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1.
Cell ; 172(5): 937-951.e18, 2018 02 22.
Artigo em Inglês | MEDLINE | ID: mdl-29456082

RESUMO

piRNAs (Piwi-interacting small RNAs) engage Piwi Argonautes to silence transposons and promote fertility in animal germlines. Genetic and computational studies have suggested that C. elegans piRNAs tolerate mismatched pairing and in principle could target every transcript. Here we employ in vivo cross-linking to identify transcriptome-wide interactions between piRNAs and target RNAs. We show that piRNAs engage all germline mRNAs and that piRNA binding follows microRNA-like pairing rules. Targeting correlates better with binding energy than with piRNA abundance, suggesting that piRNA concentration does not limit targeting. In mRNAs silenced by piRNAs, secondary small RNAs accumulate at the center and ends of piRNA binding sites. In germline-expressed mRNAs, however, targeting by the CSR-1 Argonaute correlates with reduced piRNA binding density and suppression of piRNA-associated secondary small RNAs. Our findings reveal physiologically important and nuanced regulation of individual piRNA targets and provide evidence for a comprehensive post-transcriptional regulatory step in germline gene expression.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/metabolismo , Células Germinativas/metabolismo , RNA Interferente Pequeno/metabolismo , Sequência de Aminoácidos , Animais , Pareamento de Bases , Sequência de Bases , Sítios de Ligação , Proteínas de Caenorhabditis elegans/química , Quimera/metabolismo , Inativação Gênica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
2.
Am J Transplant ; 2024 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-39094949

RESUMO

The impact of COVID-19 vaccination on clinical outcomes in solid organ transplant (SOT) recipients remains unclear. This systematic review and network meta-analysis sought to assess the efficacy and safety of COVID-19 vaccination in SOT recipients. We searched 6 databases from inception to March 1, 2024 for randomized controlled trials (RCTs) and observational studies evaluating different COVID-19 vaccination strategies in SOT recipients. Based on patient-important outcomes, we performed frequentist random-effects pairwise meta-analyses and network meta-analyses, separating RCTs and nonrandomized evidence, and used the Grading of Recommendation, Assessment, Development, and Evaluation approach to assess our certainty in the evidence. We included 6 RCTs (N = 814) and 43 observational studies (N = 125 199). Overall, there is a paucity of randomized evidence evaluating COVID-19 vaccines in SOT recipients. The nonrandomized evidence evaluating COVID-19 vaccination strategies patient-important outcomes, including COVID-19 infection, mortality, hospitalization, ICU admission, and rejection, demonstrated low to very low certainty due to the included studies' risk of bias. Throughout the COVID-19 pandemic, clinicians and SOT recipients worked with minimal, very low-quality evidence in relation to COVID-19 vaccines in this population. In the instance of future public health emergencies, clinicians and researchers should collaborate closely with patient partners to ensure there is sufficient evidence in the transplant population on patient-important outcomes.

3.
Pancreatology ; 24(6): 863-869, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39174438

RESUMO

BACKGROUND: Pseudocyst formation is common in many patients with acute pancreatitis during follow-up. Many risk factors have been proposed to be associated with the development of PP, but the predictive factors are still underexplored. The focus of this study was to investigate whether early laboratory indicators could effectively predict the occurrence of PP. METHODS: 2811 AP patients hospitalized in the Second Affiliated Hospital of Soochow University between November 2008 and September 2020 were retrospectively studied. Univariate and multivariate analyses were used to screen the risk variables. The nomograms of those risk factors were validated and evaluated by logistic analysis. RESULTS: AP patients had a 6.1 % (172/2811) incidence of PP. In a univariate analysis, the development of PP was correlated with serum lactate dehydrogenase (LDH), albumin (ALB), calcium (Ca), hemoglobin (Hb), organ dysfunction, CT severity index (CTSI), etiology, age, etc. Further logistic regression analysis showed that the risk factors were different between hyperlipidemic pancreatitis patients (LDH, ALB and Ca) and non-hyperlipidemic pancreatitis patients (LDH, Hb, ALB and Ca). A nomogram based on the identified risk factors was developed. Our model showed good discrimination ability, with a boostrap - corrected C index of 0.905 (95 % CI = 0.875-0.935), and had well-fitted calibration curves. The area under the curve (AUC) of the nomogram were 0.905 (95 % CI = 0.875-0.935) and 0.933 (95 % CI = 0.890-0.975) in the training and validation groups, respectively. The results of DCA indicated that the nomogram may have clinic usefulness. CONCLUSIONS: The nomogram that incorporates early laboratory data (LDH, Hb, ALB, and Ca) in AP patients is able to predict the incidence of PP with greater accuracy than the CTSI and AP severity.


Assuntos
Nomogramas , Pseudocisto Pancreático , Pancreatite , Humanos , Feminino , Masculino , Pessoa de Meia-Idade , Pancreatite/complicações , Pancreatite/etiologia , Fatores de Risco , Pseudocisto Pancreático/diagnóstico por imagem , Pseudocisto Pancreático/complicações , Estudos Retrospectivos , Adulto , Idoso , Incidência , Doença Aguda
4.
J Fluoresc ; 2024 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-38602590

RESUMO

In the present research, novel lanthanide coordination compounds [DyL(PhCOO)(CH3OH)](ClO4)2·(CH3OH)2 (1) were characterized by the compression of 2,6-diformyl-4-methyl-phenol (dmp) and 1,3-diamino-2-propanol using benzoate as the secondary ligand, where L indicates the deprotonated macrocyclic ligand. Through the high structural rigidity driven by the coordination of the macrocyclic ligand formed by condensation in methanol solution and sodium benzoate with Dy(ClO4)3·6H2O, compound 1 exhibits outstanding cyan-emitting fluorescence performance and potential applications as a fluorescent material. Additionally, hyaluronic acid (HA)/ carboxymethyl chitosan (CMCS) hydrogels were prepared with loaded resveratrol metal-organic complexes according to the synthetic chemical approach. In biological study, we evaluated the effect of hydrogels on oxidative stress on human dermal fibroblasts. Examined by molecular docking simulation, the results showed that the binding interactions were from the phenol group, the carboxyl group and also the "-N=" group, indicating Dy metal complex has excellent biological capability.

5.
Lipids Health Dis ; 23(1): 44, 2024 Feb 08.
Artigo em Inglês | MEDLINE | ID: mdl-38331899

RESUMO

BACKGROUND AND AIMS: To study the role of gene mutations in the development of severe hypertriglyceridemia (HTG) in patients with hyperlipidemic acute pancreatitis (HLAP), especially different apolipoprotein A5 (APOA5) mutations. METHODS: Whole-exome sequencing was performed on 163 patients with HLAP and 30 patients with biliary acute pancreatitis (BAP). The pathogenicity of mutations was then assessed by combining clinical information, predictions of bioinformatics programs, information from multiple gene databases, and residue location and conservation. The pathogenic mutations of APOA5 were visualized using the software. RESULTS: 1. Compared with BAP patients, pathogenic mutations of APOA5 were frequent in HLAP patients; among them, the heterozygous mutation of p.G185C was the most common. 2. All six pathogenic mutations of APOA5 identified in this study (p.S35N, p.D167V, p.G185C, p.K188I, p.R223C, and p.H182fs) were positively correlated with severe HTG; they were all in the important domains of apolipoprotein A-V (apoA-V). Residue 223 is strictly conserved in multiple mammals and is located in the lipoprotein lipase (LPL)-binding domain (Pro215-Phe261). When Arg 223 is mutated to Cys 223, the positive charge of this residue is reduced, which is potentially destructive to the binding function of apoA-V to LPL. 3. Four new APOA5 mutations were identified, namely c.563A > T, c.667C > T, c.788G > A, and c.544_545 insGGTGC. CONCLUSIONS: The pathogenic mutations of APOA5 were specific to the patients with HLAP and severe HTG in China, and identifying such mutations had clinical significance in elucidating the etiology and subsequent treatment.


Assuntos
Hipertrigliceridemia , Pancreatite , Humanos , Apolipoproteína A-V/genética , Apolipoproteínas A/genética , Apolipoproteínas A/metabolismo , Doença Aguda , Pancreatite/genética , Lipase Lipoproteica/genética , Hipertrigliceridemia/complicações , Hipertrigliceridemia/genética , Mutação
6.
Anal Chem ; 95(45): 16754-16760, 2023 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-37919241

RESUMO

Photocurrent polarity switchable photoelectrochemical (PEC) sensing has superior accuracy and anti-interference ability to conventional PEC sensing. The development of a novel strategy for photocurrent polarity switchable sensing is of great interest. Herein, a novel strategy for photocurrent polarity switchable sensing is reported by regulating electrostatic interactions between two semiconductor photoactive materials. Hyaluronic acid (HA)-modified CuO nanosheets show a negatively charged surface, which prevents the attachment of CuO nanosheets to negatively charged CdS nanodendrite-modified photoelectrodes because of the strong electrostatic repulsion. In the presence of hyaluronidase (HAase), the specific hydrolysis of HA on the surface of CuO by HAase can yield a positively charged surface, so CuO can be attached to a CdS-modified photoelectrode via electrostatic attraction, leading to photocurrent polarity switching. The photocurrent polarity switchable detection of HAase activity is achieved with an ultralow detection limit of 2 × 10-3 U mL-1 and a wide linear detection range between 0.01 and 100 U mL-1. This work provides a new and effective photocurrent polarity switching strategy for PEC sensing and a simple and efficient method for detecting HAase activity.

7.
Eur Arch Psychiatry Clin Neurosci ; 273(3): 687-697, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36251093

RESUMO

This case-control study was designed to examine the association between different types of miscarriage history and autism spectrum disorder (ASD), and determine whether the number of miscarriage history affects the risk of ASD. All of 2274 children with ASD and 1086 healthy controls were recruited. Sociodemographic and prenatal, perinatal, and neonatal characteristics were compared between the two groups. Multivariable logistic regression analyses were applied to investigate association between miscarriage history and ASD. Stratified analyses based on sex and types of miscarriages were similarly performed. History of miscarriage was potential risk factors for ASD ([aOR] = 2.919; 95% [CI] = 2.327-3.517). Stratified analyses revealed that induced ([aOR] = 2.763, 95% [CI] = 2.259-3.379) and spontaneous miscarriage history ([aOR] = 3.341, 95% [CI] = 1.939-4.820) were associated with high risk of ASD, respectively. A sex-biased ratio in the risk of ASD was observed between females ([aOR] = 3.049, 95% [CI] = 2.153-4.137) and males ([aOR] = 2.538, 95% [CI] = 1.978-3.251). Stratified analysis of induced miscarriage history revealed that only iatrogenic miscarriage history was associated with an increased risk ASD ([aOR] = 2.843, 95% [CI] = 1.534-4.268). Also, multiple spontaneous miscarriage histories ([aOR] = 1.836, 95% [CI] = 1.252-2.693) were associated with higher autism risk than one spontaneous miscarriages history ([aOR] = 3.016, 95% [CI] = 1.894-4.174). In conclusion, miscarriage history is related to an increased risk for ASD in offspring, which is affected by the types of miscarriage and sex of the fetus.


Assuntos
Aborto Espontâneo , Transtorno do Espectro Autista , Masculino , Gravidez , Feminino , Criança , Recém-Nascido , Humanos , Transtorno do Espectro Autista/epidemiologia , Aborto Espontâneo/epidemiologia , Estudos de Casos e Controles , Fatores de Risco
8.
Acta Pharmacol Sin ; 44(9): 1737-1747, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37076634

RESUMO

Epilepsy is not well controlled by current anti-seizure drugs (ASDs). High mobility group box 1 (HMGB1) is a DNA-binding protein in the nucleus regulating transcriptional activity and maintaining chromatin structure and DNA repair. In epileptic brains, HMGB1 is released by activated glia and neurons, interacting with various receptors like Toll-like receptor 4 (TLR4) and downstream glutamatergic NMDA receptor, thus enhancing neural excitability. But there is a lack of small-molecule drugs targeting the HMGB1-related pathways. In this study we evaluated the therapeutic potential of inflachromene (ICM), an HMGB-targeting small-molecule inhibitor, in mouse epilepsy models. Pentylenetetrazol-, kainic acid- and kindling-induced epilepsy models were established in mice. The mice were pre-treated with ICM (3, 10 mg/kg, i.p.). We showed that ICM pretreatment significantly reduced the severity of epileptic seizures in all the three epilepsy models. ICM (10 mg/kg) exerted the most apparent anti-seizure effect in kainic acid-induced epileptic status (SE) model. By immunohistochemical analysis of brain sections from kainic acid-induced SE mice, we found that kainic acid greatly enhanced HMGB1 translocation in the hippocampus, which was attenuated by ICM pretreatment in subregion- and cell type-dependent manners. Notably, in CA1 region, the seizure focus, ICM pretreatment mainly inhibited HMGB1 translocation in microglia. Furthermore, the anti-seizure effect of ICM was related to HMGB1 targeting, as pre-injection of anti-HMGB1 monoclonal antibody (5 mg/kg, i.p.) blocked the seizure-suppressing effect of ICM in kainic acid-induced SE model. In addition, ICM pretreatment significantly alleviated pyramidal neuronal loss and granule cell dispersion in kainic acid-induced SE model. These results demonstrate that ICM is an HMGB-targeting small molecule with anti-seizure potential, which may help develop a potential drug for treating epilepsy.


Assuntos
Epilepsia , Proteína HMGB1 , Camundongos , Animais , Ácido Caínico/efeitos adversos , Ácido Caínico/metabolismo , Epilepsia/induzido quimicamente , Epilepsia/tratamento farmacológico , Epilepsia/metabolismo , Hipocampo/metabolismo , Proteínas HMGB/metabolismo , Proteínas HMGB/farmacologia , Proteína HMGB1/metabolismo , Modelos Animais de Doenças
9.
BMC Public Health ; 23(1): 1254, 2023 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-37380948

RESUMO

AIMS: Neutrophils play a pivotal in immunity and inflammation. We aim to investigate the prevalence of neutropenia in the United States. METHODS: In this cross-sectional study, participants from the National Health and Nutrition Examination Survey (NHANES) (2011-2018) were enrolled. Demographic information, hematologic measurements, smoking status of all participants were collected for all participants. All statistical analyses were performed utilizing the NHANES survey weights. Covariate-adjusted linear regression was used to compare hematologic indices in different population grouped by age, sex, ethnicity, and smoking. We also employed multivariate-logistic regression to estimate the weighted odds ratio with a 95% confidence interval and predict the neutropenia risk among. RESULTS: 32,102 participants from NHANES survey were included, represented 286.6 million multiracial population in the United States. Black participants had lower mean leukocyte count (mean difference (MD): 0.71 × 109/L; P < 0.001) and lower neutrophil count (MD: 0.83 × 109/L; P < 0.001) compared with white participants after adjusting for age and sex. Furthermore, t a notable observation was the significant downward shift in the distribution curves of leukocyte count and neutrophil count among black participants. Smokers had a higher mean leukocyte count (MD: 1.10 × 109 cells/L; P < 0.001) and a higher mean neutrophil count (MD: 0.75 × 109 cells/L; P < 0.001) comparing with nonsmokers. The estimated prevalence of neutropenia was 1.24% (95% CI: 1.11 - 1.37%), which corresponds to approximately 35.5 million individuals in the United States. The prevalence of neutropenia in black participants was significantly higher than other races. Results of logistic regression analysis showed that black individuals, male individuals, and children younger than 5 years had a higher risk of neutropenia. CONCLUSIONS: Neutropenia is more common in the general population than we thought, especially in black individuals and children. More attention should be paid to neutropenia.


Assuntos
Neutropenia , Criança , Humanos , Masculino , Inquéritos Nutricionais , Estudos Transversais , Prevalência , Neutropenia/epidemiologia , Contagem de Leucócitos
10.
Int J Mol Sci ; 24(15)2023 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-37569858

RESUMO

Soybean is one of the most widely grown oilseed crops worldwide. Several unfavorable factors, including salt and salt-alkali stress caused by soil salinization, affect soybean yield and quality. Therefore, exploring the molecular basis of salt tolerance in plants and developing genetic resources for genetic breeding is important. Sucrose non-fermentable protein kinase 1 (SnRK1) belongs to a class of Ser/Thr protein kinases that are evolutionarily highly conserved direct homologs of yeast SNF1 and animal AMPKs and are involved in various abiotic stresses in plants. The GmPKS4 gene was experimentally shown to be involved with salinity tolerance. First, using the yeast two-hybrid technique and bimolecular fluorescence complementation (BiFC) technique, the GmSNF1 protein was shown to interact with the GmPKS4 protein. Second, the GmSNF1 gene responded positively to salt and salt-alkali stress according to qRT-PCR analysis, and the GmSNF1 protein was localized in the nucleus and cytoplasm using subcellular localization assay. The GmSNF1 gene was then heterologously expressed in yeast, and the GmSNF1 gene was tentatively identified as having salt and salt-alkali tolerance function. Finally, the salt-alkali tolerance function of the GmSNF1 gene was demonstrated by transgenic Arabidopsis thaliana, soybean hairy root complex plants overexpressing GmSNF1 and GmSNF1 gene-silenced soybean using VIGS. These results indicated that GmSNF1 might be useful in genetic engineering to improve plant salt and salt-alkali tolerance.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Proteínas de Soja/genética , Glycine max/metabolismo , Álcalis/metabolismo , Saccharomyces cerevisiae/metabolismo , Melhoramento Vegetal , Estresse Fisiológico/genética , Arabidopsis/metabolismo , Proteínas Quinases/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Arabidopsis/genética
11.
Analyst ; 147(24): 5747-5753, 2022 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-36413105

RESUMO

Photosensitizers and enzyme mimics are extensively used in photoelectrochemical (PEC) sensing, but few materials can be used as both photosensitizers and enzyme mimics in PEC sensing. Herein, we report Au nanoclusters (AuNCs) as both photosensitizers and peroxidase mimics for sensitive PEC sensing of Hg2+. It is found that AuNCs can act as photosensitizers to improve the PEC activity of WO3 nanorods; so the WO3/AuNCs composite material can be used as an advanced photosensitive material for PEC detection. AuNCs can also catalyze precipitate formation on the photoelectrode because of their peroxidase mimetic activity, and the interface electron transfer is hindered by the formed precipitate. Thus, the photocurrent of the WO3/AuNCs-based photoelectrode is quenched. When Hg2+ is present, the AuNCs-catalyzed precipitate formation is inhibited by Hg2+ because of the binding of Hg2+ to AuNCs through Hg2+-Au+ interactions. The photocurrent of the WO3/AuNCs-based photoelectrode increases accordingly, enabling "signal on" PEC detection of Hg2+. A broad linear range for Hg2+ detection is achieved between 1.0 pM and 50 nM with a detection limit of 0.2 pM. We have developed an advanced photosensitive material and introduced a simple method for PEC detection of Hg2+.


Assuntos
Mercúrio , Nanotubos , Fármacos Fotossensibilizantes , Catálise , Peroxidases
12.
J Nat Prod ; 85(11): 2592-2602, 2022 11 25.
Artigo em Inglês | MEDLINE | ID: mdl-36288556

RESUMO

In this work, four new cyclodepsipeptides, fusarihexins C-E (1-3) and enniatin Q (4), four new cyclopentane derivatives, fusarilins A-D (5-8), together with eight known compounds (9-16), were isolated from cultures of the endophytic fungus Fusarium sp. The structures of the isolated compounds were elucidated by analysis of HRMS and NMR spectroscopic data. The absolute configurations were determined using Marfey's method, a modified Mosher's method, single-crystal X-ray diffraction analysis, and ECD analysis. The antitumor activities of the isolated compounds in vitro were evaluated. Cyclodepsipeptides displayed cytotoxicities against the Huh-7, MRMT-1, and HepG-2 cell lines. Compounds 4, 9, 10, and 12 with IC50 values of 1.0-9.1 µM exhibited the most potent cytotoxicities against the three cell lines as compared to the positive control-5-fluorouracil. Compounds 1-3 and 11 exhibited moderate cytotoxic activities (IC50 values of 10.7-20.1 µM).


Assuntos
Antineoplásicos , Ciclopentanos , Depsipeptídeos , Fusarium , Antineoplásicos/química , Antineoplásicos/isolamento & purificação , Antineoplásicos/farmacologia , Cristalografia por Raios X , Ciclopentanos/química , Ciclopentanos/isolamento & purificação , Ciclopentanos/farmacologia , Depsipeptídeos/química , Fusarium/química , Estrutura Molecular , Células Hep G2 , Humanos
13.
Emerg Infect Dis ; 26(1): 179-180, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31855529

RESUMO

China has made remarkable progress in reducing schistosomiasis caused by Schistosoma japonicum over the past 7 decades but now faces a severe threat from imported schistosomiasis. Results from national surveillance during 2010-2018 indicate integrating active surveillance into current surveillance models for imported cases is urgently needed to achieve schistosomiasis elimination in China.


Assuntos
Esquistossomose/epidemiologia , Adulto , Idoso , Animais , China/epidemiologia , Erradicação de Doenças , Humanos , Masculino , Pessoa de Meia-Idade , Esquistossomose/etiologia , Esquistossomose/parasitologia , Esquistossomose/prevenção & controle , Migrantes , Viagem
14.
Biochem Cell Biol ; 98(2): 203-207, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-31484002

RESUMO

Human myeloid angiogenic cells (MACs), also termed early endothelial progenitor cells, play an important role in neovascularization and vascular repair. MicroRNAs (miRNAs) are a class of naturally occurring, noncoding, short (∼22 nucleotides), single-stranded RNAs that regulate gene expression post-transcriptionally. MiRNAs have been shown to regulate MAC function. A miRNA signature of MACs was described approximately a decade ago, and many new miRNAs have been discovered in recent years. In this study, we aimed to provide an up-to-date miRNA signature for human MACs. MACs were obtained by culture of human peripheral blood mononuclear cells in endothelial medium for 7 days. Using qPCR array analysis we identified 72 highly expressed miRNAs (CT value < 30) in human MACs. RT-qPCR quantification of select miRNAs revealed a strong correlation between the CT values detected by the array analysis and RT-qPCR, suggesting the miRNA signature generated by the qPCR array assay is accurate and reliable. Experimentally validated target genes of the 10 most highly expressed miRNAs were retrieved. Only a few of the targets and their respective miRNAs have been studied for their role in MAC biology. Our study therefore provides a valuable repository of miRNAs for future exploration of miRNA function in MACs.


Assuntos
Regulação da Expressão Gênica , Leucócitos Mononucleares/metabolismo , MicroRNAs/metabolismo , Células Mieloides/metabolismo , Neovascularização Patológica , Adulto , Células Endoteliais/metabolismo , Feminino , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Antígenos Comuns de Leucócito/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Masculino , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Reação em Cadeia da Polimerase , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/metabolismo
15.
Biotechnol Lett ; 42(2): 277-285, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31781926

RESUMO

OBJECTIVES: Developing a counterselective system for efficient markerless gene deletions in biocontrol strain P. protegens Pf-5. RESULTS: We successfully implemented a markerless deletion of upp in Pf-5 to obtain the 5-FU resistant strain Pf5139. With this strain, we performed markerless gene deletions for each component of Gac/Rsm system and a 17 kb DNA fragment with the deletion ratio of 20 to 50%, and efficiently constructed a strain with triple deletions based on the suicide plasmid pJQ200UPP. In addition, there is no obvious connection between the deleted fragment length and the deletion ratio. CONCLUSION: The upp-based counterselective system in this study is efficient and valuable for markerless gene deletions in Pf-5, indicating that it has great potential in the study of gene function and in the application of genome reduction for Pseudomonas strains.


Assuntos
Deleção de Genes , Genes Bacterianos , Pseudomonas/crescimento & desenvolvimento , Farmacorresistência Bacteriana , Fluoruracila/farmacologia , Técnicas Genéticas , Pseudomonas/efeitos dos fármacos , Pseudomonas/genética
16.
Mol Cell Biochem ; 462(1-2): 167-172, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31485855

RESUMO

MicroRNAs (miRNAs) regulate a wide range of cellular processes and functions. Blood mononuclear cells (BMNCs) participate in the immune response, inflammatory reaction and angiogenesis. In 2010, a total of 157 miRNAs were quantified by RT-qPCR and a miRNA signature was determined for human peripheral BMNCs. With the advent of technologies such as RNA sequencing, many new miRNAs have been identified. This study was designed to provide an up-to-date miRNA signature for human BMNCs. Peripheral BMNCs were isolated by Ficoll density gradient centrifugation. Using the qPCR array assay, we identified 108 highly expressed miRNAs (Ct value < 30) in human BMNCs. Further validation of the array results by quantifying select miRNAs with RT-qPCR revealed a strong correlation between Ct values derived from array analysis and RT-qPCR, suggesting the array results presented in this study are accurate and reliable. Of note, the function of the majority of the highly expressed miRNAs we have identified has not yet been studied. Our findings may help direct further studies of the regulatory roles of miRNAs in BMNC function.


Assuntos
Perfilação da Expressão Gênica , Leucócitos Mononucleares/metabolismo , MicroRNAs/genética , Regulação da Expressão Gênica , Humanos , MicroRNAs/metabolismo
17.
Can J Physiol Pharmacol ; 97(6): 562-569, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30512964

RESUMO

Hereditary hemorrhagic telangiectasia (HHT) is a rare vascular disorder inherited in an autosomal dominant manner. Patients with HHT can develop vascular dysplasias called telangiectasias and arteriovenous malformations (AVMs). Our objective was to profile and characterize micro-RNAs (miRNAs), short noncoding RNAs that regulate gene expression posttranscriptionally, in HHT patient-derived peripheral blood mononuclear cells (PBMCs). PBMCs, comprised mostly of lymphocytes and monocytes, have been reported to be dysfunctional in HHT. A total of 40 clinically confirmed HHT patients and 22 controls were enrolled in this study. PBMCs were isolated from 16 mL of peripheral blood and purified for total RNA. MiRNA expression profiling was conducted with a human miRNA array analysis. Select dysregulated miRNAs and miRNA targets were validated with reverse transcription-quantitative polymerase chain reaction. Of the 377 miRNAs screened, 41 dysregulated miRNAs were identified. Both miR-28-5p and miR-361-3p, known to target insulin-like growth factor 1 (IGF1), a potent angiogenic growth factor, were found to be significantly downregulated in HHT patients. Consequently, IGF1 mRNA levels were found to be significantly elevated. Our research successfully identified miRNA dysregulation and elevated IGF1 mRNA levels in PBMCs from HHT patients. This novel discovery represents a potential pathogenic mechanism that could be targeted to alleviate clinical manifestations of HHT.


Assuntos
Fator de Crescimento Insulin-Like I/genética , Leucócitos Mononucleares/metabolismo , MicroRNAs/genética , Telangiectasia Hemorrágica Hereditária/sangue , Telangiectasia Hemorrágica Hereditária/genética , Adulto , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/sangue , RNA Mensageiro/genética
18.
Can J Anaesth ; 65(8): 901-913, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29696581

RESUMO

PURPOSE: Hemodilutional anemia is associated with acute kidney injury (AKI) and mortality in patients undergoing cardiac surgery by mechanisms that may include tissue hypoxia. Our hypothesis was to assess if changes in the potential hypoxic biomarkers, including methemoglobin and erythropoietin, correlated with a decrease in hemoglobin (Hb) concentration following hemodilution on cardiopulmonary bypass (CPB). METHODS: Arterial blood samples were taken from patients (n = 64) undergoing heart surgery and CPB at baseline, during CPB, following CPB, and in the intensive care unit (ICU). Potential hypoxic biomarkers were measured, including methemoglobin, plasma Hb, and erythropoietin. Data were analyzed by repeated measures one-way analysis of variance on ranks and linear regression. RESULTS: Hemoglobin levels decreased following CPB and methemoglobin increased in the ICU (P < 0.001 for both). No correlation was observed between the change in Hb and methemoglobin (P = 0.23). By contrast, reduced Hb on CPB correlated with increased lactate, reduced pH, and increased erythropoietin levels following CPB (P ≤ 0.004 for all). Increased plasma Hb (P < 0.001) also correlated with plasma erythropoietin levels (P < 0.001). CONCLUSION: These data support the hypothesis that erythropoietin rather than methemoglobin is a potential biomarker of anemia-induced tissue hypoxia. The observed relationships between decreased Hb during CPB and the increase in lactate, reduced pH, and increase in erythropoietin levels suggest that early changes in plasma erythropoietin may be a pragmatic early biomarker of anemia-induced renal hypoxia. Further study is required to determine if anemia-induced increases in erythropoietin may predict AKI in patients undergoing cardiac surgery. TRIAL REGISTRATION: www.clinicaltrials.gov (NCT01883713). Registered 21 June 2013.


Assuntos
Injúria Renal Aguda/etiologia , Anemia/complicações , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Hemodiluição/efeitos adversos , Hipóxia/diagnóstico , Idoso , Biomarcadores/sangue , Ponte Cardiopulmonar/efeitos adversos , Eritropoetina/sangue , Feminino , Hemoglobinas/análise , Humanos , Masculino , Metemoglobina/análise , Pessoa de Meia-Idade , Estudos Prospectivos
19.
Chemistry ; 22(52): 18709-18713, 2016 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-27735092

RESUMO

Despite being widely used as electron acceptor in polymer solar cells, commercially available PC71 BM (phenyl-C71 -butyric acid methyl ester) usually has a "random" composition of mixed regioisomers or stereoisomers. Here PC71 BM has been isolated into three typical isomers, α-, ß1 - and ß2 -PC71 BM, to establish the isomer-dependent photovoltaic performance on changing the ternary composition of α-, ß1 - and ß2 -PC71 BM. Mixing the isomers in a ratio of α/ß1 /ß2 =8:1:1 resulted in the best power conversion efficiency (PCE) of 7.67 % for the polymer solar cells with PTB7:PC71 BM as photoactive layer (PTB7=poly[[4,8-bis[(2-ethylhexyl)oxy]benzo[1,2-b:4,5-b']dithiophene-2,6-diyl][3-fluoro-2-[(2-ethylhexyl)carbonyl]thieno[3,4-b]thiophenediyl]]). The three typical PC71 BM isomers, even though sharing similar LUMO energy levels and light absorption, render starkly different photovoltaic performances with average-performing PCE of 1.28-7.44 % due to diverse self-aggregation of individual or mixed PC71 BM isomers in the otherwise same polymer solar cells.

20.
Thorac Cardiovasc Surg ; 64(7): 555-560, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26366890

RESUMO

Background Acute kidney injury (AKI) is a common and serious complication of surgeries that include cardiopulmonary bypass (CPB). Currently, increases in serum creatinine levels are used to diagnose AKI, but this change may be slow to detect. Animal studies pertaining to renal hypoxia suggest a correlation between vasopressinase activity and AKI. The objective of this study is to determine if vasopressinase activity can be used as an early biomarker for renal hypoxia and CPB-associated AKI. This could potentially help improve the diagnosis and subsequent treatment of the condition. Materials and Methods We conducted a single-center, prospective observational study which analyzed serum vasopressinase activity and creatinine levels at seven time points from 31 patients undergoing CPB. We also measured urine vasopressinase activity in 19 of the 31 patients at five of the time points. Results Results show that serum and urine vasopressinase activity peak at the time of arrival to the ICU for patients undergoing CPB. This increase occurred earlier than the increase in creatinine, which generally occurred on postoperative day 2. In the five patients who were diagnosed with AKI, vasopressinase activity peaked 30 minutes into CPB while creatinine peaked on postoperative day 2. Conclusion Our findings suggest that vasopressinase might be a potential early biomarker for AKI. Further studies with other AKI biomarkers are required to determine if the vasopressinase response can be directly attributed to the presence of AKI.


Assuntos
Injúria Renal Aguda/diagnóstico , Ponte Cardiopulmonar/efeitos adversos , Ensaios Enzimáticos Clínicos , Cistinil Aminopeptidase/sangue , Cistinil Aminopeptidase/urina , Injúria Renal Aguda/sangue , Injúria Renal Aguda/urina , Idoso , Biomarcadores/sangue , Biomarcadores/urina , Creatinina/sangue , Diagnóstico Precoce , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Ontário , Valor Preditivo dos Testes , Estudos Prospectivos , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento , Regulação para Cima
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