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Artemisia herba-alba (AHA) is a traditionally used plant to treat various diseases, including diabetes and metabolic dysfunctions. Plant extracts are generally explored empirically without a deeper assessment of their mechanism of action. Here, we describe a combinatorial study of biochemical, molecular, and bioinformatic (metabolite-protein pharmacology network) analyses to elucidate the mechanism of action of AHA and shed light on its multilevel effects in the treatment of diabetes-related advanced glycation end-products (AGE)-induced liver damages. The extract's polyphenols and flavonoids content were measured and then identified via LC-Q-TOF-MS/MS. Active compounds were used to generate a metabolite-target interaction network via Swiss Target Prediction and other databases. The extract was tested for its antiglycation and aggregation properties. Next, THLE-2 liver cells were challenged with AGEs, and the mechanistic markers were measured [TNF-α, IL-6, nitric oxide, total antioxidant capacity, lipid peroxidation (LPO), and caspase 3]. Metabolite and network screening showed the involvement of AHA in diabetes, glycation, liver diseases, aging, and apoptosis. Experimental confirmation showed that AHA inhibited protein modification and AGE formation. Additionally, AHA reduced inflammatory mediators (IL-6, TNFα), oxidative stress markers (NO, LPO), and apoptosis (Caspase 3). On the other hand, cellular total antioxidant capacity was restored to normal levels. The combinatorial study showed that AHA regulates AGE-induced liver damages through MAPK-AKT and AGE-RAGE signaling pathways. This report highlights the combination of experimental and network pharmacology for the exact elucidation of AHA mechanism of action as a multitarget option in the therapy of diabetes and AGEs-related diseases.
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Artemisia , Diabetes Mellitus , Antioxidantes/farmacologia , Artemisia/metabolismo , Caspase 3/metabolismo , Diabetes Mellitus/tratamento farmacológico , Flavonoides/farmacologia , Produtos Finais de Glicação Avançada/metabolismo , Mediadores da Inflamação/metabolismo , Interleucina-6/metabolismo , Fígado/metabolismo , Óxido Nítrico/metabolismo , Extratos Vegetais/farmacologia , Polifenóis/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Transdução de Sinais , Espectrometria de Massas em Tandem , Fator de Necrose Tumoral alfa/metabolismoRESUMO
The construction of a rapid and easy immunofluorescence bioassay for SARS-CoV-2 detection is described. We report for the first time a novel one-pot synthetic approach for simultaneous photoinduced step-growth polymerization of pyrene (Py) and ring-opening polymerization of ε-caprolactone (PCL) to produce a graft fluorescent copolymer PPy-g-PCL that was conjugated to SARS-CoV-2-specific antibodies using EDC/NHS chemistry. The synthesis steps and conjugation products were fully characterized using standard spectral analysis. Next, the PPy-g-PCL was used for the construction of a dot-blot assay which was calibrated for applications to human nasopharyngeal samples. The analytical features of the proposed sensor showed a detection range of 6.03-8.7 LOG viral copy mL-1 (Ct Scores: 8-25), the limit of detection (LOD), and quantification (LOQ) of 1.84 and 6.16 LOG viral copy mL-1, respectively. The repeatability and reproducibility of the platform had a coefficient of variation (CV) ranging between 1.2 and 5.9%. The fluorescence-based dot-blot assay was tested with human samples. Significant differences were observed between the fluorescence intensity of the negative and positive samples, with an overall correct response of 93.33%. The assay demonstrated a high correlation with RT-PCR data. This strategy opens new insights into simplified synthesis procedures of the reporter molecules and their high potential sensing and diagnosis applications.
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COVID-19 , SARS-CoV-2 , Bioensaio , COVID-19/diagnóstico , Caproatos , Corantes , Humanos , Lactonas , Poli A , Poliésteres , Polimerização , Reprodutibilidade dos TestesRESUMO
INTRODUCTION: The chronicity of advanced glycation end-products (AGEs) imparts various damages resulting in metabolic dysfunction and diseases involving inflammation and oxidative stress. The use of plant extracts is of high interest in complementary medicine. Yet, extracts are multicomponent mixtures, and difficult to pinpoint their exact mechanism. OBJECTIVES: We hypothesise that network pharmacology and bioinformatics can help experimental findings depict the exact active components and mechanism of action by which they induce their effects. Additionally, the toxicity and variability can be lowered and standardised with proper encapsulation methods. METHODOLOGY: Here, we propose the formulation of phytoniosomes encapsulating two Artemisia species (Artemisia dracunculus and Artemisia absinthium) to mitigate AGEs and their induced cell redox dysregulation in the liver. Extracts from different solvents were identified via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS/MS). Phytoniosomes were explored for their anti-glycating effect and modulation of AGE-induced damages in THLE-2 liver cells. Network pharmacology tools were used to identify possible targets and signalling pathways implicated. RESULTS: Data demonstrated that A. absinthium phytoniosomes had a significant anti-AGE effect comparable to reference molecules and higher than A. dracunculus. They were able to restore cell dysfunction through the restoration of tumour necrosis alpha (TNF-α), interleukin 6 (IL-6), nitric oxide, and total antioxidant capacity. Phytoniosomes were able to protect cells from apoptosis by decreasing caspase 3 activity. Network pharmacology and bioinformatic analysis confirmed the induction of the effect via Akt-PI3K-MAPK and AGE-RAGE signalling pathways through quercetin and luteolin actions. CONCLUSION: The current report highlights the potential of Artemisia phytoniosomes as strong contenders in AGE-related disease therapy.
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Artemisia , Diabetes Mellitus , Medicamentos de Ervas Chinesas , Antioxidantes/farmacologia , Artemisia/química , Caspase 3 , Cromatografia Líquida , Interleucina-6 , Fígado/metabolismo , Luteolina , Farmacologia em Rede , Óxido Nítrico , Fosfatidilinositol 3-Quinases , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Quercetina , Solventes , Espectrometria de Massas em Tandem/métodos , Fator de Necrose Tumoral alfaRESUMO
Chemotherapy has seen great progress in the development of performant treatment strategies. Nanovesicles such as liposomes and polymersomes demonstrated great potential in cancer therapy. However, these nanocarriers deliver their content passively, which faces a lot of constraints during blood circulation. The main challenge resides in degradation and random delivery to normal tissues. Hence, targeting drug delivery using specific molecules (such as antibodies) grafted over the surface of these nanocarriers came as the answer to overcome many problems faced before. The advantage of using antibodies is their antigen/antibody recognition, which provides a high level of specificity to reach treatment targets. This review discusses the many techniques of nanocarrier functionalization with antibodies. The aim is to recognize the various approaches by describing their advantages and deficiencies to create the most suitable drug delivery platform. Some methods are more suitable for other applications rather than drug delivery, which can explain the low success of some proposed targeted nanocarriers. In here, a critical analysis of how every method could impact the recognition and targeting capacity of some nanocarriers (liposomes and polymersomes) is discussed to make future research more impactful and advance the field of biomedicine further.
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Sistemas de Liberação de Medicamentos , Lipossomos/química , Preparações Farmacêuticas/administração & dosagem , Animais , Anticorpos/química , Sistemas de Liberação de Medicamentos/métodos , Humanos , Nanopartículas/química , Polímeros/química , Propriedades de SuperfícieRESUMO
Supply shortage for the development and production of preventive, therapeutic, and diagnosis tools during the COVID-19 pandemic is an important issue affecting the wealthy and poor nations alike. Antibodies and antigens are especially needed for the production of immunological-based testing tools such as point-of-care tests. Here, we propose a simple and quick magnetic nanoparticle (MNP)-based separation/isolation approach for the repurposing of infected human samples to produce specific antibodies and antigen cocktails. Initially, an antibody cocktail was purified from serums via precipitation and immunoaffinity chromatography. Purified antibodies were conjugated onto MNPs and used as an affinity matrix to separate antigens. The characterization process was performed by ELISA, SDS-PAGE, electrochemistry, isothermal titration calorimetry, and LC-Q-TOF-MS/MS analyses. The MNP-separated peptides can be used for mass spectrometry-based as well as paper-based lateral flow assay diagnostic. The exploitation of the current workflow for the development of efficient diagnostic tools, specific treatments, and fundamental research can significantly impact the present or eventual pandemic. This workflow can be considered as a two birds, one stone-like strategy.
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Anticorpos Antivirais/isolamento & purificação , Antígenos Virais/isolamento & purificação , COVID-19/diagnóstico , Análise Custo-Benefício , Imunoensaio/economia , SARS-CoV-2/isolamento & purificação , Viremia/virologia , Anticorpos Antivirais/sangue , Antígenos Virais/sangue , COVID-19/virologia , Calorimetria , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Humanos , SARS-CoV-2/imunologia , Manejo de Espécimes , Espectrometria de Massas em Tandem , Viremia/sangue , Fluxo de TrabalhoRESUMO
Synthetic cannabinoids are one of the many substances of abuse widely spreading in modern society. Medical practitioners and law enforcement alike highly seek portable, efficient, and reliable tools for on-site detection and diagnostics. Here, we propose a colorimetric lateral flow assay (LFA) combined with dye-loaded polymersome to detect the synthetic cannabinoid JWH-073 efficiently. Rhodamine B-loaded polymersome was conjugated to antibodies and fully characterized. Two LFA were proposed (sandwich and competitive), showing a high level of sensitivity with a limit of detection (LOD) reaching 0.53 and 0.31 ng/mL, respectively. The competitive assay was further analyzed by fluorescence, where the LOD reached 0.16 ng/mL. The application of the LFA over spiked synthetic saliva or real human saliva demonstrated an overall response of 94% for the sandwich assay and 97% for the competitive LFA. The selectivity of the system was assessed in the presence of various interferents. The analytical performance of the LFA system showed a coefficient of variation below 6%. The current LFA system appears as a plausible system for non-invasive detection of substance abuse and shows promise for synthetic cannabinoid on-site sensing.
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Canabinoides/análise , Corantes Fluorescentes/química , Poliésteres/química , Polietilenoglicóis/química , Rodaminas/química , Detecção do Abuso de Substâncias/métodos , Anticorpos Imobilizados/imunologia , Canabinoides/imunologia , Colorimetria/instrumentação , Colorimetria/métodos , Humanos , Drogas Ilícitas/análise , Drogas Ilícitas/imunologia , Imunoensaio/instrumentação , Imunoensaio/métodos , Indóis/análise , Indóis/imunologia , Limite de Detecção , Naftalenos/análise , Naftalenos/imunologia , Papel , Reprodutibilidade dos Testes , Saliva/química , Detecção do Abuso de Substâncias/instrumentaçãoRESUMO
Real-time detection of substance use is an approach of high interest leading to the optimization of behavioral interventions and drug abuse intervention. The current methods in use suffer many limitations and need high logistical and laboratory requirements. Biosensors have shown a great potential in overcoming these limitations. In the present study, the electrochemical biosensor composed of a screen-printed electrode (SPE) was designed for the detection of synthetic cannabinoid (SC). Antibody-immobilized magnetic nanoparticles were also used to create a surface on the transducer with magnetic interactions in order to detect JWH-073 as a SC model. The use of immobilized magnetic nanoparticles to create working surfaces makes the electrode a reusable SPE which can be reutilized after the cleansing. To examine and observe any possible changes on the surface due to its interaction with the analyte, different electrochemical techniques such as differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectrometry were applied. Based on the obtained results, the linearity of the biosensor was found between 5 and 400 ng/mL, and the detection limit was calculated as 22 ng/mL (n = 6) using the 3 Sb/m formula. The biosensor functionality was studied in the presence of some related interferents that showed lower responses than JWH-073, thus demonstrating the good selectivity of the prepared biosensor. Finally, the sensory platform was used to test synthetic urine sample, and the results were compared with obtained results from liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), which showed that the proposed method could be utilized to identify abuse drugs.
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Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Drogas Ilícitas/análise , Indóis/análise , Nanopartículas de Magnetita/química , Naftalenos/análise , Psicotrópicos/análise , Anticorpos Imobilizados/imunologia , Técnicas Biossensoriais/instrumentação , Técnicas Eletroquímicas/instrumentação , Eletrodos , Drogas Ilícitas/imunologia , Separação Imunomagnética , Indóis/imunologia , Limite de Detecção , Naftalenos/imunologia , Propilaminas/química , Psicotrópicos/imunologia , Reprodutibilidade dos Testes , Silanos/químicaRESUMO
Artificial nanostructures using polymers to produce polymeric vesicles are inspired by the many intricate structures found in living organisms. Polymersomes are a class of self-assembled vesicles known for their great stability and application in drug delivery. They can be tuned according to their intended use by changing their components and introducing activable block copolymers that transform these polymersomes into smart nanocarriers. In this study, we propose the synthesis of a poly (ethylene oxide)-poly (ε-caprolactone)-based polymersome (PEO-PCL) loaded with GSH as a pH-responsive drug delivery molecule for cancer and protein alteration inhibition. Initially, the nanocarrier was synthesized and characterized by DLS, TEM/SEM microscopy as well as gel permeation chromatography (GPC) and 1H NMR. Their CMC formation, encapsulation efficiency, and pH responsiveness were analyzed. In addition, empty and GSH-loaded PEO-PCL polymersomes were tested for their toxicity and therapeutic effect on normal and cancer cells via an MTT test. Subsequently, protein alteration models (aggregation, glycation, and oxidation) were performed in vitro where the polymersomes were tested. Results showed that other than being non-toxic and able to highly encapsulate and release the GSH in response to acidic conditions, the nanocomposites do not hinder its content's ameliorative effects on cancer cells and protein alterations. This infers that polymeric nanocarriers can be a base for future smart biomedicine applications and theranostics.
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Portadores de Fármacos , Glutationa , Proteínas de Neoplasias/metabolismo , Neoplasias , Poliésteres , Animais , Chlorocebus aethiops , Preparações de Ação Retardada/química , Preparações de Ação Retardada/farmacocinética , Preparações de Ação Retardada/farmacologia , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Portadores de Fármacos/farmacologia , Glutationa/química , Glutationa/farmacocinética , Glutationa/farmacologia , Glicosilação/efeitos dos fármacos , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Neoplasias/patologia , Poliésteres/química , Poliésteres/farmacocinética , Poliésteres/farmacologia , Células VeroRESUMO
A quick and easy colorimetric sensor based on gold nanoparticles (GNPs) and aptamers for the detection of cocaine was developed. The sensor was named as 'GAPTA' and showed extremely interesting results regarding cocaine detection with a sensitivity to doses of 0.2 nM. The experimental approach consisted of creating a conjugate between GNPs (10 nm size) and aptamers as a sensing base with the addition of an electrolyte (NaCl) that plays the role of aggregation inducer. In the absence of the aptamer, the electrolyte was able to induce aggregation of the GNPs turning the color of the solution from red to blue while the presence of the aptamer is able to hinder the charges attraction and protects the GNPs from aggregating. The optimization of the aptamer and electrolyte concentration was determined to be 118 nM and 55 mM, respectively, and the resultant GAPTA sensor had a detection limit of 0.97 nM. Furthermore, the selectivity of the platform was tested in the presence of different interferents and showed a specific response towards cocaine while interference ranged between 20 and 40%. The applicability of the GAPTA biosensor was tested on synthetic saliva and demonstrated a sensitivity range between 0.2 and 25 nM. These results suggest the potential of the current colorimetric sensor in abuse drugs screening and creates a stable base for new routine platforms for biomedical and toxicology applications. Graphical abstract.
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Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Cocaína/análise , Cocaína/química , Limite de Detecção , Coloides , Colorimetria , Eletrólitos/química , Ouro/química , Nanopartículas Metálicas/químicaRESUMO
Age-related complications including protein alterations seen in diabetes and Alzheimer's disease are a major issue due to their accumulation and deleterious effects. This report aims to investigate the effect of zinc supplementation on the anti-glycoxidation activity of carnosine on the in vitro model of albumin-based protein modification. Besides, the therapeutic effect of this combination was tested through the addition of the molecules in tandem (co-treatment) or post initiation (post-treatment) of the protein modification process. Glycation was induced via the addition of glucose to which carnosine (5 mM) alone or with various zinc concentrations (125, 250, and 500 µM) were added either at 0 h or 24 h post-glycation induction. On the other hand, protein oxidation was induced using chloramine T (20 mM) and treated in the same way with carnosine and zinc. The different markers of glycation (advanced glycation end products (AGEs), dityrosine, and beta-sheet formation (aggregation)) and oxidation (AOPP, advanced oxidation protein products) were estimated via fluorescence and colorimetric assays. Zinc addition induced a significant enhancement of carnosine activity by reducing albumin modification that outperformed aminoguanidine both in the co- and post-treatment protocols. Zinc demonstrated a supplementary effect in combination with carnosine highlighting its potential in the protection against age-related protein modifications processes such as the ones found in diabetes.
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Carnosina/farmacologia , Modelos Biológicos , Soroalbumina Bovina/antagonistas & inibidores , Zinco/farmacologia , Animais , Bovinos , Glicosilação , Oxirredução , Soroalbumina Bovina/metabolismoRESUMO
Colon cancer is thought to develop in a stepwise fashion. In this study, the relationship between aberrant crypt foci (ACF) regional distribution and oxidative stress evolution was studied in a murine model of initial colon carcinogenesis induced by dimethylhydrazine (DMH). Mice were given 2 weekly subcutaneous injections of DMH (20 mg/kg) and killed at the 10th, 12th or 14th week. ACF was scored for number, distribution and crypt multiplicity after methylene-blue coloration and histologically analyzed afterwards. Oxidative stress evaluation was assessed through myeloperoxidase activity (MPO), nitric oxide (NO), L-ornithine and malondialdehyde (MDA) levels as well as antioxidant CAT, SOD and GSH. DMH treatment showed a shift from small to large ACF but also from distal to proximal colon between week 10 and 14 (p < 0.05). This was further illustrated histologically with crypt disruption and mucin depletion. Oxidative stress imbalance was observed in all DMH-treated groups. All markers (MPO, MDA and NO) peaked at week 12 (p < 0.01) and decreased at week 14 (p < 0.05) while L-ornithine decreased through all protocol (p < 0.01). Antioxidants decreased in all points (p < 0.05) but only GSH increased at week 14 (p < 0.05). This work provided insight to response-patterns of oxidative stress between distal and proximal colon, showing for the first time a decreasing implication during the development process and suggesting other inflammatory, immunologic or microbiota implication as factors to be considered during chemotherapy approaches.
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1,2-Dimetilidrazina/farmacologia , Focos de Criptas Aberrantes/induzido quimicamente , Carcinogênese/efeitos dos fármacos , Neoplasias do Colo/induzido quimicamente , Estresse Oxidativo/efeitos dos fármacos , Focos de Criptas Aberrantes/metabolismo , Animais , Antioxidantes/metabolismo , Biomarcadores Tumorais/metabolismo , Colo/efeitos dos fármacos , Colo/metabolismo , Neoplasias do Colo/metabolismo , Modelos Animais de Doenças , Feminino , Malondialdeído/metabolismo , Camundongos , Mucinas/metabolismo , Óxido Nítrico/metabolismo , Ornitina/metabolismo , Peroxidase/metabolismoRESUMO
We have previously shown that ethanolic extract from bark (EEB) of Salix aegyptiaca (Musk Willow) can inhibit proliferation and motility and induce apoptosis in colon cancer cells. Tandem mass spectrometry revealed EEB to be rich in catechin, catechol, and salicin. The present study investigated the chemopreventive effect of HPLC-fingerprinted EEB on 1,2-dimethylhydrazine (DMH)-induced aberrant crypt foci (ACF) formation in mice. DMH (20 mg/kg body weight) was weekly injected subcutaneously to mice for the first 2 weeks. EEB (100 and 400 mg/kg body weight) was provided orally from the 7th to 14th week, after which colon tissues were evaluated histologically and biochemically. DMH treatment induced high number of ACF; EEB significantly reduced the number and multiplicity of ACF, along with a restoration in goblet cells and mucin accumulation. EEB supplementation improved the markers of inflammation (myeloperoxidase and neutrophil infiltration) and oxidative stress. More importantly, EEB amplified apoptosis of neoplastic cells in the colon mucosa of DMH-treated mice. It also lowered levels of markers for early transformation events such as EGFR, nuclear ß-catenin, and COX-2 in colon cancer cell lines HT-29 and HCT-116. The innocuity of EEB (up to 1600 mg/kg) to mice reinforces its potential as a chemopreventive agent.
Assuntos
1,2-Dimetilidrazina/toxicidade , Focos de Criptas Aberrantes/tratamento farmacológico , Anticarcinógenos/farmacologia , Neoplasias do Colo/prevenção & controle , Extratos Vegetais/farmacologia , Salix/química , Focos de Criptas Aberrantes/induzido quimicamente , Focos de Criptas Aberrantes/patologia , Animais , Anticarcinógenos/química , Carcinógenos/toxicidade , Cromatografia Líquida de Alta Pressão , Neoplasias do Colo/induzido quimicamente , Neoplasias do Colo/patologia , Etanol/química , Células HCT116/efeitos dos fármacos , Humanos , Concentração Inibidora 50 , Masculino , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Casca de Planta/química , Extratos Vegetais/químicaRESUMO
Biosensor research has long focused on achieving the lowest possible Limits of Detection (LOD), driving significant advances in sensitivity and opening up new possibilities in analysis. However, this intense focus on low LODs may not always meet the practical needs or suit the actual uses of these devices. While technological improvements are impressive, they can sometimes overlook important factors such as detection range, ease of use, and market readiness, which are vital for biosensors to be effective in real-world applications. This review advocates for a balanced approach to biosensor development, emphasizing the need to align technological advancements with practical utility. We delve into various applications, including the detection of cancer biomarkers, pathology-related biomarkers, and illicit drugs, illustrating the critical role of LOD within these contexts. By considering clinical needs and broader design aspects like cost-effectiveness, sustainability, and regulatory compliance, we argue that integrating technical progress with practicality will enhance the impact of biosensors. Such an approach ensures that biosensors are not only technically sound but also widely useable and beneficial in real-world applications. Addressing the diverse analytical parameters alongside user expectations and market demands will likely maximize the real-world impact of biosensors.
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In the ongoing pursuit of enhancing food safety and quality through advanced technologies, silver nanoparticles (AgNPs) stand out for their antimicrobial properties. Despite being overshadowed by other nanoparticles in food sensing applications, AgNPs possess inherent qualities that make them effective tools for rapid and selective contaminant detection in food matrices. This review aims to reinvigorate the interest in AgNPs in the food industry, emphasizing their sensing mechanism and the transformative potential of integrating them with artificial intelligence (AI) for enhanced food safety monitoring. It discusses key AI tools and principles in the food industry, demonstrating their positive impact on food analytical chemistry. The interplay between AI and biosensors offers many advantages and adaptability to dynamic analytical challenges, significantly improving food safety monitoring and potentially redefining the landscape of food safety and quality assurance.
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Anti-Infecciosos , Nanopartículas Metálicas , Prata/química , Nanopartículas Metálicas/química , Análise de Alimentos , Inteligência Artificial , Anti-Infecciosos/químicaRESUMO
In contemporary society, developing dependable point-of-care (POC) biosensors for the timely detection of cancer markers is crucial. Among various sensor types, screen-printed electrode (SPE)-based sensors, particularly electrochemical ones, stand out as promising candidates for POC applications. Despite ongoing efforts to create numerous SPE-based sensors, there is a continuous pursuit to enhance their sensitivity and analytical capabilities. This study presents an advanced electrochemical sensor designed to sensitively detect the hepatocellular carcinoma (HCC) marker Alpha-fetoprotein (AFP) in saliva. The sensor employs a gold SPE modified with hydroxyapatite, TiO2 nanoparticles, 1-butyl-3-methylimidazolium bis(trifluoromethyl sulfonyl)imide ionic liquid (IL), and AFP monoclonal antibodies. After thorough characterization and optimization using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), the biosensor exhibited a broad detection range (0.01-400 ng/mL), a low limit of detection (LOD) at 0.058 ng/mL, and demonstrated high selectivity, repeatability, reproducibility, and stability. Furthermore, when tested with spiked human saliva samples, the biosensor displayed excellent recovery and robustness, showcasing its potential for noninvasive and POC diagnosis of HCC. In an environmentally conscious evaluation, the biosensor's greenness was assessed using the AGREE metric, yielding a high score of 0.85. This score indicates the biosensor's alignment with the principles of green analytical chemistry, underlining its eco-friendly attributes. This innovative electrochemical sensor contributes to the ongoing efforts for efficient and reliable POC diagnostic tools and aligns with a broader commitment to developing environmentally friendly solutions.
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BACKGROUND: Alpha-fetoprotein (AFP) is a fetal protein that can indicate congenital anomalies such as Down syndrome and spinal canal blockage when detected at abnormal levels in pregnant women. Current AFP detection methods rely on invasive blood or serum samples, which require sophisticated equipment. From the many solutions proposed, colorimetric paper-based assays excel in point-of-care settings. The concept of paper-based ELISA (p-ELISA) enhances traditional methods, aligning with the ASSURED criteria for diagnostics in resource-limited regions. Despite success in microfluidic paper-based assay devices, laser printing remains underexplored for p-ELISA. Additionally, modifying the paper surface provides an additional layer of sensitivity enhancement. RESULTS: In this study, we developed a novel laser-printed paper-based ELISA (LP-pELISA) for rapid, sensitive, and noninvasive detection of AFP in saliva samples. The LP-pELISA platform was fabricated by printing hydrophobic barriers on filter paper using a laser printer, followed by depositing hydroxyapatite (HAp) as an immobilization material for the antibodies. The colorimetric detection was achieved using AuNPs functionalized with anti-AFP antibodies and silver nitrate enhancement. The LP-pELISA exhibited a linear response for AFP detection in both buffer and saliva samples over a range of 1.0-800 ng mL-1, with a limit of detection (LOD) reaching 1.0 ng mL-1. The assay also demonstrated good selectivity, repeatability, reproducibility, and stability. The LP-pELISA was further validated by testing spiked human saliva samples, showing its potential for point-of-care diagnosis of congenital disabilities. SIGNIFICANCE: The LP-pELISA is a noninvasive platform showcasing simplicity, cost-effectiveness, and user-friendliness, utilizing laser printing, hydroxyapatite modification, and saliva samples to efficiently detect AFP. Beyond its application for AFP, this method's versatility extends to other biomarkers, positioning it as a catalyst for the evolution of paper-based biosensors. The LP-pELISA holds promise as a transformative tool for point-of-care diagnostics, fostering advancements in healthcare with its innovative technology.
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Colorimetria , Durapatita , Ensaio de Imunoadsorção Enzimática , Lasers , Papel , Saliva , alfa-Fetoproteínas , Humanos , Saliva/química , Durapatita/química , alfa-Fetoproteínas/análise , Impressão , Ouro/química , Limite de Detecção , Anticorpos Imobilizados/imunologia , Anticorpos Imobilizados/químicaRESUMO
Repurposing discarded cells stands as a groundbreaking paradigm shift in sustainable biotechnology, with profound implications across diverse industrial sectors. Our study proposes a transformative concept by harnessing histone proteins from discarded CHO cells to produce bioactive peptides. We systematically isolated and hydrolyzed histones using Trypsin and Neutrase enzymes, optimizing reaction conditions. Ultrafiltration yielded distinct peptide fractions (<3 kDa and 3-10 kDa), which we analyzed for DPP-IV inhibition, antioxidant potential, and other activities. Furthermore, LC-Q-TOF-MS analysis and in silico tools unveiled the structural composition of bioactive peptides within these fractions. Three peptide sequences with high bioactivity potential were identified: KLPFQR, VNRFLR, and LSSCAPVFL. Our findings demonstrated exceptional DPP-IV inhibition, potent antioxidant effects, and effective anti-lipid peroxidation activities, surpassing reference compounds. Hemolytic activity assessment indicated promising biocompatibility, enhancing therapeutic application prospects. Pioneering the strategic repurposing of discarded cells, this research addresses cost-efficiency in cell-based studies and promotes sustainable use of biological resources across sectors. This novel approach offers an efficient, eco-friendly method for bioactive molecule procurement and resource management, revolutionizing cell culture studies and biotechnological applications.
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Biotecnologia , Cricetulus , Peptídeos , Células CHO , Animais , Peptídeos/química , Peptídeos/farmacologia , Biotecnologia/métodos , Antioxidantes/farmacologia , Antioxidantes/química , Histonas/metabolismo , Histonas/química , Sequência de Aminoácidos , Hemólise/efeitos dos fármacosRESUMO
Water pollution caused by pesticides is a significant threat to the environment and human health. Silver and gold nanoparticle (AgNPs, AuNPs)-based biosensors are affordable tools, ideal for environmental monitoring. Microfluidic paper-based devices (µPADs) are a promising approach for on-site testing, but few studies have explored the use of laser printing (LP) for µPAD-based biosensors. This study investigates the feasibility of using laser printing to fabricate paper-based biosensors for pesticide detection in water samples. The µPAD was designed and optimized by using different filter paper porosities, patterns, and channel thicknesses. The developed LP-µPAD was used to sense the pesticide atrazine in water through colorimetric assessments using a smartphone-assisted image analysis. The analytical assessment showed a limit of detection (LOD) of 3.5 and 10.9 µM for AgNPs and AuNPs, respectively. The sensor had high repeatability and reproducibility. The LP-µPAD also demonstrated good recovery and functionality in simulated contaminated water. Furthermore, the detection of pesticides was found to be specific under the influence of interferents, such as NaCl and pH levels. By combining laser printing and nanoparticles, the proposed sensor could contribute to developing effective and low-cost solutions for monitoring water quality that are widely accessible.
RESUMO
Nanomaterials and nanotechnology offer promising opportunities in point-of-care (POC) diagnostics and therapeutics due to their unique physical and chemical properties. POC platforms aim to provide rapid and portable diagnostic and therapeutic capabilities at the site of patient care, offering cost-effective solutions. Incorporating nanomaterials with distinct optical, electrical, and magnetic properties can revolutionize the POC industry, significantly enhancing the effectiveness and efficiency of diagnostic and theragnostic devices. By leveraging nanoparticles and nanofibers in POC devices, nanomaterials have the potential to improve the accuracy and speed of diagnostic tests, making them more practical for POC settings. Technological advancements, such as smartphone integration, imagery instruments, and attachments, complement and expand the application scope of POCs, reducing invasiveness by enabling analysis of various matrices like saliva and breath. These integrated testing platforms facilitate procedures without compromising diagnosis quality. This review provides a summary of recent trends in POC technologies utilizing nanomaterials and nanotechnologies for analyzing disease biomarkers. It highlights advances in device development, nanomaterial design, and their applications in POC. Additionally, complementary tools used in POC and nanomaterials are discussed, followed by critical analysis of challenges and future directions for these technologies.
Assuntos
Nanoestruturas , Sistemas Automatizados de Assistência Junto ao Leito , Humanos , Testes Imediatos , Nanoestruturas/química , Nanotecnologia/métodos , SalivaRESUMO
Minimally invasive approaches for cancer diagnosis are an integral step in the quest to improve cancer survival. Liquid biopsies such as blood samples are matrices explored to extract valuable information about the tumor and its state through various indicators, such as proteins, peptides, tumor DNA, or circulating tumor cells. Although these markers are scarce, making their isolation and detection in complex matrices challenging, the development in polymer chemistry producing interesting structures, including molecularly imprinted polymers, branched polymers, nanopolymer composites, and hybrids, allowed the development of enhanced platforms with impressive performance for liquid biopsies analysis. This review describes the latest advances and developments in polymer synthesis and their application for minimally invasive cancer diagnosis. The polymer structures improve the operational performances of biosensors through various processes, such as increased affinity for enhanced sensitivity, improved binding, and avoidance of non-specific interactions for enhanced specificity. Furthermore, polymer-based materials can be a tremendous help in signal amplification of usually low-concentrated targets in the sample. The pros and cons of these materials, how the synthesis process affects their performance, and the device applications for liquid biopsies diagnosis will be critically reviewed to show the essentiality of this technology in oncology and clinical biomedicine.