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1.
Appl Microbiol Biotechnol ; 106(13-16): 5063-5079, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35833950

RESUMO

Beta-glucosidase (Bgl) is an enzyme with considerable food, beverage, and biofuel processing potential. However, as many Bgls are inhibited by their reaction end product glucose, their industrial applications are greatly limited. In this study, a novel Bgl gene (Bgl1973) was cloned from Leifsonia sp. ZF2019 and heterologously expressed in E. coli. Sequence analysis and structure modeling revealed that Bgl1973 was 748 aa, giving it a molecular weight of 78 kDa, and it showed high similarity with the glycoside hydrolase 3 (GH3) family Bgls with which its active site residues were conserved. By using pNPGlc (p-nitrophenyl-ß-D-glucopyranoside) as substrate, the optimum temperature and pH of Bgl1973 were shown to be 50 °C and 7.0, respectively. Bgl1973 was insensitive to most metal ions (12.5 mM), 1% urea, and even 0.1% Tween-80. This enzyme maintained 60% of its original activity in the presence of 20% NaCl, demonstrating its excellent salt tolerance. Furthermore, it still had 83% residual activity in 1 M of glucose, displaying its outstanding glucose tolerance. The Km, Vmax, and kcat of Bgl1973 were 0.22 mM, 44.44 µmol/min mg, and 57.78 s-1, respectively. Bgl1973 had a high specific activity for pNPGlc (19.10 ± 0.59 U/mg) and salicin (20.43 ± 0.92 U/mg). Furthermore, molecular docking indicated that the glucose binding location and the narrow and deep active channel geometry might contribute to the glucose tolerance of Bgl1973. Our results lay a foundation for the studying of this glucose-tolerant ß-glucosidase and its applications in many industrial settings. KEY POINTS: • A novel ß-glucosidase from GH3 was obtained from Leifsonia sp. ZF2019. • Bgl1973 demonstrated excellent glucose tolerance. • The glucose tolerance of Bgl1973 was explained using molecular docking analysis.


Assuntos
Actinomycetales , beta-Glucosidase , Actinomycetales/genética , Actinomycetales/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Glucose/metabolismo , Glicosídeo Hidrolases/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Simulação de Acoplamento Molecular , Especificidade por Substrato , beta-Glucosidase/metabolismo
2.
Int J Appl Earth Obs Geoinf ; 98: 102301, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35464667

RESUMO

The COVID-19 pandemic in China in the winter-spring of 2019-2020 has decreased and even stopped many human activities. This study investigates whether there were any changes in the water quality of the Lower Min River (China) during the lockdown period. The time-series remote sensing images from November 2019 to April 2020 was used to examine the dynamics of the river's total suspended solids (TSS) concentrations in the period. A new remote sensing-based prototype was developed to recalibrate an existing algorithm for retrieving TSS concentrations in the river. The Nechad and the Novoa algorithms were used to validate the recalibrated algorithm. The results show that the recalibrated algorithm is highly consistent with the two algorithms. All of the three algorithms indicate significant fluctuation in TSS concentrations in the Lower Min River during the study period. February (COVID-19 lockdown period) has witnessed a 48% fall in TSS concentration. The TSS in March-April showed a progressive and recovery back to normal levels of pre-COVID-19. The spatiotemporal change of TSS has worked as a good indicator of human activities, which revealed that the decline of TSS in the lockdown period was due largely to the substantially-reduced discharges from industrial estates, densely-populated city center, and river's shipping. Remote sensing monitoring of the spatiotemporal changes of TSS helps understand important contributors to the water-quality changes in the river and the impacts of anthropogenic activities on river systems.

3.
Biochem Biophys Res Commun ; 513(3): 740-745, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-30992133

RESUMO

RecFOR and RecA are key recombination factors in Deinococcus radiodurans, a bacterium that possesses robust DNA repair capability and is also naturally transformable. While RecFOR functioning as a RecA loader during DNA repair has been established, their relative roles in transformation need further exploration. Here, we constructed recFOR and recA deletion mutants of D. radiodurans, and investigated the effect of these mutations on DNA transformation. recA deletion causes defects in both plasmid and chromosomal transformation. However, it was found that recFOR is not involved in chromosomal transformation, and that only recO and recR mutations compromise plasmid transformation. How recO, recR and recA mutations influence plasmid transformation was further examined by complementation plasmids. Interestingly, the transformation process remains defective in the recA mutant, but gets restored in the recO and recR mutants. This indicates that unlike RecA, RecOR may not be essential for DNA uptake. Therefore, we provide evidence that RecFOR is dispensable for RecA to protect incoming exogenous DNA and to catalyze recombination during transformation. Instead, RecO and RecR are likely to promote later steps in plasmid transformation.


Assuntos
Proteínas de Bactérias/metabolismo , Reparo do DNA , Deinococcus/metabolismo , Recombinases Rec A/metabolismo , Transformação Genética , Proteínas de Bactérias/genética , Dano ao DNA , Deinococcus/genética , Deleção de Genes , Recombinases Rec A/genética , Recombinação Genética
4.
Mol Microbiol ; 106(4): 518-529, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28862774

RESUMO

RecF, together with the recombination mediators RecO and RecR, is required in the RecFOR homologous recombination repair pathway in bacteria. In this study, a recF-dr1088 operon, which is highly conserved in the Deinococcus-Thermus phylum, was identified in Deinococcus radiodurans. Interaction between DRRecF and DR1088 was confirmed by yeast two-hybrid and pull-down assays. DR1088 exhibited some RecO-like biochemical properties including single/double-stranded DNA binding activity, ssDNA binding protein (SSB) replacement ability and ssDNA (with or without SSB) annealing activity. However, unlike other recombination proteins, dr1088 is essential for cell viability. These results indicate that DR1088 might play a role in DNA replication and DNA repair processes.


Assuntos
Deinococcus/genética , Deinococcus/metabolismo , Proteínas de Bactérias/metabolismo , Reparo do DNA , Replicação do DNA/fisiologia , DNA Bacteriano/metabolismo , DNA de Cadeia Simples/metabolismo , Proteínas de Ligação a DNA/metabolismo , Recombinases Rec A/metabolismo , Recombinação Genética , Reparo de DNA por Recombinação/fisiologia
5.
J Bacteriol ; 197(12): 2048-61, 2015 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-25868646

RESUMO

UNLABELLED: In archaea, the NurA nuclease and HerA ATPase/helicase, together with the Mre11-Rad50 complex, function in 3' single-stranded DNA (ssDNA) end processing during homologous recombination (HR). However, bacterial homologs of NurA and HerA have not been characterized. From Deinococcus radiodurans, we identified the manganese-dependent 5'-to-3' ssDNA/double-stranded DNA (dsDNA) exonuclease/endonuclease NurA (DrNurA) and the ATPase HerA (DrHerA). These two proteins stimulated each other's activity through direct protein-protein interactions. The N-terminal HAS domain of DrHerA was the key domain for this interaction. Several critical residues of DrNurA and DrHerA were verified by site-directed mutational analysis. Temperature-dependent activity assays confirmed that the two proteins had mesophilic features, with optimum activity temperatures 10 °C to 15 °C higher than their optimum growth temperatures. Knocking out either nurA or herA affected cell proliferation by shortening the growth phase, especially for growth at a high temperature (37 °C). In addition, both mutant strains displayed almost 10-fold-reduced intermolecular recombination efficiency, indicating that DrNurA and DrHerA might be involved in homologous recombination in vivo. However, single- and double-gene deletions did not show significantly decreased radioresistance. Our results confirmed that the biochemical activities of bacterial NurA and HerA proteins were conserved with archaea. Our phenotypical results suggested that these proteins might have different functions in bacteria. IMPORTANCE: Deinococcus radiodurans NurA (DrNurA) was identified as a manganese-dependent 5'-to-3' ssDNA/dsDNA exonuclease/endonuclease, and Deinococcus radiodurans HerA (DrHerA) was identified as an ATPase. Physical interactions between DrNurA and DrHerA explained mutual stimulation of their activities. The N-terminal HAS domain on DrHerA was identified as the interaction domain. Several essential functional sites on DrNurA and DrHerA were characterized. Both DrHerA and DrNurA showed mesophilic biochemical features, with their optimum activity temperatures 10 °C to 15 °C higher than their optimum growth temperatures in vitro. Knockout of nurA or herA led to abnormal cell proliferation and reduced intermolecular recombination efficiency but no obvious effect on radioresistence.


Assuntos
Proteínas de Bactérias/metabolismo , Deinococcus/metabolismo , Adenosina Trifosfatases/genética , Adenosina Trifosfatases/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proliferação de Células , Biologia Computacional , Reparo do DNA/fisiologia , DNA Bacteriano/genética , Deleção de Genes , Regulação Bacteriana da Expressão Gênica/fisiologia , Regulação Enzimológica da Expressão Gênica , Temperatura Alta , Modelos Moleculares , Mutagênese Sítio-Dirigida , Óperon , Conformação Proteica
6.
Biochim Biophys Acta ; 1840(10): 3052-7, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25086254

RESUMO

BACKGROUND: A new G-quadruplex structure located in the B-cell CLL/lymphoma 2 (Bcl-2) P1 promoter and its physiological function related to Bcl-2 transcription have been studied to find a potential anticancer therapeutic target. METHODS: Absorption, polyacrylamide gel electrophoresis, fluorescence, circular dichroism, and nuclear magnetic resonance spectra have been employed to determine G-quadruplex structure and the interaction between G-quadruplex and phenanthrolin-dicarboxylate. Real time polymerase chain reaction and luciferase assay were done to assess the physiological function of the G-quadruplex structure. RESULTS: The UV-melting and polyacrylamide gel electrophoresis studies show that the p32 DNA sequence forms an intramolecular G-quadruplex structure. Circular dichroism and nuclear magnetic resonance spectra indicate that the G-quadruplex is a hybrid-type structure with four G-tetrads. Fluorescence spectra show that a phenanthroline derivative has a higher binding affinity for p32 G-quadruplex than duplex. Further circular dichroism and nuclear magnetic resonance studies indicate that the phenanthroline derivative can regulate p32 G-quadruplex conformation. Real time polymerase chain reaction and luciferase assays show that the phenanthroline derivative has down-modulated Bcl-2 transcription activity in a concentration-dependent manner. However, no such effect was observed when p32 G-quadruplex was denatured through base mutation. CONCLUSION: The newly identified G-quadruplex located in the P1 promoter of Bcl-2 oncogene is intimately related with Bcl-2 transcription activity, which may be a promising anticancer therapeutic target. GENERAL SIGNIFICANCE: The newly identified G-quadruplex in the Bcl-2 P1 promoter may be a novel anticancer therapeutic target.


Assuntos
Quadruplex G , Regulação Neoplásica da Expressão Gênica/efeitos da radiação , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transcrição Gênica/efeitos da radiação , Raios Ultravioleta , Linhagem Celular Tumoral , Humanos , Proteínas Proto-Oncogênicas c-bcl-2/genética
7.
Analyst ; 139(3): 581-4, 2014 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-24325001

RESUMO

A colorimetric and fluorometric dual-modal supramolecular chemosensor has been fabricated by using the H- and J-aggregates of a cyanine dye, which has been successfully applied to detect human serum albumin (HSA) in urine with high specificity.


Assuntos
Técnicas de Química Analítica/instrumentação , Fluorometria , Albumina Sérica/análise , Colorimetria , Humanos , Modelos Moleculares , Conformação Proteica , Albumina Sérica/química
8.
Acta Biochim Biophys Sin (Shanghai) ; 46(5): 368-76, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24681881

RESUMO

The RecFOR DNA repair pathway is one of the major RecA-dependent recombinatorial repair pathways in bacteria and plays an important role in double-strand breaks repair. RecO, one of the major recombination mediator proteins in the RecFOR pathway, has been shown to assist RecA loading onto single-stranded binding protein (SSB) coated single-stranded DNA (ssDNA). However, it has not been characterized whether the protein-protein interaction between RecO and SSB contributes to that process in vivo. Here, we identified the residue arginine-121 of Deinococcus radiodurans RecO (drRecO-R121) as the key residue for RecO-SSB interaction. The substitution of drRecO-R121 with alanine greatly abolished the binding of RecO to SSB but not the binding to RecR. Meanwhile, SSB-coated ssDNA annealing activity was also compromised by the mutation of the residue of drRecO. However, the drRecO-R121A strain showed only modest sensitivity to DNA damaging agents. Taking these data together, arginine-121 of drRecO is the key residue for SSB-RecO interaction, which may not play a vital role in the SSB displacement and RecA loading process of RecFOR DNA repair pathway in vivo.


Assuntos
Proteínas de Bactérias/fisiologia , Reparo do DNA , Deinococcus/fisiologia , Sequência de Bases , Primers do DNA , Ensaio de Desvio de Mobilidade Eletroforética
9.
Front Biosci (Landmark Ed) ; 29(10): 341, 2024 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-39473408

RESUMO

BACKGROUND: Glioblastoma (GBM) is an aggressive primary brain tumor. The HOX gene family has been implicated in the pathogenesis of different types of tumors. This research aimed to examine the impact of homeobox D9 (HOXD9) in GBM under hypoxic conditions, as well as to elucidate its underlying molecular mechanisms. METHODS: The study assessed the differential expression of nine HOXD genes in GBM using the Mann-Whitney U test and identified genes with high correlation with the cancer genome atlas (TCGA)-GBM dataset using receiver operating characteristic (ROC) curves. Prognostic genes of GBM patients were identified through a combination of prognostic Kaplan-Meier (KM) curve and Cox analysis. In vitro experiments were conducted using U87-MG and U251-MG cells, and an animal GBM model was constructed. The study also measured the secretion level of high mobility group box 1 (HMGB1) using enzyme-linked immunosorbent assay (ELISA). Glucose uptake and lactate production levels in cells and tissues were analyzed using kits. The expressions of HOXD9 and 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) were detected by immunofluorescence, and chromatin immunoprecipitation (ChIP) validated their relationship. RESULTS: HOXD9 was identified as the target gene, showing a significant correlation between HOXD9 expression and prognostic clinical outcomes. Overexpression of HMGB1 enhanced cell proliferation, migration, and the expression levels of HOXD9 and PFKFB3 and promoted HMGB1 secretion, glucose uptake, and lactate generation. HOXD9 bound to the PFKFB3 promoter region in U87-MG and U251-MG cells. Furthermore, PFKFB3 overexpression partially counteracted the suppressive effects of HOXD9 silencing on tumor formation. CONCLUSION: HOXD9 promoted hypoxia-induced HMGB1 secretion and glycolysis in GBM through the transcriptional activation of PFKFB3, which in turn promoted tumorigenesis.


Assuntos
Neoplasias Encefálicas , Regulação Neoplásica da Expressão Gênica , Glioblastoma , Glicólise , Proteína HMGB1 , Proteínas de Homeodomínio , Fosfofrutoquinase-2 , Fosfofrutoquinase-2/metabolismo , Fosfofrutoquinase-2/genética , Proteína HMGB1/metabolismo , Proteína HMGB1/genética , Humanos , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Glicólise/genética , Linhagem Celular Tumoral , Glioblastoma/genética , Glioblastoma/metabolismo , Glioblastoma/patologia , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Animais , Proliferação de Células/genética , Masculino , Camundongos , Feminino , Camundongos Nus , Prognóstico , Hipóxia Tumoral/genética , Pessoa de Meia-Idade , Proteínas de Neoplasias
10.
Enzyme Microb Technol ; 181: 110522, 2024 Oct 04.
Artigo em Inglês | MEDLINE | ID: mdl-39378560

RESUMO

Crocetin di/mono-glucosyl esters (crocin-4 and crocin-5) are rarely distributed in nature, limiting their potential applications in the food and pharmaceutical industries. In the present study, a novel GH3 family ß-glucosidase Lf18920 was identified from Leifsonia sp. ZF2019, which selectively hydrolyzed crocin-1 (crocetin di-gentiobiosyl ester) to crocin-5 and crocin-4, but not to its aglycone, crocetin. Under the optimal condition of 40 °C and pH 6.0 for 120 min, Lf18920 almost completely hydrolyzed crocin-1, yielding 73.50±5.66 % crocin-4 and 16.19±1.38 % crocin-5. Molecular docking and point mutation studies revealed that Lf18920 formed a narrow binding channel that facilitated crocin-1 binding. Five single amino acid variants (D50A, D53A, W274A, G420A, and Q421A) were constructed, all of which showed reduced hydrolytic activity. Mutations at D50 and D53, located distal to the active site, increased binding energy and decreased hydrolytic activity, while mutations at W274, G420, and Q421, proximal to the active site, disrupted hydrolytic function. These findings suggest that the narrow binding channel and specific enzyme-substrate interactions are crucial for Lf18920's selective hydrolytic activity. Overall, this study is the first to report a ß-glucosidase capable of selectively transforming crocin-1 to crocetin di/mono-glucosyl esters, offering potential for synthesizing crocin-4 and crocin-5.

11.
Stem Cell Rev Rep ; 19(6): 1907-1921, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37099039

RESUMO

Intracerebral hemorrhage (ICH) is a severe subtype of stroke for which there is no effective treatment. Stem cell and exosome (Exo) therapies have great potential as new approaches for neuroprotection and neurorestoration in treating ICH. We aimed to investigate whether Exo affects ICH by regulating the ecology of gut microbiota and metabolism and the mechanisms involved. First, differential miRNAs in ICH were screened by bioinformatics and verified by qRT-PCR. Then, Exo was extracted from mouse bone marrow mesenchymal stem cells (MSCs) and identified. Dual-luciferase reporter gene assay was utilized to verify the binding relationship between miR-150-3p and TRAF6. A mouse ICH model was constructed and treated with Exo. Next, we knocked down miR-150-3p and performed fecal microbiota transplantation (FMT). Then changes in gut microbiota and differential metabolites were detected by 16S rRNA sequencing and metabolomics analysis. We found that miR-150-3p expression was lowest in the brain tissue of the ICH group compared to the Sham group. Besides, low miR-150-3p level in ICH was encapsulated by MSC-derived Exo. Moreover, miR-150-3p bound to TRAF6 and was negatively correlated. With the addition of ExomiR-150-3p inhibitor, we found that MSC-derived exosomal miR-150-3p may affect ICH injury via TRAF6/NLRP3 axis. MSC-derived exosomal miR-150-3p caused changes in gut microbiota, including Proteobacteria, Muribaculaceae, Lachnospiraceae_NK4A136_group, and Acinetobacter. Moreover, MSC-derived exosomal miR-150-3p caused changes in metabolism. After further FMT, gut microbiota-mediated MSC-derived Exo affected ICH with reduced apoptosis and reduced levels of inflammatory factors. In conclusion, MSC-derived exosomal miR-150-3p affected ICH by regulating TRAF6/NF-κB axis, gut microbiota and metabolism.


Assuntos
Microbioma Gastrointestinal , Células-Tronco Mesenquimais , MicroRNAs , Animais , Camundongos , Hemorragia Cerebral/genética , Hemorragia Cerebral/metabolismo , Modelos Animais de Doenças , Microbioma Gastrointestinal/genética , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismo
12.
Food Chem ; 424: 136260, 2023 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-37244184

RESUMO

Since lipid oxidation often causes serious food safety issues worldwide, determination of oil's oxidative deterioration becomes quite significant, which still calls for efficient analytical methods. In this work, high-pressure photoionization time-of-flight mass spectrometry (HPPI-TOFMS) was firstly introduced for rapid detection of oxidative deterioration in edible oils. Through non-targeted qualitative analysis, oxidized oils with various oxidation levels were successfully discriminated for the first time by coupling HPPI-TOFMS with the orthogonal partial least squares discriminant analysis (OPLS-DA). Furthermore, by targeted interpretation of the HPPI-TOFMS mass spectra and the subsequent regression analysis (signal intensities vs TOTOX values), good linear correlations were observed for several predominant VOCs. Those specific VOCs were promising oxidation indicators, which would play important roles as TOTOX to judge the oxidation states of tested samples. The proposed HPPI-TOFMS methodology can be used as an innovative tool for accurate and effective assessment of lipid oxidation in edible oils.


Assuntos
Alimentos , Óleos , Espectrometria de Massas , Estresse Oxidativo , Óleos de Plantas/química
13.
Food Chem ; 405(Pt A): 134655, 2023 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-36345096

RESUMO

In our previous study, AI-3, a mixture of isoflavones, was obtained from Apios Americana Medik tuber and showed strong protective ability on oxidative damaged RIN-m5F cells. This study aimed to identify the main compounds of AI-3 and elucidate their activities and underlying mechanism. In results, eleven compounds were purified from AI-3. Among them, Compound 8 (2'-Hydroxy, 5-methoxy genistein-7-O-glucoside, HMG) was the most effective compound against H2O2-induced injury in RIN-m5F cells (stronger than positive control α-LA). Further RNA-seq analysis found that compared with H2O2 group, 388 differentially expressed genes (DEGs) were identified in HMG group. The enrichment analyses revealed fluid shear stress and atherosclerosis pathway and hepatocellular carcinoma pathway enriched the most DEGs, in which HOX-1, GST, NQO1, SQSTM, TrxR1 were significantly up-regulated. The finding indicated Keap1-Nrf2-ARE signaling pathway may play essential role in the protective effect of HMG on oxidative damaged RIN-m5F cells.


Assuntos
Fabaceae , Isoflavonas , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch/metabolismo , Peróxido de Hidrogênio/metabolismo , Fabaceae/metabolismo , Estresse Oxidativo , Isoflavonas/farmacologia
14.
Nat Commun ; 14(1): 3219, 2023 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-37270607

RESUMO

Extreme atmospheric rivers (EARs) are responsible for most of the severe precipitation and disastrous flooding along the coastal regions in midlatitudes. However, the current non-eddy-resolving climate models severely underestimate (~50%) EARs, casting significant uncertainties on their future projections. Here, using an unprecedented set of eddy-resolving high-resolution simulations from the Community Earth System Model simulations, we show that the models' ability of simulating EARs is significantly improved (despite a slight overestimate of ~10%) and the EARs are projected to increase almost linearly with temperature warming. Under the Representative Concentration Pathway 8.5 warming scenario, there will be a global doubling or more of the occurrence, integrated water vapor transport and precipitation associated with EARs, and a more concentrated tripling for the landfalling EARs, by the end of the 21st century. We further demonstrate that the coupling relationship between EARs and storms will be reduced in a warming climate, potentially influencing the predictability of future EARs.

15.
Biomark Res ; 11(1): 47, 2023 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-37138354

RESUMO

BACKGROUND: Urinary bladder cancer (UBC) is a common malignancy of the urinary tract; however, the mechanism underlying its high recurrence and responses to immunotherapy remains unclear, making clinical outcome predictions difficult. Epigenetic alterations, especially DNA methylation, play important roles in bladder cancer development and are increasingly being investigated as biomarkers for diagnostic or prognostic predictions. However, little is known about hydroxymethylation since previous studies based on bisulfite-sequencing approaches could not differentiate between 5mC and 5hmC signals, resulting in entangled methylation results. METHODS: Tissue samples of bladder cancer patients who underwent laparoscopic radical cystectomy (LRC), partial cystectomy (PC), or transurethral resection of bladder tumor (TURBT) were collected. We utilized a multi-omics approach to analyze both primary and recurrent bladder cancer samples. By integrating various techniques including RNA sequencing, oxidative reduced-representation bisulfite sequencing (oxRRBS), reduced-representation bisulfite sequencing (RRBS), and whole exome sequencing, a comprehensive analysis of the genome, transcriptome, methylome, and hydroxymethylome landscape of these cancers was possible. RESULTS: By whole exome sequencing, we identified driver mutations involved in the development of UBC, including those in FGFR3, KDMTA, and KDMT2C. However, few of these driver mutations were associated with the down-regulation of programmed death-ligand 1 (PD-L1) or recurrence in UBC. By integrating RRBS and oxRRBS data, we identified fatty acid oxidation-related genes significantly enriched in 5hmC-associated transcription alterations in recurrent bladder cancers. We also observed a series of 5mC hypo differentially methylated regions (DMRs) in the gene body of NFATC1, which is highly involved in T-cell immune responses in bladder cancer samples with high expression of PD-L1. Since 5mC and 5hmC alternations are globally anti-correlated, RRBS-seq-based markers that combine the 5mC and 5hmC signals, attenuate cancer-related signals, and therefore, are not optimal as clinical biomarkers. CONCLUSIONS: By multi-omics profiling of UBC samples, we showed that epigenetic alternations are more involved compared to genetic mutations in the PD-L1 regulation and recurrence of UBC. As proof of principle, we demonstrated that the combined measurement of 5mC and 5hmC levels by the bisulfite-based method compromises the prediction accuracy of epigenetic biomarkers.

16.
Analyst ; 137(4): 862-7, 2012 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-22223064

RESUMO

Recognition of different human telomeric G-quadruplex structures has been a very important task for developing anti-cancer drug design. However, it also is a very challenging question since multiple conformational isomers of telomeric G-quadruplexes coexist under some conditions. Here, three different conformations including parallel, antiparallel, and mixed-type telomeric G-quadruplex structures have been well recognized by quinacrine (QNA) through monitoring its absorption, fluorescence, and fluorescence lifetime spectra. The multiple structures of H22 G-quadruplexes under physiological K(+) conditions could also be easily determined to coexist as mixed-type and antiparallel G-quadruplexes. The recognition mechanism based on the different binding affinity and binding sites has been further elucidated by association with the nuclear magnetic resonance (NMR) results.


Assuntos
Quadruplex G , Ressonância Magnética Nuclear Biomolecular/métodos , Quinacrina , Telômero/química , Sítios de Ligação , Desenho de Fármacos , Humanos , Modelos Moleculares , Conformação Molecular , Conformação de Ácido Nucleico , Telomerase/antagonistas & inibidores , Telomerase/metabolismo
17.
Analyst ; 137(24): 5713-5, 2012 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-23108173

RESUMO

A supramolecular probe is designed for visual detection of potassium based on a novel cyanine dye aggregate recognizing the motif transition of telomeric G-quadruplexes under the Na(+) background. The practical application for colorimetric measurement of urine potassium has been tested.


Assuntos
Carbocianinas/química , Colorimetria/métodos , Corantes/química , Quadruplex G , Motivos de Nucleotídeos , Potássio/análise , Humanos , Potássio/urina , Telômero/genética
18.
Nucleic Acids Res ; 38(3): 1022-33, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19933263

RESUMO

The supramolecular assembly of a novel cyanine dye, 3,3'-di(3-sulfopropyl)-4,5,4',5'-dibenzo-9-ethyl-thiacarbocyanine triethylammonium salt (ETC) was designed to verify specific intramolecular G-quadruplexes from duplex and single-strand DNAs. Spectral results have shown that ETC presented two major distinct signatures with specific intramolecular G-quadruplexes in vitro: (i) dramatic changes in the absorption spectra (including disappearance of absorption peak around 660 nm and appearance of independent new peak around 584 nm); (ii) approximately 70 times enhancement of fluorescence signal at 600 nm. Furthermore, based on (1)H-nuclear magnetic resonance and circular dichroism results, the preferring binding of ETC to specific intramolecular G-quadruplexes probably result from end-stacking, and the loop structure nearby also plays an important role.


Assuntos
Carbocianinas/química , DNA/química , Corantes Fluorescentes/química , Quadruplex G , DNA de Cadeia Simples/química , Modelos Moleculares , Ressonância Magnética Nuclear Biomolecular , Espectrometria de Fluorescência , Espectrofotometria
19.
Int J Mol Sci ; 13(2): 2249-2262, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22408451

RESUMO

Sasa argenteastriatus, with abundant active compounds and high antioxidant activity in leaves, is a new leafy bamboo grove suitable for exploitation. To utilize it more effectively and scientifically, we investigate the seasonal variations of antioxidant composition in its leaves and antioxidant activity. The leaves of Sasa argenteastriatus were collected on the 5th day of each month in three same-sized sample plots from May 2009 to May 2011. The total flavonoids (TF): phenolics (TP) and triterpenoid (TT) of bamboo leaves were extracted and the contents analyzed by UV-spectrophotometer. Our data showed that all exhibited variations with the changing seasons, with the highest levels appearing in November to March. Antioxidant activity was measured using DPPH and FRAP methods. The highest antioxidant activity appeared in December with the lowest in May. Correlation analyses demonstrated that TP and TF exhibited high correlation with bamboo antioxidant activity. Eight bamboo characteristic compounds (orientin, isoorientin, vitexin, homovitexin and p-coumaric acid, chlorogenic acid, caffeic acid, ferulic acid) were determined by RP-HPLC synchronously. We found that chlorogenic acid, isoorientin and vitexin are the main compounds in Sasa argenteastriatus leaves and the content of isovitexin and chlorogenic acid showed a similar seasonal variation with the TF, TP and TT. Our results suggested that the optimum season for harvesting Sasa argenteastriatus leaves is between autumn and winter.


Assuntos
Antioxidantes/análise , Extratos Vegetais/química , Folhas de Planta/química , Sasa/química , Estações do Ano , Antioxidantes/metabolismo , Apigenina/análise , Apigenina/metabolismo , Ácidos Cafeicos/análise , Ácidos Cafeicos/metabolismo , Cromatografia Líquida de Alta Pressão , Ácidos Cumáricos/análise , Ácidos Cumáricos/metabolismo , Flavonoides/análise , Flavonoides/metabolismo , Glucosídeos/análise , Glucosídeos/metabolismo , Fenóis/análise , Fenóis/metabolismo , Folhas de Planta/metabolismo , Propionatos , Sasa/metabolismo , Triterpenos/análise , Triterpenos/metabolismo
20.
World J Clin Cases ; 10(31): 11555-11560, 2022 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-36387783

RESUMO

BACKGROUND: Acute myocardial infarction (AMI) can be induced by several factors. However, AMI induced by Kounis syndrome (an allergic reaction) is extremely rare and is highly susceptible to misdiagnosis. CASE SUMMARY: A 70-year-old man presented after suffering an allergic reaction that caused chest pain triggered upon eating ice cream. Troponin I was found to be elevated, and an electrocardiogram showed ST-segment elevation. The diagnosis was AMI. He underwent two coronary angiographies, with one intravascular ultrasound during hospitalization showing no evidence of atherosclerotic coronary artery disease. The final diagnosis was vasospastic myocardial infarction due to Kounis syndrome. The patient was then treated with hydrocortisone and intravenous antihistamines, and his chest pain symptoms resolved. CONCLUSION: Allergic reactions (such as Kounis syndrome) can cause serious damage to the heart. Physicians should be alert to the consequences and avoid misdiagnosis.

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