RESUMO
Here, we describe a novel mycovirus, tentatively designated as "Rhizoctonia solani fusarivirus 6" (RsFV6), which was discovered in Rhizoctonia solani AG-3 PT strain 3P-2-2. The virus has a single-stranded positive-sense RNA (+ssRNA) genome of 6141 nucleotides containing two open reading frames (ORFs) and a poly(A) tail. ORF1 encodes a large polypeptide of 1,862 amino acids (aa) with conserved RNA-dependent RNA polymerase (RdRp) and helicase (Hel) domains. ORF2 encodes a putative 167-aa protein of unknown function. BLASTp searches revealed that the ORF1-encoded polypeptide showed the highest sequence similarity (70.67% identity) to that of Rhizoctonia solani fusarivirus 3 (RsFV3), which was isolated from Rhizoctonia solani AG-2-2LP. Multiple sequence alignments and phylogenetic analysis based on RdRp and Hel sequences indicated that RsFV6 could be a novel member of the genus Alphafusarivirus family Fusariviridae.
Assuntos
Micovírus , Vírus de RNA , RNA Viral/genética , Filogenia , Genoma Viral , Rhizoctonia/genética , RNA Polimerase Dependente de RNA/genética , Micovírus/genética , Fases de Leitura AbertaRESUMO
NHC-alcohol adduct-mediated deoxygenation of alcohols under photocatalytic conditions is described. This process provides various alkyl radicals, which can react with 2-isocyanobiaryls to afford 6-substituted phenanthridine derivatives in moderate to good yields. This method offered the first example of directly using alcohols as radical sources for 6-phenanthridine synthesis.
Assuntos
Etanol , FenantridinasRESUMO
The nucleotide sequence of a viral double-stranded RNA (dsRNA) from Rhizoctonia solani AG-4 HGIII strain XMC-IF (designated as "Rhizoctonia solani mitovirus 106", RsMV-106) was determined. The complete sequence was 2794 bp in length with a 57.50% A + U content and contained a large open reading frame (ORF) when the fungal mitochondrial genetic code was used. The ORF potentially encodes a 95.76-kDa protein containing a conserved domain of an RNA-dependent RNA polymerase (RdRp). BLASTp analysis revealed that the RdRp domain of RsMV-106 shared 47.52-73.24% sequence identity with those of viruses of the genus Duamitovirus and was most similar (73.24% identity) to that of Alternaria alternata mitovirus 1 (AaMV1). Phylogenetic analysis showed that RsMV-106 is a novel member of the genus Duamitovirus, family Mitoviridae. This is the first report of the full genome sequence of a mitovirus associated with R. solani AG-4 HGIII.
Assuntos
Micovírus , Vírus de RNA , Filogenia , Genoma Viral , Vírus de RNA/genética , Rhizoctonia/genética , Fases de Leitura Aberta , RNA de Cadeia Dupla , RNA Viral/genética , Micovírus/genéticaRESUMO
The complete genome sequence of a double-stranded RNA (dsRNA) virus, Rhizoctonia solani dsRNA virus 11 (RsRV11), isolated from Rhizoctonia solani AG-1 IA strain 9-11 was determined. The RsRV11 genome is 9,555 bp in length and contains three conserved domains: structural maintenance of chromosomes (SMC) superfamily, phosphoribulokinase (PRK), and RNA-dependent RNA polymerase (RdRp). The RsRV11 genome has two non-overlapping open reading frames (ORFs). ORF1 is predicted to encode a 204.12-kDa protein that shares low but significant amino acid sequence similarity with a putative protein encoded by Rhizoctonia solani RNA virus HN008 (RsRV-HN008). ORF2 potentially encodes a 132.41-kDa protein that contains the conserved domain of the RdRp. Phylogenetic analysis indicated that RsRV11 clustered with RsRV-HN008 in a separate clade from other virus families. This implies that RsRV11 and RsRV-HN008 should be included in a new mycovirus taxon close to the family Megabirnaviridae and that RsRV11 is a new mycovirus.
Assuntos
Micovírus/genética , Genoma Viral , Filogenia , Rhizoctonia/virologia , China , Micovírus/isolamento & purificação , Fases de Leitura Aberta , RNA de Cadeia Dupla , Rhizoctonia/isolamento & purificação , Proteínas Virais/genética , Zea mays/microbiologiaRESUMO
A CuI-catalyzed coupling reaction of benzofuran-3(2H)-ones with amines has been well established for the direct synthesis of α-ketoamides. This process involves C-O bond cleavage and C[double bond, length as m-dash]O/C-N bond formation. Mechanism studies indicated that this α-ketoamide formation reaction may involve a free radical process.
RESUMO
Methods based on prussian blue nanoparticles (PBNPs) have been reported for photothermal immunoassays in analytical nanoscience fields but most suffer from low sensitivity and are not beneficial for routine use. Herein, we design an in situ amplified near-infrared (NIR) photothermal immunoassay for the quantitative screening of neuron-specific enolase (NSE) on a portable thermometer using PBNP-encapsulated nanoliposomes as photosensitive materials. Biotinylated liposomes loaded with numerous prussian blue nanoparticles were synthesized through a typical reverse-phase evaporation method. The photothermal immunoassay was carried out in an anti-NSE capture antibody-coated microplate using the biotinylated anti-NSE secondary antibody. With the sandwiched immunoreaction and the biotin-avidin linkage, the subsequent photothermal measurement of PBNPs released from the liposomes with buffered surfactant including Tween 20 was conducted on a digital thermometer under near-infrared 808 nm laser irradiation, accompanied by the convertion of NIR-light wavelength to heat. Under the optimum conditions, the photothermal immunoassay displayed a wide dynamic concentration range of 0.1-100 ng mL-1 with a low detection limit for NSE of 0.053 ng mL-1. Good reproducibility (RSD ≤ 2.78% for intra-assay; RSD ≤ 4.39% for inter-assay), high selectivity against other biomarkers, and a long-term stability (≥94.9% of the initial signal during six-month storage) were acquired in the photothermal immunoassay. Impressively, the analysis of 7 human serum specimens for target NES via the photothermal immunoassay also gave well-matched results with the referenced human NSE enzyme-linked immunosorbent assay.
Assuntos
Ferrocianetos/química , Imunoensaio/métodos , Lipossomos/química , Nanopartículas/química , Fosfopiruvato Hidratase/sangue , Ferrocianetos/efeitos da radiação , Humanos , Imunoensaio/instrumentação , Raios Infravermelhos , Limite de Detecção , Nanopartículas/efeitos da radiação , Reprodutibilidade dos Testes , TermômetrosRESUMO
An innovative signal-transduction tag based on cross-linked urease nanoparticles (CLENP) was designed for the development of a pH meter-based immunoassay of lipocalin-2 (LCN2). The CLENP was synthesized with a typical desolvation method using ethanol as desolvation agent, followed by functionalization with polyaspartic acid. The carboxylated CLENP were used as the signal-generation tags for the labelling of secondary antibodies via the carbodiimide coupling. Upon target LCN2 introduction, a sandwich-type immune reaction was performed between capture antibody-coated plate and the labeled secondary antibody on the CLENP. The conjugated CLENP in the microplate hydrolyzed urea into ammonia (NH4+) and carbonate (CO32-), resulting in the pH change of solution, which was determined with a handheld pH meter. The pH variation was proportional to target concentration in the sample. By monitoring the pH variation of the urea solution, the level of LCN2 at a concentration as low as 5.2 pg mL-1 was evaluated. The pH meter-based electrochemical immunoassay can be utilized for mass production of miniaturized lab-on-a-chip devices with handheld pH meter, thereby opening new opportunities for protein diagnostics and biosecurity. Graphical abstract An innovative signal-transduction tag based on cross-linked urease nanoparticles was designed for high-efficiency immunoassay of lipocalin-2 with pH meter readout.
Assuntos
Imunoensaio/métodos , Lipocalina-2/análise , Nanopartículas/química , Urease/química , Anticorpos Imobilizados/imunologia , Anticorpos Monoclonais/imunologia , Canavalia/enzimologia , Técnicas Eletroquímicas/métodos , Enzimas Imobilizadas/química , Humanos , Concentração de Íons de Hidrogênio , Limite de Detecção , Lipocalina-2/imunologia , Estudo de Prova de Conceito , Ureia/químicaRESUMO
A highly selective and sensitive molecularly imprinted electrochemiluminescence (MIECL) sensor was developed based on the multiwall carbon nanotube (MWCNT)-enhanced molecularly imprinted quantum dots (MIP-QDs) for the rapid determination of cyfluthrin (CYF). The MIP-QDs fabricated by surface grafting technique exhibited excellent selective recognition to CYF, resulting in a specific decrease of ECL signal at the MWCNT/MIP-QD modified electrode. Under optimal conditions, the MIECL signal was proportional to the logarithm of the CYF concentration in the range of 0.2 µg/L to 1.0 × 103 µg/L with a determination coefficient of 0.9983. The detection limit of CYF was 0.05 µg/L, and good recoveries ranging from 86.0% to 98.6% were obtained in practical samples. The proposed MIECL sensor provides a novel, rapid, high sensitivity detection strategy for successfully analyzing CYF in fish and seawater samples.
RESUMO
A high-efficiency pyrethroid-degrading bacterium, Photobacterium ganghwense strain 6046 (PGS6046), was first isolated from an offshore seawater environment. Metabolomics method was used to investigate the biotransformation pathway of PGS6046 to cyfluthrin wherein 156 metabolites were identified. The growth rates of the PGS6046 cultivated in nourishing media were much higher than those cultivated in seawater, regardless of the presence of cyfluthrin. Statistical analyses revealed that the metabolic profile of PGS6046 was associated with the culture medium, the presence of cyfluthrin, and culture time. The PGS6046 cultivated in a nourishing medium was characterized by higher levels of amino acids, a lower abundance of intermediates in the tricarboxylic acid cycle, and the presence of some fatty acids than those cultivated in seawater. The effects of cyfluthrin on PGS6046 metabolism varied based on the culture medium, whereas the cyanoalanine levels increased under both culture conditions. Culture time significantly affected the metabolism of amino acids and carbohydrates in PGS6046. The present study revealed the metabolic characteristics of PGS6046 under different culture conditions and will further facilitate the exploration of the fundamental questions regarding PGS6046 and its potential applications in environmental bioremediation.
Assuntos
Metabolômica/métodos , Nitrilas/metabolismo , Photobacterium/fisiologia , Piretrinas/metabolismo , Biodegradação Ambiental , China , Meios de Cultura/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Inseticidas/metabolismo , Photobacterium/efeitos dos fármacos , Photobacterium/isolamento & purificação , Filogenia , Água do Mar/microbiologia , Fatores de TempoRESUMO
Herein a new potentiometric immunoassay for the point-of-care detection of enterovirus 71 (EV71) was developed by using a silver (Ag+) ion-selective electrode (ISE). Initially, the carboxylated dendrimer-doped AgCl nanospheres were synthesized by the reverse micelle method. Then the synthesized nanospheres were used to label a polyclonal mouse anti-EV71 antibody via a typical carbodiimide coupling method. The immunoreaction was executed on a monoclonal anti-EV71 antibody-coated microplate by using biofunctional AgCl nanospheres as the detection antibody. With a sandwich-type immunoassay format, the carried AgCl nanospheres could be dissolved in the presence of NH3·H2O, and the released silver ions were determined with an external silver ion-selective electrode. Under optimal conditions, the shift in the potential increased with the increase in the EV71 concentration, in a wide linear range of 0.3-300 ng mL-1, with a detection limit of 0.058 ng mL-1. Intra- and inter-assay relative standard deviations with identical batches were less than 4.15% and 6.15%, respectively. By validating the spiked serum samples, our system shows consistency with the enzyme-linked immunosorbent assay (ELISA) kit.
Assuntos
Técnicas Biossensoriais , Enterovirus Humano A/isolamento & purificação , Eletrodos Seletivos de Íons , Nanosferas , Prata , Anticorpos Monoclonais , Dendrímeros , Humanos , Imunoensaio , NanoconjugadosRESUMO
The well-dispersive and superparamagnetic Fe3O4-nanocrystals (Fe3O4-NCs) which could significantly enhance the anodic electrochemiluminescence (ECL) behavior of luminol, were synthesized in this study. Compared to ZnS, ZnSe, CdS and CdTe nanoparticles, the strongest anodic ECL signals were obtained at +1.6 V on the Fe3O4-NCs coated glassy carbon electrode. The ECL spectra revealed that the strong ECL resonance energy transfer occurred between luminol and Fe3O4-NCs. Furthermore, under the optimized ECL experimental conditions, such as the amount of Fe3O4-NCs, the concentration of luminol and the pH of supporting electrolyte, BPA exhibited a stronger distinct ECL quenching effect than its structural analogs and a highly selective and sensitive ECL sensor for the determination of bisphenol A (BPA) was developed based on the Fe3O4-NCs. A good linear relationship was found between the ECL intensity and the increased BPA concentration within 0.01â»5.0 mg/L, with a correlation coefficient of 0.9972. The detection limit was 0.66 × 10-3 mg/L. Good recoveries between 96.0% and 105.0% with a relative standard deviation of less than 4.8% were obtained in real water samples. The proposed ECL sensor can be successfully employed to BPA detection in environmental aqueous samples.
RESUMO
The upregulation of reactive oxygen species (ROS) is a primary cause of cardiomyocyte apoptosis in diabetes cardiomyopathy (DCM). Mitofusin-2 (Mfn-2) is a key protein that bridges the mitochondria and endoplasmic reticulum (ER). Hydrogen sulfide (H2S)-mediated cardioprotection is related to antioxidant effects. The present study demonstrated that H2S inhibited the interaction between the ER and mitochondrial apoptotic pathway. This study investigated cardiac function, ultrastructural changes in the ER and mitochondria, apoptotic rate using TUNEL, and the expression of ER stress-associated proteins and mitochondrial apoptotic proteins in cardiac tissues in STZ-induced type I diabetic rats treated with or without NaHS (donor of H2S). Mitochondria of cardiac tissues were isolated, and MPTP opening and cytochrome c (cyt C) and Mfn-2 expression were also detected. Our data showed that hyperglycemia decreased the cardiac function by ultrasound cardiogram, and the administration of exogenous H2S ameliorated these changes. We demonstrated that the expression of ER stress sensors and apoptotic rates were elevated in cardiac tissue of DCM and cultured H9C2 cells, but the expression of these proteins was reduced following exogenous H2S treatment. The expression of mitochondrial apoptotic proteins, cyt C, and mPTP opening was decreased following treatment with exogenous H2S. In our experiment, the expression and immunofluorescence of Mfn-2 were both decreased after transfection with Mfn-2-siRNA. Hyperglycemia stimulated ER interactions and mitochondrial apoptotic pathways, which were inhibited by exogenous H2S treatment through the regulation of Mfn-2 expression.
Assuntos
Apoptose/efeitos dos fármacos , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Retículo Endoplasmático/efeitos dos fármacos , Gasotransmissores/farmacologia , Sulfeto de Hidrogênio/farmacologia , Mitocôndrias Cardíacas/efeitos dos fármacos , Miocárdio/metabolismo , Animais , Glicemia/metabolismo , Western Blotting , Citocromos c/efeitos dos fármacos , Citocromos c/metabolismo , Cardiomiopatias Diabéticas , Retículo Endoplasmático/ultraestrutura , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Imunofluorescência , GTP Fosfo-Hidrolases , Coração/efeitos dos fármacos , Coração/fisiopatologia , Marcação In Situ das Extremidades Cortadas , Masculino , Proteínas de Membrana/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Microscopia Eletrônica , Mitocôndrias Cardíacas/metabolismo , Mitocôndrias Cardíacas/ultraestrutura , Proteínas Mitocondriais/efeitos dos fármacos , Proteínas Mitocondriais/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Ratos , Ratos Wistar , Sulfetos/farmacologiaRESUMO
A simple and feasible electrochemical immunosensing platform was developed for highly efficient screening of a disease-related protein (human carbohydrate antigen 19-9, CA 19-9 used in this case) using silver-functionalized g-C3N4 nanosheets (Ag/g-C3N4) as signal-transduction tags. Initially, Ag/g-C3N4 nanohybrids were synthesized by combining thermal polymerization of the melamine precursor with the photo-assisted reduction method. Thereafter, the as-synthesized Ag/g-C3N4 nanohybrids were utilized for the labeling of the anti-CA 19-9 detection antibody by using a typical carbodiimide coupling method. The assay was carried out on a capture antibody-modified glassy carbon electrode in a sandwich-type detection mode. The detectable signal mainly derived from the voltammetric characteristics of the immobilized nanosilver particles on the g-C3N4 nanosheets within the applied potentials. Under the optimal conditions, the voltammetric peak currents increased with the increasing amount of target CA 19-9, and exhibited a wide linear range from 5.0 mU mL(-1) to 50 U mL(-1) with a detection limit of 1.2 mU mL(-1). Our strategy also displayed good reproducibility, precision and specificity. The results of the analysis of clinical serum specimens were in good accordance with the results obtained by an enzyme-linked immunosorbent assay (ELISA) method. The newly developed immunosensing system is promising for enzyme-free and cost-effective analysis of low-abundance proteins.
Assuntos
Antígeno CA-19-9/análise , Técnicas Eletroquímicas , Imunoensaio , Nanopartículas Metálicas , Transdução de Sinais , Ouro , Humanos , Limite de Detecção , Reprodutibilidade dos Testes , PrataRESUMO
AIMS: Hydrogen sulfide (H2S) inhibits the proliferation of vascular smooth muscle cells (VSMCs). However, how cystathionine-gamma-lyase (CSE), a major enzyme that produces H2S, is regulated remains unknown. Whether calcium-sensing receptor (CaSR) inhibits the proliferation of VSMCs by regulating the endogenous CSE/H2S pathway in diabetic rat has not been previously investigated. METHODS AND RESULTS: The morphological and ultrastructure alterations were tested by transmission electron microscopy, changes in the H2S concentration and the relaxation of the mesenteric secondary artery loop of diabetic rats were determined by Multiskan spectrum microplate spectrophotometer and isometric force transducer. Additionally, the expression levels of CaSR, CSE and Cyclin D1 in the mesenteric arteries of rats were examined by western blotting. The intracellular calcium concentration, the expression of p-CaMK II (phospho-calmodulin kinases II), CSE activity, the concentration of endogenous H2S and the proliferation of cultured VSMCs from rat thoracic aortas were measured by using confocal microscope, western blotting, microplate spectrophotometer, MTT and BrdU, respectively. The VSMC layer thickened, the H2S concentration dropped, the relaxation of the mesenteric secondary artery rings weakened, and the expression of CaSR and CSE decreased whereas the expression of Cyclin D1 increased in diabetic rats compared with the control group. The [Ca(2+)]i of VSMCs increased upon treatment with CaSR agonists (10 µM Calindol and 2.5 mM CaCl2), while it decreased upon administration of calhex231, U73122 and 2-APB. The expression of p-CaMK II and CSE increased upon treatment with CaSR agonists in VSMCs. CSE activity and the endogenous H2S concentration decreased in response to high glucose, while it increased with treatment of CaSR agonists. The proliferation rate increased in response to high glucose, and CaSR agonists or NaHS significantly reversed the proliferation of VSMCs caused by high glucose. CONCLUSIONS: Our results demonstrated that CaSR regulated the endogenous CSE/H2S pathway to inhibit the proliferation of VSMCs in both diabetic and high glucose models.
Assuntos
Proliferação de Células/efeitos dos fármacos , Cistationina gama-Liase/metabolismo , Diabetes Mellitus Experimental/patologia , Sulfeto de Hidrogênio/toxicidade , Receptores de Detecção de Cálcio/metabolismo , Animais , Aorta Torácica/citologia , Benzamidas/farmacologia , Cálcio/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Ciclina D1/genética , Ciclina D1/metabolismo , Cicloexilaminas/farmacologia , Cistationina gama-Liase/genética , Diabetes Mellitus Experimental/metabolismo , Estrenos/farmacologia , Glucose/farmacologia , Indóis/farmacologia , Masculino , Artérias Mesentéricas/metabolismo , Artérias Mesentéricas/patologia , Músculo Liso Vascular/citologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Naftalenos/farmacologia , Pirrolidinonas/farmacologia , Ratos , Ratos Wistar , Receptores de Detecção de Cálcio/agonistas , Receptores de Detecção de Cálcio/genéticaRESUMO
BACKGROUND: Diabetic encephalopathy is a common complication of diabetes, and it may be involved in altering intracellular calcium concentrations ([Ca(2+)]i) at its onset. The calcium sensing receptor (CaSR) is a G-protein coupled receptor, however, the functional involvement of CaSR in diabetic encephalopathy remains unclear. METHODS: In this study, diabetic rats were modeled by STZ (50 mg/kg). At the end of 4, 8 and 12 weeks, the CaSR expression in hippocampus was analyzed by Western blot. In neonatal rat hippocampal neurons, the [Ca(2+)]i was detected by laser scanning confocal microscopy, the production of reactive oxygen species (ROS) in mitochondria, the level of NO and the mitochondrial transmembrane potential were measured by MitoSOX, DAF-FM and JC-1, respectively. RESULTS: Our results showed in hippocampal neurons treated with high glucose, CaSR regulated [Ca(2+)]i through the PLC-IP3 pathway. CaSR expression was decreased and was involved in the changes in [Ca(2+)]i. Mitochondrial membrane potential, NO release and expression of p-eNOS decreased, while the production of ROS in mitochondria increased. CONCLUSION: Down-regulation of CaSR expression was accompanied by neuronal injury, calcium disturbance, increased ROS production and decreased release of NO. Up-regulation of CaSR expression attenuated these changes through a positive compensatory protective mechanism to inhibit and delay diabetic encephalopathy in rats.
Assuntos
Diabetes Mellitus Experimental/patologia , Neurônios/metabolismo , Receptores de Detecção de Cálcio/metabolismo , Animais , Cálcio/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Células Cultivadas , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/metabolismo , Regulação para Baixo/efeitos dos fármacos , Glucose/farmacologia , Hipocampo/metabolismo , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Mitocôndrias/metabolismo , Neuritos/fisiologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Estreptozocina/toxicidadeRESUMO
MAIN CONCLUSION: Successful molecular cloning and functional characterization of a high-affinity urea permease ZmDUR3 provide convincing evidence of ZmDUR3 roles in root urea acquisition and internal urea-N-remobilization of maize plants. Urea occurs ubiquitously in both soils and plants. Being a major form of nitrogen fertilizer, large applications of urea assist cereals in approaching their genetic yield potential, but due to the low nitrogen-use efficiency of crops, this practice poses a severe threat to the environment through their hypertrophication. To date, except for paddy rice, little is known about the biological basis for urea movement in dryland crops. Here, we report the molecular and physiological characterization of a maize urea transporter, ZmDUR3. We show using gene prediction, PCR-based cloning and yeast complementation, that a functional full-length cDNA encoding a 731 amino acids-containing protein with putative 15 transmembrane α-helixes for ZmDUR3 was successfully cloned. Root-influx studies using (15)N-urea demonstrated ZmDUR3 catalyzes urea transport with a K m at ~9 µM when expressed in the Arabidopsis dur3-mutant. qPCR analysis revealed that ZmDUR3 mRNA in roots was significantly upregulated by nitrogen depletion and repressed by reprovision of nitrogen after nitrogen starvation, indicating that ZmDUR3 is a nitrogen-responsive gene and relevant to plant nitrogen nutrition. Moreover, detection of higher urea levels in senescent leaves and obvious occurrence of ZmDUR3 transcripts in phloem-cells of mature/aged leaves strongly implies a role for ZmDUR3 in urea vascular loading. Significantly, expression of ZmDUR3 complemented atdur3-mutant of Arabidopsis, improving plant growth on low urea and increasing urea acquisition. As it also targets to the plasma membrane, our data suggest that ZmDUR3 functions as an active urea permease playing physiological roles in effective urea uptake and nitrogen remobilization in maize.
Assuntos
Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras/metabolismo , Ureia/metabolismo , Zea mays/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Transporte Biológico , Membrana Celular/metabolismo , Fertilizantes , Expressão Gênica , Genes Reporter , Proteínas de Membrana Transportadoras/genética , Nitrogênio/metabolismo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas , Zea mays/metabolismo , Transportadores de UreiaRESUMO
A new electrochemical immunosensor with signal enhancement was designed for sensitive detection of disease-related protein (human carbohydrate antigen 19-9, CA 19-9 used in this case). The assay was carried out on a capture antibody-modified screen-printed carbon electrode with a sandwich-type mode by using detection antibody-functionalized nanogold-encapsulated poly(amidoamine) dendrimer (AuNP-PAAD). The AuNP-PAAD was first synthesized through the in situ reduction method and functionalized with the polyclonal rabbit anti-human CA 19-9 antibody. Upon target CA 19-9 introduction, a sandwiched immunocomplex could be formed between the capture antibody and detection antibody. Accompanying the AuNP-PAAD, the electrocatalytic activity of the carried gold nanoparticles toward the hydrogen evolution reaction (HER) allowed the rapid quantification of the target analyte on the electrode. The amplified electrochemical signal mainly derived from AuNP-catalyzed HER in an acidic medium. Under optimal conditions, the immuno-HER assay displayed a wide dynamic concentration range from 0.01 to 300 U mL(-1) toward target CA 19-9 with a detection limit (LOD) of 6.3 mU mL(-1). The reproducibility, precision, specificity and stability of our strategy were acceptable. Additionally, the system was further validated by assaying 13 human serum specimens, giving well matched results obtained from the commercialized enzyme-linked immunosorbent assay (ELISA) method.
Assuntos
Dendrímeros/química , Técnicas Eletroquímicas , Ouro/química , Hidrogênio/química , Imunoensaio/métodos , Nanopartículas Metálicas/química , Poliaminas/química , Antígeno CA-19-9/sangue , Dendrímeros/síntese química , Humanos , Hidrogênio/análise , Limite de DetecçãoRESUMO
A novel silver ion (Ag(+))-assisted hairpin DNA through C-Ag(+)-C coordination chemistry was designed for immobilization-free and label-free electrochemical monitoring of human immunodeficiency virus (HIV) DNA on a negatively charged indium-tin oxide electrode, based on hybridization-induced dissociation of silver ions from the hairpin DNA.
Assuntos
Técnicas Biossensoriais/instrumentação , DNA Viral/análise , DNA Viral/química , Ácidos Nucleicos Imobilizados/química , Sequências Repetidas Invertidas , Prata/química , Sequência de Bases , DNA Viral/genética , Eletroquímica , Eletrodos , HIV , Humanos , Hibridização de Ácido Nucleico , Compostos de Estanho/químicaRESUMO
BACKGROUND: In this study, we aimed to establish a chronic intermittent hypoxia model in rats and explore the possible role of vaspin in insulin sensitivity. METHODS: Healthy male Wistar rats were randomly divided into two groups: normal control group (NC) and chronic intermittent hypoxia group (CIH). The NC group was raised under physiological conditions and the CIH group was kept in the plexiglass chamber between 9 am and 5 pm undergoing intermittent hypoxic challenge for 8 hours/day for 8 weeks. Arterial blood pressure of rats (tail cannulation) was measured before and after the study. Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), fasting insulin (FINS), vaspin, and leptin levels were measured. Vaspin mRNA expression in visceral adipose tissues was measured with Real Time-PCR. The protein levels of vaspin, Akt and phospho-Akt in visceral tissues were determined by Western-blot. RESULTS: At baseline, all the measurements in the CIH and NC groups were comparable. By the end of the experiment, the blood pressure of the CIH group was significantly higher than the NC group. The levels of FPG, FINS, TG, TC, leptin, and vaspin in the CIH group were significantly higher than in NC group. Plasma vaspin levels were correlated with FINS, HOMA-IR, and TG levels. Vaspin expression in both mRNA and protein levels in visceral adipose tissues of the CIH group were clearly higher than the NC group. Phospho-Akt protein level was decreased in visceral adipose tissues of the CIH group compared to the NC group. CONCLUSIONS: In the chronic intermittent hypoxia rat model, the expression of vaspin in visceral adipose tissues and plasma were increased, which were correlated with insulin resistance.
Assuntos
Resistência à Insulina , Gordura Intra-Abdominal/metabolismo , Serpinas/sangue , Apneia Obstrutiva do Sono/etiologia , Animais , Pressão Arterial , Biomarcadores/sangue , Glicemia/metabolismo , Colesterol/sangue , Modelos Animais de Doenças , Hipóxia/sangue , Hipóxia/complicações , Hipóxia/fisiopatologia , Insulina/sangue , Leptina/sangue , Masculino , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Mensageiro/metabolismo , Ratos Wistar , Serpinas/genética , Apneia Obstrutiva do Sono/sangue , Apneia Obstrutiva do Sono/genética , Apneia Obstrutiva do Sono/fisiopatologia , Fatores de Tempo , Triglicerídeos/sangue , Regulação para CimaRESUMO
BACKGROUND: Lipin1ß is an adipokine proposed to be associated with insulin resistance (IR). Pregnancy is a physiologic state of progressive IR. The objective of the present study was to investigate the role of lipin1ß in the development of GDM. METHODS: A total of 40 pregnant women (22 normal and 18 with GDM) who delivered healthy infants at full-term (> 37 weeks gestation) were included. The mRNA and protein levels of lipin1ß in adipose tissues were determined by real-time RT-PCR and Western-blot. Plasma glucose, lipids, insulin, and estradiol (E2) levels were measured routinely at fasting state, and HOMA-IR was calculated accordingly. RESULTS: The lipin1ß expression in both mRNA and protein levels in SAT and VAT was lower in GDM patients than controls. Lipin1ß mRNA in VAT was negatively correlated with BMI (r = -0.505, p < 0.05), FINS (r = -0.539, p < 0.05), HOMA-IR (r = -0.574, p < 0.01), TG (r = -0.471, p < 0.05), and E2 (r = -0.564, p < 0.01). Lipin1ß mRNA expression in SAT was similar with VAT. Lipin1ß mRNA was not correlated with body weight gain or blood pressure. These results indicated that the lipin1ß expression in adipose tissues is down-regulated in patients with GDM. CONCLUSIONS: Lipin1ß might play a role in the pathogenesis of insulin resistance in GDM.