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1.
Eur J Neurosci ; 59(10): 2450-2464, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38480476

RESUMO

Amphetamine (AMPH) exposure induces behavioural and neurochemical sensitization observed in rodents as hyperlocomotion and increased dopamine release in response to a subsequent dose. Brain Angiotensin II modulates dopaminergic neurotransmission through its AT1 receptors (AT1-R), positively regulating striatal dopamine synthesis and release. This work aims to evaluate the AT1-R role in the development and maintenance of AMPH-induced sensitization. Also, the AT1-R involvement in striatal dopamine reuptake was analysed. The sensitization protocol consisted of daily AMPH administration for 5 days and tested 21 days after withdrawal. An AT1-R antagonist, candesartan, was administered before or after AMPH exposure to evaluate the participation of AT1-R in the development and maintenance of sensitization, respectively. Sensitization was evaluated by locomotor activity and c-Fos immunostaining. Changes in dopamine reuptake kinetics were evaluated 1 day after AT1-R blockade withdrawal treatment, with or without the addition of AMPH in vitro. The social interaction test was performed as another behavioural output. Repeated AMPH exposure induced behavioural and neurochemical sensitization, which was prevented and reversed by candesartan. The AT1-R blockade increased the dopamine reuptake kinetics. Neither the AMPH administration nor the AT1-R blockade altered the performance of social interaction. Our results highlight the AT1-R's crucial role in AMPH sensitization. The enhancement of dopamine reuptake kinetics induced by the AT1-R blockade might attenuate the neuroadaptive changes that lead to AMPH sensitization and its self-perpetuation. Therefore, AT1-R is a prominent candidate as a target for pharmacological treatment of pathologies related to dopamine imbalance, including drug addiction and schizophrenia.


Assuntos
Anfetamina , Bloqueadores do Receptor Tipo 1 de Angiotensina II , Angiotensina II , Benzimidazóis , Compostos de Bifenilo , Corpo Estriado , Dopamina , Animais , Anfetamina/farmacologia , Masculino , Dopamina/metabolismo , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/metabolismo , Angiotensina II/farmacologia , Compostos de Bifenilo/farmacologia , Benzimidazóis/farmacologia , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Ratos Wistar , Ratos , Receptor Tipo 1 de Angiotensina/metabolismo , Tetrazóis/farmacologia , Estimulantes do Sistema Nervoso Central/farmacologia , Interação Social/efeitos dos fármacos , Atividade Motora/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/metabolismo
2.
Mikrochim Acta ; 190(2): 73, 2023 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-36695940

RESUMO

An innovative strategy is proposed to simultaneously exfoliate multi-walled carbon nanotubes (MWCNTs) and generate MWCNTs with immunoaffinity properties. This strategy was based on the non-covalent functionalization of MWCNTs with human immunoglobulin G (IgG) by sonicating 2.5 mg mL-1 MWCNTs in 2.0 mg mL-1 IgG for 15 min with sonicator bath. Impedimetric experiments performed at glassy carbon electrodes (GCE) modified with the resulting MWCNT-IgG nanohybrid in the presence of anti-human immunoglobulin G antibody (Anti-IgG) demonstrated that the immunoglobulin retains their biorecognition properties even after the treatment during the MWCNT functionalization. We proposed, as proof-of-concept, two model electrochemical sensors, a voltammetric one for uric acid quantification by taking advantages of the exfoliated MWCNTs electroactivity (linear range, 5.0 × 10-7 M - 5.0 × 10-6 M; detection limit, 165 nM) and an impedimetric immunosensor for the detection of Anti-IgG through the use of the bioaffinity properties of the IgG present in the nanohybrid (linear range, 5-50 µg mL-1; detection limit, 2 µg mL-1).


Assuntos
Técnicas Biossensoriais , Nanotubos de Carbono , Humanos , Técnicas Biossensoriais/métodos , Nanotubos de Carbono/química , Imunoensaio , Imunoglobulina G , Eletrodos
3.
Analyst ; 147(10): 2130-2140, 2022 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-35421882

RESUMO

This work presents for the first time the systematic preparation of a novel carbon nanotube-MCM-41 hybrid employing the mesoporous material MCM-41 as a successful dispersant for multiwall carbon nanotubes (MWCNTs). Relevant dispersion variables such as the amount of MWCNTs, MCM-41 concentration, and sonication time were optimized through a central composite design (CDD)/response surface methodology (RSM). Several solvents were evaluated and N,N-dimethylformamide (DMF) was selected because it allowed reaching stable dispersions with very good electrochemical response. The electrochemical performance of glassy carbon electrodes (GCE) modified with different hybrids was evaluated by cyclic voltammetry (CV) using ascorbic acid (AA) as redox marker, while their surface morphology was characterized by SEM microscopy. The optimal MWCNT-MCM-41 dispersion condition was 0.75 mg mL-1 MWCNTs, 0.25 mg mL-1 MCM-41, and 30 min sonication. Both, electrochemical results and SEM images correlate with a percolation behavior from MWCNT-MCM-41 hybrid. Electrooxidation of AA at GCE modified with the optimal hybrid occurred under diffusion control and exhibited an enhanced current response (65 µA) and a lower overvoltage (-0.005 V) compared to bare GCE (ip = 22 µA, Ep = 0.255 V). The amperometric response of AA at GCE/MWCNT-MCM-41 exhibited remarkable figures of merit, including an ultralow detection limit (1.5 nM), high sensitivity (45.4 × 103 µA M-1), excellent short- and long-term stability, and very good anti-interference ability for AA detection. The analytical applicability of the developed electrochemical sensor was evaluated by sensing AA in several real samples, showing excellent correlation with the values reported by manufacturers in both pharmaceutical and food samples.


Assuntos
Nanotubos de Carbono , Ácido Ascórbico/química , Técnicas Eletroquímicas/métodos , Eletrodos , Nanotubos de Carbono/química , Dióxido de Silício
4.
ScientificWorldJournal ; 2022: 4505349, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35837366

RESUMO

Solar energy found abundantly in nature is considered a renewable energy source. It is also of great interest as an option for energy generation and CO2 emissions reduction. Several technologies of solar concentrating systems, known internationally as CSP (concentrated solar power), are found in the industrial and scientific environment. One of the most mature and internationally known technologies is the parabolic trough solar collector (PTSC), which has several applications, such as electricity generation, desalination, steam generation, and refrigeration systems, among others. However, more research and development (R&D) has been done to improve its performance, using new materials, absorber tube geometries, solar tracking systems, and work (thermal oils, nanofluids). Thus, the present work describes the development of a low-cost PTSC for academic and research purposes. The PTSC was built with an edge angle of 120°, an opening area of 2.2 m2, and a copper absorber tube of 42 mm in outer diameter without a glass envelope. The gutter structure is composed of wooden sheets cut in a parabolic shape, where a 1.2 mm-thick galvanized steel sheet coated with a reflective film is supported, thus functioning as the reflective surface of the PTSC. The solar tracking system is one of the active types with two axes containing photoresistive sensors, which are used to determine the solar position and electric actuators to correct the positioning of the gutter. The monitoring system was developed through an interactive panel to visualize the operating parameters of the sensing elements, thermocouples that measure the inlet and outlet temperature in the absorber tube, and the flow sensor to measure the flow of the heat transport fluid. Laboratory tests were performed with deionized water as a transport fluid, establishing two testing conditions. The first test condition analyzed the efficiency of the collector at different temperatures. Thus, the inlet temperature varied, between 30 and 70°C, presenting a flow of 0.020 kg/s. The second one evaluated the collector efficiency for different flows, subjecting the collector to flows from 0.002 to 0.030 kg/s. Thus, the proposed collector obtained an efficiency as a function of the temperature represented by the expression η = 0.324-2.47443 c', where c' is a parameter that relates the inlet temperature to the ambient temperature as a function of the solar radiation available. Yet, the efficiency in function of the flow became optimal when the flow regime became turbulent. It was concluded that the proposed solar collector obtained lower efficiency when compared with other collectors in the literature, which was assumed to be due to the diffusion losses of the parabolic trough reflector and thermal losses by convection in the parabolic trough absorber tube (optical efficiency, removal factor, and heat loss coefficient).

5.
Anal Bioanal Chem ; 412(15): 3539-3546, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32239259

RESUMO

This work is focused on the development of a genosensor for microRNA-21 quantification using surface plasmon resonance (SPR) to transduce the hybridization event. The biosensing platform was built by self-assembling two bilayers of poly(diallyldimethylammonium chloride) (PDDA) and graphene oxide (GO) at a gold surface modified with 3-mercaptopropane sulfonate (MPS), followed by the covalent attachment of the DNA probe. GO was used in two directions, to allow the anchoring of the probe DNA and to increase the sensitivity of the biosensing event due to its field enhancer effect. The new bioanalytical platform represents an interesting alternative for the label-free biosensing of microRNA-21, with a linear range between 1.0 fM and 10 nM, a sensitivity of 5.1 ± 0.1 moM-1 and a detection limit of 0.3fM. The proposed sensing strategy was successfully used for the quantification of microRNA-21 in enriched urine samples. Graphical abstract.


Assuntos
Grafite/química , MicroRNAs/urina , Ressonância de Plasmônio de Superfície/métodos , Sondas de DNA/química , Ouro/química , Humanos , Limite de Detecção , MicroRNAs/análise , Polietilenos/química , Compostos de Amônio Quaternário/química
6.
Anal Bioanal Chem ; 412(21): 5089-5096, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32009193

RESUMO

This work reports the successful non-covalent functionalization of multi-walled carbon nanotubes (MWCNTs) with bathocuproinedisulfonic acid (BCS) and the analytical application of the resulting dispersion (MWCNTs-BCS) to develop an electrochemical sensor for Cu(II) quantification. The sensor was obtained by casting glassy carbon electrodes (GCEs) with MWCNTs-BCS. The sensing mechanism was based on the open circuit preconcentration of Cu(II) at the electrode surface by complexation of Cu(II) through the phenanthroline ring nitrogen of the BCS that supports the MWCNTs, the reduction of the preconcentrated Cu(II), and final differential pulse voltammetry-anodic stripping in 0.020 M acetate buffer, pH 5.00. The sensitivity of the sensor was (2.73 ± 0.08) µA µM-1, with a linear range between 5.0 × 10-7 M and 6.0 × 10-6 M, a detection limit of 0.15 µM (9.5 µg L-1), and reproducibility of 6.2% using the same dispersion and 7.1% using three different MWCNTs-BCS dispersions. The quantification of Cu(II) was highly selective even in the presence of As3+, Cr3+, Cd2+, Ni2+, Pb2+, Co2+, Zn2+, Fe2+, Hg2+, Rh, Ir, and Ru. The proposed sensor was successfully used for quantifying Cu(II) in tap water. Graphical abstract.

7.
Phys Chem Chem Phys ; 19(9): 6792-6806, 2017 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-28217769

RESUMO

The electronic density of states and its contribution to the capacitance of graphene compounds (oxidized and reduced) were investigated using an electrochemical impedance-derived capacitance spectroscopic approach. It is clearly demonstrated that graphene oxide, which is known to exhibit semiconductor electronic characteristics, has little influence on the magnitude of the measured capacitance. Moreover, when graphene oxide is electrochemically reduced to graphene, the capacitance increases dramatically by about three orders of magnitude (from microfaradays to millifaradays). This increased capacitive effect has been interpreted as being directly associated with the electrochemical non-faradaic (super- or ultracapacitive) characteristics of the interface (i.e. associated with its electroactive area, for instance). The results obtained and interpretation made in this work demonstrate that the magnitude of the measured capacitance is a consequence of an electrochemical capacitive phenomenon (mesoscopic in essence; thus, the associated capacitance is equivalently termed mesoscopic capacitance) that energetically contains, in series, both electrostatic (geometrical) and quantum effects, thus being essentially different from those exclusively related to the amount of existing interfacial sites for ions (i.e. beyond those associated with pure double-layer capacitive effects). Conceptually, it is proposed that the mesoscopic capacitance of reduced graphene can be explained mainly through quantum chemical effects, ultimately following first-principles quantum mechanics supported on density functional theory, wherein the density of states is central.

8.
Biosensors (Basel) ; 14(4)2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38667189

RESUMO

L-Lactate is an important bioanalyte in the food industry, biotechnology, and human healthcare. In this work, we report the development of a new L-lactate electrochemical biosensor based on the use of multiwalled carbon nanotubes non-covalently functionalized with avidin (MWCNT-Av) deposited at glassy carbon electrodes (GCEs) as anchoring sites for the bioaffinity-based immobilization of a new recombinant biotinylated lactate oxidase (bLOx) produced in Escherichia coli through in vivo biotinylation. The specific binding of MWCNT-Av to bLOx was characterized by amperometry, surface plasmon resonance (SPR), and electrochemical impedance spectroscopy (EIS). The amperometric detection of L-lactate was performed at -0.100 V, with a linear range between 100 and 700 µM, a detection limit of 33 µM, and a quantification limit of 100 µM. The proposed biosensor (GCE/MWCNT-Av/bLOx) showed a reproducibility of 6.0% and it was successfully used for determining L-lactate in food and enriched serum samples.


Assuntos
Avidina , Técnicas Biossensoriais , Ácido Láctico , Oxigenases de Função Mista , Nanotubos de Carbono , Nanotubos de Carbono/química , Oxigenases de Função Mista/química , Avidina/química , Técnicas Eletroquímicas , Ressonância de Plasmônio de Superfície , Enzimas Imobilizadas/química , Escherichia coli , Biotinilação , Eletrodos , Espectroscopia Dielétrica , Limite de Detecção
9.
Talanta ; 270: 125520, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38147722

RESUMO

We report a nanohybrid material obtained by non-covalent functionalization of multi-walled carbon nanotubes (MWCNTs) with the new ligand (((1E,1'E)-(naphthalene-2,3-diylbis(azaneylylidene))bis(methaneylylidenedene)) bis(4-hydroxy-3,1-phenylene))diboronic acid (SB-dBA), rationally designed to mimic some recognition properties of biomolecules like concanavalin A, for the development of electrochemical biosensors based on the use of glycobiomolecules as biorecognition element. We present, as a proof-of-concept, a hydrogen peroxide biosensor obtained by anchoring horseradish peroxidase (HRP) at a glassy carbon electrode (GCE) modified with the nanohybrid prepared by sonication of 2.0 mg mL-1 MWCNTs and 0.50 mg mL-1 SB-dBA in N,N-dimethyl formamide (DMF) for 30 min. The hydrogen peroxide biosensing was performed at -0.050 V in the presence of 5.0 × 10-4 M hydroquinone. The analytical characteristics of the resulting biosensor are the following: linear range between 0.175 µM and 6.12 µM, detection limit of 58 nM, and reproducibility of 2.0 % using the same nanohybrid (6 biosensors), and 9.0 % using three different nanohybrids. The sensor was successfully used to quantify hydrogen peroxide in enriched milk and human blood serum samples and in a commercial disinfector.


Assuntos
Técnicas Biossensoriais , Nanotubos de Carbono , Humanos , Nanotubos de Carbono/química , Ácidos Borônicos , Peróxido de Hidrogênio/química , Bases de Schiff , Reprodutibilidade dos Testes , Técnicas Biossensoriais/métodos , Peroxidase do Rábano Silvestre/química , Eletrodos , Técnicas Eletroquímicas
10.
Mem Inst Oswaldo Cruz ; 108 Suppl 1: 80-7, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24473806

RESUMO

Mosquitoes are the culprits of some of the most important vector borne diseases. A species' potential as a vector is directly dependent on their pattern of behaviour, which is known to change according to the female's physiological status such as whether the female is virgin/mated and unfed/blood-fed. However, the molecular mechanism triggered by and/or responsible for such modulations in behaviour is poorly understood. Clock genes are known to be responsible for the control of circadian behaviour in several species. Here we investigate the impact mating and blood-feeding have upon the expression of these genes in the mosquito Aedes aegypti. We show that blood intake, but not insemination, is responsible for the down-regulation of clock genes. Using RNA interference, we observe a slight reduction in the evening activity peak in the fourth day after dstim injection. These data suggest that, as in Drosophila, clock gene expression, circadian behaviour and environmental light regimens are interconnected in Ae. aegypti.


Assuntos
Aedes/genética , Proteínas CLOCK/genética , Relógios Circadianos/genética , Inseminação/genética , Fotoperíodo , Interferência de RNA/fisiologia , Animais , Ritmo Circadiano/genética , Regulação para Baixo/genética , Comportamento Alimentar/fisiologia , Feminino , Expressão Gênica , Atividade Motora/genética , Reação em Cadeia da Polimerase em Tempo Real , Comportamento Sexual Animal
11.
Plants (Basel) ; 12(11)2023 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-37299094

RESUMO

In its natural distribution, Araucaria araucana is a plant species usually exposed to extreme environmental constraints such as wind, volcanism, fires, and low rainfall. This plant is subjected to long periods of drought, accentuated by the current climate emergency, causing plant death, especially in its early growth stages. Understanding the benefits that both arbuscular mycorrhizal fungi (AMF) and endophytic fungi (EF) could provide plants under different water regimes would generate inputs to address the above-mentioned issues. Here, the effect of AMF and EF inoculation (individually and combined) on the morphophysiological variables of A. araucana seedlings subjected to different water regimes was evaluated. Both the AMF and EF inocula were obtained from A. araucana roots growing in natural conditions. The inoculated seedlings were kept for 5 months under standard greenhouse conditions and subsequently subjected to three different irrigation levels for 2 months: 100, 75, and 25% of field capacity (FC). Morphophysiological variables were evaluated over time. Applying AMF and EF + AMF yielded a noticeable survival rate in the most extreme drought conditions (25% FC). Moreover, both the AMF and the EF + AMF treatments promoted an increase in height growth between 6.1 and 16.1%, in the production of aerial biomass between 54.3 and 62.6%, and in root biomass between 42.5 and 65.4%. These treatments also kept the maximum quantum efficiency of PSII (Fv/Fm 0.71 for AMF and 0.64 for EF + AMF) stable, as well as high foliar water content (>60%) and stable CO2 assimilation under drought stress. In addition, the EF + AMF treatment at 25% FC increased the total chlorophyll content. In conclusion, using indigenous strains of AMF, alone or in combination with EF, is a beneficial strategy to produce A. araucana seedlings with an enhanced ability to tolerate prolonged drought periods, which could be of great relevance for the survival of these native species under the current climate change.

12.
Micromachines (Basel) ; 14(5)2023 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-37241682

RESUMO

Nowadays, there is no doubt about the high electrocatalytic efficiency that is obtained when using hybrid materials between carbonaceous nanomaterials and transition metal oxides. However, the method to prepare them may involve differences in the observed analytical responses, making it necessary to evaluate them for each new material. The goal of this work was to obtain for the first time Co2SnO4 (CSO)/RGO nanohybrids via in situ and ex situ methods and to evaluate their performance in the amperometric detection of hydrogen peroxide. The electroanalytical response was evaluated in NaOH pH 12 solution using detection potentials of -0.400 V or 0.300 V for the reduction or oxidation of H2O2. The results show that for CSO there were no differences between the nanohybrids either by oxidation or by reduction, unlike what we previously observed with cobalt titanate hybrids, in which the in situ nanohybrid clearly had the best performance. On the other hand, no influence in the study of interferents and more stable signals were obtained when the reduction mode was used. In conclusion, for detecting hydrogen peroxide, any of the nanohybrids studied, i.e., in situ or ex situ, are suitable to be used, and more efficiency is obtained using the reduction mode.

13.
J Pharm Biomed Anal ; 232: 115370, 2023 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-37163830

RESUMO

The COVID-19 pandemic had devastating effects throughout the world, producing a severe crisis in the health systems and in the economy of a long list of countries, even developed ones. Therefore, highly sensitive and selective analytical bioplatforms that allow the descentralized and fast detection of the severe acute respiratory síndrome coronavirus 2 (SARS-CoV-2), are extremely necessary. Since 2020, several reviews have been published, most of them focused on the different strategies to detect the SARS-CoV-2, either from RNA, viral proteins or host antibodies produced due to the presence of the virus. In this review, the most relevant biosensors for the detection of SARS-CoV-2 RNA are particularly addressed, with special emphasis on the discussion of the biorecognition layers and the different schemes for transducing the hybridization event.


Assuntos
Técnicas Biossensoriais , COVID-19 , Ácidos Nucleicos , Humanos , SARS-CoV-2 , COVID-19/diagnóstico , RNA Viral/genética , Pandemias
14.
PLoS Negl Trop Dis ; 17(4): e0010862, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-37043542

RESUMO

Phlebotomine sand flies are of global significance as important vectors of human disease, transmitting bacterial, viral, and protozoan pathogens, including the kinetoplastid parasites of the genus Leishmania, the causative agents of devastating diseases collectively termed leishmaniasis. More than 40 pathogenic Leishmania species are transmitted to humans by approximately 35 sand fly species in 98 countries with hundreds of millions of people at risk around the world. No approved efficacious vaccine exists for leishmaniasis and available therapeutic drugs are either toxic and/or expensive, or the parasites are becoming resistant to the more recently developed drugs. Therefore, sand fly and/or reservoir control are currently the most effective strategies to break transmission. To better understand the biology of sand flies, including the mechanisms involved in their vectorial capacity, insecticide resistance, and population structures we sequenced the genomes of two geographically widespread and important sand fly vector species: Phlebotomus papatasi, a vector of Leishmania parasites that cause cutaneous leishmaniasis, (distributed in Europe, the Middle East and North Africa) and Lutzomyia longipalpis, a vector of Leishmania parasites that cause visceral leishmaniasis (distributed across Central and South America). We categorized and curated genes involved in processes important to their roles as disease vectors, including chemosensation, blood feeding, circadian rhythm, immunity, and detoxification, as well as mobile genetic elements. We also defined gene orthology and observed micro-synteny among the genomes. Finally, we present the genetic diversity and population structure of these species in their respective geographical areas. These genomes will be a foundation on which to base future efforts to prevent vector-borne transmission of Leishmania parasites.


Assuntos
Leishmania , Leishmaniose Cutânea , Phlebotomus , Psychodidae , Animais , Humanos , Phlebotomus/parasitologia , Psychodidae/parasitologia , Leishmania/genética , Genômica
15.
Micromachines (Basel) ; 13(11)2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36422406

RESUMO

We report the advantages of glassy carbon electrodes (GCE) modified with multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with polyarginine (PolyArg) for the adsorption and electrooxidation of different DNAs and the analytical applications of the resulting platform. The presence of the carbon nanostructures, and mainly the charge of the PolyArg that supports them, facilitates the adsorption of calf-thymus and salmon sperm double-stranded DNAs and produces an important decrease in the overvoltages for the oxidation of guanine and adenine residues and a significant enhancement in the associated currents. As a proof-of-concept of possible GCE/MWCNTs-PolyArg biosensing applications, we develop an impedimetric genosensor for the quantification of microRNA-21 at femtomolar levels, using GCE/MWCNTs-PolyArg as a platform for immobilizing the DNA probe, with a detection limit of 3fM, a sensitivity of 1.544 × 103 Ω M-1, and a successful application in enriched biological fluids.

16.
Biosens Bioelectron X ; 12: 100222, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36118917

RESUMO

We report two novel genosensors for the quantification of SARS-CoV-2 nucleic acid using glassy carbon electrodes modified with a biocapture nanoplatform made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with avidin (Av) as a support of the biotinylated-DNA probes. One of the genosensors was based on impedimetric transduction offering a non-labelled and non-amplified detection of SARS-CoV-2 nucleic acid through the increment of [Fe(CN)6]3-/4- charge transfer resistance. This biosensor presented an excellent analytical performance, with a linear range of 1.0 × 10-18 M - 1.0 × 10-11 M, a sensitivity of (5.8 ± 0.6) x 102 Ω M-1 (r2 = 0.994), detection and quantification limits of 0.33 aM and 1.0 aM, respectively; and reproducibilities of 5.4% for 1.0 × 10-15 M target using the same MWCNTs-Av-bDNAp nanoplatform, and 6.9% for 1.0 × 10-15 M target using 3 different nanoplatforms. The other genosensor was based on a sandwich hybridization scheme and amperometric transduction using the streptavidin(Strep)-biotinylated horseradish peroxidase (bHRP)/hydrogen peroxide/hydroquinone (HQ) system. This genosensor allowed an extremely sensitive quantification of the SARS-CoV-2 nucleic acid, with a linear range of 1.0 × 10-20 M - 1.0 × 10-17 M, detection limit at zM level, and a reproducibility of 11% for genosensors prepared with the same MWCNTs-Av-bDNAp1 nanoplatform. As a proof-of-concept, and considering the extremely high sensitivity, the genosensor was challenged with highly diluted samples obtained from SARS-CoV-2 RNA PCR amplification.

17.
Curr Biol ; 31(19): 4207-4218.e4, 2021 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-34331859

RESUMO

The Drosophila circadian clock is driven by a transcriptional feedback loop in which CLOCK-CYCLE (CLK-CYC) binds E-boxes to transcribe genes encoding the PERIOD-TIMELESS (PER-TIM) repressor, which releases CLK-CYC from E-boxes to inhibit transcription. CLOCKWORK ORANGE (CWO) reinforces PER-TIM repression by binding E-boxes to maintain PER-TIM bound CLK-CYC off DNA, but also promotes CLK-CYC transcription through an unknown mechanism. To determine how CWO activates CLK-CYC transcription, we identified CWO target genes that are upregulated in the absence of CWO repression, conserved in mammals, and preferentially expressed in brain pacemaker neurons. Among the genes identified was a putative ortholog of mouse Clock Interacting Protein Circadian (Cipc), which represses CLOCK-BMAL1 transcription. Reducing or eliminating Drosophila Cipc expression shortens period, while overexpressing Cipc lengthens period, which is consistent with previous work showing that Drosophila Cipc represses CLK-CYC transcription in S2 cells. Cipc represses CLK-CYC transcription in vivo, but not uniformly, as per is strongly repressed, tim less so, and vri hardly at all. Long period rhythms in cwo mutant flies are largely rescued when Cipc expression is reduced or eliminated, indicating that increased Cipc expression mediates the period lengthening of cwo mutants. Consistent with this behavioral rescue, eliminating Cipc rescues the decreased CLK-CYC transcription in cwo mutant flies, where per is strongly rescued, tim is moderately rescued, and vri shows little rescue. These results suggest a mechanism for CWO-dependent CLK-CYC activation: CWO inhibition of CIPC repression promotes CLK-CYC transcription. This mechanism may be conserved since cwo and Cipc perform analogous roles in the mammalian circadian clock.


Assuntos
Proteínas de Drosophila , Drosophila , Fatores de Transcrição ARNTL/genética , Fatores de Transcrição ARNTL/metabolismo , Animais , Proteínas CLOCK/genética , Proteínas CLOCK/metabolismo , Clorprofam/metabolismo , Ritmo Circadiano/genética , Drosophila/fisiologia , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/fisiologia , Regulação da Expressão Gênica , Mamíferos/genética , Camundongos , Transcrição Gênica
18.
iScience ; 24(1): 101893, 2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33364582

RESUMO

The vrille (vri) gene encodes a transcriptional repressor required for Drosophila development as well as circadian behavior in adults. Alternate first exons produce vri transcripts predicted to produce a short VRI isoform during development and long VRI in adults. A vri mutant (vri Δ679) lacking long VRI transcripts is viable, confirming that short VRI is sufficient for developmental functions, yet behavioral rhythms in vri Δ679 flies persist, showing that short VRI is sufficient for clock output. E-box regulatory elements that drive rhythmic long VRI transcript expression are required for developmental expression of short VRI transcripts. Surprisingly, long VRI transcripts primarily produce short VRI in adults, apparently due to a poor Kozak sequence context, demonstrating that short VRI drives circadian behavior. Thus, E-box-driven long VRI transcripts primarily control circadian rhythms via short VRI, whereas the same E-boxes drive short VRI transcripts that control developmental functions using short VRI.

19.
J Pharm Biomed Anal ; 191: 113526, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32892085

RESUMO

We are reporting a new strategy for preparing carbon nanotubes (CNTs)-based hydrogen peroxide and glucose amperometric sensors by taking advantage of the dual role of bathocuproine disulfonic acid (BCS) as dispersing agent of multi-walled carbon nanotubes (MWCNTs) and as ligand for the preconcentration of Cu(II). The platform was obtained by casting glassy carbon electrodes (GCE) with the dispersion of MWCNTs in BCS (MWCNTs-BCS) followed by the preconcentration of Cu(II) by surface complex formation at open circuit potential (GCE/MWCNTs-BCS/Cu). The resulting electrode was used for the sensitive amperometric quantification of hydrogen peroxide at 0.400 V catalyzed by the preconcentrated copper, with a linear range between 5.0 × 10-7 and 7.4 × 10-6 M, a sensitivity of 24.3 mA.M-1, and a detection limit of 0.2 µM. The adsorption of GOx at GCE/MWCNTs-BCS/Cu followed by the immobilization of Nafion (Naf), allowed the construction of a sensitive and selective amperometric glucose biosensor with a linear range between 5.0 × 10-6 M and 4.9 × 10-4 M, a sensitivity of (477 ± 3) µA.M-1 and a detection limit of 2 µM. The proposed (bio)sensors were successfully used for the quantification of hydrogen peroxide in enriched milk samples and glucose in milk and commercial beverages without any pretreatment.


Assuntos
Técnicas Biossensoriais , Nanotubos de Carbono , Cobre , Técnicas Eletroquímicas , Eletrodos , Glucose , Peróxido de Hidrogênio , Ligantes , Fenantrolinas
20.
Biosens Bioelectron ; 148: 111764, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31707325

RESUMO

We are reporting an original supramolecular architecture based on a rationally designed new nanohybrid with enhanced peroxidase-like activity and site-specific biorecognition properties using avidin-functionalized multi-walled carbon nanotubes (MWCNTs-Av) and Ru nanoparticles (RuNPs). The nanohybrid-electrochemical interface was obtained by drop-coating of MWCNTs-Av dispersion at glassy carbon electrodes (GCE) followed by solvent evaporation and further electrodeposition of RuNPs (50 ppm RuCl2 for 15 s at -0.600 V). The simultaneous presence of MWCNTs and RuNPs produces a synergic effect on the non-enzymatic catatalytic reduction of H2O2 and allows the quantification of H2O2 in a wide linear range (from 5.0 × 10-7 M to 1.75 × 10-3 M) with a low limit of detection (65 nM). The avidin residues present in MWCNTs-Av/RuNPs hybrid nanomaterial allowed the anchoring by bioaffinity of biotinylated glucose oxidase (biot-GOx) as proof-of-concept of the analytical application of MWCNTs-Av platform for biosensors development. The resulting nanoarchitecture behaves as a bienzymatic-like glucose biosensor with a competitive analytical performance: linear range between 2.0 × 10-5 M and 1.23 × 10-3 M, sensitivity of (0.343 ±â€¯0.002) µA mM-1 or (2.60 ±â€¯0.02) µA mM-1 cm-2, detection limit of 3.3 µM, and reproducibility of 5.2% obtained with five different GCE/MWCNTs-Av/RuNPs/biot-GOx bioplatforms prepared the same day using the same MWCNTs-Av dispersion, and 9.1% obtained with nine biosensors prepared in different days with nine different MWCNTs-Av dispersions. The average concentrations of glucose in Gatorade®, Red bull® and Pepsi® with the biosensor demonstrated excellent agreement with those reported in the commercial beverages.


Assuntos
Avidina/química , Técnicas Biossensoriais/métodos , Nanopartículas/química , Nanotubos de Carbono/química , Rutênio/química , Aspergillus niger/enzimologia , Bebidas/análise , Materiais Biomiméticos/química , Biotinilação , Catálise , Técnicas Eletroquímicas/métodos , Glucose/análise , Glucose Oxidase/química , Peróxido de Hidrogênio/análise , Limite de Detecção , Nanopartículas/ultraestrutura , Nanotubos de Carbono/ultraestrutura , Peroxidase/química
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