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1.
Phytochemistry ; 180: 112522, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33010537

RESUMO

Strawberries (Fragaria × ananassa Duch.) are one of the most economically important fruit crops worldwide, several commercially viable cultivars are cultivated in the northern region of Thailand. The morphological characters at the young vegetative seedling stage can be very similar, which has hindered breeding efforts. The present study assesses the ability of random amplification of polymorphic DNA (RAPD) markers and metabolomics techniques to distinguish six strawberry cultivars. Both techniques showed congruent results for the leaf tissue and classified the cultivars into three major clusters. For the most different cultivars, Akihime and Praratchatan No.80, fruits were analysed at eight fruit ripening stages. The data highlighted a broad biological variation at the early ripening stages and less biological variation at the mature stages. Key metabolic differences included the polyphenol profile in Praratchatan No.80 and fatty acid synthesis/oxidation in Akihime. In summary, the RAPD and metabolite data can be used to distinguish strawberry cultivars and elucidate the metabolite composition of each phenotype. This approach to the characterisation of genotypes will benefit future breeding programmes.


Assuntos
Fragaria , Fragaria/genética , Frutas/genética , Marcadores Genéticos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Tailândia
2.
Vestn Oftalmol ; 136(4. Vyp. 2): 193-200, 2020.
Artigo em Russo | MEDLINE | ID: mdl-32880139

RESUMO

PURPOSE: To evaluate the effectiveness of bony orbital decompression in patients with dysthyroid optic neuropathy (DON). MATERIAL AND METHODS: The study analyzed 255 patients with thyroid eye disease (TED) and bony orbital decompression. Those among them who had DON as an indication for surgery were investigated further. Patients underwent standard ophthalmological examination, computer perimetry, color vision assessment using Ishihara tables, relative afferent pupillary defect (RAPD), computed tomography (CT) of the orbit, and in some cases optical coherence tomography (OCT) of the optic nerve. RESULTS: Final analysis included 31 patients (52 eyes). On 13 orbits, only lateral wall decompression was performed, and medial wall decompression was the only intervention in 7 orbits. In other cases, these techniques were performed either simultaneously - in 14 orbits, or alternately - in 18 orbits. In the postoperative period, all patients showed significant positive dynamics in terms of visual acuity, visual field, and proptosis. In all cases, decrease of the amount of orbital inflammation was observed. Exophthalmos significantly decreased after surgery and averaged 20.5±3.1 mm, which is 4.7 mm less than the initial one. All changes were statistically significant (p<0.01). CONCLUSION: Bony orbital decompression is an effective and safe treatment option for DON resistant to high doses of glucocorticoids. In the vast majority of cases, this intervention is the only way to improve and stabilize visual function in this severe category of patients.


Assuntos
Oftalmopatia de Graves/cirurgia , Doenças do Nervo Óptico/cirurgia , Descompressão Cirúrgica , Humanos , Nervo Óptico , Órbita/cirurgia , Técnica de Amplificação ao Acaso de DNA Polimórfico , Estudos Retrospectivos , Resultado do Tratamento
3.
Biol Res ; 53(1): 37, 2020 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-32847618

RESUMO

BACKGROUND: Soil salinity causes huge economic losses to agriculture productivity in arid and semiarid areas worldwide. The affected plants face disturbances in osmotic adjustment, nutrient transport, ionic toxicity and reduced photosynthesis. Conventional breeding approaches produce little success in combating various stresses in plants. However, non-conventional approaches, such as in vitro tissue culturing, produce genetic variability in the development of salt-tolerant plants, particularly in woody trees. RESULTS: Embryogenic callus cultures of the date palm cultivar Khalas were subjected to various salt levels ranging from 0 to 300 mM in eight subcultures. The regenerants obtained from the salt-treated cultures were regenerated and evaluated using the same concentration of NaCl with which the calli were treated. All the salt-adapted (SA) regenerants showed improved growth characteristics, physiological performance, ion concentrations and K+/Na+ ratios than the salt non-adapted (SNA) regenerants and the control. Regression between the leaf Na+ concentration and net photosynthesis revealed an inverse nonlinear correlation in the SNA regenerants. Leaf K+ contents and stomatal conductance showed a strong linear relationship in SA regenerants compared with the inverse linear correlation, and a very poor coefficient of determination in SNA regenerants. The genetic fidelity of the selected SA regenerants was also tested using 36 random amplified polymorphic DNA (RAPD) primers, of which 26 produced scorable bands. The primers generated 1-10 bands, with an average of 5.4 bands per RAPD primer; there was no variation between SA regenerants and the negative control. CONCLUSION: This is the first report of the variants generated from salt-stressed cultures and their potential adaptation to salinity in date palm cv. Khalas. The massive production of salt stress-adapted date palm plants may be much easier using the salt adaptation approach. Such plants can perform better during exposure to salt stress compared to the non-treated date palm plants.


Assuntos
Aclimatação , Phoeniceae/genética , Tolerância ao Sal/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Salinidade
4.
Niger J Clin Pract ; 23(7): 912-918, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32620719

RESUMO

Background: Investigating genetic relatedness between methicillin-resistant Staphylococcus aureus (MRSA) strains from humans and different animal species may clarify the epidemiological characteristic of MRSA infections together. Aim: The aim of the study was to perform genotypic characterization and type strains of MRSA isolated from different clinical sources, by molecular techniques. Materials and Methods: The molecular characterization of the strains was performed by polymerase chain reaction (PCR), using several specific oligonucleotides. These were as follows: S. aureus species-specific sau gene, mecA gene coding PBP2a responsible for methicillin resistance, femA gene coding for a protein, which influences the level of methicillin resistance of S. aureus, and is universally present in all MRSA strains; spa gene coding for protein A; coa gene coding for coagulase, and blaZ gene coding for the production of beta-lactamase. To determine the genetic diversity of these strains, random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) was performed. Results: Among the 415 S. aureus strains, 61 were phenotypically identified as MRSA, and confirmed as S. aureus by amplification of sau gene. However, 90.16% of the strains were mecA positive, while all were negative for femA gene. The presence and polymorphism of coa and spa genes were investigated and 83.60% and 18.03% strains were positive for coa and spa, respectively. While these strains were grouped into six coa-types by PCR, no polymorphism was found for spa gene among strains having only single 190 bp of the band. bla genes were found in 75.40% of strains. These strains were divided into 12 RAPD types. Conclusions: The results showed the relatively high heterogeneity and variation of coa gene among MRSA strains, while further studies on sequencing of these strains may identify which sequence type is predominant in this region.


Assuntos
Proteínas de Bactérias/genética , Staphylococcus aureus Resistente à Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Animais , Antibacterianos/farmacologia , Proteínas de Bactérias/isolamento & purificação , Coagulase/genética , Genótipo , Humanos , Meticilina , Resistência a Meticilina/genética , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética
5.
Sci Rep ; 10(1): 6826, 2020 04 22.
Artigo em Inglês | MEDLINE | ID: mdl-32321960

RESUMO

Five different weed plants viz. Convulvulus arvensis, Chenopodium murale, Tribulus terrestris, Trianthema portulacastrum, and Achyranthes aspera were investigated for their entomocidal and genotoxic effects against Culex quinquefasciatus mosquitoes. High mortality was observed at 72 hours in a dose dependent manner. Among all the tested plants, A. aspera was found highly significant which showed 100% mortality at 250 ppm after 72 hours with LC50 of 87.46, 39.08 and 9.22 ppm at 24, 48, respectively. In combination with Bacillus thuringiensis israelensis (Bti); A. aspera also caused 100% mortality at 250 ppm concentration after 72 hours (LC50 8.29 ppm). Phytochemical analysis of all the tested weed plants showed the presence of flavonoids, saponins, tannins, steroids, cardiac glycosides, alkaloids, anthrequinones and terpenoids. Random Amplification of Polymorphic DNA-Polymerase chain reaction (RAPD-PCR) and comet assay were performed to assess the genotoxic effect of A. aspera but no change in DNA profile was observed. Furthermore, FTIR showed the presence of phenolic compounds in A. aspera extract. It is suggested that certain phenolic compounds such as flavonoids modulate the enzymatic activity and, hence, cause the death of larvae of Cx. quinquefasciatus. Altogether, current study would serve as an initial step towards replacement of synthetic insecticides to plant-microbe based biopesticide against Culex mosquitoes in future.


Assuntos
Culex/efeitos dos fármacos , Inseticidas/toxicidade , Mutagênicos/toxicidade , Extratos Vegetais/toxicidade , Animais , Bactérias/efeitos dos fármacos , Bioensaio , Culex/enzimologia , Larva/efeitos dos fármacos , Larva/enzimologia , Compostos Fitoquímicos/análise , Plantas Daninhas/química , Técnica de Amplificação ao Acaso de DNA Polimórfico , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo
6.
Environ Sci Pollut Res Int ; 27(18): 22735-22748, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32323236

RESUMO

In this research, several biochemical variations in plant of Lemna minor L. were investigated to reflect Ag+ toxicity. Lemna minor L. changed colorless AgNO3 to colloidal brown at doses equal to and greater than 1 mg L-1. Optical and fluorescence microscopy revealed the presence of bright spots in roots of tested plant related to Ag/Ag2O-NPs. Photosynthetic pigment contents of Lemna minor L. declined upon exposure to Ag+ with an evidently higher decrease in chlorophyll a than in chlorophyll b. Similarly, Ag+ treatment caused an evident reduction in the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). The reduction in antioxidase activity was significantly higher in POD than in SOD and CAT. Ag+ treatment resulted in a significant increment in the level of malondialdehyde (MDA) content as the judging criteria of cellular injury which showed sign of dose-related. The alterations occurred in RAPD profiles of treated samples following Ag+ toxicity containing loss of normal bands, appearance of new bands, and variation in band intensities compared with the normal plants. In addition, morphological character and biomass of Lemna minor L. subjected to increasing Ag+ concentrations were evaluated to reveal Ag+ toxicity. Our study demonstrated that Lemna minor L. have a high sensitivity to indicate fluctuation of water quality. It would be beneficial that modulating the genotype of Lemna minor L. to bear high proportion of contaminates.


Assuntos
Araceae , Prata , Clorofila , Clorofila A , Biomarcadores Ambientais , Técnica de Amplificação ao Acaso de DNA Polimórfico , Superóxido Dismutase
7.
World J Microbiol Biotechnol ; 36(4): 58, 2020 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-32236741

RESUMO

The commercial production of Morchella mushrooms calls for urgent breeding of excellent varieties or strains with appropriate tools, such as protoplast fusion. However, the protoplast fusion in morels has not been studied. In this paper, interspecific hybridization between cultivated morels of M. importuna and M. sextelata by PEG-induced protoplast fusion was conducted. Apart from functional complementation of double inactivated protoplasts, the fusants were characterized by cultural and cultivated characters and molecular markers of random amplified polymorphic DNA (RAPD). The results suggested that the hybrids and their parents showed significant difference in their inoculum recovery time, mycelial growth rate, yield of cultivation and total amino acid content of ascocarps. Moreover, positive barrage reactions were observed between parental strains as well as between each parent and a hybrid line. A dendrogram created on the basis of RAPD fingerprints exhibited three major clusters, in which morel hybrids showed intra-cluster variations, M. sextelata #6 formed an out group, while M. importuna #4 was phylogenetically closer to morel hybrids. All the results demonstrated that real fusants were obtained in our study. Protoplast fusion may provide an ideal alternative for new strain selection, and thus will promote the healthy development of morel industry.


Assuntos
Agaricales/crescimento & desenvolvimento , Polietilenoglicóis/farmacologia , Protoplastos/fisiologia , Agaricales/classificação , Agaricales/genética , Quimera , DNA Fúngico/genética , Filogenia , Melhoramento Vegetal , Técnica de Amplificação ao Acaso de DNA Polimórfico
8.
Zhongguo Zhong Yao Za Zhi ; 45(6): 1272-1278, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-32281336

RESUMO

Molecular biology is a new subject that clarifies the phenomena and nature of life at the molecular level. Its development provides new biotechnology and methods for the study of traditional pharmacognosy. The formation of molecular biology has brought the development of pharmacognosy into a new era of gene research. Lonicerae Japonicae Flos is a classical Chinese medicine. Many scholars of home and abroad have carried out relevant studies on its molecular biology on the basis of the in-depth study with traditional methods, and have achieved certain results. In order to provide references on the method, technical for promoting the modernization of Lonicerae Japonicae Flos, and the development, protection, and utilization of other traditional Chinese medicine resources. This article summarized the application status of molecular biology methods and techniques on the identification, biosynthesis of active constituents, and molecular mechanism of secondary metabolite under stress conditions of Lonicerae Japonicae Flos in recent years. In hybridization technology of tag(RFLP), molecular markers based on PCR(RAPD, AFLP, SSR and ISSR), based on DNA sequence analysis of SNP and DNA barcode for the variety identification, diagnosis, identification of Lonicerae Japonicae Flos, and so forth in detail. At the same time, it is proposed that multi-omics technology can be used to build systems biology technology and platforms, and establish related models of secondary metabolite biosynthesis, so as to deepen acknowledge the molecular mechanism of the active component biosynthesis of Lonicerae Japonicae Flos and the accumulation of metabolites, life activities of other medicinal plants under adverse environment, then to regulate them.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Lonicera/química , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Cromatografia Líquida de Alta Pressão , Código de Barras de DNA Taxonômico , Medicina Tradicional Chinesa , Repetições de Microssatélites , Plantas Medicinais/química , Polimorfismo de Fragmento de Restrição , Polimorfismo de Nucleotídeo Único , Técnica de Amplificação ao Acaso de DNA Polimórfico , Metabolismo Secundário
9.
New Microbiol ; 43(2): 82-88, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32310301

RESUMO

In order to investigate molecular typing and fatty acid methyl esters (FAMEs) typing of clinical Stenotrophomonas maltophilia (S.maltophilia) isolates based on Random Amplification Polymorphic DNA (RAPD) and Gas Chromatography-Mass Spectrometer (GC-MS) methods, we collected 35 drug-resistant S. maltophilia isolates from March to December 2017 in a comprehensive hospital. The VITEK 2 Compact System was used to determine bacterial antibiotic susceptibility. The analysis of molecular typing was performed by RAPD. GC-MS was used to obtain FAMEs profiles. In total, all 35 isolates were multidrug-resistant S.maltophilia. Their resistance rates to CAZ and LEV were 21.4% and 21.1%, and to SXT up to 13.5%. S. maltophilia isolates were typed to six main clones by RAPD methods and four main clones by FAMEs fingerprint, respectively. The concordance rate of these two methods was 69.0%. Clonal typing provides evidence that multidrug-resistant isolates are prevalent among wards in the hospital. FAMEs profiles may be an easy and sensitive method for bacteria classification. The effectiveness and feasibility of different typing methods should be comprehensively considered.


Assuntos
Ácidos Graxos , Tipagem Molecular , Stenotrophomonas maltophilia , Antibacterianos/farmacologia , Ácidos Graxos/análise , Cromatografia Gasosa-Espectrometria de Massas , Infecções por Bactérias Gram-Negativas/microbiologia , Humanos , Testes de Sensibilidade Microbiana , Técnica de Amplificação ao Acaso de DNA Polimórfico , Stenotrophomonas maltophilia/química , Stenotrophomonas maltophilia/classificação , Stenotrophomonas maltophilia/efeitos dos fármacos
10.
Ecotoxicol Environ Saf ; 197: 110620, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32311615

RESUMO

The aim of this study was to evaluate the ecotoxic effect of high concentration cesium (Cs) exposure on plant root growth and its toxicological mechanism. The radicle of broad bean (Vicia faba) was selected as experimental material. The cytotoxic and genotoxic effects of plants exposed to different Cs levels (0.19-1.5 mM) for 48 h were evaluated using scanning electron microscopy (SEM), X-ray fluorescence (XRF) analysis, single-cell gel electrophoresis (SCGE) and random amplified polymorphic DNA (RAPD) assays. The results showed that radicle elongation decreased clearly after 48 h of exposure treatment with different concentrations of Cs solution. The root cell structure was obviously damaged in the Cs treatment groups (0.19-1.5 mM). At a Cs concentration of 1.5 mM, the percentages of viable non-apoptotic cells, viable apoptotic cells, non-viable apoptotic cells, and non-viable cells were 40.09%, 20.67%, 28.73%, and 10.52%, respectively. SCGE showed DNA damage in radicle cells 48 h after Cs exposure. Compared with the control group, the percentage of tail DNA in Cs exposed group (0.38-1.5 mM) increased by 0.56-1.12 times (P < 0.05). RAPD results showed that the genomic stability of V. faba radicles decreased by 4.44%-15.56%. This study confirmed that high concentration Cs exposure had cytotoxicity and genotoxicity effects on plants.


Assuntos
Césio/toxicidade , Dano ao DNA , Vicia faba/efeitos dos fármacos , Apoptose , Citotoxinas/toxicidade , Eletroforese , Instabilidade Genômica/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Análise de Célula Única , Vicia faba/genética
11.
Phytochemistry ; 174: 112330, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32146386

RESUMO

10 primers each of random amplified polymorphic DNA (RAPD), inter-simple sequence repeats (ISSR), inter primer binding site (iPBS) and start codon targeted (SCoT) were used to analyze genetic polymorphism and relationship between 50 genotypes of 5 economical important native bamboos (Bambusa cacharensis, B. mizorameana, Dendrocalamus manipureanus, D. hamiltonii and D. sikkimensis) of North-East India. The 40 different primers generated 111, 115, 116 and 138 polymorphic bands for RAPD, ISSR, iPBS and SCoT markers respectively. The comparative analysis of 4 marker systems based on polymorphic information content (PIC), effective multiplex ratio (EMR) and marker index (MI) values showed SCoT to be more informative with higher discriminating power than the other three markers. The correlation value (r) as determined by the Mantel test ranged from 0.60 (SCoT and RAPD) to 0.83 (iPBS and ISSR) indicating a high positive correlation between the markers. The close correspondence between the genetic matrices of RAPD, ISSR, iPBS and SCoT markers revealed the effectiveness of each marker system in determining the genetic relationship between bamboos. UPGMA (Unweighted Pair Group Arithmetic Mean Method) dendrograms generated from DNA marker analysis demonstrated species-specific clustering of different bamboo genotypes. Except for RAPD, the dendrograms of ISSR, iPBS and SCoT markers also showed a close association of bamboo genotypes based on geographical origin. Principal coordinate analysis (PCoA) revealed the distribution of different bamboo genotypes in accordance with the cluster analysis. The cluster grouping based on phytochemical study not only discriminated the different bamboo species but also illustrated a location-specific grouping of the genotypes. The bamboo clustering pattern derived from phytochemical analysis matched closely with the dendrograms generated by the DNA markers. The present investigation established the possibility of using a combined molecular and phytochemical marker approach to determine the genetic relationship between 5 native bamboos of North-East India with high precision.


Assuntos
Variação Genética , Repetições de Microssatélites , Sítios de Ligação , Códon de Iniciação , DNA , Índia , Filogenia , Compostos Fitoquímicos , Técnica de Amplificação ao Acaso de DNA Polimórfico
12.
Ecotoxicol Environ Saf ; 195: 110499, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32208213

RESUMO

The objective of this research was to evaluate Ag+ toxicity in Trifolium pratense L. seedlings subjected to increasing doses of Ag+ by determining photosynthetic pigment and malondialdehyde (MDA) contents, microstructure and hereditary substance alterations, changes in activities of antioxidase-superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) as well as the content of total Ag absorbed in vivo with evaluation of root growth. Doses of approximately 80 mg L-1 Ag+ severely affected photosynthetic efficiency in Trifolium pratense L. seedlings promoted by damages in photosynthetic apparatus evidenced by downward trend in photosynthetic pigment contents and obvious chlorosis. Alterations in enzymatic activity, lipid peroxidation, genic material damage and the presence of Ag+in vivo had impacted on photosynthetic machinery as well. A hormesis effect was observed at 60 mg L-1 Ag+ for the photosynthetic pigments and antioxidase for Trifolium pratense L. seedlings. Tissue changes (i.e., roots, stems and leaves) observed in fluorescence microscope with obvious chlorosis, roots blackening and formation of agglomerated black particles, were related to the lesion promoted by excessive ROS in vivo. Asynchronous change of antioxidase activity corresponded to the alteration in the MDA content, indicating the synchronization in the elimination of ROS. The changes occurred in RAPD profiles of treated samples following Ag+ toxicity containing loss of normal bands, appearance of new bands and variation in band intensity compared to the normal plants with a dose-dependent effect. On average, the roots of Trifolium pratense L. immobilized 92.20% of the total Ag absorbed as a metal exclusion response. Root growth was significantly sensitive to Ag+ stress with obvious hormesis, which corresponded to the changes in Ag uptake, demonstrating the functional alterations in plants. To sum up, we suggest that modulating the genotype of Trifolium pratense L. seedlings to bear higher proportion of pollutants is conducive to contamination site treatment.


Assuntos
Dano ao DNA , Fotossíntese/efeitos dos fármacos , Pigmentos Biológicos/metabolismo , Prata/toxicidade , Poluentes do Solo/toxicidade , Trifolium/efeitos dos fármacos , Biodegradação Ambiental , Catalase/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/farmacologia , Peroxidase/metabolismo , Fotossíntese/genética , Pigmentos Biológicos/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/metabolismo , Prata/metabolismo , Poluentes do Solo/metabolismo , Superóxido Dismutase/metabolismo , Trifolium/genética , Trifolium/metabolismo
13.
BMC Oral Health ; 20(1): 72, 2020 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-32171292

RESUMO

BACKGROUND: The aim of this study was to investigate the prevalence and genotypic profiles of Candida albicans in patients with oral lichen planus (OLP). MATERIALS AND METHODS: Positive rates and genotypic profiles of Candida albicans strains from OLP patients and healthy controls were analyzed. Random amplified polymorphic DNA and internal transcribed spacer of ribosome DNA polymerase chain reactions were used to sequence the DNA of these strains, and then their genetic similarity was measured using BLAST, UIV Band, and Vector NTI Suite Sequence Analyses Software. RESULTS: The prevalence of C. albicans strains detected from erosive-OLP, non-erosive OLP, and normal individuals was 18.87, 18.75, and 7.92%, respectively. Four different genotypes were revealed by the two methods. To be specific, type I was found only in the healthy subjects; type II a and II b were found in non-erosive OLP, and type III was identified in erosive OLP. Intragroup similarity coefficients, i.e. SAB were 100%, and inter-groups similarity coefficients, i.e. SAB were less than 30%. CONCLUSIONS: The genotypic results of C. albicans in OLP revealed an endogenous rather than exogenous infection of C. albicans. In addition, a possible pathogenic role of C. albicans in OLP, with the etiologic sense contributing to a more proper recognition on the pathogenesis, development, and progression of OLP, as well as some strategies for its diagnosis and treatment were identified.


Assuntos
Candida albicans/genética , Candida albicans/patogenicidade , Candidíase Bucal/microbiologia , Líquen Plano Bucal/diagnóstico , Adulto , Idoso , Candida albicans/isolamento & purificação , Estudos de Casos e Controles , Feminino , Genótipo , Humanos , Líquen Plano Bucal/microbiologia , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA Polimórfico , Análise de Sequência de DNA
14.
J Biotechnol ; 313: 29-38, 2020 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-32151644

RESUMO

The involvement of two extremely important signalling molecules, nitric oxide (NO) and abscisic acid (ABA) has been employed by plants to facilitate the adaptive/tolerate response during stressful conditions. However, the interactive role of exogenously applied NO and ABA is very less studied at physiological, biochemical and molecular levels. The present study therefore, evaluated the effects of individual and simultaneous addition of exogenous NO donor SNP (100µM) and ABA (10µM) on photosynthesis, Calvin-Benson cycle enzymes, S-assimilation enzymes, oxidative stress components, and genotoxicity in Brassica juncea cv. Varuna, exposed to polyethylene glycol (PEG)-induced drought stress. Results showed that a loss induced by PEG was significantly surpassed by the application of NO or/and ABA with PEG for chlorophyll content, net photosynthestic rate (Pn), internal CO2 concentration (Ci), stomatal conductance (gs), transpiration rate (Tr), maximum photosystem II (PSII) efficiency (Fv/Fm), actual PSII efficiency (ΦPSII), intrinsic PSII efficiency (Fv´/ Fm´), photochemical quenching (qP), non-photochemical quenching (NPQ), electron transport chain (ETC), ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCo), glyceraldehyde-3-phosphate dehydrogenase (GapDH), phosphoribulokinase (PRK), ATP-sulfurylase (ATP-S), and serine acetyltransferase (SAT) activities. The genomic template stability (GTS) (measured as changes in RAPD profiles) was significantly affected and showed varying degrees of DNA polymorphism, highest in PEG and lowest in PEG + NO and PEG + NO + ABA. Furthermore, the changes in RAPD profiles showed consistent results when compared with various photosynthetic and oxidative parameters. Altogether, this study concluded that supplementation of individual NO and together with ABA was more effective than individual ABA in alleviating PEG-induced drought stress in B. juncea L. seedlings.


Assuntos
Ácido Abscísico/farmacologia , Mostardeira/fisiologia , Óxido Nítrico/farmacologia , Fotossíntese/efeitos dos fármacos , Reguladores de Crescimento de Planta/farmacologia , Transdução de Sinais/efeitos dos fármacos , Clorofila/metabolismo , Secas , Fluorescência , Mostardeira/efeitos dos fármacos , Mostardeira/genética , Polietilenoglicóis/farmacologia , Técnica de Amplificação ao Acaso de DNA Polimórfico , Plântula/efeitos dos fármacos , Plântula/fisiologia , Estresse Fisiológico
15.
J Dairy Sci ; 103(5): 4056-4067, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32173014

RESUMO

Robiola di Roccaverano, from the Piedmont region of Italy, is a Protected Designation of Origin soft cheese made with raw goat milk. The peculiarity of this cheese is that during the manufacturing process, a natural starter culture (NC) is added to raw milk. This study examined the viable microorganisms of technological interest, including lactic acid bacteria and fungal populations, in samples of raw milk, NC, and fresh and ripened cheese collected from one dairy using culture-dependent techniques. First, the isolated colonies were analyzed using random amplification of polymorphic DNA (RAPD) PCR, and strains with similar fingerprints were clustered together. Further, representative isolates of each group were subjected to 16S or 26S ribosomal DNA sequencing. Finally, species-specific PCR was conducted to distinguish the Lactococcus lactis ssp. lactis and Lc. lactis ssp. cremoris. Among the studied lactic acid bacteria, 13 RAPD profiles were obtained, corresponding to 9 different bacterial species or subspecies. Concerning mold and yeast isolates, 5 species were found that coincided with 5 RAPD types. Observing the strains isolated in the study, Lc. lactis was the most prevalent species in raw milk and NC samples, and Leuconostoc mesenteroides was the predominant species identified in 5- and 15-d cheese isolates. Furthermore, whereas only these 2 species were detected in NC, Enterococcus and Lactobacillus genera were found in raw milk and cheese, respectively. Concerning the mold and yeast isolates, in NC Kluyveromyces spp. was mainly found, and in cheese samples the representative species were Geotrichum candidum and Yarrowia lipolytica. Finally, raw milk and cheese safety were evaluated, and the samples complied with the standard required by European Commission regulation number 2073/2005.


Assuntos
Bactérias/isolamento & purificação , Queijo/microbiologia , Enterococcus/isolamento & purificação , Lactobacillus/isolamento & purificação , Animais , Bactérias/classificação , Enterococcus/classificação , Microbiologia de Alimentos , Geotrichum/classificação , Geotrichum/isolamento & purificação , Cabras , Kluyveromyces/classificação , Kluyveromyces/isolamento & purificação , Leite/microbiologia , Tipagem Molecular , Técnica de Amplificação ao Acaso de DNA Polimórfico , Leveduras/classificação , Leveduras/isolamento & purificação
16.
PLoS One ; 15(2): e0228459, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32027671

RESUMO

BACKGROUND: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is emerging as a significant pathogen causing healthcare-associated infections. Matrix-assisted laser desorption/ionisation mass spectrometry time-of-flight mass spectrometry (MALDI-TOF MS) is used by clinical microbiology laboratories to address the need for rapid, cost-effective and accurate identification of microorganisms. We evaluated application of machine learning methods for differentiation of drug resistant bacteria from susceptible ones directly using the profile spectra of whole cells MALDI-TOF MS in 46 CRKP and 49 CSKP isolates. METHODS: We developed a two-step strategy for data preprocessing consisting of peak matching and a feature selection step before supervised machine learning analysis. Subsequently, five machine learning algorithms were used for classification. RESULTS: Random forest (RF) outperformed other four algorithms. Using RF algorithm, we correctly identified 93% of the CRKP and 100% of the CSKP isolates with an overall classification accuracy rate of 97% when 80 peaks were selected as input features. CONCLUSIONS: We conclude that CRKPs can be differentiated from CSKPs through RF analysis. We used direct colony method, and only one spectrum for an isolate for analysis, without modification of current protocol. This allows the technique to be easily incorporated into clinical practice in the future.


Assuntos
Algoritmos , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , Infecções por Klebsiella/diagnóstico , Klebsiella pneumoniae/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Aprendizado de Máquina Supervisionado , Antibacterianos/uso terapêutico , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Carbapenêmicos/uso terapêutico , Infecção Hospitalar/diagnóstico , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/microbiologia , DNA Bacteriano/análise , DNA Bacteriano/isolamento & purificação , Humanos , Infecções por Klebsiella/tratamento farmacológico , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , Testes de Sensibilidade Microbiana , Valor Preditivo dos Testes , Técnica de Amplificação ao Acaso de DNA Polimórfico
17.
Genome ; 63(3): 169-177, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32031004

RESUMO

Genomic DNA polymorphism and variation in biologically active components of Moringa oleifera were investigated by two different techniques: RAPD-PCR and HPLC analysis. The concentrations of phenolic compounds (cinnamic, caffeic, ferulic, and coumaric acids) and the content of flavonoids (rutin) were quantified by HPLC analysis. Among 20 RAPD primers, 13 were selected to generate polymorphic amplicons producing an average of 5028 bands, of which 83.7% were found to be polymorphic among 57 accessions of M. oleifera (MO 1 to MO 57) and one outgroup (ACB 58) from Banasthali region, India. In total, 57 accessions were clustered into five major groups within the dendrogram. The results of this analysis were further confirmed by principal coordinate analysis (PCoA). There was also high diversity in the concentration of active compounds in the collected samples as revealed by HPLC analysis. The data revealed that the content of polyphenolic compounds varied between 0.06 (sample KVKB) and 210.5 mg/kg (sample BG). The results suggest that there is a strong correlation between phytochemical variables and DNA polymorphism. The study concludes that the results of the genetic, morphological, and phytochemical diversity could be used to select the best accessions of M. oleifera for agricultural cultivation and breeding.


Assuntos
Moringa oleifera/classificação , Compostos Fitoquímicos/isolamento & purificação , Proteínas de Plantas/genética , Cromatografia Líquida de Alta Pressão , Índia , Moringa oleifera/química , Moringa oleifera/genética , Filogenia , Melhoramento Vegetal , Polimorfismo Genético , Análise de Componente Principal , Técnica de Amplificação ao Acaso de DNA Polimórfico
18.
Helicobacter ; 25(2): e12684, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32074664

RESUMO

BACKGROUND: Resistant Helicobacter pylori to commonly used antimicrobial agents are associated with severe upper gastrointestinal disorders. To provide an epidemiological picture of H pylori and characterize the resistance pattern and genetic variation of clinical isolates, stomach biopsies from patients with functional dyspepsia were evaluated in northeast of Iran. MATERIALS AND METHODS: In this study, 80 patients were recruited. Finally, fifty H pylori strains were isolated from antrum and corpus biopsies by culturing on Columbia agar. All strains were identified by standard laboratory procedures. Susceptibility testing of antibiotics was performed using minimum inhibitory concentration test. Allele-specific primer (ASP)-PCR of 23S rRNA which associated with clarithromycin resistance was done among resistant strains. Moreover, cagA gene and polymorphism in vacA were detected. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was applied to investigate the genetic variations among all strains. RESULTS: Antibiotic resistance pattern of H pylori strains was as follows: 68% (34/50) to metronidazole, 50% (25/50) to rifampicin, 30% (15/50) to amoxicillin, 28% (14/50) to levofloxacin, 22% (11/50) to clarithromycin, and 16% (8/50) to tetracycline. Multidrug-resistant strains were observed in 19 strains (38%). ASP-PCR of 23S rRNA showed four strains had A2143G mutation, six strains had A2142G mutation, and one strain had a Wt+A2143G mutation. Amplification of virulence-associated genes revealed that cagA was present in 27 isolates (54%) and vacA in 36 isolates (72%). The most common genotype of H pylori was vacA s1am2 (40%) followed by vacA s2m2 (14%), vacA s1am1 (12%), vacA s1bm1 (4%), and vacA s1bm2 (2%). DNA fingerprinting pattern indicated a high heterogeneity among isolated strains. CONCLUSION: An alarming level of resistance to metronidazole and rifampicin and high heterogeneity among H pylori isolates highlighted the importance of continued monitoring of antimicrobial susceptibility and epidemiological surveillance of this pathogen.


Assuntos
Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana Múltipla/genética , Infecções por Helicobacter , Helicobacter pylori , Virulência/genética , Adulto , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Biópsia , Feminino , Variação Genética , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/epidemiologia , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/genética , Helicobacter pylori/isolamento & purificação , Humanos , Irã (Geográfico)/epidemiologia , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , RNA Ribossômico 23S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Estômago/microbiologia
19.
Mycoses ; 63(4): 361-368, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31954083

RESUMO

AIM: During the last decade a continuous increase in non-albicans species isolation has been observed with Candida parapsilosis being one of the leading species. Aim of this study was to describe the epidemiology of candidemia, particularly of C parapsilosis, its predictors and clinical outcome. MATERIALS AND METHODS: Incidences of candidemia was evaluated analyzing data from both a prospective collection (2012-2016) and a retrospective one (2008-2011). Predictors and outcome were based only on the prospective phase. C parapsilosis potential clusters were analysed by randomly amplified polymorphic DNA (RAPD) technique. RESULTS: 1240 episodes were identified. Incidences of candidemia increased from 1.97 episodes/10 000 patient-days in 2008 to 4.59/10 000 patient-days in 2016 (P < .001), mainly due to an increase of C parapsilosis (incidence rate ratio, IRR: 1.04, P < .001). 33.0% of C parapsilosis strains were resistant to fluconazole; no resistance to echinocandins was found. Independent predictors of C parapsilosis candidemia were time of infection (P = .007), previous use of echinocandins (P < .0001) and year in which the episode was registered (P < .0001). 30 days mortality was 32.4% for C parapsilosis, with a significant difference compared to C non-parapsilosis. Potential clonal C parapsilosis strains were detected by genetic analyses, showing RAPD profile A as the most represented (72.6% of isolates). DISCUSSION: C parapsilosis candidemia is an emerging issue in our center, possibly attributed to some extent to horizontal transmission of the pathogen, as confirmed by the analysis of isolates similarities. Further microbiological and epidemiological investigations are needed in order to identify the most effective measures to reduce the rate of this infection.


Assuntos
Candida parapsilosis , Candidemia/epidemiologia , Farmacorresistência Fúngica , Fluconazol/uso terapêutico , Idoso , Idoso de 80 Anos ou mais , Antifúngicos/uso terapêutico , Candida parapsilosis/efeitos dos fármacos , Candida parapsilosis/isolamento & purificação , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Estudos Retrospectivos
20.
Methods Mol Biol ; 2102: 213-224, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31989557

RESUMO

Overwhelming evidence suggests that in addition to the genetic changes of DNA mutations, epigenetic changes of DNA methylation and histone modifications play important role in regulation of gene expression. DNA methylation is the most frequent epigenetic alteration observed in mammalian genomes, and generally it is negatively correlated with gene expression. Various methods are available for the detection of DNA methylation changes. Although the recent high-throughput methods for DNA methylation analysis have various advantages, they require high levels of technical expertise, costly equipment, and reagents. Because of these reasons, many of the global DNA methylation analysis methods are mainly performed at core facility, and laboratories with limited resources and expertise are not able to use these methods. Methylation-Sensitive-Random Amplified Polymorphic DNA-Polymerase Chain Reaction (MS-RAPD-PCR) is a restriction enzyme digestion and PCR-based method for the analysis of DNA methylation changes. This method is cost-effective, requires simple and basic instrumentation, and therefore can easily be performed in any laboratory with basic setup having a regular DNA thermal cycler and DNA gel electrophoresis system. Additional advantages of this method over other methods for DNA methylation analysis are that it requires very less amount of DNA and can screen DNA methylation changes globally at genome-wide level with high sensitivity. This method has been successfully used to detect changes in DNA methylation either occurring naturally or induced by various toxicants and environmental factors. A detail experimental protocol for MS-RAPD-PCR is described in this chapter.


Assuntos
Metilação de DNA/genética , Epigenômica/métodos , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Animais , Células Cultivadas , DNA/isolamento & purificação , DNA-Citosina Metilases , Eletroforese em Gel de Ágar , Serina Endopeptidases , Fluxo de Trabalho
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