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1.
J Endocrinol Invest ; 2024 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-39361240

RESUMO

PURPOSE: We aimed to identify differentially expressed spliceosome components in growth hormone (GH)-secreting pituitary tumors and investigate their roles in pathogenesis. METHODS: We performed transcriptome analysis of 20 somatotroph adenomas and 6 normal pituitary tissues to select dysregulated spliceosome components. Clinical characteristics were analyzed based on gene expression in 64 patients with acromegaly. Proliferation, invasion, and hormonal activity of GH secreting pituitary adenoma cells were investigated. RESULTS: TCERG1 expression was significantly higher in somatotroph adenomas than in normal pituitaries (log2 fold change 0.59, adjusted P = 0.0002*). Genotype-phenotype analysis revealed that patients with higher TCERG1 expression had lower surgical remission rates than those with lower expression (63.64% vs. 95.45%, P = 0.009*). TCERG1 expression was significantly higher in groups with cavernous sinus (CS) invasion or Ki67 index over 3 (all P>0.05*). TCERG1 overexpression led to a 29.60% increase in proliferation (P<0.001*) and a 249.47% increase in invasion after 48 h in GH3 cells (P = 0.026*). Conversely, TCERG1 silencing significantly decreased cell proliferation (25.76% at 72 h, P<0.001*) and invasion (96.87% at 48 h, P = 0.029*). E-cadherin was decreased, but vimentin was increased in both TCERG1 overexpressed GH3 cells and somatotroph adenomas. And TCERG1 silence reversed the expression of the genes (CDH2, SNAI1, ZEB2, and VIM) in GH3 cells. CONCLUSIONS: Spliceosome machinery provide novel insights into the pathogenesis of GH-secreting pituitary tumor and highlight the potential role of TCERG1 as a biomarker for tumor aggressiveness.

2.
Int J Mol Sci ; 25(3)2024 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-38338917

RESUMO

Viruses have evolved sophisticated mechanisms to manipulate host cell processes and utilize intracellular organelles to facilitate their replication. These complex interactions between viruses and cellular organelles allow them to hijack the cellular machinery and impair homeostasis. Moreover, viral infection alters the cell membrane's structure and composition and induces vesicle formation to facilitate intracellular trafficking of viral components. However, the research focus has predominantly been on the immune response elicited by viruses, often overlooking the significant alterations that viruses induce in cellular organelles. Gaining a deeper understanding of these virus-induced cellular changes is crucial for elucidating the full life cycle of viruses and developing potent antiviral therapies. Exploring virus-induced cellular changes could substantially improve our understanding of viral infection mechanisms.


Assuntos
Viroses , Replicação Viral , Humanos , Organelas/ultraestrutura , Interações Hospedeiro-Patógeno
3.
J Allergy Clin Immunol ; 147(4): 1453-1463, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32795589

RESUMO

BACKGROUND: The relationship between allergic and eosinophilic inflammation, either systemic or local, in allergic diseases remains unclear. OBJECTIVE: We performed combined genome-wide association study (GWAS) and epigenome-wide (EWAS) for atopy and tissue eosinophilia to identify both genetic and epigenetic signatures between systemic and local allergic inflammation, and to capture global patterns of gene regulation. METHODS: We included 126 subjects for atopy analysis and 147 for tissue eosinophilia analysis, as well as 18 normal nasal tissue samples. We identified differentially methylated positions (DMPs) and genes associated with atopy and tissue eosinophilia. Furthermore, we performed mendelian randomization analysis and penalized regression along with replication in an independent cohort. RESULTS: EWAS identified genes, including Musashi RNA binding protein 2 (MSI2), associated with atopy, which contained enriched DMPs that genetically affect atopy. A direct association was observed between MSI2 single-nucleotide polymorphisms and atopy, as was a causal effect of changes in MSI2 expression and methylation on atopy, which was replicated in a Costa Rican population. Regarding tissue eosinophilia, EWAS identified genes with enriched DMPs directly contributing to tissue eosinophilia at the gene level, including CAMK1D. The gene ontology terms of the identified genes for both phenotypes encompassed immune-related terms. CONCLUSION: EWAS combined with GWAS identified novel candidate genes, especially the methylation of MSI2, contributing to systemic allergic inflammation. Certain genes displayed a greater association with either systemic or local allergic inflammation; however, it is expected that a harmonized effect of these genes influences immune responses.


Assuntos
Eosinofilia/genética , Hipersensibilidade/genética , Proteínas de Ligação a RNA/genética , Adulto , Metilação de DNA , Epigênese Genética , Epigenoma , Feminino , Ontologia Genética , Estudo de Associação Genômica Ampla , Humanos , Inflamação/genética , Masculino , Pessoa de Meia-Idade , Mucosa Nasal , Polimorfismo de Nucleotídeo Único
4.
J Chem Ecol ; 47(6): 564-576, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33881708

RESUMO

Based on the hypothesis that the variation of the metabolomes of latex is a response to selective pressure and should thus be affected differently from other organs, their variation could provide an insight into the defensive chemical selection of plants. Metabolic profiling was used to compare tissues of three Euphorbia species collected in diverse regions. The metabolic variation of latexes was much more limited than that of other organs. In all the species, the levels of polyisoprenes and terpenes were found to be much higher in latexes than in leaves and roots of the corresponding plants. Polyisoprenes were observed to physically delay the contact of pathogens with plant tissues and their growth. A secondary barrier composed of terpenes in latex and in particular, 24-methylenecycloartanol, exhibited antifungal activity. These results added to the well-known role of enzymes also present in latexes, show that these are part of a cooperative defense system comprising biochemical and physical elements.


Assuntos
Euphorbia/metabolismo , Euphorbia/microbiologia , Geografia , Herbivoria , Látex/metabolismo , Metabolômica , Euphorbia/fisiologia , Especificidade da Espécie
5.
Molecules ; 26(3)2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33499348

RESUMO

Mass spectrometry-based molecular imaging has been utilized to map the spatial distribution of target metabolites in various matrixes. Among the diverse mass spectrometry techniques, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) is the most popular for molecular imaging due to its powerful spatial resolution. This unparalleled high resolution, however, can paradoxically act as a bottleneck when the bio-imaging of large areas, such as a whole plant, is required. To address this issue and provide a more versatile tool for large scale bio-imaging, direct analysis in real-time-time of flight-mass spectrometry (DART-TOF-MS), an ambient ionization MS, was applied to whole plant bio-imaging of a medicinal plant, Ephedrae Herba. The whole aerial part of the plant was cut into 10-20 cm long pieces, and each part was further cut longitudinally to compare the contents of major ephedra alkaloids between the outer surface and inner part of the stem. Using optimized DART-TOF-MS conditions, molecular imaging of major ephedra alkaloids of the whole aerial part of a single plant was successfully achieved. The concentration of alkaloids analyzed in this study was found to be higher on the inner section than the outer surface of stems. Moreover, side branches, which are used in traditional medicine, represented a far higher concentration of alkaloids than the main stem. In terms of the spatial metabolic distribution, the contents of alkaloids gradually decreased towards the end of branch tips. In this study, a fast and simple macro-scale MS imaging of the whole plant was successfully developed using DART-TOF-MS. This application on the localization of secondary metabolites in whole plants can provide an area of new research using ambient ionization mass spectroscopy and an unprecedented macro-scale view of the biosynthesis and distribution of active components in medicinal plants.


Assuntos
Alcaloides/metabolismo , Ephedra/metabolismo , Espectrometria de Massas/métodos , Plantas Medicinais/metabolismo , Efedrina/análogos & derivados , Efedrina/metabolismo , Espectrometria de Massas/instrumentação , Imagem Molecular/instrumentação , Imagem Molecular/métodos , Componentes Aéreos da Planta/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
6.
Planta Med ; 83(17): 1351-1360, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28561204

RESUMO

(S)-Allyl-l-cysteine is the major bioactive compound in garlic. (S)-Allyl-l-cysteine is metabolized to (S)-allyl-l-cysteine sulfoxide, N-acetyl-(S)-allyl-l-cysteine, and N-acetyl-(S)-allyl-l-cysteine sulfoxide after oral administration. An accurate LC-MS/MS method was developed and validated for the simultaneous quantification of (S)-allyl-l-cysteine and its metabolites in rat plasma, and the feasibility of using it in pharmacokinetic studies was tested. The analytes were quantified by multiple reaction monitoring using an atmospheric pressure ionization mass spectrometer. Because significant quantitative interference was observed between (S)-allyl-l-cysteine and N-acetyl-(S)-allyl-l-cysteine as a result of the decomposition of N-acetyl-(S)-allyl-l-cysteine at the detector source, chromatographic separation was required to discriminate (S)-allyl-l-cysteine and its metabolites on a reversed-phase C18 analytical column with a gradient mobile phase consisting of 0.1% formic acid and acetonitrile. The calibration curves of (S)-allyl-l-cysteine, (S)-allyl-l-cysteine sulfoxide, N-acetyl-(S)-allyl-l-cysteine, and N-acetyl-(S)-allyl-l-cysteine sulfoxide were linear over each concentration range, and the lower limits of quantification were 0.1 µg/mL [(S)-allyl-l-cysteine and N-acetyl-(S)-allyl-l-cysteine] and 0.25 µg/mL [(S)-allyl-l-cysteine sulfoxide and N-acetyl-(S)-allyl-l-cysteine sulfoxide]. Acceptable intraday and inter-day precisions and accuracies were obtained at three concentration levels. The method satisfied the regulatory requirements for matrix effects, recovery, and stability. The validated LC-MS/MS method was successfully used to determine the concentration of (S)-allyl-l-cysteine and its metabolites in rat plasma samples after the administration of (S)-allyl-l-cysteine or aged garlic extract.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Cisteína/análogos & derivados , Alho/química , Espectrometria de Massas/métodos , Extratos Vegetais/metabolismo , Administração Oral , Animais , Cisteína/administração & dosagem , Cisteína/química , Cisteína/metabolismo , Cisteína/farmacocinética , Masculino , Estrutura Molecular , Extratos Vegetais/administração & dosagem , Extratos Vegetais/química , Extratos Vegetais/farmacocinética , Ratos , Ratos Sprague-Dawley
7.
Phytother Res ; 31(3): 475-487, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28127806

RESUMO

In this study, we investigated the antiinflammatory effects of ethanol extracts of Potentilla. supina Linne (EPS) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages and septic mice. EPS suppressed LPS-induced nitric oxide, prostaglandin E2 , TNF-α, interleukin-6 and interleukin-1ß at production and mRNA levels in LPS-induced RAW 264.7 macrophages. Consistent with these observations, EPS attenuated the expressions of inducible nitric oxide synthase and cyclooxygenase-2 by downregulation of their promoter activities. Molecularly, EPS reduced the LPS-induced transcriptional activity and DNA-binding activity of nuclear factor-κB (NF-κB), and this was associated with a decrease of translocation and phosphorylation of p65 NF-κB by inhibiting the inhibitory κB-α degradation and IKK-α/ß phosphorylation. Furthermore, EPS inhibited the LPS-induced activation of activator protein-1 by reducing the expression of c-Fos and c-Jun in nuclear. EPS also suppressed the phosphorylation of mitogen-activated protein kinase, such as p38 mitogen-activated protein kinase and c-Jun N-terminal kinase. In an LPS-induced endotoxemia mouse model, pretreatment with EPS reduced the mRNA levels of inducible nitric oxide synthase, cyclooxygenase-2 and proinflammatory cytokines and increased the survival rate of mice. Collectively, these results suggest that the antiinflammatory effects of EPS were associated with the suppression of NF-κB and activator protein-1 activation and support its possible therapeutic role for the treatment of endotoxemia. Copyright © 2017 John Wiley & Sons, Ltd.


Assuntos
Anti-Inflamatórios/farmacologia , Etanol/química , Inflamação/prevenção & controle , Lipopolissacarídeos , Macrófagos/efeitos dos fármacos , Extratos Vegetais/farmacologia , Potentilla/química , Choque Séptico/tratamento farmacológico , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/uso terapêutico , Linhagem Celular , Citocinas/metabolismo , Endotoxinas , Etanol/farmacologia , Inflamação/induzido quimicamente , Inflamação/metabolismo , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Choque Séptico/induzido quimicamente , Choque Séptico/imunologia , Choque Séptico/metabolismo , Fator de Transcrição AP-1/antagonistas & inibidores , Fator de Transcrição AP-1/metabolismo
9.
Int J Syst Evol Microbiol ; 66(2): 1100-1103, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26585518

RESUMO

Species demarcation in Bacteria and Archaea is mainly based on overall genome relatedness, which serves a framework for modern microbiology. Current practice for obtaining these measures between two strains is shifting from experimentally determined similarity obtained by DNA-DNA hybridization (DDH) to genome-sequence-based similarity. Average nucleotide identity (ANI) is a simple algorithm that mimics DDH. Like DDH, ANI values between two genome sequences may be different from each other when reciprocal calculations are compared. We compared 63 690 pairs of genome sequences and found that the differences in reciprocal ANI values are significantly high, exceeding 1 % in some cases. To resolve this problem of not being symmetrical, a new algorithm, named OrthoANI, was developed to accommodate the concept of orthology for which both genome sequences were fragmented and only orthologous fragment pairs taken into consideration for calculating nucleotide identities. OrthoANI is highly correlated with ANI (using BLASTn) and the former showed approximately 0.1 % higher values than the latter. In conclusion, OrthoANI provides a more robust and faster means of calculating average nucleotide identity for taxonomic purposes. The standalone software tools are freely available at http://www.ezbiocloud.net/sw/oat.

10.
Int J Syst Evol Microbiol ; 65(Pt 3): 959-964, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25563916

RESUMO

A Gram-stain negative, rod-shaped, non-spore-forming, obligate aerobic bacterial strain, JC2949(T), was isolated from grassland soil in Gwanak Mountain, Seoul, Republic of Korea. Phylogenetic analysis, based on 16S rRNA sequences, indicated that strain JC2949(T) belongs to the genus Burkholderia, showing highest sequence similarities with Burkholderia grimmiae R27(T) (98.8 %), Burkholderia cordobensis LMG 27620(T) (98.6 %), Burkholderia jiangsuensis MP-1T(T) (98.6 %), Burkholderia zhejiangensis OP-1(T) (98.5 %), Burkholderia humi LMG 22934(T) (97.5 %), Burkholderia terrestris LMG 22937(T) (97.3 %), Burkholderia telluris LMG 22936(T) (97.2 %) and Burkholderia glathei ATCC 29195(T) (97.0 %). The major fatty acids of strain JC2949(T) were C18 : 1ω7c, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0. Its predominant polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and an unknown amino phospholipid. The dominant isoprenoid quinone was ubiquinone Q-8. The pairwise average nucleotide identity values between strain JC2949(T) and the genomes of 30 other species of the genus Burkholderia ranged from 73.4-90.4 %, indicating that the isolate is a novel genomic species within this genus. Based on phenotypic and chemotaxonomic comparisons, it is clear that strain JC2949(T) represents a novel species of the genus Burkholderia. We propose the name for this novel species to be Burkholderia megalochromosomata sp. nov. The type strain is JC2949(T) ( = KACC 17925(T) = JCM 19905(T)).


Assuntos
Burkholderia/classificação , Pradaria , Filogenia , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Composição de Bases , Burkholderia/genética , Burkholderia/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Dados de Sequência Molecular , Fosfolipídeos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA , Ubiquinona/química
11.
Int J Syst Evol Microbiol ; 64(Pt 2): 346-351, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24505072

RESUMO

Among available genome relatedness indices, average nucleotide identity (ANI) is one of the most robust measurements of genomic relatedness between strains, and has great potential in the taxonomy of bacteria and archaea as a substitute for the labour-intensive DNA-DNA hybridization (DDH) technique. An ANI threshold range (95-96%) for species demarcation had previously been suggested based on comparative investigation between DDH and ANI values, albeit with rather limited datasets. Furthermore, its generality was not tested on all lineages of prokaryotes. Here, we investigated the overall distribution of ANI values generated by pairwise comparison of 6787 genomes of prokaryotes belonging to 22 phyla to see whether the suggested range can be applied to all species. There was an apparent distinction in the overall ANI distribution between intra- and interspecies relationships at around 95-96% ANI. We went on to determine which level of 16S rRNA gene sequence similarity corresponds to the currently accepted ANI threshold for species demarcation using over one million comparisons. A twofold cross-validation statistical test revealed that 98.65% 16S rRNA gene sequence similarity can be used as the threshold for differentiating two species, which is consistent with previous suggestions (98.2-99.0%) derived from comparative studies between DDH and 16S rRNA gene sequence similarity. Our findings should be useful in accelerating the use of genomic sequence data in the taxonomy of bacteria and archaea.


Assuntos
Archaea/classificação , Bactérias/classificação , Genômica/métodos , RNA Ribossômico 16S/genética , Análise de Sequência de DNA/métodos , Archaea/genética , Bactérias/genética , Hibridização de Ácido Nucleico/métodos
12.
Int J Syst Evol Microbiol ; 64(Pt 2): 689-691, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24425826

RESUMO

EzEditor is a Java-based molecular sequence editor allowing manipulation of both DNA and protein sequence alignments for phylogenetic analysis. It has multiple features optimized to connect initial computer-generated multiple alignment and subsequent phylogenetic analysis by providing manual editing with reference to biological information specific to the genes under consideration. It provides various functionalities for editing rRNA alignments using secondary structure information. In addition, it supports simultaneous editing of both DNA sequences and their translated protein sequences for protein-coding genes. EzEditor is, to our knowledge, the first sequence editing software designed for both rRNA- and protein-coding genes with the visualization of biologically relevant information and should be useful in molecular phylogenetic studies. EzEditor is based on Java, can be run on all major computer operating systems and is freely available from http://sw.ezbiocloud.net/ezeditor/.


Assuntos
RNA Ribossômico/genética , Alinhamento de Sequência/métodos , Software , Sequência de Aminoácidos , Sequência de Bases , Biologia Computacional , Filogenia , Linguagens de Programação
13.
PLoS One ; 19(5): e0300607, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38787824

RESUMO

Listening to music is a crucial tool for relieving stress and promoting relaxation. However, the limited options available for stress-relief music do not cater to individual preferences, compromising its effectiveness. Traditional methods of curating stress-relief music rely heavily on measuring biological responses, which is time-consuming, expensive, and requires specialized measurement devices. In this paper, a deep learning approach to solve this problem is introduced that explicitly uses convolutional neural networks and provides a more efficient and economical method for generating large datasets of stress-relief music. These datasets are composed of Mel-scaled spectrograms that include essential sound elements (such as frequency, amplitude, and waveform) that can be directly extracted from the music. The trained model demonstrated a test accuracy of 98.7%, and a clinical study indicated that the model-selected music was as effective as researcher-verified music in terms of stress-relieving capacity. This paper underlines the transformative potential of deep learning in addressing the challenge of limited music options for stress relief. More importantly, the proposed method has profound implications for music therapy because it enables a more personalized approach to stress-relief music selection, offering the potential for enhanced emotional well-being.


Assuntos
Musicoterapia , Música , Redes Neurais de Computação , Estresse Psicológico , Humanos , Música/psicologia , Estresse Psicológico/terapia , Musicoterapia/métodos , Aprendizado Profundo , Masculino , Feminino , Adulto , Espectrografia do Som/métodos , Adulto Jovem
14.
Int J Syst Evol Microbiol ; 63(Pt 12): 4784-4789, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23996833

RESUMO

A Gram-stain-negative, rod-shaped, pale yellow, aerobic bacterial strain, JC2902(T), was isolated from a sediment sample of Ungok Lake in Gochang, Republic of Korea. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain JC2902(T) belongs to the genus Flavobacterium and forms a distinct phyletic line within a clade containing four recognized species of the genus Flavobacterium. The genomic relatedness between strain JC2902(T) and closely related strains was calculated using average nucleotide identity values of whole genome sequences, which indicated that the new isolate represents a novel genomic species. Through comparison of chemotaxonomic and other phenotypic characteristics between strain JC2902(T) and the type strains of the four phylogenetically related species, a number of characteristics differentiated strain JC2902(T) from the previously described type strains. Differential characteristics of strain JC2902(T) include fatty acid profiles, cellular motility, inability to grow on Luria-Bertani and tripticase soy agar media, and absence of N-acetyl-ß-glucosaminidase and flexirubin-type pigments. Based on data from this polyphasic taxonomic study, strain JC2902(T) is considered to represent a novel species of the genus Flavobacterium, for which the name Flavobacterium limnosediminis sp. nov. is proposed. The type strain is JC2902(T) ( = KACC 16937(T) = JCM 18661(T)).


Assuntos
Flavobacterium/classificação , Sedimentos Geológicos/microbiologia , Lagos/microbiologia , Filogenia , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Flavobacterium/genética , Flavobacterium/isolamento & purificação , Dados de Sequência Molecular , Polienos/química , RNA Ribossômico 16S/genética , República da Coreia , Análise de Sequência de DNA
15.
Front Public Health ; 11: 1203201, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37483927

RESUMO

Objective: We aimed to investigate the effect of internet-based and in-person cognitive interventions on cognition, mood, and activities of daily living (ADL) on patients with mild to moderate Alzheimer's disease (AD) and examine whether internet-based intervention is as effective as the in-person intervention. Methods: We recruited 52 patients with probable mild AD, of whom 42 completed the trial. We randomly divided participants into intervention and control groups at a 1:1 ratio and statistically compared the neuropsychological test results of the two groups. In addition, patients in the intervention group were randomly assigned to a 4 weeks internet-based or in-person intervention, with subsequent crossover to the other group for 4 weeks. We statistically analyzed and compared the neuropsychological test scores between internet-based and in-person interventions. Results: Compared with the control group, the intervention group (internet-based and in-person) showed significantly improved profile in cognition (p < 0.001), depression (p < 0.001), anxiety (p < 0.001) and ADL (p < 0.001). In addition, the effect of the internet-based intervention on cognition (p = 0.918) and depression (p = 0.282) was not significantly different from that of the in-person intervention. However, in the Beck anxiety inventory (p = 0.009) and Seoul instrumental activity of daily living (p = 0.023), in-person intervention was more effective than internet-based intervention. Conclusion: This study suggests that both types of cognitive intervention (in-person and internet-based) may be viable supplementary treatments along with approved pharmacological therapy. In terms of anxiety and ADL, the effect of the in-person interventions may be more effective than the-internet based interventions.


Assuntos
Doença de Alzheimer , Humanos , Doença de Alzheimer/terapia , Atividades Cotidianas , Cognição , Ansiedade , Internet
16.
Int J Syst Evol Microbiol ; 62(Pt 3): 716-721, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22140171

RESUMO

Despite recent advances in commercially optimized identification systems, bacterial identification remains a challenging task in many routine microbiological laboratories, especially in situations where taxonomically novel isolates are involved. The 16S rRNA gene has been used extensively for this task when coupled with a well-curated database, such as EzTaxon, containing sequences of type strains of prokaryotic species with validly published names. Although the EzTaxon database has been widely used for routine identification of prokaryotic isolates, sequences from uncultured prokaryotes have not been considered. Here, the next generation database, named EzTaxon-e, is formally introduced. This new database covers not only species within the formal nomenclatural system but also phylotypes that may represent species in nature. In addition to an identification function based on Basic Local Alignment Search Tool (blast) searches and pairwise global sequence alignments, a new objective method of assessing the degree of completeness in sequencing is proposed. All sequences that are held in the EzTaxon-e database have been subjected to phylogenetic analysis and this has resulted in a complete hierarchical classification system. It is concluded that the EzTaxon-e database provides a useful taxonomic backbone for the identification of cultured and uncultured prokaryotes and offers a valuable means of communication among microbiologists who routinely encounter taxonomically novel isolates. The database and its analytical functions can be found at http://eztaxon-e.ezbiocloud.net/.


Assuntos
Archaea/classificação , Archaea/genética , Bactérias/classificação , Bactérias/genética , Bases de Dados de Ácidos Nucleicos , Genes de RNAr , RNA Ribossômico 16S/genética , Classificação/métodos , Biologia Computacional/métodos
17.
Front Mol Biosci ; 9: 984307, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36250021

RESUMO

Aspergillus cristatus is a beneficial fungus of microbial fermented teas such as China's Fuzhuan brick tea and Pu-erh tea, and is commonly called golden flower fungus (GFF) because its cleistothecium has a yellow millet or sand grain shape. Since natural materials fermented with GFF exhibit various physiological activities, a new active cosmeceutical ingredient was developed by solid-state fermentation of ginseng, a famous active material for healthy skin, with GFF. The extract of solid-state fermented ginseng with GFF (GFFG) exhibited potent anti-aging efficacy on the skin such as the increase of hyaluronic acid synthesis, aquaporin expression, and mRNA level of filaggrin in HaCaT keratinocyte. GFFG also inhibited the expression of MMP-1 increased by TNF-α in human dermal fibroblast. Sophisticated chromatographic and spectroscopic studies have elucidated isodihydroauroglaucin and flavoglaucin as the metabolites which were not present in ginseng extract nor GFF extract alone. Bioassay of these metabolites revealed that these compounds were part of active principles of GFFG. These results suggest that GFFG would be a potential active ingredient in anti-aging cosmeceutical products.

18.
Biomed Pharmacother ; 154: 113637, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36058149

RESUMO

Parkinson's disease (PD) is the second most common neurodegenerative disease, after Alzheimer's disease, and becomes increasingly prevalent with age. α-Synuclein (α-syn) forms the major filamentous component of Lewy bodies, which are pathological hallmarks of α-synucleinopathies such as PD. We evaluated the neuroprotective effects of MT101-5, a standardized herbal formula that consists of an ethanolic extract of Genkwae Flos, Clematidis Radix, and Gastrodiae Rhizoma, against α-synuclein-induced cytotoxicity in vivo. MT101-5 protected against behavioral deficits and loss of dopaminergic neurons in human α-syn-overexpressing transgenic mice after treatment with 30 mg/kg/day for 5 months. We investigated transcriptomic changes within MT101-5 mechanisms of action (MOA) suppressing α-syn aggregation in an α-synuclein preformed fibril (α-syn PFF) mouse model of sporadic PD. We found that inhibition of α-syn fibril formation was associated with changes in transcripts in mitochondrial biogenesis, electron transport, chaperones, and proteasomes following treatment with MT101-5. These results suggest that the mixed herbal formula MT101-5 may be used as a pharmaceutical agent for preventing or improving PD.


Assuntos
Doenças Neurodegenerativas , Doença de Parkinson , Animais , Modelos Animais de Doenças , Neurônios Dopaminérgicos/patologia , Humanos , Camundongos , Camundongos Transgênicos , Doença de Parkinson/tratamento farmacológico , Doença de Parkinson/patologia , alfa-Sinucleína
19.
Food Funct ; 13(14): 7638-7649, 2022 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-35735022

RESUMO

Neuroinflammation, which occurs due to microglia, is related to the pathogenesis of neurodegenerative disorders. Recently, the development of functional foods that down-regulate over-activated microglial cells to prevent the progression of neurodegenerative disorders has been proposed, since over-activated microglia induce a chronic source of neurotoxic factors and reduce neuronal survival. Thus, the anti-neuroinflammatory effects of a functional food mixture (CCL01) including Cuscuta seeds and Lactobacillus paracasei NK112 on lipopolysaccharide (LPS)-induced experimental models were investigated. In LPS-induced in vitro models, the expression levels of inflammatory mediators (e.g., inducible nitric oxide synthase, cyclooxygenase-2, nitric oxide, and prostaglandin E2) and pro-inflammatory cytokines (e.g., tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6) were decreased upon CCL01 treatment. CCL01 showed an anti-neuroinflammatory effect in LPS-induced microglial cells via the inhibition of the mitogen-activated protein kinase (MAPK)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway and the activation of the nuclear factor erythroid-2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway. In the LPS-treated in vivo mouse models, the increased expression of ionized calcium binding adaptor molecule 1 (Iba-1), which indicates microglial activity, was markedly decreased upon treatment with CCL01 (50 and 200 mg kg-1) in the hippocampus and cortex areas of the mouse brains in comparison with the LPS-injected group. In addition, the groups to which CCL01 was administered had significantly decreased plasma levels of tumor necrosis factor-α, interleukin (IL)-1ß, and IL-6 in the LPS-injected mouse models. Our data suggest that CCL01 may be a potential anti-neuroinflammatory agent that can prevent microglia overactivation, and it could be useful for developing functional foods.


Assuntos
Cuscuta , Lacticaseibacillus paracasei , Animais , Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/farmacologia , Cuscuta/metabolismo , Alimento Funcional , Interleucina-6/metabolismo , Lacticaseibacillus paracasei/metabolismo , Lipopolissacarídeos/farmacologia , Camundongos , Microglia , NF-kappa B/genética , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Extratos Vegetais/metabolismo , Extratos Vegetais/farmacologia , Sementes/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
20.
Neurol Genet ; 8(6): e200040, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36475054

RESUMO

Background and Objectives: While somatic mutations have been well-studied in cancer, their roles in other complex traits are much less understood. Our goal is to identify somatic variants that may contribute to the formation of saccular cerebral aneurysms. Methods: We performed whole-exome sequencing on aneurysm tissues and paired peripheral blood. RNA sequencing and the CRISPR/Cas9 system were then used to perform functional validation of our results. Results: Somatic variants involved in supervillin (SVIL) or its regulation were found in 17% of aneurysm tissues. In the presence of a mutation in the SVIL gene, the expression level of SVIL was downregulated in the aneurysm tissue compared with normal control vessels. Downstream signaling pathways that were induced by knockdown of SVIL via the CRISPR/Cas9 system in vascular smooth muscle cells (vSMCs) were determined by evaluating changes in gene expression and protein kinase phosphorylation. We found that SVIL regulated the phenotypic modulation of vSMCs to the synthetic phenotype via Krüppel-like factor 4 and platelet-derived growth factor and affected cell migration of vSMCs via the RhoA/ROCK pathway. Discussion: We propose that somatic variants form a novel mechanism for the development of cerebral aneurysms. Specifically, somatic variants in SVIL result in the phenotypic modulation of vSMCs, which increases the susceptibility to aneurysm formation. This finding suggests a new avenue for the therapeutic intervention and prevention of cerebral aneurysms.

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