24 research outputs found
Post-intervention Status in Patients With Refractory Myasthenia Gravis Treated With Eculizumab During REGAIN and Its Open-Label Extension
OBJECTIVE: To evaluate whether eculizumab helps patients with anti-acetylcholine receptor-positive (AChR+) refractory generalized myasthenia gravis (gMG) achieve the Myasthenia Gravis Foundation of America (MGFA) post-intervention status of minimal manifestations (MM), we assessed patients' status throughout REGAIN (Safety and Efficacy of Eculizumab in AChR+ Refractory Generalized Myasthenia Gravis) and its open-label extension. METHODS: Patients who completed the REGAIN randomized controlled trial and continued into the open-label extension were included in this tertiary endpoint analysis. Patients were assessed for the MGFA post-intervention status of improved, unchanged, worse, MM, and pharmacologic remission at defined time points during REGAIN and through week 130 of the open-label study. RESULTS: A total of 117 patients completed REGAIN and continued into the open-label study (eculizumab/eculizumab: 56; placebo/eculizumab: 61). At week 26 of REGAIN, more eculizumab-treated patients than placebo-treated patients achieved a status of improved (60.7% vs 41.7%) or MM (25.0% vs 13.3%; common OR: 2.3; 95% CI: 1.1-4.5). After 130 weeks of eculizumab treatment, 88.0% of patients achieved improved status and 57.3% of patients achieved MM status. The safety profile of eculizumab was consistent with its known profile and no new safety signals were detected. CONCLUSION: Eculizumab led to rapid and sustained achievement of MM in patients with AChR+ refractory gMG. These findings support the use of eculizumab in this previously difficult-to-treat patient population. CLINICALTRIALSGOV IDENTIFIER: REGAIN, NCT01997229; REGAIN open-label extension, NCT02301624. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that, after 26 weeks of eculizumab treatment, 25.0% of adults with AChR+ refractory gMG achieved MM, compared with 13.3% who received placebo
Minimal Symptom Expression' in Patients With Acetylcholine Receptor Antibody-Positive Refractory Generalized Myasthenia Gravis Treated With Eculizumab
The efficacy and tolerability of eculizumab were assessed in REGAIN, a 26-week, phase 3, randomized, double-blind, placebo-controlled study in anti-acetylcholine receptor antibody-positive (AChR+) refractory generalized myasthenia gravis (gMG), and its open-label extension
Global urban environmental change drives adaptation in white clover
Urbanization transforms environments in ways that alter biological evolution. We examined whether urban environmental change drives parallel evolution by sampling 110,019 white clover plants from 6169 populations in 160 cities globally. Plants were assayed for a Mendelian antiherbivore defense that also affects tolerance to abiotic stressors. Urban-rural gradients were associated with the evolution of clines in defense in 47% of cities throughout the world. Variation in the strength of clines was explained by environmental changes in drought stress and vegetation cover that varied among cities. Sequencing 2074 genomes from 26 cities revealed that the evolution of urban-rural clines was best explained by adaptive evolution, but the degree of parallel adaptation varied among cities. Our results demonstrate that urbanization leads to adaptation at a global scale
Growth and survival of recombinant Rhodococcus sp. isolate RHA1 (fcb) in commercial peat and in PCB contaminated soil
O grupo de organoclorados, Bifenilas Policloradas (PCBs) é de difícil degradação e persistente no meio ambiente, tendo sido associado a diversos problemas nos organismos devido ao potencial toxicológico. Biodegradação constitui uma ferramenta eficaz e barata para remoção destes contaminantes do ambiente. O isolado RHA1 (fcb) de Rhodococcus sp. foi geneticamente construído com a introdução do operon de degradação hidrolítica de 4-clorobenzoato (fcb) para evitar a formação de produtos tóxicos durante a degradação de ácidos clorobenzóicos. Com o intuito de se obter informações sobre o processo adaptativo do recombinante Rhodococcus sp. isolado RHA1 (fcb) em substratos contendo PCBs, foram feitos dois ensaios avaliando-se a sobrevivência e o crescimento deste isolado. Rhodococcus sp. isolado RHA1 (fcb) foi inoculado (104 células.g-1) em substrato turfoso previamente irradiado a 50 KGy, contendo ou não 200 mg.Kg-1 de bifenilo. Em outro ensaio, além do recombinante, as bactérias Escherichia coli e Arthrobacter sp. foram inoculados em sedimento coletado na região do Estuário de Santos, contendo PHAs e PCBs, também irradiado (50 KGy). O crescimento das bactérias em ambos os substratos foi monitorado através de contagem de Unidades Formadoras de Colônias (UFCs). Algumas colônias eram selecionadas aleatoriamente para extração de DNA, detecção do operon fcb através de amplificação por PCR e sequenciamento do gene 16S rRNA. Aumento no número de UFCs nos tratamentos inoculados com o recombinante foi observado até 150 dias no ensaio com substrato turfoso e 70 dias na amostra ambiental. Entretanto, houve queda no número de UFCs após os 10 dias nos tratamentos inoculados com E. coli e Arthrobacter sp. Os genes fcbA e fcbB do operon fcb foram detectados nas colônias isoladas dos tratamentos inoculados com o isolado RHA1 (fcb) em ambos os substratos. A análise das seqüências pertencentes às colônias isoladas do tratamento inoculado com o isolado RHA1 (fcb) feita através de BLAST nos sites do NCBI e Ribossomal Database Project, apresentou 99% de identidade com a seqüência do gene ribossomal 16S de Rhodococcus sp. isolado ZC3 (AM076672.1). Somente as seqüências referentes ao tratamento inoculado com E. coli foram analisadas, as quais apresentaram 99% de identidade com a seqüência do gene ribossomal 16S de E. coli isolado K-12 MG 1655 (U00096.2). Estes resultados sugerem que Rhodococcus sp. isolado RHA1 (fcb) cresce na turfa irradiada (até 150 dias) na presença e ausência de PCB e nesta amostra de sedimento irradiada (até 70 dias), com aparente estabilidade do operon fcb durante este período e nestas condições. A possível presença dos genes fcbB e fcbA em bactérias nativas crescidas em meio K1 com ácido 4- clorobenzóico isoladas do sedimento antes da irradiação, sugere a presença de bactérias do local com potencial biodegradador deste composto.The group of organochlorates Biphenyl Polychlorates (PCBs) is of difficult degradation and persistent in the environment, being associated to several problems in the organisms due to its toxicological potential. The isolate RHA1 (fcb) from Rhodococcus sp. was genetically built with the introduction of the operon of hydrolytic degradation 4-chlorobenzoate (fcb) to avoid the formation of toxic products during the degradation of chlorobenzoic acids. In order to obtain information about the adaptative process of the recombinant Rhodococcus sp. isolate RHA1 (fcb) in substrates containing PCBs, two essays were made evaluating the survival and growth of this isolate. Rhodococcus sp. isolate RHA1 (fcb) was inoculated (104 cells.g-1) in peat substrate previously irradiated with 50 kGy, with and without 200 mg.kg-1 of biphenyl. In another essay, besides of the recombinant, the bacteria Escherichia coli and Arthrobacter sp. were inoculated in soil, also irradiated (50 kGy), from the Estuário de Santos region containing PHAs and PCBs. The growth of the bacteria in both substrates was monitorated counting the Colony Forming Units (CFUs). Some colonies were selected randomly for DNA extraction, fcb operon detection through PCR, and sequencing of the 16S rRNA gene. Rising in the number of CFUs in the recombinant inoculated treatments was observed until 150 days in the essay with peat substrate, and until 70 days in the environmental sample. Nonetheless, there was a reduction in the number of CFUs after 10 days in the treatment inoculated with E. coli and Arthrobacter sp. The genes fcbA and fcbB from the operon fcb were detected in the isolated colonies of the treatments inoculated with the isolate RHA1 (fcb) in both substrates. The analysis of the sequences belonging to the colonies isolated from the treatment inoculated with the isolate RHA1 (fcb) through BLAST in the NCBI and Ribosomal Database Project sites showed 99% identity with the sequence of the gene 16S ribosomal from Rhodococcus sp. isolate ZC-3 (AM076672.1). Only the sequences referring to the treatment inoculated with E. coli were analyzed, which showed 99% identity with the sequence of the 16S ribosomal gene from E. coli isolate K-12 MG 1655 (U00096.2). These results suggest that Rhodococcus sp. isolate RHA1 (fcb) grows in the peat irradiated (until 150 days), in the presence and absence of PCB and in this irradiated sediment sample (until 70 days), with apparent stability of the fcb operon during this period and in these conditions. The possible presence of the fcbA and fcbB genes in native bacteria grown in K1 medium with 4-chlorobenzoate acid isolated from sediment before irradiation suggests the presence of native bacteria with biodegradation potential of this compound
Growth and survival of recombinant Rhodococcus sp. isolate RHA1 (fcb) in commercial peat and in PCB contaminated soil
O grupo de organoclorados, Bifenilas Policloradas (PCBs) é de difícil degradação e persistente no meio ambiente, tendo sido associado a diversos problemas nos organismos devido ao potencial toxicológico. Biodegradação constitui uma ferramenta eficaz e barata para remoção destes contaminantes do ambiente. O isolado RHA1 (fcb) de Rhodococcus sp. foi geneticamente construído com a introdução do operon de degradação hidrolítica de 4-clorobenzoato (fcb) para evitar a formação de produtos tóxicos durante a degradação de ácidos clorobenzóicos. Com o intuito de se obter informações sobre o processo adaptativo do recombinante Rhodococcus sp. isolado RHA1 (fcb) em substratos contendo PCBs, foram feitos dois ensaios avaliando-se a sobrevivência e o crescimento deste isolado. Rhodococcus sp. isolado RHA1 (fcb) foi inoculado (104 células.g-1) em substrato turfoso previamente irradiado a 50 KGy, contendo ou não 200 mg.Kg-1 de bifenilo. Em outro ensaio, além do recombinante, as bactérias Escherichia coli e Arthrobacter sp. foram inoculados em sedimento coletado na região do Estuário de Santos, contendo PHAs e PCBs, também irradiado (50 KGy). O crescimento das bactérias em ambos os substratos foi monitorado através de contagem de Unidades Formadoras de Colônias (UFCs). Algumas colônias eram selecionadas aleatoriamente para extração de DNA, detecção do operon fcb através de amplificação por PCR e sequenciamento do gene 16S rRNA. Aumento no número de UFCs nos tratamentos inoculados com o recombinante foi observado até 150 dias no ensaio com substrato turfoso e 70 dias na amostra ambiental. Entretanto, houve queda no número de UFCs após os 10 dias nos tratamentos inoculados com E. coli e Arthrobacter sp. Os genes fcbA e fcbB do operon fcb foram detectados nas colônias isoladas dos tratamentos inoculados com o isolado RHA1 (fcb) em ambos os substratos. A análise das seqüências pertencentes às colônias isoladas do tratamento inoculado com o isolado RHA1 (fcb) feita através de BLAST nos sites do NCBI e Ribossomal Database Project, apresentou 99% de identidade com a seqüência do gene ribossomal 16S de Rhodococcus sp. isolado ZC3 (AM076672.1). Somente as seqüências referentes ao tratamento inoculado com E. coli foram analisadas, as quais apresentaram 99% de identidade com a seqüência do gene ribossomal 16S de E. coli isolado K-12 MG 1655 (U00096.2). Estes resultados sugerem que Rhodococcus sp. isolado RHA1 (fcb) cresce na turfa irradiada (até 150 dias) na presença e ausência de PCB e nesta amostra de sedimento irradiada (até 70 dias), com aparente estabilidade do operon fcb durante este período e nestas condições. A possível presença dos genes fcbB e fcbA em bactérias nativas crescidas em meio K1 com ácido 4- clorobenzóico isoladas do sedimento antes da irradiação, sugere a presença de bactérias do local com potencial biodegradador deste composto.The group of organochlorates Biphenyl Polychlorates (PCBs) is of difficult degradation and persistent in the environment, being associated to several problems in the organisms due to its toxicological potential. The isolate RHA1 (fcb) from Rhodococcus sp. was genetically built with the introduction of the operon of hydrolytic degradation 4-chlorobenzoate (fcb) to avoid the formation of toxic products during the degradation of chlorobenzoic acids. In order to obtain information about the adaptative process of the recombinant Rhodococcus sp. isolate RHA1 (fcb) in substrates containing PCBs, two essays were made evaluating the survival and growth of this isolate. Rhodococcus sp. isolate RHA1 (fcb) was inoculated (104 cells.g-1) in peat substrate previously irradiated with 50 kGy, with and without 200 mg.kg-1 of biphenyl. In another essay, besides of the recombinant, the bacteria Escherichia coli and Arthrobacter sp. were inoculated in soil, also irradiated (50 kGy), from the Estuário de Santos region containing PHAs and PCBs. The growth of the bacteria in both substrates was monitorated counting the Colony Forming Units (CFUs). Some colonies were selected randomly for DNA extraction, fcb operon detection through PCR, and sequencing of the 16S rRNA gene. Rising in the number of CFUs in the recombinant inoculated treatments was observed until 150 days in the essay with peat substrate, and until 70 days in the environmental sample. Nonetheless, there was a reduction in the number of CFUs after 10 days in the treatment inoculated with E. coli and Arthrobacter sp. The genes fcbA and fcbB from the operon fcb were detected in the isolated colonies of the treatments inoculated with the isolate RHA1 (fcb) in both substrates. The analysis of the sequences belonging to the colonies isolated from the treatment inoculated with the isolate RHA1 (fcb) through BLAST in the NCBI and Ribosomal Database Project sites showed 99% identity with the sequence of the gene 16S ribosomal from Rhodococcus sp. isolate ZC-3 (AM076672.1). Only the sequences referring to the treatment inoculated with E. coli were analyzed, which showed 99% identity with the sequence of the 16S ribosomal gene from E. coli isolate K-12 MG 1655 (U00096.2). These results suggest that Rhodococcus sp. isolate RHA1 (fcb) grows in the peat irradiated (until 150 days), in the presence and absence of PCB and in this irradiated sediment sample (until 70 days), with apparent stability of the fcb operon during this period and in these conditions. The possible presence of the fcbA and fcbB genes in native bacteria grown in K1 medium with 4-chlorobenzoate acid isolated from sediment before irradiation suggests the presence of native bacteria with biodegradation potential of this compound
Abordagem multianalítica no estudo da composição e funcionalidade da microbiota associada aos ciclos do carbono e nitrogênio em solos sob ambientes preservados e perturbados
The implementation of different agricultural management practices and land use systems promote physicochemical changes in soil associated with loss of nitrogen (N) and carbon (C) from soil and greenhouse gases (GHG) emission. In Brazil, the expansion of sugarcane production and land use changes can affect microbial groups that play essential roles in biogeochemical cycles of N and C. This thesis focused on the microbial community structure, composition, and functions associated with the N and C cycling on sugarcane-cultivated soil and soils under different land use in the Brazilian Amazon. Using a culture-independent approach based on high-throughput DNA sequencing and microarray technology, this thesis is composed by three chapters: the first and the second chapters encompass studies applying GeoChip v.5.0M microarray technology to investigate N and C functional genes as potential bioindicators of vinasse organic residue in combination with N mineral fertilizer in archaeal and acidobacterial communities inhabiting sugarcane-cultivated soil. In the third chapter, the co-occurrence of archaeal taxonomic groups was evaluated in primary and secondary forests, agricultural and cattle pasture soils in the Brazilian Amazon based on high-throughput amplicon sequencing and qPCR of the 16S rRNA gene. Our results in sugarcane-cultivated soil revealed that 87% of genes families associated with N metabolism from soil microbiota were responsive to vinasse with N fertilizer in 7 first days after application. The main gene families responsives were related to processes of nitrification (amoA and hao), ammonification (gdh and ureC), and denitrification (p450nor). The potential O2 decrease and the increase of K and P due vinasse addition can promote the growth of halophile Archaea (Natronomonas) and N2O reducing bacteria (Anaeromyxobacter), but also decrease ammonia oxidizer bacteria (AOB). Regarding the Acidobacteria, subgroups Gp13 and Gp18 revealed positive correlations with the C gene families associated with degradation, especially hemicellulose, but low abundance in vinasse presence. On the other hand, Gp4 was the most abundant acidobacterial subgroup in the vinasse treatment but was not associated with C gene families. This soil management practice can reduce the total Acidobacteria abundance, including that potentially involved with C degradation in sugarcane crops. The co-occurrence of archaeal classes analysis revealed that forest to pasture or agriculture conversion in Amazonian may reduce the syntropy between groups, an important strategy from Archaea to get energy and can promote the reduction of key groups related to N metabolism as ammonia oxidizer Archaea (AOA). These findings of the microbial functionality associated with N and C cycle in tropical soils can support public policy to mitigate N losses in sustainable agroecosystemsA implantação de diferentes sistemas de uso da terra e de práticas de manejo na agricultura promovem mudanças físico-químicas no solo associadas com perdas de nitrogênio (N) e carbono (C) provenientes da emissão de gases de efeito estufa (GEE). No Brasil, a expansão da produção da cana-de-açúcar e dos diferentes usos da terra podem afetar grupos microbianos que atuam nos ciclos biogeoquímicos do N e do C. Essa tese aborda a estrutura, composição e funções de comunidades microbiana associadas com os ciclos do N e do C em solos sob cultivo de cana-de-açúcar e sob diferentes usos da terra na Amazônia brasileira. Usando metodologia independente de cultivo, com base em sequenciamento de DNA de alto rendimento e tecnologia de microarranjo de DNA, essa tese está organizada em três capítulos: no primeiro e no segundo capítulos foi utilizada a versão de microarranjo GeoChip 5.0.M para investigar genes funcionais associados ao metabolismo de N e C como potenciais bioindicadores do uso combinado de vinhaça, um subproduto da produção de etanol, em combinação com fertilizante N mineral em comunidades de Archaea e de Acidobacteria que habitam o solo cultivado com cana-de-açúcar. No terceiro capítulo, foi avaliada a co-ocorrência entre grupos de Archaea em florestas primária e secundária, em solos agrícolas e em pastagens na Amazônia brasileira utilizando a técnica de PCR quantitativo em tempo real (qPCR) e sequenciamento de amplicon do gene 16S rRNA. Nossos resultados com cultivo de cana-de-açúcar sob adição de vinhaça e do fertilizante nitrogenado revelaram que 87% das famílias gênicas associadas com o metabolismo de N provenientes da microbiota do solo foram responsivas aos 7 dias após a aplicação. As principais famílias gênicas responsivas estão relacionadas aos processos de nitrificação (amoA e hao), amonificação (gdh e ureC) e denitrificação (p450nor). A possível redução na disponibilidade de O2 e o aumento de K e P no solo devido a adição da vinhaça podem promover o crescimento de Archaea halophilas (Natronomonas) e bactérias redutoras de N2O (Anaeromyxobacter), e reduzir AOB (bactéria oxidadora de amônia). Os subgrupos Gp13 e Gp18 de Acidobacteria foram correlacionados com famílias gênicas relacionadas com a degradação de C, principalmente hemicelulose. Entretanto, apresentaram baixa abundância no tratamento com vinhaça. O subgrupo Gp4 foi o mais abundante no tratamento com vinhaça, mas não foi correlacionado às famílias gênicas de C. Essa prática de manejo do solo pode reduzir a abundância de Acidobacteria total, incluindo aquelas potencialmente envolvidas com a degradação de C em culturas de cana-de-açúcar. Os estudos realizados nessa tese contribuem com informações sobre a dinâmica funcional da microbiota ativa nos ciclos biogeoquímicos do N e C, orientando políticas públicas que visam a redução de perdas de N e C em agroecossistema
Long-term land use in Amazon influence the dynamic of microbial communities in soil and rhizosphere
Brazil has become the world leader in soy production, leading to an increase in the conversion of the Amazon rainforest into cropland. These actions had consequences for Forest's biodiversity, including the soil. In this sense, a better understanding of how long-term land use affects soil microbial communities, and their functions is urgent. This study aimed to evaluate the long-term land-use effects over bacterial and archaeal communities in soil and soybean rhizosphere in the Amazon region. For this, mesocosms experiments were carried out with Amazon soils with a history of 2-, 8-, and 20-years of agricultural use. We then assessed the bacterial and archaeal communities based on the 16S rRNA sequencing and real-time PCR. Our results showed a distinct bacterial community structure in soils with 20-years of land use. For both, bulk soil and soybean rhizosphere with 20-years of use, there was an increase in the abundance of Gemmatimonadetes, Chloroflexi, Firmicutes, and Planctomycetes. Interestingly, the niche occupancy analysis revealed an increase of specialist microbes in these soils. Also, these soils with 20-years of use showed a more complex network for both bulk and rhizosphere samples, highlighting the importance of Actinobacteria and Chloroflexi phyla to soil network structure. Our analysis also revealed an increased abundance of total bacteria, N-fixers, and ammonia-oxidizers bacteria in rhizosphere soil with 20-years of use. In addition, based on the potential functional analysis, nitrification processes increased in those soils. However, we noticed a homogenization in the abundance of the genes between rhizosphere and bulk soil with 20-years of use. In general, the differences were associated with changes in soil chemical characteristics such as pH, Ca2+, Mg2+, and organic matter, which are a consequence of liming and no-till practices over time. Our findings demonstrate that long-term agriculture in Amazon soils affects microbial community composition and functions, bringing new insights to better understand anthropogenic actions over the soil microbiome
Innovative infrastructure to access Brazilian fungal diversity using deep learning
In the present investigation, we employ a novel and meticulously structured database assembled by experts, encompassing macrofungi field-collected in Brazil, featuring upwards of 13,894 photographs representing 505 distinct species. The purpose of utilizing this database is twofold: firstly, to furnish training and validation for convolutional neural networks (CNNs) with the capacity for autonomous identification of macrofungal species; secondly, to develop a sophisticated mobile application replete with an advanced user interface. This interface is specifically crafted to acquire images, and, utilizing the image recognition capabilities afforded by the trained CNN, proffer potential identifications for the macrofungal species depicted therein. Such technological advancements democratize access to the Brazilian Funga, thereby enhancing public engagement and knowledge dissemination, and also facilitating contributions from the populace to the expanding body of knowledge concerning the conservation of macrofungal species of Brazil