. (11) Síndrome hemolítica por ostreolisina de Pleurotus ostreatus e espécies relacionadas. São analisados os sintomas, as toxinas envolvidas, os mecanismos de ação, quando conhecidos, e as espécies de cogumelos responsáveis pelas intoxicações.

Endovascular Therapy: The Second Round Begins / 대한뇌졸중학회지

No abstract available.

Evaluation of the supplementation of a feed additive as a potential protector against the adverse effects of 2. 5 ppm T-2 toxin on growing broiler chickens / Avaliação da suplementação de um aditivo alimentar como um protetor potencial contra os efeitos adversos de 2, 5ppm de toxina T-2 em frangos em crescimento

A trial was conducted to evaluate a feed additive containing epoxidase activity from a bacterium (Mycofix-S) as a potential protection against the adverse effects of 2.5 ppm dietary T-2 toxin in male growing broiler chickens. A total of 144 one-day-old Ross 308 male chicks were individually wing-banded and allotted into each of the four experimental groups. Group 1: negative control, no T-2 toxin or additive; group 2: Mycofix-S, 2.5 g/kg; group 3: positive control, 2.5 ppm T-2 toxin; group 4: 2.5 ppm T-2 toxin + 2.5 g/kg Mycofix-S. Feed and water were provided ad libitum for 28 days (days 1 to 28 of age). Each experimental treatment was replicated 6 times, with 6 birds per replicate pen. Response variables included performance parameters, serum activity of alkaline phosphatase (ALP) and amylase, relative weight of selected organs and histology of the upper digestive system. T-2 toxin at 2.5 ppm significantly (P = 0.016) decreased the 28-day body weight gain and cumulative feed intake without affecting feed conversion. The feed additive counteracted these adverse effects. Serum enzyme activities were not significantly (P>0.05) affected for the four experimental groups but when data from the groups receiving T-2 toxin was pooled and compared against the pooled data from groups without the toxin a significant decrease in amylase activity was observed in chickens receiving T-2 toxin. The histological examination of the upper digestive system revealed lesions in mouth, esophagus, proventriculus, gizzard and duodenum in the chickens fed T-2 toxin without the additive. Chickens fed T-2 toxin plus the additive showed lesions in the same tissues except in the duodenum. The results of the present study show that the addition of 2.5 g/kg of the feed additive tested protects against adverse effects on performance and also the integrity of the duodenal mucosa.(AU)

Foi realizado um experimento com o objetivo de avaliar um aditivo alimentar contendo atividade de epoxidase de uma bactéria (Mycofix-S) como proteção potencial contra os efeitos adversos de uma dieta com 2,5ppm de toxina T-2 em frangos de corte machos. Um total de 144 pintos machos Ross 308 de um dia de idade foram marcados na asa individualmente e alocados em um de quatro grupos experimentais: grupo 1: controle negativo, sem toxina T-2 ou aditivo; grupo 2: 2,5g/kg de Mycofix-S; grupo 3: controle positivo, 2,5ppm de toxina T-2; grupo 4: 2,5ppm de toxina T-2 + 2,5g/kg de Mycofix-S. Alimento e água foram fornecidos ad libitum por 28 dias (dias um a 28 de idade). Cada tratamento experimental foi replicado seis vezes, com seis pintos por gaiola de replicação. As variáveis de resposta incluíram parâmetros de desempenho, atividade sérica de fosfatase alcalina (ALP) e amilase, peso relativo de órgãos selecionados e histologia do sistema digestivo superior. A toxina T-2 a 2,5ppm diminuiu significativamente (P = 0.016) o ganho de peso corporal aos 28 dias e o consumo de alimento acumulado, sem afetar a conversão alimentar. O aditivo diminuiu os efeitos adversos. As atividades séricas das enzimas não foram afetadas significativamente (P>0.05) nos quatro grupos experimentais, porém, quando os dados dos grupos que receberam a toxina T-2 foram combinados e comparados com o pool de dados dos grupos sem toxina, foi observado um decréscimo significativo da atividade de amilase nos frangos que receberam a toxina T-2. O exame histológico do sistema digestivo superior revelou lesões em boca, esôfago, pró-ventrículo, moela e duodeno nos frangos alimentados com toxina T-2 sem aditivo. Frangos alimentados com toxina T-2 mais aditivo mostraram lesões nos mesmos tecidos, exceto no duodeno. Os resultados do presente estudo mostram que a adição de 2,5g/kg do aditivo alimentar testado protege contra os efeitos adversos sobre o desempenho e a integridade da mucosa duodenal.(AU)

Probabilistic assessment of dietary exposure to both deoxynivalenol and zearalenone from cereal-based products in Chinese populations / 中华预防医学杂志

<p><b>OBJECTIVE</b>To elucidate the dietary exposure to deoxynivalenol (DON) and zearalenone (ZEN) from cereal-based products in Chinese populations using the probabilistic assessment approach.</p><p><b>METHODS</b>A total of 292 wheat flours and 347 corn-based products were collected from sampling sites of 107 supermarkets or farmers markets, which were randomly selected from 44 cities of 13 provinces in 2009 by the stratified cluster random sampling method. Then, DON and ZEN contamination levels in these samples above analyzed by UPLC-MS/MS in combination with the food consumption data of 68 959 respondents, who were divided into group 1 aged 3 to 13 years old, and group 2 aged 14 and over 14 years old (≥14 years old), obtained by China National Nutrition and Health Survey in 2002 were investigated. A probabilistic assessment model using Monte Carlo simulation was applied to derive the intake distribution of P(1)-P(99) percentile of dietary exposure to DON and ZEN. Meanwhile, all parameters related to dietary exposure to both toxins were compared with either the provisional maximum tolerable daily intake (PMTDI) of 1 µg·kg(-1)·d(-1) for DON, or the tolerable daily intake (TDI) of 0.25 µg·kg(-1)·d(-1) for ZEN in order to evaluate the risk of dietary intake of two toxins and find the minimum percentile of dietary exposure to these two toxins. The statistical differences of dietary exposure to these two toxins between two groups were achieved by t test.</p><p><b>RESULTS</b>The detection frequencies of DON in wheat flours and corn-based products were 100% (292/292) and 97.4% (338/347), respectively. A total of 21 out of 639 samples (wheat flours: 5/292, corn-based products: 16/347) were positive for DON at the levels exceeding the Chinese regulatory limit of 1 000 µg/kg for DON. And the detection frequencies of ZEN in wheat flours and corn-based products were 53.4% (156/292) and 87.6% (304/347), respectively.54 out of 347 corn-based products and no wheat flours were positive for ZEN at the levels exceeding the Chinese regulatory limit of 60 µg/kg for ZEN. Meanwhile, the mean values (95% CI) of the P(50), P(75), P(90), P(95), P(97.5) and P(99) percentile of dietary exposure to DON in populations of 3 to 13 years old were 0.170 (0.170-0.171), 0.762 (0.759-0.765), 2.066 (2.038-2.069), 3.515 (3.501-3.530), 5.342 (5.314-5.372), and 9.220 (9.155-9.279) µg · kg(-1)·d(-1), which were higher than those in populations of ≥14 years old (0.131 (0.130-0.131), 0.500 (0.498-0.501), 1.280 (1.276-1.285), 2.138 (2.128-2.14), 3.510 (3.494-3.527), and 5.512 (5.474-5.546) µg·kg(-1)·d(-1)), with t values of 87.19, 163.87, 164.66, 157.78, 105.47 and 96.31, and all P values less than 0.001. And the mean values (95% CI) of the P(50), P(75), P(90), P(95), P(97.5) and P(99) percentile of dietary exposure to ZEN in populations of 3 to 13 years old were 0.001 (0.001-0.001), 0.006 (0.006-0.006), 0.039 (0.038-0.039), 0.101 (0.100-0.101), 0.195 (0.194-0.197) and 0.378 (0.374-0.381) µg · kg(-1)·d(-1), which were also higher than those in populations of ≥14 years old (0.001 (0.001-0.001), 0.004 (0.004-0.004), 0.026 (0.026-0.026), 0.061 (0.060-0.061), 0.115 (0.115-0.116) and 0.232 (0.231-0.235) µg·kg(-1)·d(-1)) with T-values of 151.11, 73.80, 96.81, 100.81, 91.93 and 76.13, and all P values less than 0.001. Besides, the minimum percentile of dietary exposure to DON in populations of 3 to 13 years old and ≥14 years old exceeded the corresponding PMTDI of 1 µg·kg(-1)·d(-1) was found in the probability distribution of P(76) (99% percentile = 1.03 µg·kg(-1)·d(-1)) and P(84) (95% percentile = 1.01 µg·kg(-1)·d(-1)) percentile, respectively. And the minimum percentile of dietary exposure to ZEN in populations of 3 to 13 years old and ≥14 years old exceeded the corresponding TDI of 0.25 µg·kg(-1)·d(-1) was found in the probability distribution of P(97) (95% percentile = 0.25 µg·kg(-1)·d(-1)) and P(98) (90% percentile = 0.26 µg·kg(-1)·d(-1)) percentile, respectively.</p><p><b>CONCLUSION</b>The contamination levels of DON and ZEN in wheat flours and corn-based products and the risk of dietary exposure to both DON and ZEN in populations in Chinese populations were at relatively low levels. The dietary exposure to both DON and ZEN in populations of 3 to 13 years old was higher than those in populations of ≥14 years old . Populations of 3 to 13 years old were the populations at the high risk of dietary exposure to both mycotoxins.</p>

Development of a monoclonal antibody against deoxynivalenol for magnetic nanoparticle-based extraction and an enzyme-linked immunosorbent assay

Monoclonal antibody (mAb, NVRQS-DON) against deoxynivalenol (DON) was prepared. DON-Ag coated enzyme linked immunosorbent assay (ELISA) and DON-Ab coated ELISA were prepared by coating the DON-BSA and DON mAb. Quantitative DON calculation ranged from 50 to 4,000 ng/mL for DON-Ab coated ELISA and from 25 to 500 ng/mL for DON-Ag coated ELISA. 50% of inhibitory concentration values of DON, HT-2, 15-acetyl-DON, and nivalenol were 23.44, 22,545, 5,518 and 5,976 ng/mL based on the DON-Ab coated ELISA. Cross-reactivity levels of the mAb to HT-2, 15-acetyl-DON, and nivalenol were 0.1, 0.42, and 0.40%. The intra- and interassay precision coefficient variation (CV) were both <10%. In the mAb-coated ELISA, mean DON recovery rates in animal feed (0 to 1,000 microg/kg) ranged from 68.34 to 95.49% (CV; 4.10 to 13.38%). DON in a buffer solution (250, 500 and 1,000 ng/mL) was isolated using 300 microg of NVRQS-DON and 3 mg of magnetic nanoparticles (MNPs). The mean recovery rates of DON using this mAb-MNP system were 75.2, 96.9, and 88.1% in a buffer solution spiked with DON (250, 500, and 1,000 ng/mL). Conclusively we developed competitive ELISAs for detecting DON in animal feed and created a new tool for DON extraction using mAb-coupled MNPs.

Mushroom Poisoning by Podostroma cornu-damae: A Case Report and Review of the Literature

Podostroma cornu-damae is a rare species of fungus belonging to the Hyocreaceae family. Its fruit body is highly toxic, as it contains trichothecene mycotoxins. Unfortunately, it highly resembles Ganoderma lucidum and Cordyceps, well-known health foods; this can lead to poisoning. We experienced such a case of a 42-year old man who received mushroom poisoning by injesting Podostroma cornu-damae. The patient was presented with severe pancytopenia and infection. The patient recovered without any complications after conservative care, antibiotics therapy, and granulocyte colony stimulating factor administration. The most common complications of podostroma cornu-damae intoxication were reported pancytopenia, infection, disseminated intravascular coagulation, acute renal failure, etc. It is important to provide enough fluid therapy, use of antibiotics to infection and granulocyte colony stimulating factor administration.

Incidence and multiplex PCR based detection of trichothecene chemotypes of Fusarium culmorum isolates collected from freshly harvested Maize kernels in Southern India

Hundred Fusarium culmorum strains, isolated from freshly harvested maize grain samples from Southern parts of India, were incubated in czapek-dox medium and analyzed for trichothecene (DON/NIV) production. The mPCR assay was standardized targeting trichothecene metabolic pathway genes viz., Tri6, Tri7, Tri13 for detection of trichothecene (DON/NIV) chemotypes and rDNA gene for specific detection of F. culmorum species. Primers for targeted genes were designed and used to predict whether these isolates could produce deoxynivalenol/nivalenol, 94 isolates were able to produce DON/NIV by mPCR assay. Chemical analysis of DON/NIV was carried out for mPCR positive isolates by high performance-thin layer chromatography (HPTLC). To check the practical usefulness of developed mPCR assay, 150 field samples of maize were evaluated and results were compared with conventional HPTLC method. Out of 150 samples, 34% samples stayed as a positive for NIV contamination whereas 44% were found to have deoxynivalenol contamination. Moreover, mPCR results are equivocally matched with the HPTLC chemical analysis for field samples. Chemotyping of F. culmorum isolates were reported for the first time from India, and highlights the important potential of F. culmorum to contaminate maize with DON/NIV.

Toxigenic potential of Fusarium graminearum isolated from maize of northwest Argentina

Twenty six isolates of Fusarium graminearum from grains of maize hybrids harvested in ±west Argentina were grown on autoclaved rice grain to assess their ability to produce type B trichothecenes. Chemical analysis indicated that 38% of isolates were nivalenol (NIV) producers only, 31% were major NIV producers with high DON(deoxynivalenol)/NIV ratios, 8% were major DON producers with minor NIV production, and 23% were DON producers only. Isolates showed a high variability in their toxigenic potential which was not related to fungal biomass. The distribution of the different chemotypes as well as the high and the low trichothecene-producing Fusarium isolates could not be associated to a geographical origin. Our results confirmed for the first time that isolates of Fusarium graminearum from maize of northwest Argentina are able to produce DON and NIV. A substancial contamination with both NIV and DON is likely in maize from northwest Argentina. Their contents should be quantified in regional surveillances for mycotoxin contamination.

Effects of barley and barley bran contaminated with Fusarium spp. on the growth and feed efficiency of fattening and growing pigs / 대한수의학회지

The present study was carried out to investigate the effect of barley and barley bran contaminated with Fusarium spp on growth performance and feed efficiency of fattening and growing pigs. In experiment 1, total 48 fattening Landrace pigs were used in a fattening trial for 71 days. Pigs weighing around 75 kg were allocated into different substitution groups containing 0, 10, 20 and 30% of barley contaminated Fusarium spp. In experiment 2, total 16 growing Landrace pigs were used in a growing trial for 45 days. Pigs weighing around 29.4 kg were allocated into different substitution groups containing 0, 5, 10 and 20% of barley bran contaminated Fusarium spp. Mycotoxin concentrations of barley and barley bran contaminated with 30% Fusarium spp were 0.452 and 1.049 ppm for deoxynivalenol, 8.125 and 17.646 ppm for nivalenol and 0.023 and 0.029 ppm for zearalenone, respectively. In experiment 1, no differences were found in weight gain and feed intake between control group (0%) and 10 or 20% substitution groups, but in 30% substitution group, weight gain and feed intake were significantly lower (p < 0.05) than those in control group. After slaughtering, the extended haemorrhage of the fundus region in stomach was observed in 20 or 30% substitution groups. In experiment 2, weight gain and feed intake were not significantly different among treatment groups. After slaughtering of experimental pigs, the extended haemorrhage of the fundus region in stomach was observed in pigs fed diet with 20% substitution group. These results suggest that the feeding of diet with contaminated highly levels of Fusarium spp was negative effect on growth and feed efficiency in growing and fattening pig.

Natural occurrence of masked deoxynivalenol and multi-mycotoxins in cereals from China harvested in 2007 and 2008 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To elucidate the natural occurrence of masked deoxynivalenol (DON-3-G) and other multi-mycotoxins in cereals from parts of China.</p><p><b>METHODS</b>A total of 446 corn and wheat samples harvested in 2007 and 2008 collected from Henan, Hebei, Guangxi, Anhui, Sichuan, Chongqing and Jiangsu provinces were analyzed for DON-3-G and other multi-mycotoxins (including deoxynivalenol (DON), zearalenone (ZEN), nivalenol (NIV), et al) by UPLC-MS/MS.</p><p><b>RESULTS</b>Corn and wheat samples were mainly contaminated by DON and its derivatives as well as ZEN.88% (169/192) of wheat samples were positive for DON (range: 1.5 - 590.7 µg/kg; median: 30.8 µg/kg); 22.9% (44/192) of wheat samples were contaminated with ZEN (range: 1.7 - 3425.0 µg/kg; median: 8.0 µg/kg) and six samples contained ZEN concentration higher than the ZEN tolerance limit of 60 µg/kg. DON was detected in 50.5% (103/204) corn samples (range: 1.6 - 4374.4 µg/kg; median: 94.9 µg/kg); Seven samples contained DON exceeding the tolerance limit of 1000 µg/kg for DON. Additionally, ZEN was found in 41.7% (85/204) corn samples with the concentration between 1.6 µg/kg and 4808.7 µg/kg (median: 48.5 µg/kg) and there were 37 corn samples with ZEN level in the excess of tolerance limit for ZEN (60 µg/kg). DON-3-G was detected in corn and wheat samples for the first time in China with the median level of 21.4 µg/kg and 34.6 µg/kg for wheat and corn, respectively. Wheat was more heavily contaminated with DON-3-G than both 3-acetyl-DON (3-A-DON, median: 4.1 µg/kg) and 15-acetyl-DON (15-A-DON, median: 3.1 µg/kg) (t values were 5.111 and 5.966, respectively, both P values < 0.01). While, the level of 15-A-DON (median: 48.6 µg/kg) in corn was higher than 3-A-DON (median: 6.8 µg/kg) (t = -3.579, P < 0.01). The concentration of DON, DON-3-G, 3-A-DON, 15-A-DON and ZEN in corn were higher than that in wheat (Z values were -3.492, -1.960, -2.467, -8.711 and -6.272, respectively, all P values < 0.05). Wheat (median: 29.0 µg/kg) contained higher NIV in comparison with corn (median: 18.2 µg/kg, Z = -2.086, P < 0.05).</p><p><b>CONCLUSION</b>Wheat and corn samples from parts of China were contaminated with multi-mycotoxins and DON was the predominant;in comparison of wheat, corn was more heavily contaminated with DON, DON-3-G, 3-A-DON, 15-A-DON and ZEN.</p>

The cytotoxic effect and injury mechanism of deoxynivalenol on articular chondrocytes in human embryo / 中华预防医学杂志

<p><b>OBJECTIVE</b>This study was to explore the cytotoxic effect and the related injury mechanism of deoxynivalenol (DON) on articular chondrocytes in human embryo.</p><p><b>METHODS</b>Articular cartilage cells were isolated from knees of human embryo and cultured in DMEM/F12 medium. The cells of the 4th generation were divided into five groups and incubated with varying concentrations of DON as the followings: control group and group with DON of 0.1, 0.2, 0.4, 1.0 µg/ml. The effects of DON were observed 72 hours after incubation. Cell apoptosis was assayed by flow cytometry (FCM) with Annexin V-FITC/PI staining; MMP-13 and PGE2 were detected by ELISA kits; NO was measured by Griess assay with spectrophotometer. Inducible nitric oxide synthase (iNOS) and collagen II in cells were detected by FCM. The expression levels of iNOS, mRNA and collagen II mRNA were measured with RT-PCR.</p><p><b>RESULTS</b>The rates of cell apoptosis in DON groups were 6.78% - 19.05%, which were significantly higher than that in control (1.20%, F = 174.761, P < 0.05). The levels of NO in DON groups were 20.8 - 40.7 µmol/L, which were significantly higher than that in control (10.2 µmol/L, F = 91.966, P < 0.05). The levels of MMP-13 in DON groups were 0.25 - 0.56 µmol/L, which were significantly higher than that in control (0 µmol/L, F = 78.420, P < 0.05). The levels of PGE2 in DON groups were 3.2-20.6 µmol/L, which were significantly higher than that in control (11.6 µmol/L, F = 276.453, P < 0.05). The proportions of cells with positive iNOS in DON groups were 14.8% - 56.8% which were significantly higher than that in controls (7.1%, F = 214.614, P < 0.05). The proportions of cells with positive collagen II in groups with DON of 0.4 µg/ml and 1.0 µg/ml were 56.7% and 52.7%, which were significantly lower than that in control (62.2%, F = 5.134, P < 0.05). The relative absorbance values of iNOS mRNA in DON groups were 1.07 - 1.33, which were significantly higher than that in control (0.62, F = 8.358, P < 0.05). The levels of collagen II mRNA in groups with DON of 0.4 µg/ml and 1.0 µg/ml were 0.83 and 0.82, which were significantly lower than that in control (1.14, F = 7.887, P < 0.05).</p><p><b>CONCLUSION</b>DON could promote anabolism of NO in articular cartilage cells by which up-regulated the expression of PGE2 and MMP-13, which both promoted resolution of articular cartilage matrix such as collagen II. DON induced apoptosis in articular cartilage cells.</p>

Enhancing effect of deoxynivalenol-mediated GRP78 down-regulation on heavy chain secretion and bioactivity of two-chain FVIII gene co-transfected cells / 药学学报

Although two chain transfering separately could be used to overcome the volume limitation of adeno-associated virus vectors (AAV) in coagulation factor VIII (FVIII) gene delivery, it leads to chain imbalance for inefficient heavy chain secretion. In this study we aimed to improve the efficacy of two chain strategy in FVIII gene delivery through the degradation of glucose-regulated protein 78 (GRP78) known as a protein chaperone in endoplasmic reticulum (ER) by deoxynivalenol (DON) to decrease GRP78-bound FVIII heavy chain. By treating the two-chain gene transduced 293 cells with DON, the heavy chain (HC) secretion and FVIII bioactivity were observed. Data showed that 293 cells after three hours post-treatment with DON at a concentration of 500 ng mL(-1) resulted in obvious decrease the level of GRP78 but no effect on the cell proliferation. The HC secreted from DON-treated cells transfected with HC gene alone was 59 +/- 11 ng mL(-1), higher than that secreted by control cells (15 +/- 4 ng mL(-1)), and the HC secretion was further increasing to 146 +/- 34 ng mL(-1) in light chain (LC) gene co-transfected cells with an activity measured up to 0.66 +/- 0.15 U mL(-1), also greater than control cells (76 +/- 17 ng mL(-1) and 0.35 +/- 0.09 U mL(-1)). Taken together, these data suggest that DON-mediated GRP78 down-regulation could improve the efficacy of two-chain FVIII gene transfering by facilitating HC secretion, providing an experimental basis for in vivo dual-AAV application in FVIII gene delivery.

Induction of apoptotic lesions in liver and lymphoid tissues and modulation of cytokine mRNA expression by acute exposure to deoxynivalenol in piglets

Six 1-month-old piglets were intravenously injected with deoxynivalenol (DON) at the concentration of 1 mg/kg body weight, with three pigs each necropsied at 6 and 24 h post-injection (PI) for investigation of hepatotoxicity and immunotoxicity with special attention to apoptotic changes and cytokine mRNA expression. Histopathological examination of the DON-injected pigs revealed systemic apoptosis of lymphocytes in lymphoid tissues and hepatocytes. Apoptosis of lymphocytes and hepatocytes was confirmed by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method and immunohistochemical staining against single-stranded DNA and cleaved caspase-3. The number of TUNEL-positive cells in the thymus and Peyer's patches of the ileum was increased at 24 h PI compared to 6 h PI, but the peak was at 6 h PI in the liver. The mRNA expression of interleukin (IL)-1beta, IL-6, IL-18, and tumor necrosis factor (TNF)-alpha in the spleen, thymus and mesenteric lymph nodes were determined by semi-quantitative RT-PCR, and elevated expression of IL-1beta mRNA at 6 h PI and a decrease of IL-18 mRNA at 24 h PI were observed in the spleen. IL-1beta and IL-6 mRNA expressions increased significantly at 6 h PI in the thymus, but TNF-alpha decreased at 6 h PI in the mesenteric lymph nodes. These results show the apoptosis of hepatocytes suggesting the hepatotoxic potential of DON, in addition to an immunotoxic effect on the modulation of proinflammatory cytokine genes in lymphoid organs with extensive apoptosis of lymphocytes induced by acute exposure to DON in pigs.

Natural occurrence of nivalenol and mycotoxigenic potential of Fusarium graminearum strains in wheat affected by head blight in Argentina

The principal agents of Fusarium head blight in the main cropping area of Argentina were investigated in heavily infected samples. The ability of the isolates to produce trichothecenes was determined by GC and HPLC. Fusarium graminearum was the predominant species and of 33 isolates, 10 produced deoxinivalenol (DON) (0.1- 29 mg kg-1), 13 produced both deoxinivalenol (1.0- 708 mg kg-1) and nivalenol (0.1- 6.2mg kg-1), 12 produced 3-acetyldeoxinivalenol (0.1- 14 mg kg-1), 13 produced 15-acetyldeoxinivalenol (0.1- 1.9 mg kg-1), 10 produced Fusarenone X (0.1- 2.4 mg kg-1) and 7 produced zearalenone (0.1- 0.6 mg kg-1). These results suggest that F. graminearum strains isolated from the wheat growing regions in Argentina belong to DON chemotype. Although some strains produced both deoxinivalenol and nivalenol, nivalenol was produced in lower levels. The natural occurrence of nivalenol in wheat affected by head-blight collected in the main production area during two years (2001-2002) was also determined. From 19 samples 13 were contaminated with deoxinivalenol in a range of 0.3 to 70 mg kg-1and 2 samples with both deoxinivalenol (7.5 and 6.7 mg kg-1) and nivalenol (0.05 and 0.1 mg kg-1), respectively. This is the first report of natural occurrence of nivalenol in wheat cultivate in Argentina.

O principal causador de giberela no trigo na Argentina e sua capacidade de produzir tricotecenos foram estudados por GC e HPLC em amostras altamente infectadas. A espécie predominante foi Fusarium graminearum, sendo que de um total de 33 isolados, 10 produziram deoxinivalenol (0,1-29 mg kg -1), 13 produziram deoxinivalenol (1,0-708 mg kg-1) e nivalenol (0,1-6,2 mg kg-1), 12 produziram 3-acetildeoxinivalenol (0,1-14 mg kg-1), 13 produziram 15-acetildeoxinivalenol (0,1-1,9 mg kg-1), 10 produziram fusarenona X (0,1- 2,4 mg kg-1) e 7 produziram zearalenona (0,1- 0,6 mg kg-1). Esses resultados sugerem que as cepas de F. graminearum isoladas de trigo cultivado na Argentina pertencem ao quimiotipo DON. Embora algumas cepas tenham produzido tanto DON quanto NIV, NIV foi produzido em quantidade inferior ao DON. A ocorrência natural de nivalenol em trigo afetado pela giberela coletado na principal área de produção durante dois anos (2001-2002) foi também determinada. De 19 amostras, 13 estavam contaminadas com deoxinivalenol na faixa de 0,3 a 70 mg kg-1 e 2amostras continham tanto deoxinivalenol (7,5 e 6,7 mg kg-1) quanto nivalenol (0,05 e 0,1 mg kg-1), respectivamente. Esse é o primeiro relato da ocorrência de nivalenol em trigo cultivado na Argentina.

The combination of deoxynivalenol and zearalenone at permitted feed concentrations causes serious physiological effects in young pigs

This study was to investigate the effects of the combination of deoxynivalenol (DON) and zearalenone (ZON) on pigs. Twenty-four weaning piglets were divided into a control group fed a diet free of mycotoxins and a toxin group fed a diet containing 1 mg/kg DON and 250 microgram/kg ZON. The results showed that supplementation of DON and ZON in diets had extensive effects on pigs. More specifically, DON and ZON caused levels of total protein, albumin, and globulin in sera to decrease (p < 0.05) by 14.5%, 6.5% and 11.3%, respectively, and at the same time increased (p < 0.05) the serum enzyme activities of gamma-glutamyltransferase, aspartate aminotransferase and alanine aminotransferase by 72.0%, 32.6% and 36.6%, respectively. In addition, DON and ZON decreased (p < 0.05) the level of anticlassical swine fever antibody titers by 14.8%. Real-time PCR showed that DON and ZON caused the mRNA expression levels of IFN-gamma, TNF-alpha, IL-2, to decrease (p < 0.05) by 36.0%, 29.0% and 35.4%, respectively. Histopathological studies demonstrated that DON and ZON caused abnormalities in the liver, spleen, lymph nodes, uterus, and kidney. The concentrations of DON and ZON used in this study are in line with the published critical values permitted by BML. Our study clearly put the standard and adequacy of safety measures for these toxins into question. The authors suggest that with the increasing availability of cellular and molecular technologies, it is time to revisit the safety standards for toxins in feeds so as to make feeds safer, providing consumers with safer products.

Plasma haptoglobin and immunoglobulins as diagnostic indicators of deoxynivalenol intoxication

This study aimed to discover potential biomarkers for dioxynivalenol (DON) intoxication. B6C3F1 male mice were rally exposed to 0.83, 2.5 and 7.5 mg/kg body weight (bw) DON for 8 days and the differential protein expressions in their blood plasma were determined by SELDI - Time-of-Flight/Mass Spectrometry (TOF/MS) and the immunoglobulins (Igs) G, A, M and E in the serum were investigated. 11.7 kDa protein was significantly highly expressed according to DON administration and this protein was purified by employing a methyl ceramic HyperD F column with using optimization buffer for adsorption and desorption. The purified protein was identified as a haptoglobin precursor by peptide mapping with using LC/Q-TOF/MS and MALDI-TOF/MS and this was confirmed by western blotting and ELISA. IgG and IgM in serum were decreased in a dose-dependent manner and IgA was decreased at 7.5 mg/kg bw DON administration, but the IgE level was not changed. To compare the expressions of haptoglobin and the Igs patterns between aflatoxin B1 (AFB1), zearalenone (ZEA) and DON intoxications, rats were orally administered with AFB1 1.0, ZEA 240 and DON 7.5 mg/kg bw for 8 days. Haptoglobin was increased only at DON 7.5 mg/kg bw, while it was slightly decreased at ZEA 240 mg/kg bw and it was not detected at all at AFB1 1.0 mg/kg bw. IgG and IgA were decreased by DON, but IgG, IgA, IgM and IgE were all increased by AFB1. No changes were observed by ZEA administration. These results show that plasma haptoglobin could be a diagnostic biomarker for DON intoxication when this is combined with examining the serum Igs.

Desoxinivalenol(DON) em trigo e farinha de trigo comercializados na cidade de São Paulo / Desoxynivalenol(DON)in wheat: occurrence in samples commercialize in the São Paulo City

Vinte e oito amostras de farinha de trigo e 14 amostras de trigo em grão foram adquiridos na cidade de SãoPaulo e analisados para determinação de desoxinivalenol (DON). As amostras foram extraidas comacetonitrila-água (84+16) seguidas de limpeza dos extratos com colunas MycoSep. A separação e aquantificação foram pela técnica da Cromatografia em Camada Delgada (CCD) e a toxina visualizadacom solução de AlCl3. A eficiência das colunas MycoSep 225 e 227 foi avaliada com amostras de farinhade trigo e trigo em grão, contaminadas com DON em dois níveis, 80,0 e 100,0 »g/kg. As recuperaçõesmédias das colunas 225 e 227 foram, respectivamente, 72% e 107% para farinha de trigo e 90% e 125%para trigo em grão. Os desvios padrão relativos foram 11% e 18% para farinha de trigo e 8% e 20% paratrigo em grão, respectivamente. DON foi detectado em 19 (45%) das 42 amostras analisadas em níveisque variaram de 82-1500µg/kg.Algumas amostras (8) foram confirmadas por CLAE-UV.

Effects of vitamin C on apoptosis and proliferation inhibition of human peripheral blood mononuclear cells induced by deoxynivalenol in vitro / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the effects of Vitamin C (Vit C) on the apoptosis and proliferation inhibition of human peripheral blood mononuclear cells (HPBMCs) induced by deoxynivalenol (DON) in vitro.</p><p><b>METHODS</b>The effects of Vit C pretreatment at different dosages (25 micromol/L and 100 micromol/L) on apoptosis, apoptosis related genes expression and proliferation inhibition of HPBMCs induced by DON were evaluated with cell culture, flow cytometric DNA analysis and Western blotting.</p><p><b>RESULTS</b>Flow cytometry (FCM) analysis showed that the apoptosis rate of HPBMCs in 2000 microg/L DON group was (28.82 +/- 1.67)%, which was significantly higher than that in control group (14.07 +/- 0.70, P < 0.05). Compared with DON group, the apoptosis rate of HPBMCs in 25 micromol/L Vit C pretreatment group was significantly decreased (28.82 +/- 1.67)% vs (22.39 +/- 1.05)%, P < 0.05, while that in 100 micromol/L Vit C pretreatment group was obviously increased (36.07 +/- 2.92)%, P < 0.05. Western blotting analysis showed that the expression of Bax and Caspase-3 up-regulated by DON was markedly decreased, while the expression of Bcl-2 down-regulated by DON was increased by 25 micromol/L Vit C pretreatment (P < 0.05). 100 micromol/L Vit C pretreatment could further increase the expression of Bax and Caspase-3 of HPBMCs induced by DON, while no significant effects on the Bcl-2 expression induced by DON were seen. FCM analysis showed that the proliferation index of HPBMCs in Vit C pretreatment groups at different dosages was all dramatically increased as compared with that in DON groups (P < 0.05).</p><p><b>CONCLUSION</b>25 micromol/L Vit C pretreatment could at certain extent inhibit the apoptosis and reverse the abnormal expression of apoptosis related genes of HPBMCs induced by DON in vitro, while 100 micromol/L Vit C pretreatment could further increase the apoptosis rate of HPBMCs induced by DON. Vit C pretreatment could reverse the proliferation inhibition of HPBMCs induced by DON in vitro.</p>

Effects of vitamin C on the inhibition of human leucocyte antigen class I (HLA-I) expression of human peripheral blood mononuclear cells induced by deoxynivalenol in vitro / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the putative effects of Vitamin C (Vit C) on inhibition of human leucocyte antigen class I (HLA-I) expression of human peripheral blood mononuclear cells (HPBMCs) induced by deoxynivalenol (DON) in vitro.</p><p><b>METHODS</b>The effects of Vit C on the changes of HLA-I expression of HPBMCs induced by DON in vitro were evaluated with cell culture, flow cytometry (FCM), Western blotting and immunocytochemical methods.</p><p><b>RESULTS</b>FCM analysis showed that HLA-I expression of HPBMCs in DON treated cells was significantly lower than that in controls (FI 0.88 +/- 0.02 vs 1.00 +/- 0.03, P < 0.05). As compared with DON group, the HLA-I expressions of HPBMCs in the two Vit C (25 micromol/L and 100 micromol/L) pretreatment groups were all significantly increased (1.15 +/- 0.06 and 1.10 +/- 0.02 vs 0.88 +/- 0.02, P < 0.05). Exposure to different dosage of Vit C alone could dramatically increase the expression of HLA-I of HPBMCs in vitro as compared with that in the normal control (FI for 25 micromol/L and 100 micromol/L Vit C treatment group was 1.28 +/- 0.03 and 1.25 +/- 0.05 respectively, P < 0.05). Immunocytochemical results showed that the percentages of HLA-I positive expression of HPBMCs in Vit C pretreatment groups at different dosages were significantly higher than those in DON group (70.10 +/- 6.90)%, (64.50 +/- 5.50)% vs (42.20 +/- 4.30)%, P < 0.05. Western blotting confirmed the results of FCM and immunocytochemistry.</p><p><b>CONCLUSIONS</b>Vitamin C pretreatment at different dosages could reverse at some extent the inhibitive effects of DON on HLA-I expression of HPBMCs.</p>

Avaliação intralaboratorial e otimização de método para determinação de tricotecenos em milho por cromatografia a gás-espectrometria de massas / Intralaboratorial evaluation and otimization of a method for determination of trichothecenes in corn by gas chromatography-mass spectrometry

Tricotecenos são micotoxinas produzidas por diferentes espécies de Fusarium. Estas micotoxinas formam um grupo de cerca de mais de cem compostos caracterizados pela presença em suas estruturas do mesmo sistema de anéis tetracíclicos cirpenol.Alguns deles são contaminantes naturais em trigo e milho,como o desoxinivalenol(DON),o nivalenol (NIV),a toxina T-2 (T2),o diacetoxiscirpenol (DAS) e a toxina HT-2(HT2).No presente trabalho foram avaliados e otimizados sistemas de limpeza, extração e derivação para determinação simultânea de DON, NIV, DAS, HT2 e T2 em amostras de milho e seus produtos por cromatografia a gás associada à espectrometria de massas (CG/EM).A extração com acetonitrila: água (84:16) seguida de limpeza com coluna MycoSep 227 apresentou os melhores resultados para as cinco toxinas estudadas. A derivação com anidrido trifluoroacético/ bicabornato de sódio mostrou-se melhor tanto para a detecção como para quantificação dos tricotecenos estudados por CG/EM. As recuperações obtidas variaram de 83,3 a 113,3% para DON, 84,6 a 114,5% para NIV,52,1 a 122,7% para DAS,71,1 a 95,9% para T2 e de 81,8 a 155,4% para HT2.Os limites de detecção variaram de 20 a 50 ng/g para DON,de 10 a 40 ng/g para NIV,de 30 a 120 ng/g para DAS,de 20 a 100 ng/g para T2 e de 20 a 50 ng/g para HT2.A repetividade do método mostrou desvios padrão relativos variando de 4,5 a 18,1 para DON,2,9 a 15,4 para NIV,3,4 a 48,1 para DAS, 3,1 a 14,4 para HT2 e de 6,4 a 47,3 para T2