Amazonian soil fungi are efficient degraders of glyphosate herbicide; novel isolates of Penicillium, Aspergillus, and Trichoderma / Os fungos do solo amazônico são degradadores eficientes do herbicida glifosato; novos isolados de Penicillium, Aspergillus e Trichoderma

Abstract Pesticide residues that contaminate the environment circulate within the hydrological cycle can accumulate within the food chain and cause problems to both environmental and human health. Microbes, however, are well known for their metabolic versatility and the ability to degrade chemically stable substances, including recalcitrant xenobiotics. The current study focused on bio-prospecting within Amazonian rainforest soils to find novel strains fungi capable of efficiently degrading the agriculturally and environmentally ubiquitous herbicide, glyphosate. Of 50 fungal strains isolated (using culture media supplemented with glyphosate as the sole carbon-substrate), the majority were Penicillium strains (60%) and the others were Aspergillus and Trichoderma strains (26 and 8%, respectively). All 50 fungal isolates could use glyphosate as a phosphorous source. Eight of these isolates grew better on glyphosate-supplemented media than on regular Czapek Dox medium. LC-MS revealed that glyphosate degradation by Penicillium 4A21 resulted in sarcosine and aminomethylphosphonic acid.

Resumo Resíduos de agrotóxicos que contaminam o meio ambiente circulam no ciclo hidrológico, podendo se acumular na cadeia alimentar e causar problemas tanto à saúde ambiental quanto humana. Por sua vez, microrganismos são bem conhecidos por sua versatilidade metabólica e capacidade de degradar substâncias quimicamente estáveis, incluindo xenobióticos recalcitrantes. O estudo atual se concentrou na bioprospecção nos solos da floresta amazônica para encontrar novas linhagens de fungos capazes de degradar com eficiência o herbicida onipresente na agricultura e no meio ambiente, o glifosato. Entre os 50 fungos isolados (usando meio de cultura suplementado com glifosato como única fonte de carbono), a maioria eram isolados do gênero Penicillium (60%) e os outros eram isolados de Aspergillus e Trichoderma (26 e 8%, respectivamente). Todos os 50 isolados de fungos foram capazes de usar glifosato como fonte de fósforo. Oito desses isolados cresceram melhor em meio suplementado com glifosato do que em meio Czapek Dox regular. LC-MS revelou que a degradação do glifosato por Penicillium 4A21 resultou nos metabólitos sarcosina e ácido aminometilfosfônico.

Normalized difference vegetation index for desiccation evaluation with glyphosate + 2, 4-D in magnetized spray solution / Índice de vegetação por diferença normalizada para avaliação de dessecação com glyphosate + 2, 4-D em calda magnetizada

Abstract Water magnetization and geoprocessing are increasingly utilized tools in weed management. Our objective was to study the influence of water magnetization on herbicide efficiency and to verify whether there is a relationship between control scores and the normalized difference vegetation index (NDVI). In the laboratory experiment, water was subjected to magnetization and evaluated with respect to four characteristics. In the field experiment, plots of Brachiaria grass were subjected to treatments in a factorial scheme (6 × 2 + 1). Six herbicidal factors (doses of glyphosate and glyphosate + 2,4-D) and the magnetization or absence of magnetization of the spray solution were evaluated and compared against the control treatment (without spraying). Weed control assessments were carried out six times. Images were obtained using an embedded multispectral camera to determine the NDVI values. Data related to water characteristics were analyzed using the t test. Weed control and NDVI data were subjected to analysis of variance and are presented in regression graphs. Dispersion analysis of NDVI data was performed according to the control scores. The magnetization process decreased the pH of the water and increased the surface tension, but it did not influence the control scores or the NDVI. As the glyphosate dose was increased, the control scores were higher and the NDVI values were lower. Magnetized water did not affect the biological efficiency of the herbicides, and there was a strong correlation between the control scores and the NDVI values.

Resumo A magnetização da água e o geoprocessamento são ferramentas cada vez mais utilizadas no manejo de ervas daninhas. Nosso objetivo foi estudar a influência da magnetização da água na eficiência do herbicida e verificar se existe uma relação entre os escores de controle e o índice de vegetação por diferença normalizada (NDVI). No experimento de laboratório, a água foi submetida à magnetização e avaliada em relação a quatro características. No experimento de campo, parcelas de capim-braquiária foram submetidas a tratamentos em esquema fatorial (6 × 2 + 1). Seis fatores herbicidas (doses de glyphosate e glyphosate + 2,4-D) e a magnetização ou ausência de magnetização da calda foram avaliados e comparados com o tratamento controle (sem pulverização). Avaliações de controle de plantas daninhas foram realizadas seis vezes. As imagens foram obtidas usando uma câmera multiespectral incorporada para determinar os valores de NDVI. Os dados relacionados às características da água foram analisados ​​por meio do teste t. O controle de plantas daninhas e os dados de NDVI foram submetidos à análise de variância e são apresentados em gráficos de regressão. A análise de dispersão dos dados NDVI foi realizada de acordo com os escores de controle. O processo de magnetização diminuiu o pH da água e aumentou a tensão superficial, mas não influenciou nos escores de controle ou no NDVI. Conforme a dose de glyphosate foi aumentada, os escores de controle foram maiores e os valores de NDVI foram menores. A água magnetizada não afetou a eficiência biológica dos herbicidas, e houve uma forte correlação entre os escores de controle e os valores de NDVI.

Ixazomib-Based Regimen Followed by Autologous Hematopoietic Stem Cell Transplantation in the Treatment of POEMS Syndrome / 中国实验血液学杂志

OBJECTIVE@#To investigate the safety and efficacy of a new proteasome inhibitor Ixazomib followed by autologous hematopoietic stem cell transplantation (AHSCT) in the treatment of POEMS syndrome.@*METHODS@#The clinical manifestations, diagnosis and treatment process and follow-up results of 4 patients with POEMS syndrome who were treated with Ixazomib-based regimen combined with AHSCT in Wuhan No.1 Hospital from February 2018 to July 2020 were analyzed retrospectively. All patients were male, aged from 37-54 years old, with varying degrees of peripheral neuropathy, organ enlargement (liver, spleen or lymph nodes), circulatory overload (peripheral edema and/or pleural effusion), osteosclerosis, endocrine diseases (thyroid, gonads, etc.), skin changes (pigmentation, hemangioma, white nails, etc.), M protein, papilledema and other clinical manifestations and characteristics at the time of initial treatment. Two patients were pathologically diagnosed as hyaline vascular Castleman disease by lymph node biopsy. Three patients underwent lumbar puncture examinations and all showed elevated cerebrospinal fluid protein. All patients received at least 2 cycles of sequential AHSCT after induction chemotherapy based on ixazomib. The follow-up time was 10-28 months, and the median follow-up time was 16 months.@*RESULTS@#All cases survived. The complications were controllable during the treatment. Moreover, the clinical symptoms related to the disease were improved to a certain extent after the treatment. The levels of vascular endothelial growth factor (VEGF) showed a gradual decline.@*CONCLUSION@#Ixazomib combined with AHSCT is safe and effective in the treatment of POEMS syndrome.

Effects of glyphosate on soil fungal communities: A field study / Efecto del glifosato en las comunidades fúngicas del suelo: estudio a campo

ABSTRACT The driving forces behind many soil processes are microorganisms and they are able to respond immediately to environmental changes. The soil microbial community impacts on many soil properties. More than one-third of the terrestrial ecosystems are semiarid. However, a limited number of studies have been conducted to characterize soil fungal communities in semiarid grasslands, in particular those of agricultural fields. The aim of this study was to explore changes in the diversity and structure of soil fungal communities in semiarid grasslands, after different doses of glyphosate were applied under field conditions. Changes in soil fungal communities were examined using different approaches including culturing, calcofluor white stain and denaturing gradient gel electrophoresis (DGGE). The different approaches complement each other, revealing different aspects of the effect of glyphosate on soil fungal communities. We demonstrated a negative effect of glyphosate on soil fungal biomass at high doses and an early and transitory stimulatory effect on soil fungal biomass. We also found a negative effect of glyphosate on the species richness of cultivable fungi and changes in the molecular structure of soil fungal communities after double doses or long-term glyphosate application. In summary, our findings demonstrate an overall negative effect of glyphosate on soil fungal communities.

RESUMEN Los microorganismos del suelo son los responsables de llevar a cabo la mayoría de los procesos biológicos que ocurren en el suelo, y son capaces de reaccionar ante el estrés ambiental. Más de un tercio de los ecosistemas terrestres son semiáridos. Sin embargo, son escasos los estudios realizados para caracterizar las comunidades fúngicas en suelos agrícolas en ecosistemas semiáridos. El objetivo del presente trabajo fue estudiar los cambios que se producen en la biomasa, la diversidad y la estructura de las comunidades fúngicas del suelo, luego de la aplicación de distintas dosis de glifosato en condiciones de campo. Se emplearon diferentes técnicas incluidas el cultivo, la tinción directa con blanco de calcoflúor y PCR acoplada a electroforesis en geles de gradiente desnaturalizante (DGGE). Las distintas metodologías empleadas se complementan entre sí al detectar cada una distintos aspectos del efecto del glifosato en las comunidades fúngicas del suelo. Se encontró que el glifosato produce un efecto negativo sobre la biomasa fúngica, también se encontró un efecto transitorio estimulante inmediatamente posterior a la aplicación del herbicida. Además, se vio un efecto negativo sobre la riqueza de hongos cultivables, así como también cambios en la estructura molecular de las comunidades luego de aplicaciones repetidas. En conclusión, se demostró un efecto negativo generalizado sobre las comunidades fúngicas del suelo.

Study of muscle fibers of the extensor digitorium longus and soleus muscles of C57BL/6 females exposed to glyphosate during pregnancy and lactation / Estudo das fibras musculares dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e a lactação

ABSTRACT Objective To evaluate the morphology and morphometry of the muscles extensor digitorium longus and soleus of C57BL/6 females, who were exposed to glyphosate during pregnancy and lactation. Methods Twelve female mice from the C57BL/6 lineage were used. After detection of pregnancy, they were divided into a Control Group, which received only water, and a Glyphosate Group, which received water with 0.5% glyphosate during pregnancy and lactation. Both groups received ad libitum standard diet. After weaning, the females were euthanized and weighed; naso-anal length was measured, and fats were collected and weighed. The muscles extensor digitorium longus and soleus were collected, and their length and weight were measured. Then, the muscles were fixed in Methacarn to perform the histological study of muscle fibers. Results Glyphosate Group presented lower weight gain during pregnancy and also lower final body weight and naso-anal length; however, the other body parameters evaluated did not present a significant difference in relation to the Control Group. Significant differences were also not observed in the analysis of muscle fibers and connective tissue. Conclusion Exposure to 0.5% glyphosate during pregnancy and lactation resulted in lower weight gain during pregnancy, final weight, and naso-anal length. Despite not directly altering the morphology of muscle tissue, these results may indicate enough exposure to interfere with animal metabolism.

RESUMO Objetivo Avaliar a morfologia e a morfometria dos músculos extensor longo dos dedos e sóleo de fêmeas C57BL/6 expostas ao glifosato durante a prenhez e lactação. Métodos Foram utilizados 12 camundongos fêmeas da linhagem C57BL/6. Após detecção da prenhez, foram separadas em Grupo Controle, que recebeu somente água, e Grupo Glifosato, que recebeu água com 0,5% de glifosato durante a prenhez e lactação. Ambos os grupos receberam dieta padrão ad libitum. Após o desmame, as fêmeas foram eutanasiadas e pesadas; o comprimento nasoanal foi mensurado, e as gorduras foram coletadas e pesadas. Os músculos extensor longo dos dedos e sóleo foram coletados, e seu comprimento e peso foram mensurados. Em seguida, os músculos foram fixados em Methacarn para a realização do estudo histológico das fibras musculares. Resultados O Grupo Glifosato apresentou menor ganho de peso durante a prenhez e também menor peso corporal final e comprimento nasoanal, entretanto os demais parâmetros corporais avaliados não apresentaram diferença significativa em relação ao Grupo Controle. Na análise das fibras musculares e do tecido conjuntivo, também não foram observadas diferenças significativas. Conclusão A exposição a 0,5% de glifosato durante a prenhez e lactação resultou em menor ganho de peso na gestação, peso final e comprimento nasoanal, o que pode indicar que, apesar de não alterar a morfologia do tecido muscular diretamente, a exposição foi suficiente para interferir no metabolismo dos animais.

Synthesis of (S)-4-fluorophenylglycine by using immobilized amidase based on metal-organic framework / 生物工程学报

A stable Zr-based metal-organic framework (MOF, UiO-66-NH2) synthesized via micro-water solvothermal method was used to immobilize amidase by using the glutaraldehyde crosslinking method. The effect of immoblization conditions on enzyme immoblization efficiency was studied. An activity recovery rate of 86.4% and an enzyme loading of 115.3 mg/g were achieved under the optimal conditions: glutaraldehyde concentration of 1.0%, cross-linking time of 180 min, and the weight ratio of MOF to enzyme of 8:1. The optimal temperature and optimal pH of the immobilized amidase were determined to be 40 °C and 9.0, respectively, and the Km, Vmax and kcat of the immoblized amidase were 58.32 mmol/L, 16.23 μmol/(min·mg), and 1 670 s⁻¹, respectively. The immobilized enzyme was used for (S)-4-fluorophenylglycine synthesis and the optimal reaction conditions were 300 mmol/L of N-phenylacetyl-4-fluorophenylglycine, 10 g/L of immobilized enzyme loading, and reacting for 180 min at pH 9.0 and 40 °C. A conversion rate of 49.9% was achieved under the optimal conditions, and the conversion rate can be increased to 99.9% under the conditions of enantiomeric excess. The immobilized enzyme can be repeatedly used, 95.8% of its original activity can be retained after 20 cycles.

Isolation of culturable mycota from Argentinean soils exposed or not-exposed to pesticides and determination of glyphosate tolerance of fungal species in media supplied with the herbicide / Aislamiento de la microbiota cultivable de suelos argentinos expuestos y no expuestos a plaguicidas y determinación de la tolerancia a glifosato de especies fúngicas en medios suplementados con el herbicida

Abstract The current agricultural system has led to the development of glyphosate (GP)-resistant weeds, causing an increase in GP doses and applications. Native mycota of pesticide-contaminated sites are the major source of pesticide-degrading microorganisms. The aims of the present study were to isolate the GP-tolerant culturable mycota in two soils with different pesticide exposure from Córdoba, Argentina, and to evaluate the growth parameters in native fungal isolates in the presence of GP and the effective dose that caused 50% growth reduction. The results showed that the genera Fusarium, Aspergillus, Mucor, Penicil-lium and Sterilia were the prevalent fungi isolated from soils both exposed and not-exposed to pesticides. The highest value (>100 mM) of effective concentration of herbicide that caused 50% growth inhibition (EC50), was found for Trichoderma isolates. Sterilia spp. had EC50 val-ues of 100mM, while Aspergillus spp. and Mucor had EC50 values between 50 and 100mM. The growth rate evaluation varied according to the isolates and GP concentrations. The data showed that all Aspergillus spp., Trichoderma spp., Mucor and three Sterilia spp. had the best growth performance in media supplied with GP after a variable acclimation period. This study provides valuable data for further studies that would allow to know the metabolic capacity of these fungal species that can be potential candidates for GP removal from contaminated environments.

Resumen El sistema agrícola actual condujo ai desarrollo de malezas resistentes a glifos-ato (GP), lo que deriva en el aumento de las dosis y el número de aplicaciones requeridas. La microbiota nativa de sitios contaminados con plaguicidas constituye la fuente principal de microorganismos capaces de degradar este herbicida. Los objetivos del presente estudio fueron aislar la microbiota cultivable tolerante a GP a partir de dos suelos agrícolas con diferente grado de exposición a plaguicidas, ambos de la provincia de Córdoba, Argentina, y evaluar en los hongos nativos aislados los parámetros de crecimiento en presencia de GP y la dosis efectiva que produce un 50% de reducción del crecimiento (EC50). Los géneros Fusarium, Aspergillus, Mucor, Penicillium y Sterilia fueron los de mayor prevalencia, tanto en suelos expuestos como no expuestos a plaguicidas. La mayor EC50 hallada (> 100 mM) correspondió a dos aislamientos de Trichoderma. En dos aislamientos de Sterilia, los valores de EC50 fueron de 100 mM, mientras que en los dos de Aspergillus y en uno de Mucor fueron de entre 50 y 100 mM. La velocidad de crecimiento varió según la concentración de GP empleada y el aislamiento considerado. Todos los aislamientos de Aspergillus, Trichoderma y Mucor, y tres aislamientos de Sterilia mostraron el mejor rendimiento en cuanto al crecimiento en los medios con GP después de un período de aclimatación variable. Este trabajo proporciona datos preliminares valiosos referidos a la capacidad metabólica de estos aislados y su potencial para remover GP en ambientes contaminados.

Tolerance of dark septate endophytic fungi (DSE) to 1 agrochemicals in vitro / Tolerancia de hongos endofíticos septados oscuros a agroquímicos in vitro

Abstract Dark septate endophytes (DSE) are a heterogeneous group of fungi, mostly belonging to the Phylum Ascomycota, that are involved in a mutualistic symbiosis with plant roots. The aim of this study is to evaluate the behavior of two strains of DSE isolated from wheat roots of two cropping areas in the province of Buenos Aires, Argentina, against some agrochemicals. Of all the isolates obtained, two strains were identified as Alternaria alternata and Cochliobolus sp. These DSE were found to be tolerant to glyphosate, carbendazim and cypermethrin when evaluated at the recommended agronomic dose (AD), 2 AD and, in some cases, 10 AD. This work contributes to the study of the biology of this group of fungi and their tolerance in the presence of xenobiotics widely used in agriculture.© 2019 Asociaci´on Argentina de Microbiolog´ıa. Published by Elsevier Espa˜na, S.L.U. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/bync-nd/4.0/).

Resumen Los endófitos septados oscuros (DSE) son un grupo heterogéneo de hongos que participan de una simbiosis mutualista con raíces de plantas, perteneciendo principalmente al Phylum Ascomycota. El objetivo de este estudio fue aislar DSE de raíces de trigo proveniente de dos áreas de cultivo de la provincia de Buenos Aires y evaluar el comportamiento de dos cepas de DSE aisladas de raíces de trigo frente a algunos agroquímicos en dos áreas de cultivo de la provincia de Buenos Aires. De todos los aislamientos obtenidos se seleccionaron dos cepas que se identificaron como Alternaria alternata y Cochliobolus sp. Se encontró que estos DSE son tolerantes al glifosato, el carbendazim y la cipermetrina, evaluados a las dosis agronómicas recomendadas (AD), a 2x AD y, en algunos casos, a 10x AD. Este trabajo contribuye al conocimiento de la biología de este grupo de hongos y su tolerancia a xenobióticos ampliamente utilizados en la agricultura.

Efecto del glifosato sobre la microbiota, calidad del suelo y cultivo de frijol biofortificado en el departamento del Cesar, Colombia / Effect of glyphosate on microbiota, soil quality and biofortified bean crop in Codazzi, department of Cesar, Colombia

Resumen Las estrategias en seguridad alimentaria con cultivos de alto contenido nutricionaldeben enmarcarse en prácticas agrícolas sostenibles, orientadas a la conservación del suelo, elalto rendimiento y la inocuidad. Esta última característica implica la producción de alimentos sintrazas detectables de agroquímicos, los que podrían amenazar la salud del consumidor. Se des-arrolló un estudio con el objetivo de evaluar el efecto del herbicida glifosato sobre la fertilidadquímica y microbiológica del suelo, así como su residualidad en la semilla de frijol biofortificadocultivado en el departamento de Cesar, Colombia. La metodología comprendió un análisis corre-lacional de indicadores de calidad de suelo, rendimiento del cultivo y residualidad en los granos.Los tratamientos evaluados incluyeron la aplicación o no de glifosato, el uso de coberturas sin-téticas (mulch) o naturales, frente al control manual de las malezas. Se hallaron diferenciasde rendimiento y de la respuesta de los indicadores químicos y microbiológicos en función deltratamiento y las condiciones iniciales de la rizósfera de frijol. La aplicación del herbicida glifo-sato en suelo sin mulch generó una disminución del 29% en el rendimiento, asociada a la mayorprevalencia de plagas y enfermedades fúngicas. En ninguno de los tratamientos con aplicaciónde glifosato se observó residualidad de este herbicida en semillas, pero sí de otras moléculasderivadas de insecticidas usados en el sistema local de producción. De este estudio se concluyóque en lotes de frijol biofortificado con alta presión de la maleza Cyperus rotundus (coquito), se recomienda el uso del herbicida glifosato acompañado de mulch como alternativa para mantener la productividad en el tiempo. Esto constituye un sistema de protección frente al efectonegativo del herbicida sobre ambos, el sistema radical de la planta y la microbiota del suelo.© 2019 Asociacion Argentina de Microbiologıa. Publicado por Elsevier España, S.L.U. Este es un artıculo Open Access bajo la licencia CC BY-NC-ND (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Abstract Strategies aimed at achieving food safety in crops of high nutritional value shouldbe carried out through sustainable agricultural practices aimed at soil conservation, high yieldand food safety. This latter characteristic implies food production without detectable tracesof agrochemicals that threaten health. The objective of this study was to assess the effect ofthe herbicide glyphosate on the chemical and biological fertility of the soil and to determineits residual activity on biofortified bean seeds in Codazzi, Department of Cesar, Colombia. Themethod included a correlational analysis of soil quality, crop yield and residuality in bean grains.The treatments included glyphosate application and synthetic and natural mulches, comparedto manual control. The results showed differences in the response of chemical, microbiologicaland yield indicators between treatments and the initial conditions of the bean rhizosphere. Theuse of the herbicide glyphosate in mulch-free soil generates yield losses of 29% associated witha higher incidence of pests and fungal diseases; in all treatments, no glyphosate residualitywas detected in seeds; however, residuality was detected in other molecules derived frominsecticides associated with the local production system. In conclusion, in plots with high weedpressure by Cyperus rotundus, the use of mulch is recommended as an alternative to maintainbean productivity over time; by offering protection against the effect of the herbicide on boththe root system of the plant and the soil microbiota of the biofortified bean crop.© 2019 Asociaci´on Argentina de Microbiolog´ıa. Published by Elsevier España, S.L.U. This is anopen access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Morphological effects on helminth parasites caused by herbicide under experimental conditions / Efeitos morfológicos em helminto parasito causado por herbicida em condições experimentais

Abstract Helminth parasites have been studied as potential accumulators for different pollutants. Echinostoma paraensei is a foodborne trematode whose vertebrate host, the rodent Nectomys squamipes, is naturally exposed to environmental pesticides. However, little information exists regarding the pesticide's effects on helminths. This study investigated the morphological effects on the trematode, E. paraensei, after experimental Roundup® herbicide exposure, in concentrations below those recommended for agricultural use. After two hours of exposure, scanning electron microscopy (SEM) showed changes to the tegument, such as furrowing, shrinkage, peeling, spines loss on the peristomic collar, and histopathological evidence of altered cells in the cecum and acinus vitelline glands with vacuoles and structural changes to the muscular layers. Glycidic content was decreased, primarily in the connective tissue. As E. paraensei is an intestinal parasite of the semi-aquatic wild rodent, N. squamipes, it is predisposed to pesticide exposure resulting from agricultural practices. Therefore, we emphasize the need to evaluate its impact on helminth parasites, due to their pivotal role in regulating host populations.

Resumo Helmintos parasitos tem sido estudados como acumuladores potenciais para diferentes poluentes. O trematódeo E. paraensei tem como hospedeiro vertebrado o roedor Nectomys squamipes naturalmente exposto a pesticidas no meio ambiente. No entanto, pouca informação está disponível sobre os efeitos dos pesticidas em helmintos parasitos. O presente estudo investigou, em condições experimentais, os efeitos morfológicos no trematódeo E. paraensei após a exposição ao herbicida Roundup®, em concentrações abaixo das recomendadas para a utilização agrícola. A microscopia eletrônica de varredura (MEV) mostrou após duas horas de exposição, alterações no tegumento, como enrugamento, contração e descamação com perda de espinhos no colar peristômico e análise histopatológica evidenciou células do ceco alteradas, as glândulas vitelínicas com vacúolos e mudanças estruturais nas camadas musculares. Diminuição do conteúdo glicídico, principalmente no tecido conjuntivo, também foi observado. Considerando a predisposição à exposição a pesticidas agrícolas de N. squamipes infectado por E. paraensei, são necessários estudos para avaliar o impacto de tais resíduos frente aos helmintos e seus hospedeiros.

Thidiazuron (TDZ) increases fruit set and yield of 'Hosui' and 'Packham's Triumph' pear trees

ABSTRACT The low fruit set is one of the main factors leading to poor yield of pear orchards in Brazil. The exogenous application of thidiazuron (TDZ) and aminoethoxyvinilglycine (AVG) has shown promising results in some pear cultivars and other temperate fruit trees. The objective of this study was to evaluate the effect of TDZ and AVG on fruit set, yield, and fruit quality of 'Hosui' and 'Packham's Triumph' pears. The study was performed in a commercial orchard located in São Joaquim, SC. Plant material consisted of 'Hosui' and 'Packham's Triumph' pear trees grafted on Pyrus calleryana. Treatments consisted on different rates of TDZ (0 mg L-1, 20 mg L-1, 40 mg L-1 and 60 mg L-1) sprayed at full bloom for both cultivars. An additional treatment of AVG 60 mg L-1 was sprayed one week after full bloom in 'Hosui'. The fruit set, number of fruit per tree, yield, fruit weight, seed number, and fruit quality attributes were assessed. Fruit set and yield of both cultivars are consistently increased by TDZ, within the rates of 20 to 60 mg L-1. Besides, its application increased fruit size of 'Hosui' and did not negatively affect fruit quality attributes of both cultivars.

Assessment of growth of Aspergillus spp. from agricultural soils in the presence of glyphosate / Evaluación del crecimiento de Aspergillus spp. aislados de suelos agrícolas en presencia de glifosato

Agriculture is one of the bases of the Argentine economy. Glyphosate is undoubtedly one of the most important herbicides used. The increasing consumption and the efficiency of glyphosate-based herbicides have encouraged several studies on their persistence in soils, their effects on soil microbiota and their degradation processes. Fungi have been reported as being the main herbicide-degrading microorganisms as well as the most tolerant to environmental stress conditions. This study evaluated the growth performance of Aspergillus section Flavi and Aspergillus niger aggregate strains on Czapek Dox media supplied with a commercial glyphosate formulation as sole source of carbon (CZC), phosphorus (CZP) or nitrogen (CZN). Six Aspergillus spp. strains were evaluated. Each medium was stab-inoculated with fungal spores from 7-day old cultures. Two measures of colony radii were taken daily. All of the Aspergillus section Flavi strains showed a significant increase (from 24 to 44%) in growth rate on the CZN medium, as compared to controls. The A. niger aggregate strains exhibited the same behavioral pattern under all the conditions tested, except on the CZN medium. Velutinous or slightly floccose colonies with abundant sporulation were observed on CZP. Moreover, the colonies produced sparse sporulation on CZC or CZN media, being their appearances completely different from those on the CZP medium. This study establishes that A. section Flavi and A. niger aggregate strains can grow in vitro in the presence of glyphosate, especially when it is used as a sole source of phosphorus or nitrogen.

La agricultura es una de las bases de la economía argentina. El glifosato es, indudablemente, uno de los herbicidas más utilizados. El incremento en el consumo y la eficiencia de estos herbicidas han estimulado investigaciones sobre su persistencia en el suelo, sus efectos sobre la microbiota edáfica y su proceso de degradación. Los hongos son conocidos por su importancia como degradadores de herbicidas y por su tolerancia a condiciones de estrés ambiental. El objetivo del presente estudio fue evaluar el crecimiento de cepas de Aspergillus sección Flavi y del agregado Aspergillus niger en el medio Czapeck Dox suplementado con una solución comercial de glifosato como única fuente de carbono (CZC), de fósforo (CZP) o de nitrógeno (CZN). Seis cepas de Aspergillus fueron evaluadas. Cada medio fue inoculado con esporas fúngicas provenientes de cultivos de 7 días. Diariamente se tomaron 2 medidas del radio de las colonias. Todas las cepas de Aspergillus sección Flavi mostraron un incremento significativo (del 24 al 44%) en la velocidad de crecimiento en el medio CZN comparado con los controles. Las cepas del agregado A. niger mostraron el mismo comportamiento bajo todas las condiciones ensayadas, excepto en el medio CZN. Se observaron colonias velutinosas o ligeramente flocosas con abundante esporulación en los medios CZP. En los medios CZC o CZN, las colonias produjeron una esporulación escasa. Este estudio establece que cepas de Aspergillus sección Flavi y del agregado A. niger pueden desarrollar in vitro en presencia de glifosato, especialmente cuando es usado como única fuente de fósforo o nitrógeno.

Asn336 is involved in the substrate affinity of glycine oxidase from Bacillus cereus

Background: Glycine oxidase (GO), a type of D-amino acid oxidase, is of biotechnological interest for its potential in several fields. In our previous study, we have characterized a new glycine oxidase (BceGO) from Bacillus cereus HYC-7. Here, a variant of N336K with increased the affinity against all the tested substrate was obtained by screening a random mutant library of BceGO. It is observed that the residue N336 is invariable between its homogeneous enzymes. This work was aimed to explore the role of the residue N336 in glycine oxidase by site-directed mutagenesis, kinetic assay, structure modeling and substrate docking. Results: The results showed that the affinity of N336H, N336K and N336R increased gradually toward all the substrates, with increase in positive charge on side chain, while N336A and N336G have not shown a little significant effect on substrate affinity. The structure modeling studies indicated that the residue Asn336 is located in a random coil between -J-18 and a-10. Also, far-UV CD spectra-analysis showed that the mutations at Asn336 do not affect the secondary structure of enzyme. Conclusion: Asn336 site was located in a conserved GHYRNG loop which adjoining to substrate and the isoalloxazine ring of FAD, and involved in the substrate affinity of glycine oxidase. This might provide new insight into the structure-function relationship of GO, and valuable clue to redesign its substrate specificity for some biotechnological application.

Neutrophil elastase ameliorates matrix metalloproteinase-9 to promote lipopolysaccharide-induced acute lung injury in mice 1

ABSTRACT PURPOSE: To investigate the regulatory roles of neutrophil elastase (NE) and matrix metalloproteinase-9 (MMP-9) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS: To construct LPS-induced ALI mouse models, wild-type C57BL/6 mice were administered 5.0 mg/kg of LPS through endotracheal, and/or 1.0 mg/kg of ONO-5046, and/or 20.0 mg/kg of chemically modified tetracycline-3 (CMT-3) by gavage. The levels of MMP-9, tissue inhibitor of metalloprotease-1, interleukin (IL)-6 were detected by real time RT-PCR at 6 h, 24 h and 48 h, and tumor necrosis factor (TNF), lung wet-dry weight ratio, white blood cell (WBC) count and polymorphonuclear (PMN) count in bronchoalveolar lavage fluid (BALF) were tested at 48 h after administration. The 5-day survival analysis of the ALI mice was also performed. RESULTS: Both ONO-5046 and CMT-3, regardless of being used individually or combined, significantly reduced the levels of MMP-9, IL-6, and TNF in lung tissue as well as in BALF, and the WBC and PMN count in BALF. Combined treatment with ONO-5046 and CMT-3 remarkably improved the survival rate of ALI mice. CONCLUSION: Neutrophil elastase synergizes with matrix metalloproteinase-9 to promote and regulate the release of inflammatory mediators and the infiltration of inflammatory cells, consequently affecting the survival of lipopolysaccharide-induced acute lung injury mice.

The whole-cell immobilization of D-hydantoinase-engineered Escherichia coli for D-CpHPG biosynthesis

Background: D-Hydroxyphenylglycine is considered to be an important chiral molecular building-block of antibiotic reagents such as pesticides, and β-lactam antibiotics. The process of its production is catalyzed by D-hydantoinase and D-carbamoylase in a two-step enzyme reaction. How to enhance the catalytic potential of the two enzymes is valuable for industrial application. In this investigation, an Escherichia coli strain genetically engineered with D-hydantoinase was immobilized by calcium alginate with certain adjuncts to evaluate the optimal condition for the biosynthesis of D-carbamoyl-p-hydroxyphenylglycine (D-CpHPG), the compound further be converted to D-hydroxyphenylglycine (D-HPG) by carbamoylase. Results: The optimal medium to produce D-CpHPG by whole-cell immobilization was a modified Luria-Bertani (LB) added with 3.0% (W/V) alginate, 1.5% (W/V) diatomite, 0.05% (W/V) CaCl2 and 1.00 mM MnCl2.The optimized diameter of immobilized beads for the whole-cell biosynthesis here was 2.60 mm. The maximized production rates of D-CpHPG were up to 76%, and the immobilized beads could be reused for 12 batches. Conclusions: This investigation not only provides an effective procedure for biological production of D-CpHPG, but gives an insight into the whole-cell immobilization technology.

Acute toxicity and sublethal effects of the mixture glyphosate (Roundup® Active) and Cosmo-Flux®411F to anuran embryos and tadpoles of four Colombian species / Toxicidad aguda y efectos subletales de la mezcla glifosato (Roundup® Activo) y Cosmo-Flux®411F en embriones y renacuajos de cuatro especies de anuros colombianos

Glyphosate is the most widely used herbicide in the world with application in agriculture, forestry, industrial weed control, garden and aquatic environments. However, its use is highly controversial for the possible impact on not-target organisms, such as amphibians, which are vanishing at an alarming and rapid rate. Due to the high solubility in water and ionic nature, the glyphosate requires of surfactants to increase activity. In addition, for the control of coca (Erythroxylum coca) and agricultural weeds in Colombia, formulated glyphosate is mixed and sprayed with the adjuvant Cosmo-Flux®411F to increase the penetration and activity of the herbicide. This study evaluates the acute toxic and sublethal effects (embryonic development, tadpole body size, tadpole swimming performance) of the mixture of the formulated glyphosate Roundup® Active and Cosmo-Flux®411F to anuran embryos and tadpoles of four Colombian species under 96h laboratory standard tests and microcosms, which are more similar to field conditions as they include soil, sand and macrophytes. In the laboratory, embryos and tadpoles of Engystomops pustulosus were the most tolerant (LC50=3 904µg a.e./L; LC50=2 799µg a.e./L, respectively), while embryos and tadpoles of Hypsiboas crepitans (LC50=2 203µg a.e./L; LC50=1 424µg a.e./L, respectively) were the most sensitive. R. humboldti and R. marina presented an intermediate toxicity. Embryos were significantly more tolerant to the mixture than tadpoles, which could be likely attributed to the exclusion of chemicals by the embryonic membranes and the lack of organs, such as gills, which are sensitive to surfactants. Sublethal effects were observed for the tadpole body size, but not for the embryonic development and tadpole swimming performance. In microcosms, no toxicity (LC50 could not be estimated), or sublethal responses were observed at concentrations up to fourfold (14.76kg glyphosate a.e./ha) the highest field application rate of 3.69kg glyphosate a.e./ha. Thus, toxicity was less in the microcosms than in laboratory tests, which may be attributed to the presence of sediments and organic matter which rapidly adsorb glyphosate and surfactants such as POEA. It is concluded that the mixture of glyphosate (Roundup® Active) and Cosmo-Flux®411F, as used in the field, has a negligible toxic effect to embryos and tadpoles of the species tested in this study.

El glifosato es el herbicida más usado en el mundo con aplicaciones para la agricultura, control de malezas forestales, industriales, en jardines y ambientes acuáticos. Sin embargo, su uso es altamente controversial por el posible impacto sobre organismos no blanco, como los anfibios, los cuales están desapareciendo a una tasa alarmantemente rápida. Debido a su alta solubilidad en agua y naturaleza iónica, el glifosato requiere de surfactantes para incrementar su actividad. Además, para el control de Erythroxylum coca y de malezas en la agricultura en Colombia, el glifosato formulado es mezclado y rociado con el coadyuvante Cosmo-Flux®411F para incrementar la penetración y actividad del herbicida. Este estudio evalúa los efectos tóxicos agudos y subletales (desarrollo embrionario, tamaño corporal y desempeño natatorio de los renacuajos) de la mezcla del glifosato formulado Roundup® Activo con el Cosmo-Flux®411F en embriones y renacuajos de cuatro especies de anuros colombianos, bajo pruebas de 96h en condiciones estándar de laboratorio y microcosmos, que son más similares a las condiciones de campo al incluir tierra, arena y macrófitas. En laboratorio, los embriones y renacuajos de Engystomops pustulosus fueron los más tolerantes (CL50=3 904µg a.e./L; CL50=2 799µg a.e./L, respectivamente), mientras que los embriones y renacuajos de Hypsiboas crepitans fueron los más sensibles (CL50=2 203µg a.e./L; CL50=1 424µg a.e./L, respectivamente). R. humboldti y R. marina presentaron una toxicidad intermedia. Los embriones fueron más tolerantes a la mezcla que los renacuajos, lo cual podría ser atribuido a la exclusión de los químicos por las membranas embrionarias y a la falta de órganos, como las branquias, que son más sensibles a los surfactantes. Se observaron efectos subletales en el tamaño corporal de los renacuajos, pero no en el desarrollo embrionario ni el desempeño natatorio de los renacuajos. En microcosmos no se observaron efectos tóxicos ni respuestas subletales a concentraciones hasta cuatro veces (14.67kg glifosato a.e./ha) la tasa de aplicación más alta de 3.69kg glifosato a.e./ha. Por lo tanto, la toxicidad fue menor en los microcosmos que en las pruebas de laboratorio, lo que puede ser atribuido a la presencia de sedimentos y materia orgánica que absorbe rápidamente el glifosato y surfactantes como el POEA. Se concluye que la mezcla del glifosato (Roundup® Activo) y Cosmo-Flux®411F, como se aplica en campo, tiene un efecto tóxico bajo en los embriones y renacuajos de las especies estudiadas.

Conformational behavior of phenylglycines and hydroxyphenylglycines and non-planarity of phenyl rings.

The non-proteinogenic amino acids — phenylglycine (PG) and hydroxyphenylglycine (HPG) are crucial components of certain peptidic natural products and are important for the preparation of various medicines. In this, study, the conformation of model dipeptides Ac-X-NHMe of PG, p-HPG and 3, 5-di-hydroxyphenylglycine (3, 5-DHPG) was studied both in R and S form by quantum mechanical (QM) and molecular dynamics approaches. On the energy scale, the conformational states of these molecules in both the R and S were found to be degenerate by QM studies, stabilized by non-covalent interactions like carbonyl--carbonyl interactions, carbonyl-lp··π (aromatic ring) interactions etc. These interactions disappeared/weakened due to interaction of water molecules with carbonyl groups of backbone in simulation and water was found to interact with the aromatic ring through Ow-H··π or Owlp··π interactions. The degeneracy of conformational states was lifted in favor of R-form of PG and DHPG and water molecules interactions with aromatic ring led to non-planarity of the aromatic ring. In simulation studies, irrespective of the starting geometry, the , values for the R form correspond to inverse b/inverse collagen region and for the S-form, the , values correspond to b/collagen region i.e., adopt single conformation. The obtained results were in conformity with the CD spectroscopic data on D-PG and D-p-HPG. The conformational behavior of the unusual amino acids might be of great help in designing of bioactive peptides/peptide based drugs to be realized in single conformation – an essential requirement.

Influence of glyphosate in planktonic and biofilm growth of Pseudomonas aeruginosa

This study evaluated the impact of different concentrations of glyphosate (Rondup®) on planktonic and biofilm growth of P. aeruginosa. Aerobic and anaerobic cultures of P. aeruginosa ATCC®15442 inoculated in MHB + glyphosate (0.845 ppm, 1.690 ppm, 8.45 ppm, 16.90 ppm, 84.50 ppm, 169 ppm, 845 ppm, and 1690 ppm) and cultured in normoxia and anoxia, following their OD560nm every hour for 24 h. Biofilms of adapted cells were formed in the presence of glyphosate (0.845 to 1690 ppm) in normoxia and anoxia for 36 h. Glyphosate at concentrations higher than 84.5 ppm reduces the cell density of planktonic aerobic cultures (p < 0.05). However, these same concentrations favor the planktonic anaerobic growth (p < 0.05). On the other hand, the herbicide favors a slight growth of biofilms in a concentration-dependent manner up to 84.5 ppm (p > 0.05), and more pronounced over 169 ppm. Anaerobic biofilms have their growth more readily favored (p < 0.05), regardless of concentration. In a concentration-dependent manner, glyphosate interferes with the growth ability of P. aeruginosa ATCC®15442.

Phase 1 dose escalation study of rigosertib by 2-, 4-, or 8-hour infusion twice-weekly in patients with advanced cancer.

CONTEXT: Rigosertib, a potent, multi-kinase inhibitor that selectively induces mitotic arrest and apoptosis in cancer cells and is non-toxic to normal cells, is being developed for the treatment of solid tumors and hematological malignancies. AIMS: To determine the safety, doselimiting toxicities, and clinical activity of rigosertib administered by 2-, 4-, or 8-hour continuous IV infusion twice-a-week for 3 weeks out of a 4-week cycle in patients with advanced solid tumor or hematological malignancies; and to confirm the safety and tolerability of the recommended phase 2 dose (RPTD). SETTINGS AND DESIGN: Phase 1, open-label, dose-escalation study in men and women ≥18 years of age. MATERIALS AND METHODS: An escalation phase optimized the duration of infusion (2, 4, or 8 hours) of 3200 mg rigosertib twice-a-week for 3 weeks of a 4-week cycle; an expansion phase confirmed the maximum tolerated dose (MTD). STATISTICAL ANALYSIS USED: All data summaries were descriptive. PK parameters were estimated using compartmental analysis. RESULTS: 25 patients (16 male, 9 female, 26- 66 years, all Asian) were treated with rigosertib, 16 in the escalation phase; 9 in the expansion phase. MTD was determined to be 3200 mg as a 4-hour infusion and 2400 mg over 4 hours was declared to be the RPTD. Best response was stable disease in 5 of 14 evaluable patients, with a mean (range) of 90 (43-108) days. CONCLUSIONS: 2400 mg rigosertib as a 4-hour infusion was identified as the RPTD. Five patients achieved stable disease lasting 6-16 weeks.