RESUMO
The role of oxidative stress (OS) in cancer is a matter of great interest due to the implication of reactive oxygen species (ROS) and their oxidation products in the initiation of tumorigenesis, its progression, and metastatic dissemination. Great efforts have been made to identify the mechanisms of ROS-induced carcinogenesis; however, the validation of OS byproducts as potential tumor markers (TMs) remains to be established. This interventional study included a total of 80 colorectal cancer (CRC) patients and 60 controls. By measuring reduced glutathione (GSH), its oxidized form (GSSG), and the glutathione redox state in terms of the GSSG/GSH ratio in the serum of CRC patients, we identified significant changes as compared to healthy subjects. These findings are compatible with the effectiveness of glutathione as a TM. The thiol redox state showed a significant increase towards oxidation in the CRC group and correlated significantly with both the tumor state and the clinical evolution. The sensitivity and specificity of serum glutathione levels are far above those of the classical TMs CEA and CA19.9. We conclude that the GSSG/GSH ratio is a simple assay which could be validated as a novel clinical TM for the diagnosis and monitoring of CRC.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Colorretais/patologia , Glutationa/química , Glutationa/metabolismo , Estresse Oxidativo , Espécies Reativas de Oxigênio/metabolismo , Idoso , Estudos de Casos e Controles , Neoplasias Colorretais/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , OxirreduçãoRESUMO
Many important human diseases, and especially cancer, have been related to the overproduction of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG). This molecule is a product of oxidative stress processes over nucleophilic bases in DNA. In this work, an aptasensor for the rapid, selective and accurate detection of this oncomarker is presented. The aptasensor consists of a nanoporous anodic alumina material loaded with a dye and is functionalized with an aptamer-based "molecular gate". In the presence of target 8-oxo-dG, the capping aptamer displaces from the surface due to the high affinity of the analyte with the capping aptamer, thus inducing delivery of the preloaded fluorescent dye. In contrast, in the absence of 8-oxo-dG, a poor payload delivery is accomplished. This aptamer-based nanodevice has great sensitivity for 8-oxo-dG, resulting in a LOD of 1 nM and a detection time of ca. 60 min. Moreover, the aptasensor is able to accurately detect 8-oxo-dG in unmodified urine and serum without pre-concentration treatments. This diagnostic tool is validated in a set of 38 urine and serum samples from patients diagnosed of colorectal cancer and control patients. These samples are also analyzed using a standardized and specific ELISA kit. The aptasensor displays excellent sensitivity (95.83/100%) and specificity (80/100%) for 8-oxo-dG detection in serum and urine samples, respectively. Our results may serve as a basis for the development of generalized fluorogenic diagnostic platforms for the easy diagnosis of cancer in biofluids as well as for monitoring therapeutic treatments and detection of relapses without the use of expensive equipment or trained personnel.
Assuntos
Neoplasias Colorretais , Nanoporos , 8-Hidroxi-2'-Desoxiguanosina , Óxido de Alumínio , Neoplasias Colorretais/diagnóstico , Desoxiguanosina , HumanosRESUMO
Circulating microRNAs have emerged as potential diagnostic biomarkers. The deregulation of the microRNA miR-99a-5p has been previously described as an effective biomarker of early breast cancer. Herein, we present a new nanoporous anodic alumina (NAA)-based biosensor that can detect plasma miR-99a-5p with high sensitivity and selectivity. NAA pores are loaded with rhodamine B and capped with a specific oligonucleotide that is able to block cargo release until the target is present. In the presence of miR-99a-5p, the capping oligonucleotide recognizes the miR-99a-5p sequence and displaces it allowing the release of the encapsulated dye. This method is able to successfully distinguish healthy controls from breast cancer patients, even at early stages with high efficiency, showing the presented system as a promising tool for breast cancer detection.
Assuntos
Neoplasias da Mama , MicroRNAs , Nanoporos , Óxido de Alumínio , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/genética , Eletrodos , Humanos , MicroRNAs/genéticaRESUMO
Celiac disease is a complex and autoimmune disorder caused by the ingestion of gluten affecting almost 1% of global population. Nowadays an effective treatment does not exist, and the only way to manage the disease is the removal of gluten from the diet. Owing the key role played by gluten, clear and regulated labelling of foodstuff and smart methods for gluten detection are needed to fight frauds on food industry and to avoid the involuntary ingestion of this protein by celiac patients. On that scope, the development of a novel detection system of gluten is here presented. The sensor consists of nanoporous anodic alumina films loaded with a fluorescent dye and capped with an aptamer that recognizes gliadin (gluten's soluble proteins). In the presence of gliadin, aptamer sequences displace from the surface of anodic alumina resulting in pore opening and dye delivery. The dispositive shows a limit of detection (LOD) of 100 µg kg-1 of gliadin, good selectivity and a detection time of approximately 60 min. Moreover, the sensor is validated in real food samples. This novel probe allows fast gluten detection through a simple signalling process with potential use for food control.
Assuntos
Doença Celíaca , Glutens , Eletrodos , Gliadina , Humanos , Limite de DetecçãoRESUMO
Candida auris has arisen as an important multidrug-resistant fungus because of several nosocomial outbreaks and elevated rates of mortality. Accurate and rapid diagnosis of C. auris is highly desired; nevertheless, current methods often present severe limitations and produce misidentification. Herein a sensitive, selective, and time-competitive biosensor based on oligonucleotide-gated nanomaterials for effective detection of C. auris is presented. In the proposed design, a nanoporous anodic alumina scaffold is filled with the fluorescent indicator rhodamine B and the pores blocked with different oligonucleotides capable of specifically recognize C. auris genomic DNA. Gate opening modulation and cargo delivery is controlled by successful DNA recognition. C. auris is detected at a concentration as low as 6â CFU/mL allowing obtaining a diagnostic result in clinical samples in one hour with no prior DNA extraction or amplification steps.
Assuntos
Técnicas Biossensoriais/métodos , Candida/isolamento & purificação , Candidíase/diagnóstico , Oligonucleotídeos/genética , Óxido de Alumínio , Candida/genética , Diagnóstico Precoce , Humanos , Técnicas de Diagnóstico Molecular , Nanoporos , Oligonucleotídeos/química , Rodaminas/química , Sensibilidade e Especificidade , Fatores de TempoRESUMO
Pneumocystis pneumonia (PcP) is a disease produced by the opportunistic infection of the fungus Pneumocystis jirovecii. As delayed or unsuitable treatments increase the risk of mortality, the development of rapid and accurate diagnostic tools for PcP are of great importance. Unfortunately, current standard methods present severe limitations and are far from adequate. In this work, a time-competitive, sensitive and selective biosensor based on DNA-gated nanomaterials for the identification of P. jirovecii is presented. The biosensor consists of a nanoporous anodic alumina (NAA) scaffold which pores are filled with a dye reporter and capped with specific DNA oligonucleotides. In the presence of P. jirovecii genomic DNA, the gated biosensor is open, and the cargo is delivered to the solution where it is monitored through fluorescence spectroscopy. The use of capping oligonucleotides able to form duplex or triplex with P. jirovecii DNA is studied. The final diagnostic tool shows a limit of detection (LOD) of 1 nM of target complementary DNA and does not require previous amplification steps. The method was applied to identify DNA from P. jirovecii in unmodified bronchoalveolar lavage, nasopharyngeal aspirates, and sputum samples in 60 min. This is a promising alternative method for the routinely diagnosis of Pneumocystis pneumonia.
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This review includes examples of silica-based, chromo-fluorogenic nanosensors with the aim of illustrating the evolution of the discipline in recent decades through relevant research developed in our group. Examples have been grouped according to the sensing strategies. A clear evolution from simply functionalized materials to new protocols involving molecular gates and the use of highly selective biomolecules such as antibodies and oligonucleotides is reported. Some final examples related to the evolution of chromogenic arrays and the possible use of nanoparticles to communicate with other nanoparticles or cells are also included. A total of 64 articles have been summarized, highlighting different sensing mechanisms.
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The consumption of illicit drugs has increased exponentially in recent years and has become a problem that worries both governments and international institutions. The rapid emergence of new compounds, their easy access, the low levels at which these substances are able to produce an effect, and their short time of permanence in the organism make it necessary to develop highly rapid, easy, sensitive, and selective methods for their detection. Currently, the most widely used methods for drug detection are based on techniques that require large measurement times, the use of sophisticated equipment, and qualified personnel. Chromo- and fluorogenic methods are an alternative to those classical procedures.
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A nanoporous anodic alumina (NAA)-based sensor system for the detection of Mycoplasma was developed through the implementation of "molecular gates" selective to the presence of this bacterium. The capped support showed a negligible cargo release, while presence of Mycoplasma genomic DNA resulted in the release of rhodamineâ B fluorescent dye. This sensor system presents a limit of detection of 20â genomic DNA copiesâ µL-1 and was applied to the detection of Mycoplasma bacteria in competitive environments, such as culture cell media.
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El objetivo del trabajo fue evaluar y pronosticar el efecto reductor de la mortalidad por cáncer del cérvix por parte de su programa. Se compararon las pendientes de sus tasas de mortalidad, tanto específicas por edades como estandarizadas, y se observó un estancamiento en las tendencias a partir de 1971 y un ligero descenso a partir de 1975, en contraposición a la evolución ascendente entre 1964 y 1971. Se estima que el programa ha preservado la vida a más de 1 200 mujeres hasta 1985. En otros cánceres de útero se observó un franco decrecimiento de la mortalidad desde 1971. Se extrapoló la tendencia del período 1975 a 1985 hasta el 2000 y se mostró que si ésta continúa a un ritmo de descenso de la tasa promedio anual (estandarizada con la población del año 2000) de 0,09 cada año, en los próximos 15 años, es probable que se reduzca en 39 ó 14
con respecto a la estandarizada o específica de 1975, o aún a mucho menos. Esto será posible si se fortalece óptimamente el programa en diferentes aspectos, abordados en el pronóstico elaborado
Assuntos
Humanos , Feminino , Neoplasias do Colo do Útero/mortalidade , Programas Nacionais de Saúde , PrognósticoRESUMO
El objetivo del trabajo fue evaluar y pronosticar el efecto reductor de la mortalidad por cáncer del cérvix por parte de su programa. Se compararon las pendientes de sus tasas de mortalidad, tanto específicas por edades como estandarizadas, y se observó un estancamiento en las tendencias a partir de 1971 y un ligero descenso a partir de 1975, en contraposición a la evolución ascendente entre 1964 y 1971. Se estima que el programa ha preservado la vida a más de 1 200 mujeres hasta 1985. En otros cánceres de útero se observó un franco decrecimiento de la mortalidad desde 1971. Se extrapoló la tendencia del período 1975 a 1985 hasta el 2000 y se mostró que si ésta continúa a un ritmo de descenso de la tasa promedio anual (estandarizada con la población del año 2000) de 0,09 cada año, en los próximos 15 años, es probable que se reduzca en 39 ó 14 % con respecto a la estandarizada o específica de 1975, o aún a mucho menos. Esto será posible si se fortalece óptimamente el programa en diferentes aspectos, abordados en el pronóstico elaborado