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1.
Chemosphere ; 241: 125006, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31590016

RESUMO

The use of ornamental plant will increase with the improvement in living standards in green and blue-green infrastructure of urban settings. Nicotiana alata is an ornamental plant, frequently grown as a model plant for horticulture, medicine, and scientific research studies throughout the world. Despite its popularity, little is known about the response of N. alata against heavy metals (HMs). This work is based on the hydroponic study to identify the impacts of selected HMs (Cd, Cr, Cu, Ni and Pb) on N. alata, at 0, 50 and 100 µM concentration, with the co-application of EDTA, at 0 and 2.5 mM in hydroponics system. The HMs uptake was found to be dose dependent, with significant higher uptake at 100 µM of respective HM. Highest cumulative uptake (mg kg-1 of HMs in root, shoot, and leaf dried weight) noted were 767.50 ±â€¯50.83, 862.30 ±â€¯23.83, 271.29 ±â€¯18.68, 1117.49 ±â€¯46.10 and 2166.81 ±â€¯102.09, for Cd, Cr, Cu, Ni, and Pb at 100 µM, respectively. It was identified that EDTA co-application with HMs resulted in boosted HMs uptake, with cumulative uptake percentage increment of 41.62, 54.67, 53.98, 34.48 and 19.92% for 100 µM of Cd, Cr, Cu, Ni, and Pb, respectively. Higher uptake led to negative impact on plant physiology, photosynthetic pigments, and higher lipid peroxidation, H2O2 contents, and electrolyte leakage that increased the stress. Higher HMs uptake induced higher antioxidant enzymatic response. It is recommended to incorporate appropriate soil modification to grow N. alata in sustainable infrastructures.


Assuntos
Ácido Edético/farmacologia , Metais Pesados/farmacocinética , Poluentes do Solo/farmacocinética , Tabaco/crescimento & desenvolvimento , Tabaco/metabolismo , Peróxido de Hidrogênio/análise , Metais Pesados/análise , Oxirredutases/análise , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Poluentes do Solo/análise
2.
Arch Virol ; 165(2): 367-375, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31845151

RESUMO

The genus Tobravirus comprises three species: Tobacco rattle virus, Pea early-browning virus and Pepper ringspot virus. The genomes of tobraviruses consist of two positive-sense single-stranded RNA segments (RNA1 and RNA2). Infectious clones of TRV are extensively used as virus-induced gene-silencing (VIGS) vectors for studies of virus-host interactions and functions of plant genes. Complete infectious clones of pepper ringspot virus (PepRSV), the only tobravirus present in Brazil, however, have not yet been reported. Infectious clones will help to identify unique features of PepRSV RNA2 and provide another option for development of VIGS vectors. We constructed infectious clones based on two PepRSV isolates, CAM (RNA1 and RNA2) and LAV (RNA2). The cDNA constructs for both homologous (RNA1 and RNA2 of the CAM isolate) and heterologous (RNA1/CAM and RNA2/LAV) combinations were infectious in Nicotiana benthamiana plants. VIGS vector constructs with green fluorescent protein or phytoene desaturase genes inserted in RNA2 silenced the target genes. The systemic translocation of the PepRSV RNA1 construct alone (nonmultiple infection) was also confirmed in an N. benthamiana plant. These results are similar to those reported for tobacco rattle virus.


Assuntos
Vetores Genéticos , Doenças das Plantas/virologia , Vírus de RNA/crescimento & desenvolvimento , Vírus de RNA/genética , Brasil , Genes Reporter , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Oxirredutases/análise , Oxirredutases/genética , Vírus de RNA/isolamento & purificação , Genética Reversa , Coloração e Rotulagem , Tabaco/virologia
3.
Environ Monit Assess ; 191(9): 544, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31392486

RESUMO

The development of regional or local maximum permissible concentration (MPC) for a pollutant in the soil requires the field or laboratory simulation of pollution. The experimental design should include the control (uncontaminated soil with the background concentration of the pollutant) and at least three treatments with different pollutant concentrations in the range between 2 and 10 background values. Experiments are performed in at least three replicates. Soil samples are taken 30 days after contamination. In each soil sample, six biological parameters are determined: total bacterial abundance, Azotobacter abundance, catalase activity, dehydrogenase activity, cellulolytic activity, and radish root length. Analyses are made in at least six replicates. From these biological parameters, the integrated biological index (IBI) of soil is calculated. For this purpose, the value of each parameter in the uncontaminated soil is taken as 100%, and its values in the contaminated soils are expressed as percentages. The mean values of six parameters for the contaminated treatments are determined. The obtained IBI values are expressed in percentages of the background. Then, a regression equation describing the decrease in IBI values as a function of pollutant concentration in the soil is derived. The pollutant concentration corresponding to the IBI decrease by 10% of the control, which indicates a disturbance of the holistic biogeocenotic functions of soil, is calculated from this equation.


Assuntos
Monitoramento Ambiental/normas , Poluição Ambiental/análise , Metais Pesados/análise , Poluentes do Solo/análise , Solo/química , Azotobacter/isolamento & purificação , Carga Bacteriana , Catalase/análise , Oxirredutases/análise , Microbiologia do Solo
4.
BMC Complement Altern Med ; 19(1): 227, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31438933

RESUMO

BACKGROUND: Oxidative stress is an imbalance between the levels of reactive oxygen species (ROS), reactive nitrogen species (RNS) and endogenous antioxidants. The aetiology and pathogenesis of several oral diseases are attributed to this process. The antioxidant enzymes secreted in the saliva by submandibular glands maintain oral health through the scavenging of ROS. The objective of this work was to study the capacity of an aqueous extract of L. divaricata (AE), and its majority compound, nordihydroguariaretic acid (NDGA), to modulate the pro-oxidant/antioxidant status in submandibular glands in a model of oxidative stress induced by streptozotocin (STZ) in rats. METHODS: To induce oxidative stress with STZ, a group of animals was treated i.p. with 1 X PBS (control group) and other group was injected i.p. once with STZ (60 mg/kg). Ten days after the treatment, blood samples were taken from the tail vain to determine the glucose levels. Animals with glucose values ≥300 mg/ml were selected. The submandibular glands of control and STZ treated animals were incubated with either the AE (500 µg/ml) or with NDGA (1.5 µg/ml), and the content of malondialdehyde (MDA), protein carbonyl groups, ROS and RNS, and the activity and expression of peroxidase (Px), superoxide dismutase (SOD) and inducible nitric oxide synthase (iNOS) were assayed. RESULTS: AE decreased the levels of MDA (##P < 0.01) and protein carbonyl groups (#P < 0.05), and modulated the levels of ROS such as hydrogen peroxide (H2O2)(##P < 0.01), superoxide anion (O2.-) (#P < 0.05) and nitric oxide (NO) (#P < 0.05) in relation to the modulation of Px and iNOS expression. NDGA was found to be involved in these effects. CONCLUSIONS: The antioxidant activity of the AE in the submandibular glands would allow the maintenance of the antioxidant pool to prevent oral oxidative diseases.


Assuntos
Diabetes Mellitus Experimental/metabolismo , Larrea/química , Masoprocol/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Glândula Submandibular/efeitos dos fármacos , Animais , Antioxidantes/farmacologia , Feminino , Malondialdeído/análise , Oxirredutases/análise , Ratos , Ratos Wistar , Glândula Submandibular/química , Glândula Submandibular/enzimologia
5.
BMC Biochem ; 20(1): 4, 2019 04 08.
Artigo em Inglês | MEDLINE | ID: mdl-30961528

RESUMO

BACKGROUND: The reduction of tetrazolium salts by NAD(P)H to formazan product has been widely used to determine the metabolic activity of cells, and as an indicator of cell viability. However, the application of a WST-8 based assay for the quantitative measurement of dehydrogenase enzyme activity has not been described before. In this study, we reported the application of an assay based on the tetrazolium salt WST-8 for the quantitative measurement of dehydrogenase activity. The assay is performed in a microplate format, where a single endpoint is measured at 450 nm. RESULTS: The optimized dehydrogenase-WST-8 assay conditions, the limit of detection (LOD), accuracy, and precision for measuring NAD(P)H, were demonstrated. The sensitivity of the WST-8 assay for detecting NAD(P)H was 5-fold greater than the spectrophotometric measurement of NAD(P)H absorption at 340 nm (LOD of 0.3 nmole vs 1.7 nmole, respectively). In the dehydrogenase assay, the colorimetric WST-8 method exhibits excellent assay reproducibility with a Z' factor of 0.9. The WST-8 assay was also used to determine dehydrogenase activity in biological samples, and for screening the substrate of uncharacterized short-chain dehydrogenase/oxidoreductase from Burkholderia pseudomallei. CONCLUSION: The results suggest that the WST-8 assay is a sensitive and rapid method for determining NAD(P)H concentration and dehydrogenase enzyme activity, which can be further applied for the high-throughput screening of dehydrogenases.


Assuntos
Colorimetria/métodos , Oxirredutases/análise , Sais de Tetrazólio/química , Burkholderia pseudomallei/enzimologia , Glucosefosfato Desidrogenase , Humanos , Limite de Detecção , NAD/análise , NAD/metabolismo , NADP/análise , NADP/metabolismo , Oxirredutases/metabolismo , Espectrofotometria
6.
Int J Mol Sci ; 20(8)2019 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-31018495

RESUMO

Plant cell walls mostly comprise polysaccharides and proteins. The composition of monocots' primary cell walls differs from that of dicots walls with respect to the type of hemicelluloses, the reduction of pectin abundance and the presence of aromatic molecules. Cell wall proteins (CWPs) differ among plant species, and their distribution within functional classes varies according to cell types, organs, developmental stages and/or environmental conditions. In this review, we go deeper into the findings of cell wall proteomics in monocot species and make a comparative analysis of the CWPs identified, considering their predicted functions, the organs analyzed, the plant developmental stage and their possible use as targets for biofuel production. Arabidopsis thaliana CWPs were considered as a reference to allow comparisons among different monocots, i.e., Brachypodium distachyon, Saccharum spp. and Oryza sativa. Altogether, 1159 CWPs have been acknowledged, and specificities and similarities are discussed. In particular, a search for A. thaliana homologs of CWPs identified so far in monocots allows the definition of monocot CWPs characteristics. Finally, the analysis of monocot CWPs appears to be a powerful tool for identifying candidate proteins of interest for tailoring cell walls to increase biomass yield of transformation for second-generation biofuels production.


Assuntos
Brachypodium/metabolismo , Parede Celular/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Saccharum/metabolismo , Brachypodium/química , Metabolismo dos Carboidratos , Parede Celular/química , Metabolismo dos Lipídeos , Oryza/química , Oxirredutases/análise , Oxirredutases/isolamento & purificação , Oxirredutases/metabolismo , Peptídeo Hidrolases/análise , Peptídeo Hidrolases/isolamento & purificação , Peptídeo Hidrolases/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/isolamento & purificação , Proteômica , Saccharum/química , Transdução de Sinais
7.
Appl Microbiol Biotechnol ; 103(10): 4229-4240, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30923872

RESUMO

Extremely acidic soils of natural forests in Nanling National Nature Reserve have been previously investigated and revisited in two successive years to reveal the active ammonia oxidizers. Ammonia-oxidizing archaea (AOA) rather than ammonia-oxidizing bacteria (AOB) were found more functionally important in the extremely acidic soils of the natural forests in Nanling National Nature Reserve. The relative abundances of Nitrosotalea, Nitrososphaera sister group, and Nitrososphaera lineages recovered by ammonia monooxygenase subunit A (amoA) transcripts were reassessed and compared to AOA communities formerly detected by genomic DNA. Nitrosotalea, previously found the most abundant AOA, were the second-most-active lineage after Nitrososphaera sister group. Our field study results, therefore, propose the acidophilic AOA, Nitrosotalea, can better reside in extremely acidic soils while they may not contribute to nitrification proportionately according to their abundances or they are less functionally active. In contrast, the functional importance of Nitrososphaera sister group may be previously underestimated and the functional dominance further extends their ecological distribution as little has been reported. Nitrososphaera gargensis-like AOA, the third abundant lineage, were more active in summer. The analyses of AOA community composition and its correlation with environmental parameters support the previous observations of the potential impact of organic matter on AOA composition. Al3+, however, did not show a strong adverse correlation with the abundances of functional AOA unlike in the DNA-based study. The new data further emphasize the functional dominance of AOA in extremely acidic soils, and unveil the relative contributions of AOA lineages to nitrification and their community transitions under the environmental influences.


Assuntos
Amônia/metabolismo , Archaea/classificação , Archaea/metabolismo , Florestas , Microbiota , Microbiologia do Solo , Archaea/enzimologia , Archaea/genética , Concentração de Íons de Hidrogênio , Oxirredução , Oxirredutases/análise , Oxirredutases/genética , RNA Mensageiro/análise , RNA Mensageiro/genética , Solo/química
8.
World J Microbiol Biotechnol ; 35(3): 47, 2019 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-30834977

RESUMO

The present study was aimed to assess the scope of native potential endophyte Pseudomonas aeruginosa (LSE-2) strain (KX925973) with recommended Bradyrhizobium sp. (LSBR-3) (KF906140) for synergistic effect to develop as consortium biofertilizer of soybean. A total of 28 non-rhizobial endophytic bacteria were isolated from cultivated and wild sp. of soybean. All isolates were screened for multifarious PGP traits viz. Indole-3-acetic acid (IAA), phosphate (P) and zinc (Zn) solubilization, siderophore, cell wall degrading enzymes and pathogenicity. Compatible of LSBR-3 and LSE-2 enhanced IAA, P-solubilization, 1-aminocyclopropane-carboxylate deaminase and biofilm formation over the single inoculant treatment. Further, consortium was evaluated in vivo for growth, symbiotic traits, nutrient acquisition, soil quality parameters and yield attributes of soybean. Improvement in growth parameters were recorded with dual inoculant LSBR-3 + LSE-2 as compared to LSBR-3 alone and un-inoculated control treatments. Significantly (p ≥ 0.05) high symbiotic and soil quality parameters (phosphatase and soil dehydrogenase activity) was recorded with LSBR-3 + LSE-2 at vegetative and flowering stage as compared to LSBR-3 alone and un-inoculated control treatments. Single inoculation of LSBR-3 improved grain yield by 4.25% over the un-inoculated control treatment, further, enhancement in yield was recorded with consortium inoculant (LSBR-3 and LSE-2) by 3.47% over the LSBR-3 alone. Application of consortium inoculant (LSBR-3 + LSE-2) gave an additional income of Rs. 5089/ha over the un-inoculated control treatment. The results, thus strongly suggest that endophytic diazotroph LSE-2 can be used as potent bio-inoculant along with LSBR-3 as bio-enhancer for improving soybean productivity in a sustainable system.


Assuntos
Bradyrhizobium/fisiologia , Endófitos , Nutrientes , Desenvolvimento Vegetal , Pseudomonas aeruginosa/fisiologia , Soja/crescimento & desenvolvimento , Soja/microbiologia , Simbiose , Biofilmes/crescimento & desenvolvimento , Bradyrhizobium/isolamento & purificação , Índia , Ácidos Indolacéticos/metabolismo , Nitrogênio/análise , Oxirredutases/análise , Fosfatos/metabolismo , Monoéster Fosfórico Hidrolases/análise , Filogenia , Raízes de Plantas/química , Raízes de Plantas/microbiologia , Brotos de Planta/química , Brotos de Planta/microbiologia , Potássio/análise , Pseudomonas aeruginosa/classificação , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/isolamento & purificação , RNA Ribossômico 16S/genética , Sideróforos/metabolismo , Solo/química , Microbiologia do Solo , Solubilidade , Virulência , Zinco/metabolismo
9.
Arch Microbiol ; 201(5): 673-678, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30798341

RESUMO

A Gram-stain-negative, aerobic, mobile, and rod-shaped bacterium, designated JJ3T, was isolated from peanut rhizospheric soil in Qingdao, Shandong Province, China, and was characterized using a polyphasic approach. Strain JJ3T grew at 4-40 °C, at pH 5.0-9.0 and 0-4% NaCl. The strain was positive for both catalase and oxidase tests, and was able to degrade aflatoxin B1. According to the 16S rRNA gene sequence comparisons, the strain JJ3T was identified as a member of the genus Pseudomonas and was most closely related to Pseudomonas japonica JCM 21532T and Pseudomonas alkylphenolica JCM 16553T with sequence similarity of 99.0% and 98.9%, respectively. A multilocus sequence analysis (MLSA) of concatenating 16S rRNA, gyrB and rpoD gene sequences showed that strain JJ3T belonged to the Pseudomonas putida subcluster. Genomic comparison of strain JJ3T with its closest phylogenetic type strain using average nucleotide index (ANI) and digital DNA-DNA relatedness revealed 76.7-82.9% and 20.2-37.1%, respectively. All values were distinctly lower than the thresholds established for species differentiation. The predominant cellular fatty acids of strain JJ3T were C17:0 cyclo (24.0%), C16:0 (21.4%), summed features 3 (C16:1ω7c and/or C16:1ω6c) (11.5%) and summed features 8 (C18:1ω7c and/or C18:1ω6c) (10.5%). The major polar lipids of strain JJ3T were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The physiological, biochemical, and genetic characteristics support the assignment of JJ3T to the genus Pseudomonas, but are different to those of phylogenetically neighboring species to represent a novel species. The name Pseudomonas qingdaonensis sp. nov. is proposed, with JJ3T (= JCM 32579T = KCTC 62384T = CGMCC 1.16493T) as the type strain.


Assuntos
Aflatoxina B1/metabolismo , Arachis/microbiologia , Pseudomonas/classificação , Pseudomonas/metabolismo , Técnicas de Tipagem Bacteriana , Composição de Bases/genética , Catalase/análise , China , DNA Bacteriano/genética , Ácidos Graxos/análise , Genes Bacterianos , Tipagem de Sequências Multilocus , Hibridização de Ácido Nucleico , Oxirredutases/análise , Fosfolipídeos/análise , Filogenia , Pseudomonas/genética , Pseudomonas/isolamento & purificação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Solo , Microbiologia do Solo
10.
Sci Total Environ ; 649: 760-769, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30176486

RESUMO

The effect of ammonia on methanogenic biomass from a full-scale agricultural digester treating nitrogen-rich materials was characterized in batch activity assays subjected to increasing concentrations of total ammonia N. Acetotrophic and methanogenic profiles displayed prolonged lag phases and reduced specific activity rates at 6.0 gN-TAN L-1, though identical methane yields were ultimately reached. These results agreed with the expression levels of selected genes from bacteria and methanogenic archaea (qPCR of 16S rRNA and mrcA cDNA transcripts). Compound-specific isotope analysis of biogas indicated that ammonia exposure was associated to a transition in methanogenic activity from acetotrophy at 1.0 gN-TAN L-1 to intermediate and complete hydrogenotrophy at 3.5 and 6.0 gN-TAN L-1. Such pattern matched the results of 16S-Illumina sequencing of genes and transcripts in that predominant methanogens shifted, along with increasing ammonia, from the obligate acetotroph Methanosaeta to the hydrogenotrophic Methanoculleus and the poorly understood methylotrophic Methanomassiliicoccus. The underlying bacterial community structure remained rather stable but, at 6.0 gN-TAN L-1, the expression level increased considerably for a number of ribotypes that are related to potentially syntrophic genera (e.g. Clostridium, Bellilinea, Longilinea, and Bacteroides). The predominance of hydrogenotrophy at high ammonia levels clearly points to the occurrence of the syntrophic acetate oxidation (SAO), but known SAO bacteria were only found in very low numbers. The potential role of the identified bacterial and archaeal taxa with a view on SAO and on stability of the anaerobic digestion process under ammonia stress has been discussed.


Assuntos
Agricultura , Amônia/metabolismo , Archaea/metabolismo , Bactérias/metabolismo , Biomassa , Resíduos Industriais/análise , Microbiota , Anaerobiose , Archaea/genética , Proteínas Arqueais/análise , Bactérias/genética , Reatores Biológicos , Genes Bacterianos , Oxirredutases/análise , RNA Ribossômico 16S/análise
11.
Curr Med Chem ; 26(21): 3923-3957, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-29446729

RESUMO

Enzymes are a class of macromolecules that function as highly efficient and specific biological catalysts requiring only mild reaction conditions. Enzymes are essential to maintaining life activities, including promoting metabolism and homeostasis, and participating in a variety of physiological functions. Accordingly, enzymatic levels and activity are closely related to the health of the organism, where enzymatic dysfunctions often lead to corresponding diseases in the host. Due to this, diagnosis of certain diseases is based on the levels and activity of certain enzymes. Therefore, rapid real-time and accurate detection of enzymes in situ are important for diagnosis, monitoring, clinical treatment and pathological studies of disease. Fluorescent probes have unique advantages in terms of detecting enzymes, including being simple to use in highly sensitive and selective real-time rapid in-situ noninvasive and highly spatial resolution visual imaging. However, fluorescent probes are most commonly used to detect oxidoreductases, transferases and hydrolases due to the processes and types of enzyme reactions. This paper summarizes the application of fluorescent probes to detect these three types of enzymes over the past five years. In addition, we introduce the mechanisms underlying detection of these enzymes by their corresponding probes.


Assuntos
Corantes Fluorescentes/química , Fluorometria , Hidrolases/análise , Oxirredutases/análise , Transferases/análise , Humanos , Hidrolases/metabolismo , Oxirredutases/metabolismo , Transferases/metabolismo
12.
Theranostics ; 8(19): 5289-5306, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30555547

RESUMO

Molecule interacting with CasL2 (MICAL2), a microtubule-associated monooxygenase, is highly expressed in various cancers and is involved in cancer pathogenesis, but the mechanisms underlying its regulation in carcinogenesis are unclear. In this study, we aim to clarify the mechanism by which MICAL2 participates in colorectal cancer (CRC) and identify novel markers for predicting prognosis of CRC patients. Methods: The value of MICAL2 in CRC prognosis was determined by immunohistochemical analysis of a CRC biopsy array. A short hairpin RNA target MICAL2 (shMICAL2) was designed to knock down MICAL2 expression and observe MICAL2's function on CRC cell growth. mRNA expression array was used to screen target molecules of MICAL2. HCT116 p53+/+ and HCT116 p53-/- cells were used to confirm whether MICAL2 exerts its oncogenic effect through p53. The in vivo effect of MICAL2 on CRC growth was assessed by subcutaneously injecting MICAL2-knockout CRC cells into the dorsal flank of each mouse. Immunofluorescence was used to observe the effect of MICAL2 on p53 cellular location. Reverse-phase nano ESI-LCMS analysis was used to investigate if MICAL2 mediates p53 oxidation. Results: MICAL2 was found to be highly expressed in CRC tissues, and its expression was associated with CRC carcinogenesis and poor patient outcome. MICAL2-knockdown decreased growth and colony formation of CRC cells, which was linked with cell cycle arrest and apoptosis. MICAL2 physically interacted with p53 and retained p53 in the cytoplasm. MICAL2 shortened the half-life of p53, and ectopic MICAL2 expression decreased p53 protein stability through ubiquitin degradation. MICAL2 was also found to oxidize p53 at methionine 40 and 160, which mediated p53 ubiquitin degradation. MICAL2-promoted CRC growth in vivo was confirmed in nude mice. Conclusion: MICAL2 binds to p53, retains p53 in the cytoplasm and oxidizes it at Met 40 and 160, promotes p53 ubiquitination, and decreases p53 function. MICAL2-reduced p53 promotes CRC development.


Assuntos
Neoplasias Colorretais/patologia , Metionina/metabolismo , Proteínas dos Microfilamentos/análise , Oxirredutases/análise , Processamento de Proteína Pós-Traducional , Proteólise , Proteína Supressora de Tumor p53/metabolismo , Adulto , Idoso , Animais , Biópsia , Carcinogênese , Proliferação de Células , Cromatografia Líquida , Modelos Animais de Doenças , Feminino , Técnicas de Silenciamento de Genes , Técnicas de Inativação de Genes , Células HCT116 , Xenoenxertos , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Nus , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo , Pessoa de Meia-Idade , Transplante de Neoplasias , Oxirredução , Oxirredutases/genética , Oxirredutases/metabolismo , Ligação Proteica , Espectrometria de Massas por Ionização por Electrospray , Ubiquitinação
13.
PLoS One ; 13(11): e0204081, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30457998

RESUMO

Apical periodontitis is frequently associated with the presence of bacteria biofilm, which has an indisputable impact on the prognosis of endodontic therapy due to the high resistance to adverse environmental conditions, chemicals, and antibiotic therapy that characterize bacteria within biofilm. The biofilm matrix acts as a protective shield over the encased microorganisms. The aim of this investigation was to identify the main biochemical components of biofilm matrix from endodontic mono- and dual-species biofilms. Enterococcus faecalis and Actinomyces naeslundii were cultured as mono- and dual-species biofilms for 14 days. Crude extracellular polymeric substances (EPSs) from biofilm matrices were extracted using chemical and physical methods. High-performance liquid chromatography, gas chromatography, and mass spectrometry were used to determine the carbohydrate, protein, and fatty acid components. Chemical analysis of the biofilm matrices revealed that they were mainly composed of stachyose, maltose, and mannose carbohydrates. The protein profile in all biofilm samples showed abundant oxidoreductases and chaperone proteins and some virulence- associated proteins mainly located in the membrane surface. High percentages of saturated and monounsaturated fatty acids were identified in all biofilm matrices, with a major prevalence of palmitic, stearic, and oleic acids. Based on the results, it was possible to obtain for the first time a general overview of the biochemical profile of endodontic biofilm matrices.


Assuntos
Actinomyces/fisiologia , Biofilmes/crescimento & desenvolvimento , Enterococcus faecalis/fisiologia , Periodontite Periapical/microbiologia , Proteínas de Bactérias/análise , Proteínas de Bactérias/metabolismo , Metabolismo dos Carboidratos , Carboidratos/análise , Ácidos Graxos Monoinsaturados/análise , Ácidos Graxos Monoinsaturados/metabolismo , Humanos , Chaperonas Moleculares/análise , Chaperonas Moleculares/metabolismo , Oxirredutases/análise , Oxirredutases/metabolismo
14.
J Food Sci ; 83(10): 2432-2438, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30300467

RESUMO

The relationship between glycolytic dehydrogenase, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH), lactic dehydrogenase (LDH), and meat color stability was studied in this study using ovine muscle. Three different ovine muscles, including M. longissimus lumborum (LL), M. semimembranosus (SM), and M. psoas major (PM), were obtained (n = 10, respectively), and then displayed for 7 days at 4 °C. The LL and SM muscle had higher surface redness, higher (P < 0.05) GAPDH activity, nicotinamide adenine dinucleotide (NADH) content, and lower (P < 0.05) LDH-B activity than PM muscles during display. The PM muscle had the worst color stability and lowest NADH content. These results suggest that variation in color stability of physiologically different muscles may be affected by glycolysis dehydrogenases. Comparatively, our data showed that GAPDH may play a more important role than LDH-B to maintain meat color stability.


Assuntos
Gliceraldeído-3-Fosfato Desidrogenases/análise , Carne/análise , Músculo Esquelético/enzimologia , Oxirredutases/análise , Animais , Cor , Glicólise , Músculo Esquelético/química , Oxirredutases/metabolismo , Ovinos
15.
Ecotoxicology ; 27(9): 1188-1202, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30173333

RESUMO

As one of the most widely used pesticides in agriculture, chlorothalonil can pose threat to soil ecosystems. Therefore, the impact of this substance on the development of microbiological and biochemical properties of the soil as well as on the growth of spring wheat was evaluated. The study was conducted with two soils (loamy sand with pHKCl 5.6 and sandy loam with pHKCl 7.00), to which fungicide was used in the following doses: 0.00, 0.166 (recommended dose), 1.660, and 16.60 mg kg-1 dry matter of soil (DM of soil). In addition, we determined the effectiveness of fertilizing substances (Lignohumat Super and Bioilsa N 12.5) in the restoration of soil homeostasis and chlorothalonil degradation in the soil. Chlorothalonil caused modifications in the count and biological diversity of soil microorganisms. It stimulated the growth of heterotrophic bacteria and actinobacteria, and inhibited the growth of fungi. This pesticide was a potent inhibitor of dehydrogenase, catalase and acid phosphatase activities. It showed variable effects on urease and alkaline phosphatase. The fungicide also a reduction the yield of dry matter of the aboveground parts of spring wheat. It should, however, be noted that these changes in the soil environment occurred after the introduction of higher doses of chlorothalonil. The fertilizing substances used contributed to enhanced microbial and biochemical activities of soils, while they did not significantly affect plant yields. The Bioilsa N 12.5 preparation was effective in chlorothalonil degradation, while Lignohumat Super reduced the degradation rate of the tested fungicide. Based on the conducted experiment, an ecological risk assessment of chlorothalonil was made by estimating the changes occurring in the soil environment evaluated through the microbiological and biochemical analyses of the soil.


Assuntos
Fungicidas Industriais/toxicidade , Nitrilos/toxicidade , Microbiologia do Solo , Poluentes do Solo/toxicidade , Oxirredutases/análise , Solo/química
16.
Methods Mol Biol ; 1853: 207-221, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30097946

RESUMO

Zymography on peroxidase-entrapped gels enables the determination of the intrinsic specific activity (ISA) of H2O2-producing oxidases in the presence of interfering enzymes including catalase (decomposing hydrogen peroxide). To reveal the searched oxidase, the zymography gel is incubated first in the developing solution containing the appropriate substrate of the oxidase and an oxidizable chromogen such as o-phenylenediamine (o-PDA) peroxidase substrate. The zymographic gel, after image analysis, can be restained by Coomassie Blue for molecular weights, for the whole electrophoretic pattern and for the protein concentration, thus allowing for the determination of oxidase ISA.


Assuntos
Eletroforese em Gel de Poliacrilamida , Oxirredutases , Proteínas , Animais , Catalase/análise , Catalase/química , Bovinos , Ativação Enzimática , Peróxido de Hidrogênio/química , Oxirredutases/análise , Oxirredutases/química , Proteínas/análise , Proteínas/química , Corantes de Rosanilina , Espectrofotometria/métodos , Coloração e Rotulagem , Especificidade por Substrato
17.
Acta Cir Bras ; 33(7): 565-576, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30110058

RESUMO

PURPOSE: To investigate the possible effects of argan oil on the healing of colorectal anastomoses. METHODS: n Group 1 (sham), laparotomy was performed and the colon was mobilized. In the control (Group 2) and argan oil (Group 3) groups, colonic resection and anastomosis were applied. To the control and sham groups, 2 mL of 0.9% NaCl was administred rectally, and in the argan oil group, 2 mL/day argan oil was applied rectally for 7 days. RESULTS: The mean bursting pressures of the argan oil and sham groups were significantly higher than the values in the control group. A significant difference was determined between the tissue hydroxyproline and prolidase levels of control group and other groups. Histopathologically, argan oil showed significant beneficial effects on colonic wound healing. In the argan oil and sham groups, the tissue malondialdehyde and fluorescent oxidation product levels were found to be lower and total sulfhydryl levels were higher than the control group. CONCLUSIONS: The rectally administered argan oil was observed to have significantly ameliorated wound healing parameters and exerted a significant antioxidant effect. This is the first study in the literature about the beneficial effects of argan oil on colorectal anastomoses.


Assuntos
Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Colo/cirurgia , Óleos Vegetais/uso terapêutico , Reto/cirurgia , Cicatrização/efeitos dos fármacos , Anastomose Cirúrgica , Animais , Colágeno/análise , Colo/patologia , Dipeptidases/análise , Feminino , Hidroxiprolina/análise , Malondialdeído/análise , Estresse Oxidativo/efeitos dos fármacos , Oxirredutases/análise , Distribuição Aleatória , Ratos Wistar , Reto/patologia , Reprodutibilidade dos Testes , Espectrofotometria , Ferida Cirúrgica/tratamento farmacológico , Ferida Cirúrgica/patologia , Resultado do Tratamento
18.
Saudi J Kidney Dis Transpl ; 29(3): 545-553, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29970730

RESUMO

This study was performed to investigate the protective effect of combined use of Vitamins E and C on colistin-induced tubular damage in rat. Animals were treated with sterile saline, colistin methanesulfonate (CMS), CMS + Vitamin E + Vitamin C, and Vitamin E + vitamin C, respectively, for seven days. Thereafter, animals were sacrificed and the urine N-acetyl-b-D-glucosaminidase (NAG) and gamma-glutamyl transferase (GGT) levels, plasma level of creatinine (Cr), vitamin E and vitamin C, and renal tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), as well as renal histology were performed. CMS induced acute tubular necrosis, increased the NAG, GGT, and MDA levels, and reduced the Vitamin E, Vitamin C, SOD, CAT, and GPx activities. Co-treatment with vitamins E and C restored all biochemical parameters cited above and improved the histopathological damage. Tubular damage induced by colistin is at least partly due to oxidative stress. Nephroprotective effect of Vitamins E and C is partially mediated through its antioxidant properties, and the higher protection by the combination of these vitamins is related to its synergistic effects.


Assuntos
Antioxidantes/farmacologia , Ácido Ascórbico/farmacologia , Colistina/toxicidade , Rim/efeitos dos fármacos , Vitamina E/farmacologia , Lesão Renal Aguda/induzido quimicamente , Lesão Renal Aguda/patologia , Animais , Rim/química , Rim/enzimologia , Rim/patologia , Masculino , Malondialdeído/análise , Oxirredutases/análise , Ratos , Ratos Wistar
19.
Acta cir. bras ; 33(7): 565-576, July 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-949365

RESUMO

Abstract Purpose: To investigate the possible effects of argan oil on the healing of colorectal anastomoses. Methods: I n Group 1 (sham), laparotomy was performed and the colon was mobilized. In the control (Group 2) and argan oil (Group 3) groups, colonic resection and anastomosis were applied. To the control and sham groups, 2 mL of 0.9% NaCl was administred rectally, and in the argan oil group, 2 mL/day argan oil was applied rectally for 7 days. Results: The mean bursting pressures of the argan oil and sham groups were significantly higher than the values in the control group. A significant difference was determined between the tissue hydroxyproline and prolidase levels of control group and other groups. Histopathologically, argan oil showed significant beneficial effects on colonic wound healing. In the argan oil and sham groups, the tissue malondialdehyde and fluorescent oxidation product levels were found to be lower and total sulfhydryl levels were higher than the control group. Conclusions: The rectally administered argan oil was observed to have significantly ameliorated wound healing parameters and exerted a significant antioxidant effect. This is the first study in the literature about the beneficial effects of argan oil on colorectal anastomoses.


Assuntos
Animais , Feminino , Reto/cirurgia , Cicatrização/efeitos dos fármacos , Óleos Vegetais/uso terapêutico , Colo/cirurgia , Anti-Inflamatórios/uso terapêutico , Antioxidantes/uso terapêutico , Oxirredutases/análise , Reto/patologia , Espectrofotometria , Anastomose Cirúrgica , Distribuição Aleatória , Reprodutibilidade dos Testes , Colágeno/análise , Resultado do Tratamento , Ratos Wistar , Colo/patologia , Estresse Oxidativo/efeitos dos fármacos , Dipeptidases/análise , Ferida Cirúrgica/patologia , Ferida Cirúrgica/tratamento farmacológico , Hidroxiprolina/análise , Malondialdeído/análise
20.
Microbiologyopen ; 7(5): e00575, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-29900700

RESUMO

Eggerthella timonensis strain Marseille-P3135 is a new bacterial species, isolated from the stool sample of a healthy 8-year-old pygmy female. This strain (LT598568) showed a 16S rRNA sequence similarity of 96.95% with its phylogenetically closest species with standing in nomenclature Eggerthella lenta strain DSM 2243 (AF292375). This bacterium is a nonspore forming, Gram-positive, nonmotile rod with catalase but no oxidase activity. Its genome is 3,916,897 bp long with 65.17 mol% of G + C content. Of the 3,371 predicted genes, 57 were RNAs and 3,314 were protein-coding genes. Here, we report the main phenotypic, biochemical, and genotypic characteristics of E. timonensis strain Marseille-P3135 (=CSUR P3135, =CCUG 70327); ti.mo.nen'sis, N.L. masc. adj., with timonensis referring to La Timone, which is the name of the hospital in Marseille (France) where this work was performed). Strain is a nonmotile Gram-positive rod, unable to sporulate, oxidase negative, and catalase positive. It grows under anaerobic conditions between 25°C and 42°C but optimally at 37°C.


Assuntos
Actinobacteria/classificação , Actinobacteria/isolamento & purificação , Fezes/microbiologia , Actinobacteria/genética , Anaerobiose , Técnicas de Tipagem Bacteriana , Composição de Bases , Catalase/análise , Criança , Análise por Conglomerados , Congo , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Feminino , Genótipo , Humanos , Locomoção , Espectrometria de Massas , Microscopia Eletrônica de Transmissão , Oxirredutases/análise , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Temperatura , Sequenciamento Completo do Genoma
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