Characterization of QoI-Fungicide Resistance in Cercospora Isolates Associated with Cercospora Leaf Blight of Soybean from the Southern United States.

Cercospora leaf blight (CLB) of soybean, caused by Cercospora cf. flagellaris, C. kikuchii, and C. cf. sigesbeckiae, is an economically important disease in the southern United States. Cultivar resistance to CLB is inconsistent; therefore, fungicides in the quinone outside inhibitor (QoI) class have been relied on to manage the disease. Approximately 620 isolates from plants exhibiting CLB were collected between 2018 and 2021 from 19 locations in eight southern states. A novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay based on two genes, calmodulin and histone h3, was developed to differentiate between the dominant species of Cercospora, C. cf. flagellaris, and C. cf. sigesbeckiae. A multilocus phylogenetic analysis of actin, calmodulin, histone h3, ITS rDNA, and transcription elongation factor 1-α was used to confirm PCR-RFLP results and identify remaining isolates. Approximately 80% of the isolates collected were identified as C. cf. flagellaris, while 15% classified as C. cf. sigesbeckiae, 2% as C. kikuchii, and 3% as previously unreported Cercospora species associated with CLB in the United States. PCR-RFLP of cytochrome b (cytb) identified QoI-resistance conferred by the G143A substitution. Approximately 64 to 83% of isolates were determined to be QoI-resistant, and all contained the G143A substitution. Results of discriminatory dose assays using azoxystrobin (1 ppm) were 100% consistent with PCR-RFLP results. To our knowledge, this constitutes the first report of QoI resistance in CLB pathogen populations from Alabama, Arkansas, Kentucky, Mississippi, Missouri, Tennessee, and Texas. In areas where high frequencies of resistance have been identified, QoI fungicides should be avoided, and fungicide products with alternative modes-of-action should be utilized in the absence of CLB-resistant soybean cultivars.

Silicon Alleviates Changes in the Source-Sink Relationship of Wheat Plants Infected by Pyricularia oryzae.

Blast, caused by Pyricularia oryzae, has become a devastating disease on wheat in several countries worldwide. Growers need alternative methods for blast management, and silicon (Si) stands out for its potential to decrease the intensity of important diseases in several crops. This study investigated the effect of Si on improving photoassimilate production on flag leaves of wheat plants and their partitioning to spikes in a scenario where blast symptoms decreased as a result of potentiation of defense mechanisms by Si. Wheat plants (cultivar BRS Guamirim) were grown in hydroponic culture with 0 or 2 mM Si and inoculated with P. oryzae at 10 days after anthesis. The Si concentration on flag leaves and spikes of Si-supplied plants increased and resulted in lower blast symptoms. High concentrations of total soluble phenols and lignin-thioglycolic acid derivatives and greater peroxidase, polyphenoloxidase, phenylalanine ammonia-lyase, ß-1,3-glucanase, and chitinase activity occurred on flag leaves and spikes of Si-supplied plants and increased their resistance to blast. The concentration of photosynthetic pigments decreased and the photosynthetic performance of infected flag leaves and spikes from plants not supplied with Si was impaired for chlorophyll a fluorescence parameters including maximal photosystem II quantum efficiency, fraction of energy absorbed used in photochemistry, quantum yield of nonregulated energy dissipation, and quantum yield of regulated energy dissipation. The concentration of soluble sugars was lower on infected flag leaves and spikes from plants not supplied with Si, whereas the hexose-to-sucrose ratio increased on infected flag leaves. Sucrose-phosphate synthase activity was lower and acid invertase activity was higher on flag leaves and spikes of plants not supplied with Si, respectively, compared with Si-supplied plants. The starch concentration on spikes of Si-supplied plants increased. In conclusion, Si showed a beneficial effect in improving the source-sink relationship of infected flag leaves and spikes by preserving alterations in assimilate production and partitioning during the grain filling process.

Quantitative intact proteomics investigations of alveolar macrophages in sarcoidosis.

Alveolar macrophages are important for granuloma formation, which is the histological hallmark in sarcoidosis. Their function as antigen-presenting cells in sarcoidosis is also believed to be relevant to the outcome of disease, resulting either in remission or a prolonged chronic inflammation in the lungs. Our aim was to study alterations in the expression levels of the soluble proteome of alveolar macrophages in pulmonary sarcoidosis as compared with healthy controls, with the goal of identifying specific proteins and pathways important for the mechanisms of disease and/or disease phenotype. Quantitative proteomics approach using two-dimensional difference gel electrophoresis coupled to mass spectrometry was applied. Data was evaluated using multivariate modelling and pathway analyses. 69 protein spots were found to be significantly altered between sarcoidosis and healthy controls. Among these, 25 unique proteins were identified. Several of the identified proteins were related to key alveolar macrophage functionality, including the Fcγ-mediated phagocytosis and clathrin-mediated endocytosis pathways. Global proteomics analysis provided identification of alterations of a subset of proteins not previously reported in sarcoidosis. These alterations primarily affect biological pathways related to phagocytic macrophage functionality. These findings provide important insights into the role of macrophages in sarcoidosis pathogenesis.

[Copper intoxication decreases lifespan and induces neurologic alterations in Drosophila melanogaster]. / La intoxicación con cobre disminuye la sobrevida e induce alteraciones neurológicas en Drosophila melanogaster.

Wilson disease is a hereditary disorder caused by mutations of the ATP7B gene, which leads to intoxication with copper as a result of an unbalance of copper homeostasis. The clinical manifestations resulting from this intoxication are related to the affectation of liver and the encephalon in most cases. Several animal models are currently available for the study of the malady. However, in such models no neurological symptoms are observed, which limits their use for the study of pathogenic effects of this disease on the central nervous system. The aim of the present study was to evaluate if copper feeding could induce a disease state in Drosophila melanogaster to model Wilson disease. The effect of the feeding of copper at the doses of 31 microM and 47 microM on the survival was initially evaluated. Next, behavioral experiments were conducted to determine whether the motor performance was altered by the 47 microM concentration. The results suggest that copper treatment decreases the viability of the flies. In addition, the decrease of viability was associated to an increase and decrease of spontaneous motor activity at early and late stages of the intoxication, respectively. Finally, the role of the dopaminergic neurotransmission system on the observed motor alterations was evaluated. The dopamine precursor L-dopa increased motor activity. In contrast, D2 receptor antagonist, Fluphenazine, was able to block both the increase and decrease of motor activity scores induced by copper. These results suggest that Drosophila melanogaster could be used as a model organism for the study of possible interventions with potential neuroprotective effects in Wilson disease.

In the eye of the beholder: does the master see the SameSpots as the novice?

Historically, the use of two-dimensional electrophoresis (2-DE) in quantitative proteomics has been hampered by significant technical variance. Over the past decade, a range of technological leaps have reduced the overall variance of 2-DE, thus turning the technology into a robust platform for quantitative intact proteomics. However, as the confounding gel-to-gel variation improves, the variance arising from the subsequent image analysis becomes more prominent. Limitations in image alignment and spot detection of previous generations of 2-DE analysis software have demanded considerable user-intervention and manual editing, resulting in introduction of a large degree of subjectivity and software-induced variance. We evaluated the performance of SameSpots, representing a new generation of 2-DE image analysis software, using both DIGE and traditional single-stain 2-DE approaches. Evaluations of the software-induced variance in relation to other sources of variance, as well as the subjectivity through comparison of analyses performed by an expert user and a novice lab-user, were performed. In terms of statistical power, the less-experienced user achieved the better results, but no discernible difference was detected in multivariate comparisons between the users. In conclusion, we found that SameSpots represents improvements both in reproducibility and objectivity in relation to previous generations of 2-DE analysis software.

A quantitative proteomic analysis of soluble bronchoalveolar fluid proteins from patients with sarcoidosis and chronic beryllium disease.

BACKGROUND: Sarcoidosis and chronic beryllium disease (CBD) are granulomatous disorders which can lead to development of chronic inflammation and fibrosis. These diseases have several similarities from their clinical aspects. The aim of this study was to compare the protein profile at the site of active inflammation i.e. the lungs of patients with sarcoidosis and CBD. METHODS: Bronchoalveolar lavage (BAL) proteins from patients having sarcoidosis or CBD were studied using two dimensional gel based proteomics. In this study, we used Difference Gel Electrophoresis (DIGE) proteomics approach to analyse the protein expression profiles from sarcoidosis patients (n=4), CBD (n=4) and healthy controls (n = 5). Subsequently, differentially expressed proteins were identified by using mass spectrometry. RESULTS: We found 37 protein-spots with statistically different expression levels, and identified 14 of these proteins. The protein expression levels of peroxiredoxin 5, heat shock protein 70, complement C3, annexin A2 and transthyretin were significantly different in sarcoidosis versus control group. The proteins; hemopexin, beta2-microglobulin, alpha-1 antitrypsin, cystatin B, IgG kappa chain, apolipoprotein A1, and albumin were significantly different between CBD versus control group. When comparing CBD versus sarcoidosis, we found superoxide dismutase and hemoglobin upregulated in the CBD group. CONCLUSION: By using quantitative proteomics, we were able to find proteins with different expression levels in both diseases.

[Not Available]. / Composición corporal de niños de entre 8 y 10 años de edad, con y sin antecedente de bajo peso al nacer, residentes en el estado de Querétaro, México.

Introducción: el bajo peso al nacer (BPN) es un factor de riesgo para desarrollar obesidad en la vida adulta.Objetivo: evaluar diferencias en la composición corporal de niños de entre 8 y 10 años de edad con y sin antecedente de BPN.Métodos: fue un estudio observacional, transversal comparativo. Participaron 112 niños (95 con adecuado peso al nacer [APN] y 17 con BPN). Se realizó antropometría (peso, talla, circunferencias de cintura y cadera, pliegue cutáneo de tríceps [PCT] y subescapular [PCSE]).Resultados: se encontró una prevalencia combinada del 41% para sobrepeso y obesidad en ambos grupos de estudio. El porcentaje de grasa corporal total fue menor en las niñas con BPN (no significativo); sin embargo, el indicador PCT-PCSE fue significativamente más alto (p = 0,04) que el de las niñas con APN. En contra de lo esperado, al estratificar según porcentaje de grasa y peso al nacer, se encontró que el grupo con BPN presentó un porcentaje de grasa bajo (p < 0,05) en comparación con el grupo de APN, siendo 6 veces mayor la posibilidad de que un niño con BPN presente porcentaje de grasa total bajo a esta edad.Conclusiones: a estas edades no se encontró mayor porcentaje de grasa en el grupo con BPN en comparación con el de APN; sin embargo, las niñas con BPN presentaron mayor deposición de grasa troncal que las de APN. La deposición de grasa es un indicador que hay que considerar, y no únicamente el índice de masa corporal, en la evaluación nutricia infantil.

Alterations in the membrane-associated proteome fraction of alveolar macrophages in sarcoidosis.

BACKGROUND: Alveolar macrophages are implicated in the pathogenesis of lung sarcoidosis. Their interaction with T-cells leads to an inflammatory response that may either resolve within 2 years, or become chronic with an increased risk to develop lung fibrosis. OBJECTIVE: To perform quantitative profiling of the membrane-associated proteome of alveolar macrophages in sarcoidosis patients and healthy individuals to identify specific proteins and pathways involved in sarcoidosis pathology. METHODS: Differential proteomic analysis was performed on iTRAQ (isobaric Tag for Relative and Absolute Quantitation) labeled samples using tandem mass spectrometry. Subsequently, uni- and multivariate statistical analyses and pathway- and network analyses were performed. RESULTS: Eighty proteins were differentially expressed between healthy and sarcoidosis patients. Down-stream pathway analysis confirmed our recent reports of up-regulation of two phagocytotic pathways: Fcγ receptor-mediated phagocytosis and clathrin-mediated endocytosis signaling. An additional pathway, pyruvate metabolism, was found to be up-regulated in sarcoidosis patients. The oxidative phosphorylation pathway was differentially expressed in subgroups of sarcoidosis, with up-regulation in Löfgren's patients and down-regulation in non-Löfgren's patients. CONCLUSION: This unprecedented proteome profiling of the membrane-associated fraction of alveolar macrophages confirmed previous findings of alterations in phagocytotic pathways due to sarcoidosis, as well as indicated a differential dysregulation of the oxidative phosphorylation pathway related to disease outcome in sarcoidosis.

Clinical, endoscopic and manometric features of the primary motor disorders of the esophagus.

BACKGROUND: Significant incidence, diagnostic difficulties, clinical relevance and therapeutic efficacy associated with the small number of publications on the primary esophageal motor disorders, motivated the present study. AIM: To determine the manometric prevalence of these disorders and correlate them to the endoscopic and clinical findings. METHODS: A retrospective study of 2614 patients, being 1529 (58.49%) women and 1085 (41.51%) men. From 299 manometric examinations diagnosed with primary esophageal motor disorder, were sought-clinical data (heartburn, regurgitation, dysphagia, odynophagia, non-cardiac chest pain, pharyngeal globe and extra-esophageal symptoms) and/or endoscopic (hiatal hernia, erosive esophagitis, food waste) that motivated the performance of manometry. RESULTS: Were found 49 cases of achalasia, 73 diffuse spasm, 89 nutcracker esophagus, 82 ineffective esophageal motility, and six lower esophageal sphincter hypertension. In relation to the correlations, it was observed that in 119 patients clinical conditions were associated with dysphagia, found in achalasia more than in other conditions; in relationship between endoscopic findings and clinical conditions there was no statistical significance between data. CONCLUSION: The clinical and endoscopic findings have little value in the characterization of the primary motor disorders of the esophagus, showing even more the need for manometry, particularly in the preoperative period of gastroesophageal reflux disease.

[Leptin]. / Leptina.

Leptin is a hormone secreted by adipocytes, stomach and placenta. Age, sex, puberty onset and food intake are the most important physiological factors that determines leptin concentration. It has been shown that leptin secretion is regulated by other hormones such as insulin, glucocorticoids and sex steroids. Leptin has an important role in hungry and satiety regulation as well as in normal sexual maturation. There have been some studies in obese subjects showing promising results with leptin administration.

Clinical, endoscopic and manometric features of the primary motor disorders of the esophagus / Aspectos clínicos, manométricos e endoscópicos dos distúrbios motores primários do esôfago

BACKGROUND: Significant incidence, diagnostic difficulties, clinical relevance and therapeutic efficacy associated with the small number of publications on the primary esophageal motor disorders, motivated the present study. AIM: To determine the manometric prevalence of these disorders and correlate them to the endoscopic and clinical findings. METHODS: A retrospective study of 2614 patients, being 1529 (58.49%) women and 1085 (41.51%) men. From 299 manometric examinations diagnosed with primary esophageal motor disorder, were sought-clinical data (heartburn, regurgitation, dysphagia, odynophagia, non-cardiac chest pain, pharyngeal globe and extra-esophageal symptoms) and/or endoscopic (hiatal hernia, erosive esophagitis, food waste) that motivated the performance of manometry. RESULTS: Were found 49 cases of achalasia, 73 diffuse spasm, 89 nutcracker esophagus, 82 ineffective esophageal motility, and six lower esophageal sphincter hypertension. In relation to the correlations, it was observed that in 119 patients clinical conditions were associated with dysphagia, found in achalasia more than in other conditions; in relationship between endoscopic findings and clinical conditions there was no statistical significance between data. CONCLUSION: The clinical and endoscopic findings have little value in the characterization of the primary motor disorders of the esophagus, showing even more the need for manometry, particularly in the preoperative period of gastroesophageal reflux disease. .

RACIONAL: A incidência significante, a dificuldade diagnóstica, a relevância clínica e a eficácia terapêutica associada ao pequeno número de publicações sobre os distúrbios motores primários do esôfago, motivou a realização do presente estudo. OBJETIVO: Verificar a prevalência desses distúrbios em manometrias e relacioná-las aos achados endoscópicos e clínicos. MÉTODOS: Estudo retrospectivo de 2614 pacientes sendo 1529 (58,49%) do gênero feminino e 1085 (41,51%) do masculino. A partir de 299 exames manométricos com diagnóstico de distúrbio motor esofagiano primário, procuraram-se os dados clínicos (pirose, regurgitação, disfagia, odinofagia, dor torácica não cardíaca, globo faríngeo e sintomas extra-esofageanos) e/ou endoscópicos (hérnia de hiato, esofagite erosiva, resíduos alimentares) que motivaram a realização da manometria. RESULTADOS: Foram encontrados 49 casos de acalásia, 73 de espasmo difuso, 89 de esôfago em quebra-nozes, 82 de motilidade esofagiana ineficaz, e seis de esfíncter esofagiano inferior hipertensivo. Em relação às correlações, observou-se em 119 pacientes analisados que, na clínica associada às afecções, a disfagia foi mais encontrada na acalásia do que nas outras afecções; na relação entre os achados endoscópicos e as afecções não houve relevância estatística entre os dados. CONCLUSÃO: Os achados clínicos e endoscópicos têm pequeno valor na caracterização das doenças motoras primárias do esôfago, evidenciando ainda mais a necessidade da manometria, particularmente no pré-operatório da doença do refluxo gastroesofágico. .

La intoxicación con cobre disminuye la sobrevida e induce alteraciones neurológicas en Drosophila melanogaster / Copper intoxication decreases lifespan and induces neurologic alterations in Drosophila melanogaster

La enfermedad de Wilson, es un trastorno hereditario autosómico recesivo causado por mutaciones del gen de la trifosfatasa de adenosina (ATP7B). Dicha mutación ocasiona intoxicación con cobre, generando manifestaciones clínicas por los efectos tóxicos del metal, principalmente a nivel del hígado y el encéfalo. Recientemente se han desarrollado modelos genéticos de la enfermedad para su estudio clínico. Sin embargo, la utilidad de los mismos es limitada por el hecho de que en tales modelos no se observan manifestaciones neurológicas. El presente estudio tuvo como objetivo desarrollar un modelo de la enfermedad de Wilson en Drosophila melanogaster. Inicialmente se evaluó el efecto de la suplementación con concentraciones de 31 µM y 47 µM de cobre en la sobrevida. Posteriormente se realizaron estudios de conducta para determinar si existían alteraciones en el desempeño motor asociadas al tratamiento con la dosis de 47 µM de cobre. Los resultados obtenidos sugieren que el tratamiento con cobre disminuye la viabilidad de la Drosophila. La disminución de la sobrevida estuvo asociada a un aumento y una disminución de los registros de actividad motora en las etapas tempranas y tardías de la intoxicación respectivamente. Por último, se evaluó el papel del sistema de neurotransmisión dopaminérgico sobre las alteraciones conductuales inducidas por el cobre. El tratamiento con el precursor de la dopamina, L-dopa, indujo un aumento de la actividad motora similar al inducido por el cobre. Por el contrario, el tratamiento con Flufenazina, un antagonista de los receptores dopaminérgicos D2, fue capaz de impedir las alteraciones conductuales en todas las edades evaluadas. Estos resultados sugieren que la Drosophila melanogaster podría ser empleada como modelo para el estudio de posibles intervenciones con potencial terapéutico en la enfermedad de Wilson.

Wilson disease is a hereditary disorder caused by mutations of the ATP7B gene, which leads to intoxication with copper as a result of an unbalance of copper homeostasis. The clinical manifestations resulting from this intoxication are related to the affectation of liver and the encephalon in most cases. Several animal models are currently available for the study of the malady. However, in such models no neurological symptoms are observed, which limits their use for the study of pathogenic effects of this disease on the central nervous system. The aim of the present study was to evaluate if copper feeding could induce a disease state in Drosophila melanogaster to model Wilson disease. The effect of the feeding of copper at the doses of 31 µM and 47 µM on the survival was initially evaluated. Next, behavioral experiments were conducted to determine whether the motor performance was altered by the 47 µM concentration. The results suggest that copper treatment decreases the viability of the flies. In addition, the decrease of viability was associated to an increase and decrease of spontaneous motor activity at early and late stages of the intoxication, respectively. Finally, the role of the dopaminergic neurotransmission system on the observed motor alterations was evaluated. The dopamine precursor L-dopa increased motor activity. In contrast, D2 receptor antagonist, Fluphenazine, was able to block both the increase and decrease of motor activity scores induced by copper. These results suggest that Drosophila melanogaster could be used as a model organism for the study of possible interventions with potential neuroprotective effects in Wilson disease.

Leptina / Leptin

Leptin is a hormone secreted by adipocytes, stomach and placenta. Age, sex, puberty onset and food intake are the most important physiological factors that determines leptin concentration. It has been shown that leptin secretion is regulated by other hormones such as insulin, glucocorticoids and sex steroids. Leptin has an important role in hungry and satiety regulation as well as in normal sexual maturation. There have been some studies in obese subjects showing promising results with leptin administration.