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1.
Mol Phylogenet Evol ; 167: 107338, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34757168

RESUMO

Africa is known for its rich legume diversity with a significant number of endemic species originating in South Africa. Many of these legumes associate with rhizobial symbionts of the genus Bradyrhizobium, of which most represent new species. Yet, none of the Bradyrhizobium species from South Africa have been described. In this study, phylogenetic analysis of 16S rRNA gene sequences of fourteen strains isolated in southern Africa from root nodules of diverse legumes (i.e., from the tribes Crotalarieae, Acacieae, Genisteae, Phaseoleae and Cassieae) revealed that they belong to the Bradyrhizobium elkanii supergroup. The taxonomic position and possible novelty of these strains were further interrogated using genealogical concordance of five housekeeping genes (atpD, dnaK, glnII, gyrB and rpoB). These phylogenies consistently recovered four monophyletic groups and one singleton within Bradyrhizobium. Of these groups, two were conspecific with Bradyrhizobium brasilense UFLA 03-321T and Bradyrhizobium ivorense CI-1BT, while the remaining three represented novel taxa. Their existence was further supported with genome data, as well as metabolic and physiological traits. Analysis of nodA gene sequences further showed that the evolution of these bacteria likely involved adapting to local legume hosts and environmental conditions through the acquisition, via horizontal gene transfer, of optimal symbiotic loci. We accordingly propose the following names Bradyrhizobium acaciae sp. nov. 10BBT (SARCC 730T = LMG 31409T), Bradyrhizobium oropedii sp. nov. Pear76T (SARCC 731T = LMG 31408T), and Bradyrhizobium altum sp. nov. Pear77T (SARCC 754T = LMG 31407T) to accommodate three novel species, all of which are symbionts of legumes in South Africa.


Assuntos
Bradyrhizobium , Fabaceae , DNA Bacteriano/genética , Fabaceae/genética , Fabaceae/microbiologia , Fixação de Nitrogênio , Filogenia , RNA Ribossômico 16S/genética , Nódulos Radiculares de Plantas/microbiologia , Análise de Sequência de DNA , África do Sul , Simbiose/genética
2.
Int J Mol Sci ; 23(19)2022 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-36232842

RESUMO

Rhizobia, which enter into symbiosis with legumes, can also interact with non-legumes and promote plant growth. In this paper, we explored the effects of nickel (Ni, 200 µM) on Arabidopsis thaliana (Col-0) inoculated with plant growth-promoting (PGP) rhizobia nodulating ultramafic Anthyllis vulneraria. The isolated PGP strains tolerant to Ni were identified as Rhizobium sp. and Bradyrhizobium sp. The isolates highly differed in their PGP abilities and Ni resistance. Without Ni-stress, the plants inoculated with most isolates grew better and had higher photosynthetic efficiency than non-inoculated controls. Nickel treatment increased Ni concentration in inoculated plants. Plant growth, leaf anatomy, chloroplast ultrastructure, efficiency of photosynthesis, and antioxidant defense system activity were significantly impaired by Ni, however, the majority of these effects were diminished in plants inoculated with the most effective PGP rhizobia. Real-time PCR revealed an increased expression level of genes involved in auxin and gibberellin biosynthesis in the inoculated, Ni-treated plants, and this may have improved shoot and root growth after inoculation with effective isolates. Our results also suggest a positive correlation between Ni-stress parameters and antioxidant defense system activity, and also between the effectiveness of photosynthesis and plant growth parameters. We showed that the selected rhizobia, naturally nodulating Anthyllis on Ni-rich ultramafic soils can promote Arabidopsis growth and increase plant tolerance to Ni by improving different physiological and biochemical mechanisms.


Assuntos
Arabidopsis , Lotus , Rhizobium , Antioxidantes/metabolismo , Bactérias , Giberelinas/metabolismo , Ácidos Indolacéticos/metabolismo , Níquel/metabolismo , Níquel/farmacologia , Rhizobium/metabolismo , Solo/química , Microbiologia do Solo , Simbiose
3.
Int J Mol Sci ; 22(11)2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34071406

RESUMO

Coralyne is a synthetic analog of berberine related to protoberberine-isoquinoline alkaloids. Isoquinoline derivatives and analogs are renowned as potent radiosensitizers with potential medical application. In the present study, we investigated the effect of coralyne on the cell death, cytoskeletal changes and cell cycle progression of irradiated A549 cells. A clonogenic assay revealed that coralyne pretreatment decreased the viability of A549 cells in a time- and dose-dependent manner. Moreover, exposure to coralyne and ionizing radiation (IR) markedly altered the filamentous actin cytoskeletal architecture and integrin-ß binding sites of A549 cells. Treatment with 1-25 µM coralyne in combination with 2 Gy of IR significantly reduced the percentage of cells in G2/M phase compared with 2 Gy IR alone. These results indicate that coralyne is a potent radiosensitizing agent that may find an application in medicine.


Assuntos
Alcaloides de Berberina/farmacologia , Inibidor de Quinase Dependente de Ciclina p21/genética , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Células A549 , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Relação Dose-Resposta a Droga , Relação Dose-Resposta à Radiação , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos da radiação , Humanos , Microscopia Confocal , Radiação Ionizante , Radiossensibilizantes/farmacologia
4.
Int J Syst Evol Microbiol ; 69(7): 1852-1863, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31140963

RESUMO

Herein the members of the Subcommittee on Taxonomy of Rhizobia and Agrobacteria of the International Committee on Systematics of Prokaryotes review recent developments in rhizobial and agrobacterial taxonomy and propose updated minimal standards for the description of new species (and genera) in these groups. The essential requirements (minimal standards) for description of a new species are (1) a genome sequence of at least the proposed type strain and (2) evidence for differentiation from other species based on genome sequence comparisons. It is also recommended that (3) genetic variation within the species is documented with sequence data from several clearly different strains and (4) phenotypic features are described, and their variation documented with data from a relevant set of representative strains. Furthermore, it is encouraged that information is provided on (5) nodulation or pathogenicity phenotypes, as appropriate, with relevant gene sequences. These guidelines supplement the current rules of general bacterial taxonomy, which require (6) a name that conforms to the International Code of Nomenclature of Prokaryotes, (7) validation of the name by publication either directly in the International Journal of Systematic and Evolutionary Microbiology or in a validation list when published elsewhere, and (8) deposition of the type strain in two international culture collections in separate countries.


Assuntos
Agrobacterium/classificação , Rhizobium/classificação , Terminologia como Assunto , Guias como Assunto
5.
Cell Mol Neurobiol ; 37(6): 1055-1066, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27832395

RESUMO

Perturbations in the transport of mitochondria and their quality control in neuronal cells underlie many types of neurological pathologies, whereas systems enabling convenient analysis of mitochondria behavior in cellular models of neurodegenerative diseases are limited. In this study, we present a modified version of lund human mesencephalic cells, mitoLUHMES, expressing GFP and mitochondrially targeted DsRed2 fluorescent proteins, intended for in vitro analysis of mitochondria trafficking by real-time fluorescence microscopy. This cell line can be easily differentiated into neuronal phenotype and allows us to observe movements of single mitochondria in single cells grown in high-density cultures. We quantified the perturbations in mitochondria morphology and dynamics in cells treated with model neurotoxins: carbonyl cyanide m-chlorophenylhydrazone and 6-hydroxydopamine. For the first time we filmed the processes of fission, fusion, pausing, and reversal of mitochondria movement direction in LUHMES cells. We present a detailed analysis of mitochondria length, velocity, and frequency of movement for static, anterograde, and retrograde motile mitochondria. The observed neurotoxin treatment-mediated decreases in morphological and kinetic parameters of mitochondria provide foundation for the future studies exploiting mitoLUHMES as a new model for neurobiology.


Assuntos
Engenharia Celular , Mitocôndrias/metabolismo , Movimento , Neurônios/citologia , Transporte Biológico/efeitos dos fármacos , Carbonil Cianeto m-Clorofenil Hidrazona/farmacologia , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Forma Celular/efeitos dos fármacos , Proteínas de Fluorescência Verde/metabolismo , Humanos , Imageamento Tridimensional , Proteínas Luminescentes/metabolismo , Mitocôndrias/efeitos dos fármacos , Dinâmica Mitocondrial/efeitos dos fármacos , Movimento/efeitos dos fármacos , Neuritos/efeitos dos fármacos , Neuritos/metabolismo , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Oxidopamina/farmacologia
6.
Mol Phylogenet Evol ; 100: 206-218, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27068839

RESUMO

The genus Bradyrhizobium contains predominantly nitrogen-fixing legume symbionts. Phylogenetic analysis of the genes responsible for their symbiotic abilities (i.e., those encoded on the nodulation [nod] and nitrogen-fixation [nif] loci) has facilitated the development of an extensive phylogeographic framework for the genus. This framework however contains only a few nodulating isolates from Africa. Here we focused on nodulating Bradyrhizobium isolates associated with native southern African legumes in the tribes Genisteae and Crotalarieae found along the Great Escarpment in the Mpumalanga Province of South Africa. The aims of this study were to: (1) obtain rhizobial isolates from legumes in the Genisteae and Crotalarieae; (2) verify their nodulation ability; (3) characterize them to species level based on phylogenetic analyses of several protein coding gene regions (atpD, dnaK, glnII, recA, rpoB and gyrB) and (4) determine their placement in the phylogeographic framework inferred from the sequences of the symbiotic loci nodA and nifD. Twenty of the 21 Bradyrhizobium isolates belonged to six novel species, while one was conspecific with the recently described B. arachidis. Among these isolates, the nodA phylogeny revealed several new clades, with 18 of our isolates found in Clades XIV and XV, and only three forming part of the cosmopolitan Clade III. These strains formed predominantly the same groups in the nifD phylogeny although with slight differences; indicating that both vertical and horizontal inheritance of the symbiotic loci occurred. These findings suggest that the largely unexplored diversity of indigenous African rhizobia are characterized by unique ancestries that might mirror the distribution of their hosts and the environmental factors driving their evolution.


Assuntos
Bradyrhizobium/classificação , Fabaceae/microbiologia , Simbiose , Proteínas de Bactérias/genética , Bradyrhizobium/genética , Bradyrhizobium/isolamento & purificação , DNA Bacteriano/química , DNA Bacteriano/isolamento & purificação , DNA Bacteriano/metabolismo , Fabaceae/genética , Tipagem de Sequências Multilocus , Fixação de Nitrogênio , Filogenia , RNA Ribossômico 16S/química , RNA Ribossômico 16S/isolamento & purificação , RNA Ribossômico 16S/metabolismo , Análise de Sequência de DNA , África do Sul
7.
Cell Mol Neurobiol ; 35(8): 1137-47, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25986246

RESUMO

LUHMES cells, a recently established line of immortalized embryonic mesencephalic cells, are the novel in vitro model for studying Parkinson's disease (PD) and dopaminergic neuron biology. Phosphoglyceromutase 5 (PGAM5) is a mitochondrial protein involved in mitophagy, mitochondria dynamics, and other processes important for PD pathogenesis. We tested the impact of lentiviral overexpression of PGAM5 protein in LUHMES cells on their differentiation and expression of 84 PD-related genes. LUHMES cells were transduced with PGAM5 or mock and treated with 100 µM 6-hydroxydopamine (6-OHDA), a model PD neurotoxin. Real-Time PCR analysis revealed that the treatment with 6-OHDA-induced changes in expression of 44 PD-related genes. PGAM5 transduction alone did not cause alternations in PD-related genes expression, nor it affected changes in gene expression mediated by 6-OHDA. The 6-OHDA-induced PD-related gene expression profile of LUHMES cells is presented for the first time and widely discussed.


Assuntos
Proteínas de Transporte/biossíntese , Diferenciação Celular/efeitos dos fármacos , Mesencéfalo/efeitos dos fármacos , Mesencéfalo/metabolismo , Proteínas Mitocondriais/biossíntese , Oxidopamina/toxicidade , Transtornos Parkinsonianos/metabolismo , Diferenciação Celular/fisiologia , Linhagem Celular Transformada , Regulação da Expressão Gênica , Humanos , Mesencéfalo/embriologia , Transtornos Parkinsonianos/induzido quimicamente , Fosfoproteínas Fosfatases
8.
Mutagenesis ; 30(3): 431-9, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25681789

RESUMO

Silver nanoparticles (AgNPs) are one of the most widely used nanomaterials due to their antibacterial properties. Owing to the recent boost in the usage of AgNPs-containing products, human exposure to AgNPs is increasing, highlighting the need for careful evaluation of AgNPs toxicity in humans. We used two cellular models, hepatic HepG2 and epithelial A549 cell lines, to study the mechanism of AgNPs-induced toxicity at the cellular level. These two cell lines differ significantly in their response to AgNPs treatment. In the case of A549 cells, a minor decrease in viability and increase in the extent of DNA breakage were observed. A markedly different response to AgNPs was observed in HepG2 cells. In short term, a massive induction of DNA breakage was observed, suggesting that the basal activity of antioxidant defence in these cells was not sufficient to effectively protect them from the nanoparticle-induced oxidative stress. After prolonged exposure, the extent of DNA breakage decreased to the level observed in the control cells proving that a successful adaptation to the new conditions had taken place. The cells that were unable to adapt must have died, as revealed by the Neutral Red assay that indicated less than half viable cells after 24-h treatment with 100 µg/ml of 20nm AgNPs. The gene expression analysis revealed that the observed adaptation was underlain by a pro-proliferative, anti-apoptotic signal leading to up-regulation of the genes promoting proliferation and inflammatory response (EGR1, FOS, JUN, HK2, IL4, MMP10, VEGFA, WISP1, CEBPB, IL8, SELPLG), genes coding the anti-apoptotic proteins (BCL2A1, CCL2) and factors involved in the response to stress (HSPB1, GADD45A). Such a selection of highly resistant population of cells should be taken into account in the case of medical applications of nanoparticles since the sustained proliferative signalling and resistance to cell death are hallmarks of cancer, acquired by the cells in the process of carcinogenesis.


Assuntos
Apoptose/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Adaptação Fisiológica , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA , Células Hep G2 , Humanos
9.
Mol Cell Biochem ; 389(1-2): 99-111, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24390086

RESUMO

Pirin, a product of the PIR gene, is an iron-binding protein acting as a transcriptional coregulator implicated in the regulation of the NF-κB-related transcription via interaction with RelA (p65), as well as BCL3 and NF-κB1 (p50) proteins. Alterations in pirin expression were observed in various tumors and under oxidative stress conditions. The aim of the present work was to analyze the regulation of the transcription of the human PIR gene. Using constructs containing a different sized PIR promoter and the luciferase reporter genes we found that in HeLa cells PIR transcription is mostly dependent on a highly conserved antioxidant response element located 281 bp downstream of the transcription start site. We have proved that the NRF2 transcription factor binds to this element in vivo and drives the basal PIR expression. We hypothesize that regulation of the PIR expression may constitute a mechanism by which NRF2 is able to modulate the activity of NF-κB and possibly other signaling pathways.


Assuntos
Elementos de Resposta Antioxidante/genética , Proteínas de Transporte/genética , Regulação da Expressão Gênica/genética , Fator 2 Relacionado a NF-E2/genética , Proteínas Nucleares/genética , Sequência de Aminoácidos , Linhagem Celular Tumoral , Dioxigenases , Células HeLa , Humanos , Dados de Sequência Molecular , NF-kappa B/genética , Regiões Promotoras Genéticas/genética , Alinhamento de Sequência , Transdução de Sinais/genética , Fatores de Transcrição/genética , Sítio de Iniciação de Transcrição , Transcrição Gênica/genética
10.
Pol Merkur Lekarski ; 36(214): 229-32, 2014 Apr.
Artigo em Polonês | MEDLINE | ID: mdl-24868893

RESUMO

The incidence of ischemic heart disease (IHD) in patients with OSAS is estimated at around 20%. This greatly affect a common risk factors for both diseases: male gender, obesity, age and diabetes and hypertension. Attention is drawn to the possibility of genetic determinants of IHD. The aim of study was to answer the question whether the presence of polymorphisms of selected genes possibly related to IHD may be useful to isolate the group of patients with OSAS, especially vulnerable as a complication of IHD. Materials and methods. The study included 600 people with OSAS, which was isolated in patients with IHD (127 people). The remaining 473 individuals were observed as a control group. The polymorphism of three genes were evaluated to find possible influence on the occurrence of IHD or myocardial infarction as follows: SREBF1 (sterol regulatory element binding transcription factor 1), REBF2 (sterol regulatory element binding transcription factor 2) and HIF1 (hypoxia inducible factor 1, alpha subunit). Results. Analysis of relationship between polymorphisms of selected genes and the diagnosis of IHD in the whole group of patients with OSAS showed a relationship only for the gene SREBF1 finding the lowest frequency of its occurrence in AA homozygotes (at 13.6%) and twice with GG homozygotes (26.1%). Conclusions. Rating polymorphisms studied genes did not reveal their relationship to the occurrence of IHD in patients with OSAS, both in the whole group as well as separate subgroups.


Assuntos
Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Isquemia Miocárdica/epidemiologia , Isquemia Miocárdica/genética , Apneia Obstrutiva do Sono/epidemiologia , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 2/genética , Adulto , Idoso , Estudos de Casos e Controles , Comorbidade , Feminino , Predisposição Genética para Doença , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Polimorfismo Genético , Fatores de Risco
11.
Syst Appl Microbiol ; 46(5): 126452, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37634485

RESUMO

A genealogical concordance approach was used to delineate strains isolated from Acacia dealbata and Acacia mearnsii root nodules in South Africa. These isolates form part of Bradyrhizobium based on 16S rRNA sequence similarity. Phylogenetic analysis of six housekeeping genes (atpD, dnaK, glnII, gyrB, recA and rpoB) confirmed that these isolates represent a novel species, while pairwise average nucleotide identity (ANIb) calculations with the closest type strains (B. cosmicum 58S1T, B. betae PL7HG1T, B. ganzhouense CCBAU 51670 T, B. cytisi CTAW11T and B. rifense CTAW71T) resulted in values well below 95-96%. We further performed phenotypic tests which revealed that there are high levels of intraspecies variation, while an additional analysis of the nodA and nifD loci indicated that the symbiotic loci of the strains are closely related to those of Bradyrhizobium isolates with an Australian origin. Strain 14ABT (=LMG 31415 T = SARCC-753 T) is designated as the type strain of the novel species for which we propose the name Bradyrhizobium xenonodulans sp. nov.


Assuntos
Acacia , Bradyrhizobium , Acacia/genética , Genes Bacterianos/genética , Filogenia , RNA Ribossômico 16S/genética , África do Sul , Nódulos Radiculares de Plantas , DNA Bacteriano/genética , Hibridização de Ácido Nucleico , Austrália , Análise de Sequência de DNA
12.
Nat Commun ; 14(1): 4092, 2023 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-37433777

RESUMO

Perturbed cellular protein homeostasis (proteostasis) and mitochondrial dysfunction play an important role in neurodegenerative diseases, however, the interplay between these two phenomena remains unclear. Mitochondrial dysfunction leads to a delay in mitochondrial protein import, causing accumulation of non-imported mitochondrial proteins in the cytosol and challenging proteostasis. Cells respond by increasing proteasome activity and molecular chaperones in yeast and C. elegans. Here, we demonstrate that in human cells mitochondrial dysfunction leads to the upregulation of a chaperone HSPB1 and, interestingly, an immunoproteasome-specific subunit PSMB9. Moreover, PSMB9 expression is dependent on the translation elongation factor EEF1A2. These mechanisms constitute a defense response to preserve cellular proteostasis under mitochondrial stress. Our findings define a mode of proteasomal activation through the change in proteasome composition driven by EEF1A2 and its spatial regulation, and are useful to formulate therapies to prevent neurodegenerative diseases.


Assuntos
Cisteína Endopeptidases , Complexo de Endopeptidases do Proteassoma , Proteostase , Humanos , Citoplasma , Mitocôndrias , Fator 1 de Elongação de Peptídeos , Cisteína Endopeptidases/metabolismo
13.
Mol Phylogenet Evol ; 63(2): 265-77, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22230030

RESUMO

Geographic isolation and growing climate aridity played major roles in the evolution of Australian legumes. It is likely that these two factors also impacted on the evolution of their root-nodule bacteria. To investigate this issue, we applied a multilocus sequence analysis (MLSA) approach to examine Bradyrhizobium isolates originating from temperate areas of Western Australia (WA) and the tropical-monsoon area of the Northern Territory (NT). The isolates were mostly collected from the nodules of legumes belonging to tribes, genera and species endemic or native to Australia. Phylogenetic analyses of sequences for the housekeeping atpD, dnaK, glnII, gyrB, recA and 16S rRNA genes and nodulation nodA gene revealed that most isolates belonged to groups that are distinct from non-Australian Bradyrhizobium isolates, which is in line with earlier studies based on 16S rRNA gene sequence analyses. Phylogenetic analysis of the nodA data allowed identification of five major Clades among the WA and NT isolates. All WA isolates grouped in a subgroup I.1 of Clade I with strains originating from temperate eastern Australia. In contrast, the NT isolates formed part of Clades I (subgroup I.2), III (subgroup III.3), IV, V and X. Of these nodA clades, Clade I, Clade IV, Clade X presumably have an Australian origin. Overall, these data demonstrate that the impact of geographic isolation of the Australian landmass is manifested by the presence of numerous unique clusters in housekeeping and nodulation gene trees. In addition, the intrinsic climate characteristics of temperate WA and tropical-monsoon NT were responsible for the formation of distinct legume communities selecting for unrelated Bradyrhizobium groups.


Assuntos
Bradyrhizobium/classificação , Bradyrhizobium/genética , Fabaceae/microbiologia , Filogenia , Aciltransferases/genética , Austrália , Proteínas de Bactérias/genética , Sequência de Bases , DNA Bacteriano/genética , DNA Ribossômico/genética , Consórcios Microbianos , Família Multigênica/genética , Tipagem de Sequências Multilocus , RNA Ribossômico 16S/genética , Nódulos Radiculares de Plantas/microbiologia , Análise de Sequência de DNA , Simbiose/genética
14.
Bioorg Med Chem ; 20(22): 6732-8, 2012 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-23063520

RESUMO

Formation of dinitrosyl iron complexes (DNICs) was observed in a wide spectrum of pathophysiological conditions associated with overproduction of NO. To gain insight into the possible genotoxic effects of DNIC, we examined the interaction of histidinyl dinitrosyl iron complexes (HIS-DNIC) with DNA by means of circular dichroism. Formation of DNIC was monitored by EPR and FT/IR spectroscopy. Vibrational bands for aquated HIS-DNIC are reported. Dichroism results indicate that HIS-DNIC changes the conformation of the DNA in a dose-dependent manner in 10mM phosphate buffer (pH 6). Increase of the buffer pH or ionic strength decreased the effect. Comparison of HIS-DNIC DNA interaction with the effect of hydrated Fe(2+) ion revealed many similarities. The importance of iron ions in HIS-DNIC induced genotoxicity is confirmed by plasmid nicking assay. Treatment of pUC19 plasmid with 1µM HIS-DNIC did not affect the plasmid supercoiling. Higher concentrations of HIS-DNIC induced single strand breaks. The effect was completely abrogated by addition of deferoxamine, a specific strong iron chelator. Our data reveal that formation of HIS-DNIC does not prevent DNA from iron-induced damage and imply that there is no direct interrelationship between iron-NO coordination and their mutual toxicity modulation.


Assuntos
Compostos Ferrosos/química , Histidina/química , Ferro/química , Óxidos de Nitrogênio/química , Dicroísmo Circular , DNA/química , DNA/metabolismo , Quebras de DNA de Cadeia Simples , Desferroxamina/farmacologia , Espectroscopia de Ressonância de Spin Eletrônica , Compostos Ferrosos/toxicidade , Íons/química , Ferro/toxicidade , Óxidos de Nitrogênio/toxicidade , Testes de Toxicidade
15.
Syst Appl Microbiol ; 44(3): 126203, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33857759

RESUMO

In this work, we investigated Bradyrhizobium strains isolated from soils collected from the rhizosphere of native and exotic legumes species inhabiting two ecoclimatic zones - asubtropical-lowland pasture (Pampa Biome) and a volcanic plateau covered by Araucaria Moist Forests (Atlantic Forest Biome). The rhizobial strains were isolated from the nodules of seven native and one exotic legume species used as rhizobium traps. Single-gene (recA, glnII, dnaK) and combined-gene MLSA analyses (dnaK-glnII-gyrB-recA-rpoB) revealed that nearly 85% of the isolates clustered in B. elkanii supergroup, while the remaining (except for two isolates) in B. japonicum supergroup, albeit, in most cases, separately from the type strains of Bradyrhizobium species. As a symbiotic gene marker, a portion of nifD gene was sequenced for 194 strains. In the nifD-tree, an American branch III.3D (104 isolates), was the most numerous among the isolates. A significant portion of the isolates clustered in American groups; subclade III.4 (40 strains), Clade VII (3 strains), and a new Clade XX (4 strains). Most of the remaining strains belonged to a pantropical III.3C branch (39 isolates). On the other hand, identification of isolates belonging, respectively, to Clade I and Clade II may result of spreading of the Australian (Clade I) and European (Clade II) bradyrhizobia following the introduction of their legume hosts. Our study indicated that the American groups predominated in the symbiotic Bradyrhizobium communities in southern Brazil. However, there is a significant component of exotic lineages, resulting from the dispersal of pantropical Fabaceae taxa and the introduction of exotic legumes.


Assuntos
Bradyrhizobium , Fabaceae , Florestas , Pradaria , Filogenia , Bradyrhizobium/classificação , Bradyrhizobium/isolamento & purificação , Brasil , DNA Bacteriano/genética , Fabaceae/microbiologia , Genes Bacterianos , RNA Ribossômico 16S/genética , Rizosfera , Nódulos Radiculares de Plantas/microbiologia , Análise de Sequência de DNA , Simbiose
16.
Syst Appl Microbiol ; 44(4): 126228, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34265499

RESUMO

The isolation of rhizobial strains from the root and stem nodules remains a commonly used method despite its limitations as it enables the identification of mainly dominant symbiotic groups within rhizobial communities. To overcome these limitations, we used genus-specific nifD primers in a culture-independent assessment of Bradyrhizobium communities inhabiting soils in southern Brazil. The majority of nifD sequences were generated from DNA isolated from tropical-lowland pasture soils, although some soil samples originated from the Campos de Cima da Serra volcanic plateau. In the nifD tree, all the bradyrhizobial sequences comprised 38 clades, including 18 new clades. The sequences generated in this study were resolved into 22 clades and 21 singletons. The nifD bradyrhizobial assemblage contained Azorhizobium and α-proteobacterial methylotrophic genera, suggesting that these genera may have acquired their nif loci from Bradyrhizobium donors. The most common in the lowland pasture soils subclade III.3D branch comprises the isolates of mainly an American origin. On the other hand, subclade III.4, which was earlier detected in Brazil among Bradyrhizobium isolates nodulating native lupins, appears more common in the Campos de Cima da Serra soils. The second-largest group, Clade XXXVIII, has not yet been reported in culture-dependent studies, while another common group called Clade I represents a symbiovar predominating in Australia. The identification of the diverse nifD Clade I haplotypes in the tropical-lowland pastures infested by Australian Acacia spp implies that the introduction of these legumes to southern Brazil has resulted in the dissemination of their bradyrhizobial symbionts.


Assuntos
Bradyrhizobium , Lupinus , Filogenia , Bradyrhizobium/classificação , Bradyrhizobium/isolamento & purificação , Brasil , DNA Bacteriano/genética , Florestas , Lupinus/microbiologia , RNA Ribossômico 16S/genética , Nódulos Radiculares de Plantas , Análise de Sequência de DNA , Microbiologia do Solo , Simbiose
17.
Materials (Basel) ; 13(14)2020 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-32698417

RESUMO

Microglial cells clear the brain of pathogens and harmful debris, including amyloid-ß (Aß) deposits that are formed during Alzheimer's disease (AD). We studied the expression of Msr1, Ager and Cd36 receptors involved in Aß uptake and expression of Cd33 protein, which is considered a risk factor in AD. The effect of silver nanoparticles (AgNPs) and cadmium telluride quantum dots (CdTeQDs) on the expression of the above receptors and Aß uptake by microglial cells was investigated. Absorption of Aß and NPs was confirmed by confocal microscopy. AgNPs, but not CdTeQDs, caused a decrease in Aß accumulation. By using a specific inhibitor-polyinosinic acid-we demonstrated that Aß and AgNPs compete for scavenger receptors. Real-time PCR showed up-regulation of Cd33 and Cd36 gene expression after treatment with CdTeQDs for 24 h. Analysis of the abundance of the receptors on the cell surface revealed that AgNP treatment significantly reduced the presence of Msr1, Cd33, Ager and Cd36 receptors (6 and 24 h), whereas CdTeQDs increased the levels of Msr1 and Cd36 (24 h). To summarize, we showed that AgNP uptake competes with Aß uptake by microglial cells and consequently can impair the removal of the aggregates. In turn, CdTeQD treatment led to the accumulation of proinflammatory Cd36 protein on the cell surface.

18.
Pol Arch Intern Med ; 129(1): 6-11, 2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30600310

RESUMO

INTRODUCTION Although the coexistence of type 2 diabetes mellitus (T2DM) and obstructive sleep apnea (OSA) may be attributed to environmental risk factors common for both diseases, a genetic background should also be considered. Data on the role of genetic factors in the development of T2DM in patients with OSA are lacking. OBJECTIVES The study was aimed to evaluate the prevalence of polymorphisms of selected genes that are known to be associated with diabetes or obesity in patients with OSA and concomitant T2DM and to assess these polymorphisms in the context of OSA severity. PATIENTS AND METHODS Consecutive patients with newly diagnosed OSA confirmed by polysomnography underwent genotyping for the following single nucleotide polymorphisms (SNPs): SREBF1 rs11868035, HIF1A rs11549465, APOA5 rs3135506, TCF7L2 rs7903146, and FTO rs16945088. The frequency of genotypes was compared between patients with and without concomitant T2DM and was analyzed with regard to OSA severity. RESULTS A total of 600 patients with newly diagnosed OSA were enrolled to the study. Of these, 121 patients (20.2%) were diagnosed with T2DM (97 men and 24 women; median age, 58 years; range, 52-64 years). The prevalence of T2DM was significantly lower in APOA5 rs3135506 GG homozygotes than in CG heterozygotes (18.8% vs 33.3%, P = 0.02). APOA5 rs3135506 CG heterozygotes were at higher risk for developing T2DM (adjusted odds ratio, 2.64; 95% confidence interval,1.38-5.04; P = 0.003). No significant differences were found for the genotype distribution of the other investigated SNPs. CONCLUSIONS Our study shows a possible link between the polymorphism of the gene encoding APOA5 and T2DM in patients with OSA.


Assuntos
Apolipoproteína A-V , Diabetes Mellitus Tipo 2/diagnóstico , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Apneia Obstrutiva do Sono/complicações , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
19.
Syst Appl Microbiol ; 42(4): 427-439, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31031014

RESUMO

Bradyrhizobium is thought to be the largest and most diverse rhizobial genus, but this is not reflected in the number of described species. Although it was one of the first rhizobial genera recognised, its taxonomy remains complex. Various contemporary studies are showing that genome sequence information may simplify taxonomic decisions. Therefore, the growing availability of genomes for Bradyrhizobium will likely aid in the delineation and characterization of new species. In this study, we addressed two aims: first, we reviewed the availability and quality of available genomic resources for Bradyrhizobium. This was achieved by comparing genome sequences in terms of sequencing technologies used and estimated level of completeness for inclusion in genome-based phylogenetic analyses. Secondly, we utilized these genomes to investigate the taxonomic standing of Bradyrhizobium in light of its diverse lifestyles. Although genome sequences differed in terms of their quality and completeness, our data indicate that the use of these genome sequences is adequate for taxonomic purposes. By using these resources, we inferred a fully resolved, well-supported phylogeny. It separated Bradyrhizobium into seven lineages, three of which corresponded to the so-called supergroups known for the genus. Wide distribution of key lifestyle traits such as nodulation, nitrogen fixation and photosynthesis revealed that these traits have complicated evolutionary histories. We present the first robust Bradyrhizobium species phylogeny based on genome sequence information for investigating the evolution of this important assemblage of bacteria. Furthermore, this study provides the basis for using genome sequence information as a resource to make important taxonomic decisions, particularly at the species and genus levels.


Assuntos
Bradyrhizobium/classificação , Classificação/métodos , Genoma Bacteriano/genética , Filogenia , Sequência de Bases , Bradyrhizobium/genética , DNA Bacteriano/genética , Bases de Dados Genéticas , Genes Bacterianos/genética , Fixação de Nitrogênio/genética , Fotossíntese/genética , Nodulação/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
20.
Mol Phylogenet Evol ; 48(3): 1131-44, 2008 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-18539053

RESUMO

Cowpea (Vigna unguiculata) and peanut (Arachis hypogaea) in southern Africa are nodulated by a genetically diverse group of Bradyrhizobium strains. To determine the identity of these bacteria, a collection of 22 isolates originating from the root nodules of both hosts in Botswana and South Africa was investigated using the combined sequences for the core genome genes rrs, recA, and glnII. These data separated the majority of the isolates into one of three unique lineages that most likely represent novel Bradyrhizobium species. Some isolates were also conspecific with B. yuanmingense and with B. elkanii, although none grouped with B. japonicum, B. canariense or B. liaoningense. To study the evolution of nodulation genes in these bacteria, the common nodulation gene, nodA, and host-specific nodulation genes, nodZ, noeE, and noeI, were analyzed. The nodA phylogeny showed that the cowpea and peanut Bradyrhizobium isolates represent various locally adapted groups or ecotypes that form part of Clade III of the seven known BradyrhizobiumnodA clades. This large and highly diverse clade comprises all strains from sub-Saharan Africa, as well as some originating from the Americas, Australia, Indonesia, China and Japan. Some similar groupings were supported by the other nodulation genes, although the overall phylogenies for the nodulation genes were incongruent with that inferred from the core genome genes, suggesting that horizontal gene transfer significantly influences the evolution of cowpea and peanut root-nodule bacteria. Furthermore, identification of the nodZ, noeI, and noeE genes in the isolates tested indicates that African Bradyrhizobium species may produce highly decorated nodulation factors, which potentially represent an important adaptation enabling nodulation of a great variety of legumes inhabiting the African continent.


Assuntos
Arachis/genética , Bradyrhizobium/genética , Fabaceae/genética , Genes de Plantas/genética , África , Técnicas de Transferência de Genes , Transferência Genética Horizontal , Variação Genética , Geografia , Modelos Genéticos , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Especificidade da Espécie
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