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1.
Nature ; 572(7770): 461-466, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31340216

RESUMO

Effective ocean management and the conservation of highly migratory species depend on resolving the overlap between animal movements and distributions, and fishing effort. However, this information is lacking at a global scale. Here we show, using a big-data approach that combines satellite-tracked movements of pelagic sharks and global fishing fleets, that 24% of the mean monthly space used by sharks falls under the footprint of pelagic longline fisheries. Space-use hotspots of commercially valuable sharks and of internationally protected species had the highest overlap with longlines (up to 76% and 64%, respectively), and were also associated with significant increases in fishing effort. We conclude that pelagic sharks have limited spatial refuge from current levels of fishing effort in marine areas beyond national jurisdictions (the high seas). Our results demonstrate an urgent need for conservation and management measures at high-seas hotspots of shark space use, and highlight the potential of simultaneous satellite surveillance of megafauna and fishers as a tool for near-real-time, dynamic management.


Assuntos
Migração Animal , Pesqueiros/estatística & dados numéricos , Mapeamento Geográfico , Oceanos e Mares , Tubarões/fisiologia , Análise Espaço-Temporal , Animais , Densidade Demográfica , Medição de Risco , Tubarões/classificação , Navios , Fatores de Tempo
4.
J Fish Biol ; 101(6): 1591-1594, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36070183

RESUMO

Despite being one of the most abundant, economically significant, reef-associated shark species, little is known about the reproductive aspects of the Caribbean reef shark (Carcharhinus perezi). In the present study the authors report the first evidence of mating wounds and scars in female Caribbean reef sharks at the Fernando de Noronha Archipelago, a remote marine protected area in the South Atlantic Ocean. Data from four females suggest this species mates mainly during the austral summer, between February and March. Given that the archipelago has been previously described as a nursery ground for the Caribbean reef shark, these results add information about the reproductive cycle of this species in the equatorial Atlantic Ocean.


Assuntos
Tubarões , Feminino , Animais , Oceano Atlântico , Reprodução , Estações do Ano , Região do Caribe
5.
J Fish Biol ; 93(6): 1121-1129, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30281159

RESUMO

The present study aimed to analyse occurrence patterns, relative abundance and habitat use by the sicklefin devil ray Mobula tarapacana in the Saint Peter and Saint Paul Archipelago (SPSPA), Brazil, located in the Mid-Atlantic Ridge. Sampling was conducted between December 2008 and December 2012 by surface surveys and tagging with conventional tags and acoustic transmitters. Mobula tarapacana were sighted in the SPSPA year round but the highest frequency of occurrence was observed from January to June, probably due to greater food availability during this season. However, M. tarapacana might also use the area for reproductive purposes based on evidence of mating behaviour. The residency period of tagged rays measured by three autonomous acoustic receivers deployed around SPSPA spanned from 1 to 74 days with most of the detections occurring during daytime. These results suggest that the SPSPA could be an ecologically-important aggregation area for M. tarapacana, possibly providing the species with a feeding or resting habitat while transiting in the equatorial Atlantic Ocean.


Assuntos
Comportamento Animal , Estações do Ano , Rajidae/fisiologia , Migração Animal , Animais , Oceano Atlântico , Brasil , Ecossistema , Comportamento Alimentar , Densidade Demográfica , Reprodução
6.
Fish Physiol Biochem ; 43(1): 165-178, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27549099

RESUMO

Sharks are very sensitive to stress and prone to a high mortality rate after capture. Since approximately 50 million of sharks are caught as bycatch every year, and current recommendations to reduce the impact of commercial fishing strongly support immediate release, it is imperative to better understand post-release mortality caused by the stress of capture and handling. Blood samples allow the assessment of stress levels which are valuable tools to reduce mortality in commercial, recreational and scientific fishing, being essential for the improvement in those conservation measures. Biochemical analyses are widely used for sharks as stress indicators, with secondary plasma parameters (lactate, glucose and ions) being the most often employed assays. However, it is virtually impossible to determine baseline plasma parameters in free-ranging sharks, since blood withdrawal involves animal capture and restrain, which are stressful procedures. This study aims at analyzing secondary parameters of five healthy tiger sharks captured with circular hooks and handlines in Fernando de Noronha (Northeastern Brazil) and comparing them with secondary parameters of three dead tiger sharks caught off Recife (also Northeastern Brazil). The results showed that the analysis of some plasma constituents in dead animals may be an efficient tool to assess stress and lethality. However, traditional parameters such as glucose and calcium, need to be used with caution. The results also demonstrated the extreme importance of urea and phosphorus for assessing stress response and mortality in tiger sharks, both parameters frequently neglected and of utmost importance for shark's homeostasis.


Assuntos
Tubarões/sangue , Estresse Fisiológico , Estresse Psicológico/sangue , Animais , Autopsia , Glicemia/análise , Proteínas Sanguíneas/análise , Cloretos/sangue , Feminino , Proteínas de Peixes/sangue , Concentração de Íons de Hidrogênio , Ácido Láctico/sangue , Masculino , Metais/sangue , Concentração Osmolar , Fósforo/sangue , Ureia/sangue
7.
An Acad Bras Cienc ; 85(3): 1053-62, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24068092

RESUMO

An abnormally high shark attack rate verified off Recife could be related to migratory behavior of tiger sharks. This situation started after the construction of the Suape port to the south of Recife. A previous study suggested that attacking sharks could be following northward currents and that they were being attracted shoreward by approaching vessels. In this scenario, such northward movement pattern could imply a higher probability of sharks accessing the littoral area of Recife after leaving Suape. Pop-up satellite archival tags were deployed on five tiger sharks caught off Recife to assess their movement patterns off northeastern Brazil. All tags transmitted from northward latitudes after 7-74 days of freedom. The shorter, soak distance between deployment and pop-up locations ranged between 33-209 km and implied minimum average speeds of 0.02-0.98 km.h-1. Both pop-up locations and depth data suggest that tiger shark movements were conducted mostly over the continental shelf. The smaller sharks moved to deeper waters within 24 hours after releasing, but they assumed a shallower (< 50 m) vertical distribution for most of the monitoring period. While presenting the first data on tiger shark movements in the South Atlantic, this study also adds new information for the reasoning of the high shark attack rate verified in this region.


Assuntos
Migração Animal/fisiologia , Tubarões/fisiologia , Animais , Mordeduras e Picadas/epidemiologia , Brasil , Comunicações Via Satélite , Tubarões/classificação , Movimentos da Água
8.
Mar Pollut Bull ; 194(Pt A): 115326, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37527571

RESUMO

Plastic waste is currently a major threat to marine ecosystems, and the ever-growing production of plastic materials suggests that this scenario will not change soon. Understanding cryptic effects of plastic debris on keystone marine species is warranted to address ecosystem-level impacts caused by plastic pollution. This study reports on plastic entanglement in top predator tiger sharks, Galeocerdo cuvier, from the western South Atlantic Ocean. Circular plastic straps produced severe trauma in 3 % of all tiger sharks sampled and led to abnormal anatomical development, indicating worrisome incidence and consequences of plastic entanglement in this species. Removing the plastic strap from one shark which had been entangled for at least 9 months was probably the cause for the shark having experienced post-release mortality. This suggests that, in some circumstances, strap removal is not effective to promote the survival of entangled sharks. Eliminating the circular integrity of plastic materials at first use should contribute to mitigate some of the impacts of plastic pollution on marine megafauna since this shape might be determinant in the process of shark entanglement.


Assuntos
Ecossistema , Tubarões , Animais , Plásticos , Oceano Atlântico , Poluição Ambiental
9.
Sci Adv ; 8(33): eabo1754, 2022 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-35984887

RESUMO

Knowledge of the three-dimensional movement patterns of elasmobranchs is vital to understand their ecological roles and exposure to anthropogenic pressures. To date, comparative studies among species at global scales have mostly focused on horizontal movements. Our study addresses the knowledge gap of vertical movements by compiling the first global synthesis of vertical habitat use by elasmobranchs from data obtained by deployment of 989 biotelemetry tags on 38 elasmobranch species. Elasmobranchs displayed high intra- and interspecific variability in vertical movement patterns. Substantial vertical overlap was observed for many epipelagic elasmobranchs, indicating an increased likelihood to display spatial overlap, biologically interact, and share similar risk to anthropogenic threats that vary on a vertical gradient. We highlight the critical next steps toward incorporating vertical movement into global management and monitoring strategies for elasmobranchs, emphasizing the need to address geographic and taxonomic biases in deployments and to concurrently consider both horizontal and vertical movements.

10.
Phys Chem Chem Phys ; 13(48): 21233-42, 2011 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-22025281

RESUMO

The paper reports on the successful use of the quartz crystal microbalance technique to assess accurate kinetics and equilibrium parameters regarding the investigation of in situ adsorption of nanosized cobalt ferrite particles (CoFe(2)O(4)--10.5 nm-diameter) onto two different surfaces. Firstly, a single layer of nanoparticles was deposited onto the surface provided by the gold-coated quartz resonator functionalized with sodium 3-mercapto propanesulfonate (3-MPS). Secondly, the layer-by-layer (LbL) technique was used to build multilayers in which the CoFe(2)O(4) nanoparticle-based layer alternates with the sodium sulfonated polystyrene (PSS) layer. The adsorption experiments were conducted by modulating the number of adsorbed CoFe(2)O(4)/PSS bilayers (n) and/or by changing the CoFe(2)O(4) nanoparticle concentration while suspended as a stable colloidal dispersion. Adsorption of CoFe(2)O(4) nanoparticles onto the 3-MPS-functionalized surface follows perfectly a first order kinetic process in a wide range (two orders of magnitude) of nanoparticle concentrations. These data were used to assess the equilibrium constant and the adsorption free energy. Alternatively, the Langmuir adsorption constant was obtained while analyzing the isotherm data at the equilibrium. Adsorption of CoFe(2)O(4) nanoparticles while growing multilayers of CoFe(2)O(4)/PSS was conducted using colloidal suspensions with CoFe(2)O(4) concentration in the range of 10(-8) to 10(-6) (moles of cobalt ferrite per litre) and for different numbers of cycles n = 1, 3, 5, and 10. We found the adsorption of CoFe(2)O(4) nanoparticles within the CoFe(2)O(4)/PSS bilayers perfectly following a first order kinetic process, with the characteristic rate constant growing with the increase of CoFe(2)O(4) nanoparticle concentration and decreasing with the rise of the number of LbL cycles (n). Additionally, atomic force microscopy was employed for assessing the LbL film roughness and thickness. We found the film thickness increasing from about 20 to 120 nm while shifting from 3 to 10 CoFe(2)O(4)/PSS bilayers, using the 8.9 × 10(-6) (moles of cobalt ferrite per litre) suspension.


Assuntos
Cobalto/química , Compostos Férricos/química , Nanopartículas Metálicas/química , Técnicas de Microbalança de Cristal de Quartzo , Adsorção , Ácidos Alcanossulfônicos/química , Coloides/química , Cinética , Poliestirenos/química
11.
Mar Pollut Bull ; 158: 111391, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32753177

RESUMO

Sewage is among the largest components of coastal pollution, showing a variable scale and size when causing an impact. In this study, temporal and spatial sewage-related gradients were identified using univariate and multivariate methods. Phosphates and nitrogen-based nutrients, except nitrate, were associated to sewage. Abundance and species richness decrease with distance from the sewage outfall while evenness increased. Filter-feeder bivalves and grazer crustaceans showed preference for the sewage discharge site. Gastropods were more abundant at an intermediate distance, likely reflecting primary production enhancement by sewage. Beta diversity was lower at the location and time with highest sewage loadings. The turnover component of beta diversity expressed as an absolute or proportional value was also useful to detect temporal and spatial sewage-related gradients. Highly energetic hydrodynamics contributes to ameliorate sewage impacts, yet not enough to veil the effect of pollution in this study.


Assuntos
Bivalves , Esgotos/análise , Animais , Ecossistema , Monitoramento Ambiental
13.
Biosens Bioelectron ; 99: 156-162, 2018 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-28755608

RESUMO

A novel fully disposable microfluidic electrochemical array device (µFED) was developed and successfully applied for detection of the biomarker estrogen receptor alpha (ERα). The µFED was constructed using low-cost materials and an inexpensive home cutter printer enabled the manufacture of dozens of µFEDs in less than 2h, at a cost of less than US$ 0.20 in material per device. The µFED incorporates counter and reference electrodes and eight carbon-based working electrodes, which were modified with DNA sequences known as estrogen response elements (DNA-ERE), where ERα binds specifically. Paramagnetic particles heavily decorated with anti-ERα antibody and horseradish peroxidase (MP-Ab-HRP) were used to efficiently capture ERα from the sample solution. The ERα-MP-Ab-HRP bioconjugate formed was injected into the µFED and incubated with the DNA-ERE-modified electrodes, followed by amperometric detection with application of -0.2V vs. Ag|AgCl while a mixture of H2O2 and hydroquinone was injected into the microfluidic device. An ultralow limit of detection of 10.0 fg mL-1 was obtained with the proposed method. The performance of the assay, in terms of sensitivity and reproducibility, was studied using undiluted calf serum, and excellent recoveries in the range of 94.7-108% were achieved for the detection of ERα in MCF-7 cell lysate. The µFED system can be easily constructed and applied for multiplex biomarker detection, making the device an excellent cost-effective alternative for cancer diagnosis, especially in developing countries.


Assuntos
Biomarcadores Tumorais/genética , Técnicas Biossensoriais , Neoplasias da Mama/diagnóstico , Receptor alfa de Estrogênio/isolamento & purificação , Neoplasias da Mama/genética , Receptor alfa de Estrogênio/genética , Humanos , Limite de Detecção , Células MCF-7
14.
PLoS One ; 12(9): e0184763, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28926627

RESUMO

Increasing our knowledge about the spatial ecology of apex predators and their interactions with diverse habitats and fisheries is necessary for understanding the trophic mechanisms that underlie several aspects of marine ecosystem dynamics and for guiding informed management policies. A preliminary assessment of tiger shark (Galeocerdo cuvier) population structure off the oceanic insular system of Fernando de Noronha (FEN) and the large-scale movements performed by this species in the equatorial Atlantic Ocean was conducted using longline and handline fishing gear and satellite telemetry. A total of 25 sharks measuring 175-372 cm in total length (TL) were sampled. Most sharks were likely immature females ranging between 200 and 260 cm TL, with few individuals < 200 cm TL being caught. This contrasts greatly with the tiger shark size-distribution previously reported for coastal waters off the Brazilian mainland, where most individuals measured < 200 cm TL. Also, the movements of 8 individuals measuring 202-310 cm TL were assessed with satellite transmitters for a combined total of 757 days (mean = 94.6 days∙shark-1; SD = 65.6). These sharks exhibited a considerable variability in their horizontal movements, with three sharks showing a mostly resident behavior around FEN during the extent of the respective tracks, two sharks traveling west to the South American continent, and two sharks moving mostly along the middle of the oceanic basin, one of which ending up in the northern hemisphere. Moreover, one shark traveled east to the African continent, where it was eventually caught by fishers from Ivory Coast in less than 474 days at liberty. The present results suggest that young tiger sharks measuring < 200 cm TL make little use of insular oceanic habitats from the western South Atlantic Ocean, which agrees with a previously-hypothesized ontogenetic habitat shift from coastal to oceanic habitats experienced by juveniles of this species in this region. In addition, this study adds evidence that tiger sharks are able to connect marine trophic webs from the neritic provinces of the eastern and western margins of the Atlantic Ocean across the equatorial basin and that they may experience mortality induced by remote fisheries. All this information is extremely relevant for understanding the energetic balance of marine ecosystems as much as the exposure of this species to fishing pressure in this yet poorly-known region.


Assuntos
Ecossistema , Tubarões/fisiologia , Animais , Oceano Atlântico , Comportamento Animal , Tamanho Corporal , Feminino , Pesqueiros , Sistemas de Informação Geográfica , Masculino
15.
Vet J ; 171(1): 177-80, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16427595

RESUMO

The occurrence of mycobacteriosis caused by Mycobacterium marinum in a commercial breeding farm of bullfrogs (Rana catesbeiana) in Rio de Janeiro, Brazil is described. Ten animals presented skin lesions on the head and extremities. These and 38 other asymptomatic adult animals from various tanks were killed and at necropsy disseminated granulomatous lesions were observed in the 10 clinically affected animals and in 16 (42.1%) of the asymptomatic frogs. Acid-fast bacilli were observed in all smears of the 10 symptomatic frogs and in all but one from the 16 asymptomatic animals with visceral lesions. Ten samples from the 25 positive animals were randomly selected for culture which yielded four isolates of fast-growing (<7 days) mycobacteria. Those purified isolates were characterised by biochemical traditional means as M. marinum. Identification of the strains was confirmed using reverse-phase high-performance liquid chromatography and a polymerase chain reaction (PCR) restriction enzyme analysis assay. It is suggested that M. marinum is an important agent of granulomatous disease in bullfrogs and that infected animals, even when asymptomatic, could act as reservoirs spreading the disease and contaminating other frogs in the farm.


Assuntos
Infecções por Mycobacterium/veterinária , Mycobacterium/isolamento & purificação , Rana catesbeiana , Animais , Brasil/epidemiologia , Reservatórios de Doenças/veterinária , Evolução Fatal , Mycobacterium/classificação , Infecções por Mycobacterium/epidemiologia , Rana catesbeiana/microbiologia
16.
Talanta ; 146: 381-7, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26695279

RESUMO

This work describes the construction of an all-plastic disposable carbon-based electrochemical cell (DCell) using a simple procedure based on the use of a home cutter printer for prototyping and laminating. The cutter printer and adhesive vinyl films were used to produce three electrodes in an electrochemical cell layout, and a laminating process was then used to define the geometric area and insulate the electrodes. The DCell showed excellent performance in several applications including the determination of toxic metals in water samples, the immobilization of DNA and the detection of Salmonella. An unmodified DCell was applied for Pb and Cd detection in the range of 100-300 ng mL(-1) with a limit of detection of 50 and 39 ng mL(-1) for Cd and Pb, respectively. DNA was successfully immobilized on a DCell and used for studies of interaction between bisphenol A and DNA. The square wave voltammetry of a DNA modified DCell presented a guanine oxidation current 2.5 times greater after exposure of the electrode to bisphenol A and no current variation for the adenine moiety indicating that bisphenol A showed a preference for DNA interaction sites. A magneto-immunoassay was developed using a DCell for Salmonella detection in milk samples. The system presented a linear range from 100 to 700 cells mL(-1) with a limit of detection of 100 cells mL(-1) and good recovery values between 93% and 101% in milk samples, with no interference from Escherichia coli. Using the proposed method, hundreds of DCells can be assembled in less than two hours, at a material cost of less than US $0.02 per cell. The all-plastic disposable electrochemical cell developed was successfully applied as an electrochemical sensor and biosensor. The feasibility of the developed all-plastic disposable electrochemical cell was demonstrated in applications as both sensor and biosensor.


Assuntos
Técnicas Biossensoriais/instrumentação , Carbono/química , Equipamentos Descartáveis , Equipamentos e Provisões Elétricas , Compostos Benzidrílicos/química , Cádmio/análise , Cádmio/química , DNA/química , Eletroquímica , Desenho de Equipamento , Chumbo/análise , Chumbo/química , Fenóis/química , Salmonella/isolamento & purificação , Fatores de Tempo
17.
Braz. arch. biol. technol ; 64: e21210030, 2021. graf
Artigo em Inglês | LILACS | ID: biblio-1339317

RESUMO

Abstract Nowadays, a prompt and reliable diagnosis is one of the most critical measures for leprosy control. The current diagnostic is based on clinical exams by a health care professional, and it may not recognize early signs of the disease. Therefore, other leprosy diagnosis methods are needed that are sensitive, disease-specific, and easy to deliver to the end-user. This study describes the construction of an electrochemical DNA biosensor to detect PCR products of Mycobacterium leprae using methylene blue as an indicator of the hybridization. The capture probe was immobilized on the graphite electrode modified with poly(4-aminophenol). The electrode surface was morphologically characterized by atomic force microscopy. Linear voltammetry was used to monitor the concentration of methylene blue on the DNA biosensor, which indicated a limit detection of 1 x 10-10 mol/L. The biosensor showed selective when placed to hybridize with a non-complementary sequence. This study suggests that the electrochemical DNA biosensor developed is promising for detecting DNA of Mycobacterium leprae.


Assuntos
Técnicas Biossensoriais , Aminofenóis , Azul de Metileno , Mycobacterium leprae
18.
Sci Rep ; 6: 31406, 2016 08 09.
Artigo em Inglês | MEDLINE | ID: mdl-27503274

RESUMO

The conversion of solar energy into hydrogen fuel by splitting water into photoelectrochemical cells (PEC) is an appealing strategy to store energy and minimize the extensive use of fossil fuels. The key requirement for efficient water splitting is producing a large band bending (photovoltage) at the semiconductor to improve the separation of the photogenerated charge carriers. Therefore, an attractive method consists in creating internal electrical fields inside the PEC to render more favorable band bending for water splitting. Coupling ferroelectric materials exhibiting spontaneous polarization with visible light photoactive semiconductors can be a likely approach to getting higher photovoltage outputs. The spontaneous electric polarization tends to promote the desirable separation of photogenerated electron- hole pairs and can produce photovoltages higher than that obtained from a conventional p-n heterojunction. Herein, we demonstrate that a hole inversion layer induced by a ferroelectric Bi4V2O11 perovskite at the n-type BiVO4 interface creates a virtual p-n junction with high photovoltage, which is suitable for water splitting. The photovoltage output can be boosted by changing the polarization by doping the ferroelectric material with tungsten in order to produce the relatively large photovoltage of 1.39 V, decreasing the surface recombination and enhancing the photocurrent as much as 180%.


Assuntos
Bismuto/química , Compostos de Cálcio/química , Hidrogênio/química , Óxidos/química , Titânio/química , Vanadatos/química , Técnicas Eletroquímicas , Processos Fotoquímicos , Semicondutores , Energia Solar , Água
19.
PLoS One ; 10(1): e0116720, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25629732

RESUMO

Sharks are top predators in many marine ecosystems and can impact community dynamics, yet many shark populations are undergoing severe declines primarily due to overfishing. Obtaining species-specific knowledge on shark spatial ecology is important to implement adequate management strategies for the effective conservation of these taxa. This is particularly relevant concerning highly-mobile species that use wide home ranges comprising coastal and oceanic habitats, such as tiger sharks, Galeocerdo cuvier. We deployed satellite tags in 20 juvenile tiger sharks off northeastern Brazil to assess the effect of intrinsic and extrinsic factors on depth and temperature usage. Sharks were tracked for a total of 1184 d and used waters up to 1112 m in depth. The minimum temperature recorded equaled 4°C. All sharks had a clear preference for surface (< 5 m) waters but variability in depth usage was observed as some sharks used mostly shallow (< 60 m) waters whereas others made frequent incursions into greater depths. A diel behavioral shift was detected, with sharks spending considerably more time in surface (< 10 m) waters during the night. Moreover, a clear ontogenetic expansion in the vertical range of tiger shark habitat was observed, with generalized linear models estimating a ~4-fold increase in maximum diving depth from 150- to 300-cm size-classes. The time spent in the upper 5 m of the water column did not vary ontogenetically but shark size was the most important factor explaining the utilization of deeper water layers. Young-of-the-year tiger sharks seem to associate with shallow, neritic habitats but they progressively move into deeper oceanic habitats as they grow larger. Such an early plasticity in habitat use could endow tiger sharks with access to previously unavailable prey, thus contributing to a wider ecological niche.


Assuntos
Comportamento Animal , Tubarões , Animais , Tamanho Corporal , Brasil , Mergulho , Ecossistema , Feminino , Masculino , Oceanos e Mares , Fatores Sexuais , Temperatura
20.
Anal Chim Acta ; 862: 86-93, 2015 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-25682432

RESUMO

The hemoglobin (Hb) released from erythrocytes is a primary nutritive component for many blood-feeding parasites. The aspartic protease cathepsin D is a hemoglobinase that is involved in the Hb degradation process and is considered an interesting target for chemotherapy intervention. However, traditional enzymatic assays for studying Hb degradation utilize spectrophotometric techniques, which do not allow real-time monitoring and can present serious interference problems. Herein, we describe a biosensor using simple approach for the real-time monitoring of Hb hydrolysis as well as an efficient screening method for natural products as enzymatic inhibitors using a quartz crystal microbalance (QCM) technique. Hemoglobin was anchored on the quartz crystal surface using mixed self-assembled monolayers. The addition of the enzyme caused a mass change (frequency shift) due to Hb hydrolysis, which was monitored in real time. From the frequency change patterns of the Hb-functionalized QCM, we evaluated the enzymatic reaction by determining the kinetic parameters of product formation (k(cat)). The QCM enzymatic assay using immobilized human Hb was shown to be an excellent approach for screening possible inhibitors in complex mixtures, opening up a new avenue for the discovery of novel inhibitors.


Assuntos
Produtos Biológicos/análise , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/métodos , Hemoglobinas/metabolismo , Inibidores de Proteases/análise , Técnicas de Microbalança de Cristal de Quartzo , Produtos Biológicos/farmacologia , Catepsina D/antagonistas & inibidores , Catepsina D/metabolismo , Hemoglobinas/análise , Humanos , Hidrólise/efeitos dos fármacos , Inibidores de Proteases/farmacologia , Relação Estrutura-Atividade , Fatores de Tempo
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