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1.
Talanta ; 271: 125648, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38219324

RESUMO

The high toxicity and occurrence of ochratoxin A (OTA) in grains and foods has been a growing concern due to the impacts on health and the economy in many countries. In this sense, simplified devices with high sensitivity and specificity for local monitoring are enthusiastically pursued. In this work, we report for the first time the detection of ochratoxin A in coffee samples using a spoon-shaped waveguide immunosensor. The biosensor was built with the surface of the spoon-shaped waveguide covered by a 60 nm layer of gold to enable the SPR phenomenon. The measurements indicated a linear relationship between the change in the SPR phenomenon values and the OTA concentration in the range from 0.2 ppt to 5 ppt. When analyzed in coffee samples, the biosensor was highly selective and did not suffer matrix interference. The developed biosensor represents a promising analytical device for coffee quality analyses, as it is portable, simple, and suitable for onsite detection of target analytes.


Assuntos
Técnicas Biossensoriais , Ocratoxinas , Café , Imunoensaio , Ocratoxinas/análise
2.
Biosensors (Basel) ; 13(4)2023 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-37185514

RESUMO

The global need for accurate and efficient cancer cell detection in biomedicine and clinical diagnosis has driven extensive research and technological development in the field. Precision, high-throughput, non-invasive separation, detection, and classification of individual cells are critical requirements for successful technology. Lab-on-a-chip devices offer enormous potential for solving biological and medical problems and have become a priority research area for microanalysis and manipulating cells. This paper reviews recent developments in the detection of cancer cells using the microfluidics-based lab-on-a-chip method, focusing on describing and explaining techniques that use optical phenomena and a plethora of probes for sensing, amplification, and immobilization. The paper describes how optics are applied in each experimental method, highlighting their advantages and disadvantages. The discussion includes a summary of current challenges and prospects for cancer diagnosis.


Assuntos
Técnicas Biossensoriais , Neoplasias , Dispositivos Lab-On-A-Chip , Óptica e Fotônica , Fenômenos Ópticos , Análise Espectral Raman , Técnicas Biossensoriais/métodos , Neoplasias/diagnóstico
3.
Biosensors (Basel) ; 13(3)2023 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-36979582

RESUMO

The correct detection and quantification of pollutants in water is key to regulating their presence in the environment. Biosensors offer several advantages, such as minimal sample preparation, short measurement times, high specificity and sensibility and low detection limits. The purpose of this review is to explore the different types of optical biosensors, focusing on their biological elements and their principle of operation, as well as recent applications in the detection of pollutants in water. According to our literature review, 33% of the publications used fluorescence-based biosensors, followed by surface plasmon resonance (SPR) with 28%. So far, SPR biosensors have achieved the best results in terms of detection limits. Although less common (22%), interferometers and resonators (4%) are also highly promising due to the low detection limits that can be reached using these techniques. In terms of biological recognition elements, 43% of the published works focused on antibodies due to their high affinity and stability, although they could be replaced with molecularly imprinted polymers. This review offers a unique compilation of the most recent work in the specific area of optical biosensing for water monitoring, focusing on both the biological element and the transducer used, as well as the type of target contaminant. Recent technological advances are discussed.


Assuntos
Técnicas Biossensoriais , Poluentes Ambientais , Poluentes da Água , Técnicas Biossensoriais/métodos , Ressonância de Plasmônio de Superfície/métodos , Polímeros Molecularmente Impressos
4.
Biosensors (Basel) ; 13(1)2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36671925

RESUMO

Aquaculture is an expanding economic sector that nourishes the world's growing population due to its nutritional significance over the years as a source of high-quality proteins. However, it has faced severe challenges due to significant cases of environmental pollution, pathogen outbreaks, and the lack of traceability that guarantees the quality assurance of its products. Such context has prompted many researchers to work on the development of novel, affordable, and reliable technologies, many based on nanophotonic sensing methodologies. These emerging technologies, such as surface plasmon resonance (SPR), localised SPR (LSPR), and fibre-optic SPR (FO-SPR) systems, overcome many of the drawbacks of conventional analytical tools in terms of portability, reagent and solvent use, and the simplicity of sample pre-treatments, which would benefit a more sustainable and profitable aquaculture. To highlight the current progress made in these technologies that would allow them to be transferred for implementation in the field, along with the lag with respect to the most cutting-edge plasmonic sensing, this review provides a variety of information on recent advances in these emerging methodologies that can be used to comprehensively monitor the various operations involving the different commercial stages of farmed aquaculture. For example, to detect environmental hazards, track fish health through biochemical indicators, and monitor disease and biosecurity of fish meat products. Furthermore, it highlights the critical issues associated with these technologies, how to integrate them into farming facilities, and the challenges and prospects of developing plasmonic-based sensors for aquaculture.


Assuntos
Técnicas Biossensoriais , Animais , Técnicas Biossensoriais/métodos , Ressonância de Plasmônio de Superfície/métodos , Aquicultura , Controle de Qualidade , Tecnologia de Fibra Óptica
5.
Biosensors (Basel) ; 12(12)2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36551068

RESUMO

A new transmission route of SARS-CoV-2 through food was recently considered by the World Health Organization (WHO), and, given the pandemic scenario, the search for fast, sensitive, and low-cost methods is necessary. Biosensors have become a viable alternative for large-scale testing because they overcome the limitations of standard techniques. Herein, we investigated the ability of gold spherical nanoparticles (AuNPs) functionalized with oligonucleotides to detect SARS-CoV-2 and demonstrated their potential to be used as plasmonic nanobiosensors. The loop-mediated isothermal amplification (LAMP) technique was used to amplify the viral genetic material from the raw virus-containing solution without any preparation. The detection of virus presence or absence was performed by ultraviolet-visible (UV-Vis) absorption spectroscopy, by monitoring the absorption band of the surface plasmonic resonance (SPR) of the AuNPs. The displacement of the peak by 525 nm from the functionalized AuNPs indicated the absence of the virus (particular region of gold). On the other hand, the region ~300 nm indicated the presence of the virus when RNA bound to the functionalized AuNPs. The nanobiosensor system was designed to detect a region of the N gene in a dynamic concentration range from 0.1 to 50 × 103 ng·mL-1 with a limit of detection (LOD) of 1 ng·mL-1 (2.7 × 103 copy per µL), indicating excellent sensitivity. The nanobiosensor was applied to detect the SARS-CoV-2 virus on the surfaces of vegetables and showed 100% accuracy compared to the standard quantitative reverse transcription polymerase chain reaction (RT-qPCR) technique. Therefore, the nanobiosensor is sensitive, selective, and simple, providing a viable alternative for the rapid detection of SARS-CoV-2 in ready-to-eat vegetables.


Assuntos
COVID-19 , Nanopartículas Metálicas , Humanos , SARS-CoV-2/genética , COVID-19/diagnóstico , Ouro , Ressonância de Plasmônio de Superfície , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade
6.
Pharmaceutics ; 14(10)2022 Sep 20.
Artigo em Inglês | MEDLINE | ID: mdl-36297421

RESUMO

Tetanus toxin (TeNT) is produced by C. tetani, a spore-forming bacillus broadly spread in the environment. Although an inexpensive and safe vaccine is available, tetanus persists because of a lack of booster shots and variable responses to vaccines due to immunocompromised status or age-decreased immune surveillance. Tetanus is most prevalent in low- and medium-income countries, where it remains a health problem. Neutralizing monoclonal antibodies (mAbs) can prevent the severity of illness and death caused by C. tetani infection. We identified a panel of mAbs that bind to TeNT, some of which were investigated in a preclinical assay, showing that a trio of mAbs that bind to different sites of TeNT can neutralize the toxin and prevent symptoms and death in mice. We also identified two mAbs that can impair the binding of TeNT to the GT1b ganglioside receptor in neurons. In this work, to generate a series of cell lines, we constructed vectors containing sequences encoding heavy and light constant regions that can receive the paired variable regions resulting from PCRs of human B cells. In this way, we generated stable cell lines for five mAbs and compared and characterized the antibody produced in large quantities, enabling the characterization experiments. We present the results regarding the cell growth and viability in a fed-batch culture, titer measurement, and specific productivity estimation. The affinity of purified mAbs was analyzed by kinetics and under steady-state conditions, as three mAbs could not dissociate from TeNT within 36,000 s. The binding of mAbs to TeNT was confirmed by ELISA and inhibition of toxin binding to GT1b. The use of the mAbs mixture confirmed the individual mAb contribution to inhibition. We also analyzed the binding of mAbs to FcγR by surface plasmon resonance (SPR) and the glycan composition. Molecular docking analyses showed the binding site of an anti-tetanus mAb.

7.
Colloids Surf B Biointerfaces ; 219: 112794, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36162180

RESUMO

Although toxic and dangerous, Phenylmethane (PhM) dyes have a variety of medicinal functions. To optimize the use of these dyes, it is essential to understand their interaction mechanism with proteins. Through surface plasmon resonance, we investigated the kinetics and thermodynamics of interaction between bovine lactoferrin (BLF) and PhM dyes at pH 7.4, which allowed elucidate the effect of the dyes' functional groups on the binding process. Negative ΔG° revealed that at thermodynamic equilibrium the formed [BLF-PhM]° complex was more stable than the free BLF and PhM molecules. The increase in the number of methyl groups in the PhM structure led to an increase in the rates of association (ka) and dissociation (kd) and the binding constant (Kb). A similar effect was observed when comparing methyl violet B (MVB) and methyl violet 6 B (MV6B), in which the charged MV6B structure promoted an increase in the ka, kd, and Kb values. By contrast, an increase in the number of phenyl groups (2-3 rings) led to a decrease in the Kb values. The [BLF-PhM]° formation was entropically driven, indicating that hydrophobic interactions are critical for stabilizing these complexes These results are beneficial for understanding the molecular dynamics of protein-dye interactions.

8.
Front Immunol ; 13: 871874, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35898497

RESUMO

The COVID-19 pandemic caused by the severe acute syndrome virus 2 (SARS-CoV-2) has been around since November 2019. As of early June 2022, more than 527 million cases were diagnosed, with more than 6.0 million deaths due to this disease. Coronaviruses accumulate mutations and generate greater diversity through recombination when variants with different mutations infect the same host. Consequently, this virus is predisposed to constant and diverse mutations. The SARS-CoV-2 variants of concern/interest (VOCs/VOIs) such as Alpha (B.1.1.7), Beta (B.1.351), Gamma (B.1.1.28/P.1), Delta (B.1.617.2), and Omicron (B.1.1.529) have quickly spread across the world. These VOCs and VOIs have accumulated mutations within the spike protein receptor-binding domain (RBD) which interacts with the angiotensin-2 converting enzyme (ACE-2) receptor, increasing cell entry and infection. The RBD region is the main target for neutralizing antibodies; however, other notable mutations have been reported to enhance COVID-19 infectivity and lethality. Considering the urgent need for alternative therapies against this virus, an anti-SARS-CoV-2 equine immunoglobulin F(ab')2, called ECIG, was developed by the Butantan Institute using the whole gamma-irradiated SARS-CoV-2 virus. Surface plasmon resonance experiments revealed that ECIG binds to wild-type and mutated RBD, S1+S2 domains, and nucleocapsid proteins of known VOCs, including Alpha, Gamma, Beta, Delta, Delta Plus, and Omicron. Additionally, it was observed that ECIG attenuates the binding of RBD (wild-type, Beta, and Omicron) to human ACE-2, suggesting that it could prevent viral entry into the host cell. Furthermore, the ability to concomitantly bind to the wild-type and mutated nucleocapsid protein likely enhances its neutralizing activity of SARS-CoV-2. We postulate that ECIG benefits COVID-19 patients by reducing the infectivity of the original virus and existing variants and may be effective against future ones. Impacting the course of the disease, mainly in the more vulnerable, reduces infection time and limits the appearance of new variants by new recombination.


Assuntos
COVID-19 , SARS-CoV-2 , Enzima de Conversão de Angiotensina 2/genética , Animais , Anticorpos Antivirais , Cavalos , Humanos , Proteínas do Nucleocapsídeo , Pandemias , Receptores Virais/metabolismo , Glicoproteína da Espícula de Coronavírus
9.
Pharmaceutics, v. 14 ,10 , 1985, set. 2022
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4671

RESUMO

Tetanus toxin (TeNT) is produced by C. tetani, a spore-forming bacillus broadly spread in the environment. Although an inexpensive and safe vaccine is available, tetanus persists because of a lack of booster shots and variable responses to vaccines due to immunocompromised status or age-decreased immune surveillance. Tetanus is most prevalent in low- and medium-income countries, where it remains a health problem. Neutralizing monoclonal antibodies (mAbs) can prevent the severity of illness and death caused by C.tetani infection. We identified a panel of mAbs that bind to TeNT, some of which were investigated in a preclinical assay, showing that a trio of mAbs that bind to different sites of TeNT can neutralize the toxin and prevent symptoms and death in mice. We also identified two mAbs that can impair the binding of TeNT to the GT1b ganglioside receptor in neurons. In this work, to generate a series of cell lines, we constructed vectors containing sequences encoding heavy and light constant regions that can receive the paired variable regions resulting from PCRs of human B cells. In this way, we generated stable cell lines for five mAbs and compared and characterized the antibody produced in large quantities, enabling the characterization experiments. We present the results regarding the cell growth and viability in a fed-batch culture, titer measurement, and specific productivity estimation. The affinity of purified mAbs was analyzed by kinetics and under steady-state conditions, as three mAbs could not dissociate from TeNT within 36,000 s. The binding of mAbs to TeNT was confirmed by ELISA and inhibition of toxin binding to GT1b. The use of the mAbs mixture confirmed the individual mAb contribution to inhibition. We also analyzed the binding of mAbs to FcγR by surface plasmon resonance (SPR) and the glycan composition. Molecular docking analyses showed the binding site of an anti-tetanus mAb.

10.
Front Immunol, v. 13, 871874, jul. 2022
Artigo em Inglês | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP | ID: bud-4442

RESUMO

The COVID-19 pandemic caused by the severe acute syndrome virus 2 (SARS-CoV-2) has been around since November 2019. As of early June 2022, more than 527 million cases were diagnosed, with more than 6.0 million deaths due to this disease. Coronaviruses accumulate mutations and generate greater diversity through recombination when variants with different mutations infect the same host. Consequently, this virus is predisposed to constant and diverse mutations. The SARS-CoV-2 variants of concern/interest (VOCs/VOIs) such as Alpha (B.1.1.7), Beta (B.1.351), Gamma (B.1.1.28/P.1), Delta (B.1.617.2), and Omicron (B.1.1.529) have quickly spread across the world. These VOCs and VOIs have accumulated mutations within the spike protein receptor-binding domain (RBD) which interacts with the angiotensin-2 converting enzyme (ACE-2) receptor, increasing cell entry and infection. The RBD region is the main target for neutralizing antibodies; however, other notable mutations have been reported to enhance COVID-19 infectivity and lethality. Considering the urgent need for alternative therapies against this virus, an anti-SARS-CoV-2 equine immunoglobulin F(ab’)2, called ECIG, was developed by the Butantan Institute using the whole gamma-irradiated SARS-CoV-2 virus. Surface plasmon resonance experiments revealed that ECIG binds to wild-type and mutated RBD, S1+S2 domains, and nucleocapsid proteins of known VOCs, including Alpha, Gamma, Beta, Delta, Delta Plus, and Omicron. Additionally, it was observed that ECIG attenuates the binding of RBD (wild-type, Beta, and Omicron) to human ACE-2, suggesting that it could prevent viral entry into the host cell. Furthermore, the ability to concomitantly bind to the wild-type and mutated nucleocapsid protein likely enhances its neutralizing activity of SARS-CoV-2. We postulate that ECIG benefits COVID-19 patients by reducing the infectivity of the original virus and existing variants and may be effective against future ones. Impacting the course of the disease, mainly in the more vulnerable, reduces infection time and limits the appearance of new variants by new recombination.

11.
Mikrochim Acta ; 189(1): 11, 2021 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-34866167

RESUMO

Microfluidic devices that generate stable concentration gradients are efficient instruments for automated calibration for analytical and bioanalytical systems. However, little attention has been paid to the development of reusable microfluidic concentration gradient generators, which can be useful for a range of species through mathematical characterization. In this work, we develop a microfluidic device based on three steps of serial dilution that were able to generate nonlinear concentration gradient for dyes and biomolecules. The microfluidic device was described mathematically, statistically and was suitable for reusable analytical and bioanalytical analysis. The device reproducibility was assessed by experimental tests, which have shown the same gradient concentration profile for different dyes and statistical reproducibility with 95% confidence interval for bovine serum albumin (BSA). Moreover, the experimental data converged well with those  obtained by computational fluid dynamics simulation. Applicability was verified by coupling the microfluidic device to a surface plasmon resonance (SPR) biosensor, based on nanohole arrays with sensitivity of 358.7 nm RIU-1 determined by white-light SPR excitation exposed to different D-(+)-glucose aqueous solutions with 1.3361-1.4035 refractive index interval. The transmission light intensities obtained by the array of images allowed to quantify a pseudo-unknown BSA sample (160 µg mL-1) at 138 µg mL-1. The SPR analysis has been validated in parallel by fluorescence emissions, which showed a concentration of 154.8 ± 16.6 µg mL-1.

12.
Biosensors (Basel) ; 11(2)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572259

RESUMO

Chlorophene is an important antimicrobial agent present in disinfectant products which has been related to health and environmental effects, and its detection has been limited to chromatographic techniques. Thus, there is a lack of research that attempts to develop new analytical tools, such as biosensors, that address the detection of this emerging pollutant. Therefore, a new biosensor for the direct detection of chlorophene in real water is presented, based on surface plasmon resonance (SPR) and using a laccase enzyme as a recognition element. The biosensor chip was obtained by covalent immobilization of the laccase on a gold-coated surface through carbodiimide esters. The analytical parameters accomplished resulted in a limit of detection and quantification of 0.33 mg/L and 1.10 mg/L, respectively, fulfilling the concentrations that have already been detected in environmental samples. During the natural river's measurements, no significant matrix effects were observed, obtaining a recovery percentage of 109.21% ± 7.08, which suggested that the method was suitable for the fast and straightforward analysis of this contaminant. Finally, the SPR measurements were validated with an HPLC method, which demonstrated no significant difference in terms of precision and accuracy, leading to the conclusion that the biosensor reflects its potential as an alternative analytical tool for the monitoring of chlorophene in aquatic environments.


Assuntos
Técnicas Biossensoriais , Diclorofeno/análogos & derivados , Anti-Infecciosos , Carbodi-Imidas/análise , Diclorofeno/análise , Ouro , Limite de Detecção , Ressonância de Plasmônio de Superfície
13.
Molecules ; 25(20)2020 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-33065967

RESUMO

We investigate the plasmonic behavior of a fractal photonic crystal fiber, with Sierpinski-like circular cross-section, and its potential applications for refractive index sensing and multiband polarization filters. Numerical results were obtained using the finite element method through the commercial software COMSOL Multiphysics®. A set of 34 surface plasmon resonances was identified in the wavelength range from λ=630 nm to λ=1700 nm. Subsets of close resonances were noted as a consequence of similar symmetries of the surface plasmon resonance (SPR) modes. Polarization filtering capabilities are numerically shown in the telecommunication windows from the O-band to the L-band. In the case of refractive index sensing, we used the wavelength interrogation method in the wavelength range from λ=670 nm to λ=790 nm, where the system exhibited a sensitivity of S(λ)=1951.43 nm/RIU (refractive index unit). Due to the broadband capabilities of our concept, we expect that it will be useful to develop future ultra-wide band optical communication infrastructures, which are urgent to meet the ever-increasing demand for bandwidth-hungry devices.


Assuntos
Óptica e Fotônica/instrumentação , Ressonância de Plasmônio de Superfície/instrumentação , Desenho de Equipamento , Análise de Elementos Finitos , Fractais , Refratometria , Sensibilidade e Especificidade , Ressonância de Plasmônio de Superfície/métodos
14.
Lima; Instituto Nacional de Salud; mayo 2020.
Não convencional em Espanhol | BRISA/RedTESA, LILACS | ID: biblio-1116129

RESUMO

ANTECEDENTES: La inmunización es una de las intervenciones en salud pública más costo efectivas y rentables. Sarampión, parotiditis y rubeola (SPR) son enfermedades virales, que pueden causar complicaciones y consecuencias graves, especialmente en niños desnutridos e inmunodeprimidos; siendo importante destacar, que estas enfermedades son prevenibles mediante la vacunación. El resurgimiento de las infecciones por el virus de las paperas entre personas previamente vacunados con dos dosis, ha planteado preocupaciones en el mundo, sobre la ausencia de inmunidad a largo plazo después de la vacunación contra esta enfermedad y ha aperturado discusiones sobre nuevas estrategias para mitigar el riesgo de brotes futuros, incluyendo la posibilidad de implementar una tercera dosis de la vacuna SPR como respuesta a un brote epidémico, frente al cual, además surge la necesidad de estudios adicionales que evalúen la protección a largo plazo proporcionada por tres dosis de las vacunas SPR, así como la rentabilidad de la implementación de ésta intervención. OBJETIVO: El objetivo de la presente revisión sistemática es sintetizar evidencias científicas sobre la seguridad y efectividad frente a parotiditis de la vacuna Sarampión, Rubéola, Paperas (SPR) en personas mayores de 5 años. METODOLOGÍA: La búsqueda sistemática se realizó en la base de datos de Medline (PubMed), Lilacs y Cochrane Library fueron formuladas una estrategia de búsqueda para la pregunta PICO de la revisión, no se aplicaron filtros de fecha ni idiomas, la búsqueda abordó la evidencia publicada hasta 12 de marzo del 2020. La selección de título y resumen y extracción de datos fue realizada por un solo revisor. RESULTADOS: La búsqueda identificó 9 estudios: 1 revisión sistemática, 1 ensayo clínico y 7 estudios observacionales. La revisión incluyó tres estudios en niños y adolescentes. El ensayo clínico se realizó en adultos y los estudios observacionales fueron en adultos y en niños. CONCLUSIONES: No se observan diferencias estadísticamente significativas entre los niños que reciben una tercera dosis con los que reciben dos dosis. La vacuna SPR en niños mayores de 5 años presenta pocos y leves reacciones adversas. En adultos sanos, la tercera dosis de SPR no presenta reacciones adversas graves o largo plazo. En población militar, la aplicación de vacuna SPR no se asocia con aparición de diabetes mellitus tipo 1. (AU)


Assuntos
Humanos , Pré-Escolar , Criança , Vacina contra Sarampo-Caxumba-Rubéola/administração & dosagem , Sarampo/prevenção & controle , Caxumba/prevenção & controle , Avaliação da Tecnologia Biomédica , Avaliação em Saúde
15.
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
16.
Rev. chil. infectol ; Rev. chil. infectol;36(6): 774-777, dic. 2019.
Artigo em Espanhol | LILACS | ID: biblio-1058110

RESUMO

Resumen En Chile, la cepa del virus parotídeo utilizada en la vacuna es Leningrad-Zagreb (L-Z). Aunque la relación entre meningitis y la cepa L-Z sigue siendo controvertida, la mayoría de los casos reportados han presentado un cuadro de curso benigno y sin secuelas neurológicas. Presentamos el caso de una paciente que tres semanas post-inmunización con la vacuna tresvírica evolucionó con una meningitis aséptica de predominio mononuclear, con serología para IgM positiva contra el virus parotídeo. En este caso clínico, existió una relación temporal entre la vacunación, el inicio del cuadro parotídeo y posteriormente el meníngeo; la curva de inmunoglobulinas demostró una infección aguda posterior a la vacuna. Si bien no se logró aislar el virus en LCR, es razonable atribuir el cuadro a una infección post-vacunal.


In Chile, the strain of the mumps virus used in the vaccine is Leningrad-Zagreb (L-Z). Although the relationship between meningitis and the L-Z strain remains controversial, most of the reported cases have shown a benign course without permanent neurological sequelae. We present a case of a patient who presented an aseptic meningitis three weeks after immunization with a mumps vaccine; and laboratory confirmation showed positive serum mumps IgM antibody. In this clinical case, there was a temporal relationship between vaccination and the onset of the mumps and subsequently the meningeal involvement; the immunoglobulin curve demonstrates acute infection after vaccination. Although it was not possible to isolate the virus in CSF, it is reasonable to attribute the picture to a post-vaccinal infection.


Assuntos
Humanos , Meningite , Caxumba , Vacina contra Caxumba/efeitos adversos , Chile , Vírus da Caxumba
17.
Biomolecules ; 9(9)2019 09 12.
Artigo em Inglês | MEDLINE | ID: mdl-31547423

RESUMO

Trypanosoma cruzi, which causes Chagas disease, is a significant health threat in many countries and affects millions of people. Given the magnitude of this disease, a broader understanding of trypanocidal mechanisms is needed to prevent and treat infection. Natural endoperoxides, such as ergosterol peroxide, have been shown to be toxic to parasites without causing harm to human cells or tissues. Although prior studies have demonstrated the trypanocidal activity of ergosterol peroxide, the cellular and molecular mechanisms remain unknown. The results of this study indicate that a free-radical reaction occurs in T. cruzi following ergosterol peroxide exposure, leading to cell death. Using a combination of biochemical, microscopic and in silico experimental approaches, we have identified, for the first time, the cellular and molecular cytotoxic mechanism of an ergosterol peroxide obtained from Pleurotus ostreatus (Jacq) P. Kumm. f. sp. Florida.


Assuntos
Sistema Enzimático do Citocromo P-450/metabolismo , Ergosterol/análogos & derivados , Pleurotus/química , Trypanosoma cruzi/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Cristalografia por Raios X , Sistema Enzimático do Citocromo P-450/química , Citoplasma/efeitos dos fármacos , Citoplasma/metabolismo , Ergosterol/química , Ergosterol/farmacologia , Modelos Moleculares , Simulação de Acoplamento Molecular , Estrutura Molecular , Trypanosoma cruzi/metabolismo
18.
Food Chem ; 297: 125022, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31253281

RESUMO

Lactoferrin (LF) is a glycoprotein that serves as a potential vehicle for small bioactive molecules in food. In an effort to improve this functionality, the kinetic and thermodynamic interaction of LF with naringin (NR) was studied by surface plasmon resonance (SPR). The results demonstrated that the association rate constant between LF and NR was 5.00 × 104 M-1 s-1, while the dissociation rate of the complex was 0.36 s-1, at 25 °C. The stable complex predominated over free molecules (ΔG25°C0=-29.35 kJ mol-1), and the binding constant was 1.39 × 105 M-1, at 25 °C. The association of LF and NR to form an intermediate complex occurred in multi-steps. Nevertheless, the intermediate complex formation from the dissociation of the stable complex occurred in a single step with the activation energy independent of temperature. This study provides an important basis to explore LF as a vehicle for bioactive molecules.


Assuntos
Flavanonas/química , Lactoferrina/química , Ressonância de Plasmônio de Superfície , Animais , Bovinos , Flavanonas/metabolismo , Cinética , Lactoferrina/metabolismo , Ligação Proteica , Temperatura , Termodinâmica
19.
Sensors (Basel) ; 19(3)2019 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-30704098

RESUMO

The resonant wave modes in monomodal and multimodal planar Surface Plasmon Resonance (SPR) sensors and their response to a bidimensional array of gold nanoparticles (AuNPs) are analyzed both theoretically and experimentally, to investigate the parameters that rule the correct nanoparticle counting in the emerging metal nanoparticle-amplified surface plasmon resonance (PA-SPR) spectroscopy. With numerical simulations based on the Finite Element Method (FEM), we evaluate the error performed in the determination of the surface density of nanoparticles σ when the Maxwell-Garnett effective medium theory is used for fast data processing of the SPR reflectivity curves upon nanoparticle detection. The deviation increases directly with the manifestations of non-negligible scattering cross-section of the single nanoparticle, dipole-dipole interactions between adjacent AuNPs and dipolar interactions with the metal substrate. Near field simulations show clearly the set-up of dipolar interactions when the dielectric thickness is smaller than 10 nm and confirm that the anomalous dispersion usually observed experimentally is due to the failure of the effective medium theories. Using citrate stabilized AuNPs with a nominal diameter of about 15 nm, we demonstrate experimentally that Dielectric Loaded Waveguides (DLWGs) can be used as accurate nanocounters in the range of surface density between 20 and 200 NP/µm², opening the way to the use of PA-SPR spectroscopy on systems mimicking the physiological cell membranes on SiO2 supports.


Assuntos
Técnicas Biossensoriais/métodos , Nanopartículas Metálicas/química , Ressonância de Plasmônio de Superfície/métodos , Ácido Cítrico/química , Análise de Elementos Finitos , Ouro/química , Tamanho da Partícula , Dióxido de Silício/química
20.
ACS Appl Mater Interfaces ; 10(28): 23501-23508, 2018 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-29985579

RESUMO

We report the first optical biosensor for the novel and important cardiac biomarker, galectin-3 (Gal3), using the anti-Gal3 antibody as a biorecognition element and surface plasmon resonance (SPR) for transducing the bioaffinity event. The immunosensing platform was built at a thiolated Au surface modified by self-assembling four bilayers of poly(diallyldimethylammonium chloride) and graphene oxide (GO), followed by the covalent attachment of 3-aminephenylboronic acid (3ABA). The importance of GO, both as the anchoring point of the antibody and as a field enhancer for improving the biosensor sensitivity, was critically discussed. The advantages of using 3ABA to orientate the anti-Gal3 antibody through the selective link to the Fc region were also demonstrated. The new platform represents an interesting alternative for the label-free biosensing of Gal3 in the whole range of clinically relevant concentrations (linear range between 10.0 and 50.0 ng mL-1, detection limit of 2.0 ng mL-1) with successful application for Gal3 biosensing in enriched human serum samples.


Assuntos
Ressonância de Plasmônio de Superfície , Biomarcadores , Técnicas Biossensoriais , Galectina 3 , Ouro , Grafite , Humanos , Imunoensaio
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