Factores extrínsecos e intrínsecos asociados a poblaciones fúngicas micotoxigénicas de granos de maíz (Zea mays L.) almacenados en silos bolsa en Argentina

ResumenCon el objeto de caracterizar las poblaciones fúngicas, en particular las especies potencialmente micotoxigénicas, que pueden contaminar los granos de maíz almacenados en silos bolsa con un contenido de humedad superior al recomendado como seguro, se evaluaron 270 muestras extraídas al inicio, a los 90 días y al final de un período de almacenamiento de 5 meses. En dichas muestras se cuantificó e identificó la biota fúngica y se determinó la contaminación con fumonisinas y aflatoxinas. Asimismo, se evaluó el efecto de factores extrínsecos (ambiente), intrínsecos (granos) y tecnológicos (ubicación de los granos en el perfil del silo bolsa) sobre las poblaciones totales y micotoxigénicas. El pH de los granos y el nivel de O2 se redujeron significativamente a los 5 meses, mientras que la concentración de CO2 se incrementó en igual período. Los recuentos totales de la micobiota fueron significativamente mayores en los granos ubicados en el estrato superior del silo bolsa. Se identificaron especies micotoxigénicas de Fusarium, Aspergillus, Penicillium y Eurotium. La frecuencia de aislamiento de Fusarium verticillioides se redujo al final del almacenamiento y Aspergillus flavus solo se aisló en el inicio del almacenamiento. Los recuentos de Penicillium spp. y Eurotium spp. se incrementaron al final del almacenamiento. El 100% de las muestras presentaron contaminación con fumonisinas, con niveles máximos de 5,707mg/kg, mientras que las aflatoxinas contaminaron el 40% de las muestras con niveles máximos de 0,0008mg/kg. Las condiciones ambientales y de sustrato generadas durante el almacenamiento produjeron cambios en la composición de las poblaciones fúngicas y limitaron el desarrollo de hongos micotoxigénicos y la producción de micotoxinas.AbstractIn order to determine the behavior of mycotoxin-producing fungal populations linked with silobags stored corn grains with a moisture content greater at the recommended as safe, 270 samples taken in three times (beginning, 90 days, final) over a five month period of storage were evaluated. The fungal biota was quantified and identified and the contamination with fumonisin and aflatoxin was determined. Extrinsic factors (environment), intrinsic factors (grains) and technological factors (location of the grains in the profile of silobag) were taken into account to evaluate the presence and quantity of total and mycotoxigenic fungal populations. The pH of grains and O2 levels were significantly reduced after five months, while CO2 concentration increased in the same period. The total counts of mycobiota were significantly higher in grains located in the top layer of silobag. Mycotoxigenic species of Fusarium, Aspergillus, Penicillium and Eurotium were identified. The frequency of isolation of Fusarium verticillioides decreased at the end of storage and Aspergillus flavus was isolated only at the beginning of storage. The counts of the Penicillium spp. and Eurotium spp. were increased at the end of storage. Fumonisin contamination was found in all the samples (100%) with maximum levels of 5.707mg/kg whereas aflatoxin contaminated only 40% with maximum levels of 0.0008mg/kg. The environmental and substrate conditions generated during the storage limited the development of mycotoxigenic fungi and mycotoxin production

Stability of DON and OTA during the breadmaking process and determination of process and performance criteria

The fate of deoxynivalenol (DON) and ochratoxin A (OTA) during the breadmaking process was studied. In particular, toxin content was analysed in mixed baking ingredients before kneading, after fermentation and proofing, and finally after baking. Fermentation and proofing were carried out at 30 C for 1 h, while baking was performed at different temperature levels (from 170 to 210 C) and baking times from 45 to 135 min, in a full factorial design. DON increased from unkneaded mix to fermented dough, and decreased due to baking; this trend depended on the initial concentration of DON in the flour. The level in the bread was significantly lower than in the initial mix of ingredients. In contrast, deoxynivalenol-3-glucoside (DON-3-G) content increased both during kneading and fermentation, and also during baking. Moreover, the results confirmed the high stability of OTA as no significant change in its content could be observed as a result of the breadmaking process. As conclusion, the design of bakery product processes may help to control DON in final products, because although quite stable, its levels can be reduced to some extent. However, high levels of DON-3-G were released during baking, and this point should be further investigated. Mycotoxins have been always considered as stable compounds; however, in depth knowledge of the processing steps that may lead to some reduction (although limited) and those which can stimulate their release from conjugated forms, will definitely help in their control in finished foodstuffs.The authors are grateful to the Spanish government (projects AGL2010-22182-C04-04 and AGL2011-24862) for the financial support. A. Vidal thanks the Spanish Government (Ministry of Education) for the pre-doctoral grant. H. Morales is grateful to the Portuguese Government. (Ministerio da Ciecia, Tecnologia e Ensino Superior; FCT Fundacao para a Ciencia e a Tecnologia) Grant ref. SFRH/BPD/38011/2007

Hepatitis B Virus Variants with Multiple Insertions and/or Deletions in the X Open Reading Frame 3 ' End: Common Members of Viral Quasispecies in Chronic Hepatitis B Patients

Hepatitis B virus; Insertions; Next-generation sequencingVirus de l'hepatitis B; Insercions; Seqüenciació de nova generacióVirus de la hepatitis B; Inserciones; Secuenciación de próxima generaciónDeletions in the 3′ end region of the hepatitis B virus (HBV) X open reading frame (HBX) may affect the core promoter (Cp) and have been frequently associated with hepatocellular carcinoma (HCC). The aim of this study was to investigate the presence of variants with deletions and/or insertions (Indels) in this region in the quasispecies of 50 chronic hepatitis B (CHB) patients without HCC. We identified 103 different Indels in 47 (94%) patients, in a median of 3.4% of their reads (IQR, 1.3–8.4%), and 25% (IQR, 13.1–40.7%) of unique sequences identified in each quasispecies (haplotypes). Of those Indels, 101 (98.1%) caused 44 different altered stop codons, the most commonly observed were at positions 128, 129, 135, and 362 (putative position). Moreover, 39 (37.9%) Indels altered the TATA-like box (TA) sequences of Cp; the most commonly observed caused TA2 + TA3 fusion, creating a new putative canonical TATA box. Four (8%) patients developed negative clinical outcomes after a median follow-up of 9.4 (8.7–12) years. In conclusion, we observed variants with Indels in the HBX 3′ end in the vast majority of our CHB patients, some of them encoding alternative versions of HBx with potential functional roles, and/or alterations in the regulation of transcription.This research was funded by Instituto de Salud Carlos III and co-financed by the European Regional Development Fund (ERDF), grant number PI18/01436; PI19/00301; and by the Centro para el Desarrollo Tecnológico Industrial (CDTI) from the Spanish Ministry of Economy and Business, grant number IDI-20200297. The APC was funded by the grant PI18/01436

Standardized Hepatitis B Virus RNA Quantification in Untreated and Treated Chronic Patients: a Promising Marker of Infection Follow-Up.

The measurement and interpretation of HBV DNA and RNA levels in HBV infected patients treated with antiviral therapy supports the objective of HBV disease management. Here, we quantified circulating HBV RNA through a standardized and sensitive assay in follow-up samples from both naive and treated patients as a marker of infection evolution. HBV DNA (HBV DNA for use in Cobas 6800/8800 Automated Roche Molecular Systems), RNA (Roche HBV RNA Investigational Assay for use in the Cobas 6800/8800; Roche), HBeAg and HBsAg (Elycsys HBsAg chemiluminescence immunoassay by Cobas 8000; Roche), and core-related antigen (Lumipulse G chemiluminescence assay; Fujirebio) levels were measured in cohorts of untreated or nucleos(t)ide treated, HBV-infected subjects in an outpatient hospital setting. HBV DNA levels in untreated people were 3.6 log10 higher than corresponding RNA levels and were stable over 5 years of observation. While only five of 52 treated patients had DNA levels below the lower limit of quantification (10 IU/mL) at the end of follow-up, 13 had HBV RNA levels persistently above this limit, including eight with undetectable DNA. In samples with undetectable core-related antigen we observed a median HBsAg titer 2.7-fold higher than in samples with undetectable RNA (adjusted P = 0.012). Detectable HBV RNA with undetectable HBV DNA was a negative predictor of HBsAg decrease to a level ≤100 IU/mL (P = 0.03). In naive patients the difference between HBV DNA and RNA was higher than previously reported. HBV RNA rapidly decreased during treatment. However, in some cases, it was detectable even after years of effective therapy, being a negative predictor of HBsAg decrease. The investigational RNA assay for use on the Cobas 6800/8800 instruments is a sensitive and standardized method that could be applied in general management of HBV infection. IMPORTANCE This study focused on the quantification of circulating HBV RNA by using a standardized and sensitive assay. Thanks to this system we observed a higher difference between circulating HBV DNA and RNA than previously reported. In treated patients, HBV RNA decreased together with DNA, although some patients presented detectable levels even after years of successful antiviral treatment, suggesting a persistent viral transcription. Of note, the detection of viral RNA when HBV DNA is undetectable was a negative predictor of HBsAg decrease to a level ≤100 IU/mL. This assay could be extremely helpful in HBV patients management to study viral transcription and to identify those treated patients that may achieve sustained viral suppression

Critical care in South America - The new tradition

This article offers a brief discussion of some of the aspects of clinical and academic realities of critical and intensive care medicine in South America. Organizational efforts of collaborating physician and nursing intensivists from South American countries, Spain, and Portugal are outlined. Discussion includes the issues of funding and support of health care delivery of the critically ill, and some of the clinical syndromes not commonly seen in North America and Europe, but seen by intensivists in South America.132377

Critical Care In South America. The New Tradition.

This article offers a brief discussion of some of the aspects of clinical and academic realities of critical and intensive care medicine in South America. Organizational efforts of collaborating physician and nursing intensivists from South American countries, Spain, and Portugal are outlined. Discussion includes the issues of funding and support of health care delivery of the critically ill, and some of the clinical syndromes not commonly seen in North America and Europe, but seen by intensivists in South America.13377-8

Mycobiota and Potential Mycotoxin Contamination of Soybean RR in Different Production Areas in Argentina

A total of 348 freshly harvested soybean samples from a multi-environment trial, conducted in experimental fields belonging to INTA (Instituto Nacional de Tecnología Agropecuaria) in 12 localities, were analyzed for mycotoxin natural occurrence and for determining the associated endogenous mycoflora. Aflatoxins (AFs) and zearalenone (ZEN) were analyzed by HPLC and deoxynivalenol (DON) by CG.Samples from Reconquista, Tres Pozos, Rafaela and Manfredi were the most infested by fungi and those from Barrow and Balcarce the less contaminated. A total of 13,316 fungal isolates were identified from the seeds of 32 Roundup® Ready (transgenic) soybean cultivars. All fungi isolated were mitosporic fungi and ascomycetes. The most common fungi identified included species that belong to Alternaria, Fusarium, Sclerotinia, Phomopsis, Rhizoctonia and Cladosporium genera. The isolation frequencies and relative densities of species were calculated. Alternaria alternata, Fusarium equiseti, Sclerotinia sclerotiorum, Phomopsis spp., Fusarium semitectum, Cladosporium cladosporioides and Rhizoctonia solani were the predominant fungal species identified as endogenous mycoflora. No soybean samples were naturally contaminated with AFs, DON or ZEN. This is the first report on contaminant mycoflora and mycotoxin natural occurrence in transgenic soybean seeds from an extensive production area in Argentina.Fil: Zelaya, Manuel J.. Instituto Nacional de Tecnología Agropecuaria; ArgentinaFil: Gonzalez, Hector Horacio Lucas. Universidad de Buenos Aires. Facultad de Ingeniería; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Resnik, Silvia Liliana. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Pacin, Ana Maria. Fundación de Investigaciones Científicas "Teresa Benedicta de la Cruz"; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Salas, María P.. Universidad de Buenos Aires. Facultad de Ingeniería; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Martínez, María J.. Instituto Nacional de Tecnología Agropecuaria; Argentin

Trichothecenes and Mycoflora in Wheat Harvested in Nine Locations in Buenos Aires Province, Argentina

A total of 120 freshly harvested wheat samples from the 2004 season in nine locations from Northern Buenos Aires Province, Argentina, were analysed for trichothecene natural occurrence and associated mycoflora, and for determining the influence of commonly used fungicide field treatment and the cultivar type on trichothecene contamination. The trichothecenes T-2 tetraol, T-2 triol, HT-2 and T-2 toxin (HT-2, T-2), diacetoxyscirpenol (DAS), nivalenol (NIV), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON) were analysed by gas chromatography and electron capture detection. Detection limits ranged from 4 to 20 microg/kg. The isolation frequencies of species were calculated. Alternaria alternata, Fusarium graminearum, Fusarium poae and Fusarium semitectum were the predominant fungal species identified as endogenous mycoflora. The type of cultivar and the fungicide field treatment did not affect significantly the trichothecene contamination. The trichothecenes type A detected were HT-2 and T-2 triol toxins and the type B were DON, NIV and 3-ADON. Based on 120 samples the incidences were 21.7% for 3-ADON, 22.5% for HT-2, 27.5% for T-2 triol and 85% for DON. NIV was confirmed in one sample. Mean levels of trichothecene positive samples were between 7 and 2788 microg/kg.Fil: Gonzalez, Hector Horacio Lucas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ingeniería. Departamento de Ingeniería Química; ArgentinaFil: Moltó, G. A.. Universidad de Buenos Aires. Facultad de Ingeniería. Departamento de Ingeniería Química; ArgentinaFil: Pacin, Ana Maria. Universidad Nacional de Luján; Argentina. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; ArgentinaFil: Resnik, Silvia Liliana. Universidad de Buenos Aires. Facultad de Ingeniería. Departamento de Ingeniería Química; ArgentinaFil: Zelaya, M. J.. Fundación de Investigaciones Científicas "Teresa Benedicta de la Cruz"; ArgentinaFil: Masana, M.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación de Agroindustria. Instituto de Tecnología de Alimentos; ArgentinaFil: Martínez, Eric Javier. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Calculo. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Calculo; Argentin

von Hippel-Lindau mutants in renal cell carcinoma are regulated by increased expression of RSUME

Abstract Renal cell carcinoma (RCC) is the major cause of death among patients with von Hippel-Lindau (VHL) disease. Resistance to therapies targeting tumor angiogenesis opens the question about the underlying mechanisms. Previously we have described that RWDD3 or RSUME (RWD domain-containing protein SUMO Enhancer) sumoylates and binds VHL protein and negatively regulates HIF degradation, leading to xenograft RCC tumor growth in mice. In this study, we performed a bioinformatics analysis in a ccRCC dataset showing an association of RSUME levels with VHL mutations and tumor progression, and we demonstrate the molecular mechanism by which RSUME regulates the pathologic angiogenic phenotype of VHL missense mutations. We report that VHL mutants fail to downregulate RSUME protein levels accounting for the increased RSUME expression found in RCC tumors. Furthermore, we prove that targeting RSUME in RCC cell line clones carrying missense VHL mutants results in decreased early tumor angiogenesis. The mechanism we describe is that RSUME sumoylates VHL mutants and beyond its sumoylation capacity, interacts with Type 2 VHL mutants, reduces HIF-2α-VHL mutants binding, and negatively regulates the assembly of the Type 2 VHL, Elongins and Cullins (ECV) complex. Altogether these results show RSUME involvement in VHL mutants deregulation that leads to the angiogenic phenotype of RCC tumors