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In this study, the effects of the ripening process and fruit powder addition on the physical, chemical, total phenolic content, antioxidant capacity, volatile compounds, and sensory properties of Kashar cheese were determined. Total phenol content, antioxidant capacity, volatile compounds, and fatty acid esters were determined by Folin Ciocalteu, DPPH, SPME, and GC-MS, GC-FID, respectively. Of the 27 fatty acids identified in cheeses, palmitic, oleic, myristic, and stearic acids were found to have the highest ratios, respectively. While the total amount of phenol substance was 144.44 mg GAE/L in fresh, it increased to 374.84 mg GAE/L with ripening and 520.26 mg GAE/L with ripening + plant. A total of 58 volatile compounds, including 14 alcohols, 10 acids, 9 ketones, 9 hydrocarbons, 7 esters, 7 aldehydes,and 2 sulfur compounds, were detected in cheese. Alcohol (27.20%) in fresh kashar, acid (61.40%) in ripened kashar, and ketone (43.73%) in ripened + plant kashar were the volatile compounds groups determined at the highest rate. The ripening process and plant addition did not contribute positively to the sensory properties of the cheeses.
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Biofilm is a complex consortium of microorganisms attached to biotic or abiotic surfaces and live in self-produced or acquired extracellular polymeric substances (EPSs). EPSs are mainly formed by lipids, polysaccharides, proteins, and extracellular DNAs. The adherence to the surface of microbial communities is seen in food, medical, dental, industrial, and environmental fields. Biofilm development in food processing areas challenges food hygiene, and human health. In addition, bacterial attachment and biofilm formation on medical implants inside human tissue can cause multiple critical chronic infections. More than 30 years of international research on the mechanisms of biofilm formation have been underway to address concerns about bacterial biofilm infections. Antibiofilm strategies contain cold atmospheric plasma, nanotechnological, phage-based, antimicrobial peptides, and quorum sensing inhibition. In the last years, the studies on environmentally-friendly techniques such as essential oils and bacteriophages have been intensified to reduce microbial growth. However, the mechanisms of the biofilm matrix formation are still unclear. This review aims to discuss the latest antibiofilm therapeutic strategies against biofilm-forming bacteria.
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Infecções Bacterianas , Biofilmes , Humanos , Percepção de Quorum , Bactérias , Infecções Bacterianas/tratamento farmacológico , PolissacarídeosRESUMO
The incidence of cancer has exhibited an increasing trend in recent years because of many reasons such as environmental and nutritional factors. There is a great need for the development of new and natural molecules with lower side effects in the therapy of cancer. It was aimed to evaluate the antiproliferative effect of semi-purified triterpene glycosides of Holothuria poli on different human cancer cell lines. The body walls of H. poli as the main sources of saponins were used and the saponin content of the extract was characterized by MALDI-TOF/MS. The antiproliferation activity of the characterized extract was tested on cancer cell lines. The extract showed antiproliferative effect on the studied cancer cell lines. The mass analysis results reveal that Holothurin A is one of the saponins within the extract. The measured IC50 values were found as 31.41 ± 2.20, 77.45 ± 0.23, and 34.79 ± 0.90 µg mL-1 for HT-29, UPCI-SCC-131, and T84 cell lines, respectively. H. poli secretes not only specific saponins but also a cocktail of them. Specific versus. cocktails of the saponins and by also applying organic modification must be studied in further research to understand their mechanisms in the antiproliferation studies since this paper reveals promising results.
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Holothuria , Saponinas , Triterpenos , Animais , Linhagem Celular Tumoral , Holothuria/metabolismo , Humanos , Estrutura Molecular , Extratos Vegetais/metabolismo , Saponinas/farmacologia , Triterpenos/farmacologiaRESUMO
Organ-on-a-chip systems are miniaturized microfluidic 3D human tissue and organ models designed to recapitulate the important biological and physiological parameters of their in vivo counterparts. They have recently emerged as a viable platform for personalized medicine and drug screening. These in vitro models, featuring biomimetic compositions, architectures, and functions, are expected to replace the conventional planar, static cell cultures and bridge the gap between the currently used preclinical animal models and the human body. Multiple organoid models may be further connected together through the microfluidics in a similar manner in which they are arranged in vivo, providing the capability to analyze multiorgan interactions. Although a wide variety of human organ-on-a-chip models have been created, there are limited efforts on the integration of multisensor systems. However, in situ continual measuring is critical in precise assessment of the microenvironment parameters and the dynamic responses of the organs to pharmaceutical compounds over extended periods of time. In addition, automated and noninvasive capability is strongly desired for long-term monitoring. Here, we report a fully integrated modular physical, biochemical, and optical sensing platform through a fluidics-routing breadboard, which operates organ-on-a-chip units in a continual, dynamic, and automated manner. We believe that this platform technology has paved a potential avenue to promote the performance of current organ-on-a-chip models in drug screening by integrating a multitude of real-time sensors to achieve automated in situ monitoring of biophysical and biochemical parameters.
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Automação/métodos , Técnicas Biossensoriais/métodos , Avaliação Pré-Clínica de Medicamentos/métodos , Organoides/fisiologia , Automação/instrumentação , Técnicas Biossensoriais/instrumentação , Avaliação Pré-Clínica de Medicamentos/instrumentação , Coração/fisiologia , Humanos , Fígado/química , Fígado/fisiologia , Microfluídica , Modelos Biológicos , Miocárdio , Organoides/química , Organoides/efeitos dos fármacosRESUMO
We studied dynamic thiol/disulphide homeostasis, an indicator of oxidative stress, to investigate the effects of newly initiated exercise training on sedentary obese adults. Seventeen sedentary obese adults and 15 normal-weight controls were included in the sample for this study. The obese adults were given a physical exercise training program that lasted twelve weeks. Before and after the exercise training program, blood samples were collected, and serum thiol/disulphide parameters were measured by using a novel technique. Before the start of the exercise training, it was observed that thiol/disulphide homeostasis was impaired, and this impairment was positively correlated with body mass index in sedentary obese adults because of the higher reactive oxygen species production in adipose tissue. However, while the obese participants' body mass index significantly decreased, the thiol/disulphide homeostasis parameters in the obese adults did not change over time as calculated at the baseline and compared to the calculation after the twelve weeks of exercise training. Despite a decrease in body mass index that occurred after the twelve weeks of exercise training, there was a lack of improvement in the obesity-induced impairment of thiol/disulphide homeostasis, which suggests that a newly initiated exercise training program may lead to oxidative stress.
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Dissulfetos/metabolismo , Exercício Físico , Homeostase , Obesidade/reabilitação , Estresse Oxidativo , Compostos de Sulfidrila/metabolismo , Adulto , Índice de Massa Corporal , Peso Corporal , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade/metabolismo , Comportamento SedentárioRESUMO
In the post-genome age, proteomics is receiving significant attention because they provide an invaluable source of biological structures and functions at the protein level. The search for disease-specific biomarkers for diagnostic and/or therapeutic applications is one of the areas that proteomics is having a significant impact. Thus, the identification of a "good" biomarker enables a more accurate early diagnosis and prognosis of disease. Rapid advancements in mass spectrometry (MS) instrumentation, liquid chromatography MS (LCMS), protein microarray technology, and other protein profiling methodologies have a substantial expansion of our toolbox to identify disease-specific protein and peptide biomarkers. This review covers a selection of widely used proteomic technologies for biomarker discovery. In addition, we describe the most commonly used approaches for diagnosis based on proteomic biomarkers and further discuss trends and critical challenges during development of cost-effective rapid diagnostic tests and microfluidic diagnostic systems based on proteomic biomarkers.
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Biomarcadores/análise , Testes Diagnósticos de Rotina/métodos , Proteômica/métodosRESUMO
The ability of Aeribacillus pallidus E334 to produce pellicle and form a biofilm was studied. Optimal biofilm formation occurred at 60 °C, pH 7.5 and 1.5% NaCl. Extra polymeric substances (EPS) were composed of proteins and eDNA (21.4 kb). E334 formed biofilm on many surfaces, but mostly preferred polypropylene and glass. Using CLSM analysis, the network-like structure of the EPS was observed. The A. pallidus biofilm had a novel eDNA content. DNaseI susceptibility (86.8% removal) of eDNA revealed its importance in mature biofilms, but the purified eDNA was resistant to DNaseI, probably due to its extended folding outside the matrix. Among 15 cleaning agents, biofilms could be removed with alkaline protease and sodium dodecyl sulphate (SDS). The removal of cells from polypropylene and biomass on glass was achieved with combined SDS/alkaline protease treatment. Strong A. pallidus biofilms could cause risks for industrial processes and abiotic surfaces must be taken into consideration in terms of sanitation procedures.
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Bacillaceae/crescimento & desenvolvimento , Biofilmes/crescimento & desenvolvimento , Desinfetantes/farmacologia , Saneamento/métodos , Bacillaceae/efeitos dos fármacos , Bacillaceae/genética , Bacillaceae/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes/efeitos dos fármacos , Biomassa , DNA Bacteriano , Temperatura Alta , Concentração de Íons de Hidrogênio , Propriedades de SuperfícieRESUMO
The biofilm characteristics of many endospore-forming bacilli, especially the thermophiles are still unclear. In this study, a detailed identification and description of biofilm production characteristics of totally 145 isolates and reference strains belonging to Bacillaceae family, displaying thermophilic (n = 115), facultative thermophilic (n = 24) and mesophilic (n = 6) growth from genera Anoxybacillus, Geobacillus, Thermolongibacillus, Aeribacillus, Brevibacillus, Paenibacillus and Bacillus were presented. The incubation temperatures were adjusted to 37, 45 and 55-65 °C for mesophiles, facultative thermophiles, and thermophiles, respectively. The bacilli were evaluated based on their colony morphotypes on Congo red (CR) agar, their complex exopolysaccharide production on calcofluor supplemented tryptic soy agar, and as well as their pellicle formation at the liquid-air surface in tryptic soy broth cultures. Their biofilm production capabilities were also tested on abiotic surfaces of both polystyrene and stainless steel by crystal violet binding assay and viable biofilm cell enumerations, respectively. As a result, the biofilm production capacities of Bacillaceae members from genera to species level, the effects of osmolarity, temperature, incubation time and abiotic surfaces on biofilm formation as well as the CR morphotypes associated with the biofilm production were able to reveal in a wide group of bacilli. Besides, general enrichment-inoculation approaches and methodologies were also offered, which allow and facilitate the screening and determining the biofilm producing endospore forming bacilli.
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Bacillaceae/classificação , Bacillaceae/fisiologia , Biofilmes/crescimento & desenvolvimento , Bacillaceae/crescimento & desenvolvimento , Bacillaceae/isolamento & purificação , Meios de Cultura/química , Temperatura Alta , Concentração Osmolar , Poliestirenos , Esporos Bacterianos/crescimento & desenvolvimento , Esporos Bacterianos/metabolismo , Aço Inoxidável , Temperatura , Fatores de TempoRESUMO
Continual monitoring of secreted biomarkers from organ-on-a-chip models is desired to understand their responses to drug exposure in a noninvasive manner. To achieve this goal, analytical methods capable of monitoring trace amounts of secreted biomarkers are of particular interest. However, a majority of existing biosensing techniques suffer from limited sensitivity, selectivity, stability, and require large working volumes, especially when cell culture medium is involved, which usually contains a plethora of nonspecific binding proteins and interfering compounds. Hence, novel analytical platforms are needed to provide noninvasive, accurate information on the status of organoids at low working volumes. Here, we report a novel microfluidic aptamer-based electrochemical biosensing platform for monitoring damage to cardiac organoids. The system is scalable, low-cost, and compatible with microfluidic platforms easing its integration with microfluidic bioreactors. To create the creatine kinase (CK)-MB biosensor, the microelectrode was functionalized with aptamers that are specific to CK-MB biomarker secreted from a damaged cardiac tissue. Compared to antibody-based sensors, the proposed aptamer-based system was highly sensitive, selective, and stable. The performance of the sensors was assessed using a heart-on-a-chip system constructed from human embryonic stem cell-derived cardiomyocytes following exposure to a cardiotoxic drug, doxorubicin. The aptamer-based biosensor was capable of measuring trace amounts of CK-MB secreted by the cardiac organoids upon drug treatments in a dose-dependent manner, which was in agreement with the beating behavior and cell viability analyses. We believe that, our microfluidic electrochemical biosensor using aptamer-based capture mechanism will find widespread applications in integration with organ-on-a-chip platforms for in situ detection of biomarkers at low abundance and high sensitivity.
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The limited knowledge about the inheritance of traits in roses makes the efficient development of rose varieties challenging. In order to achieve breeding goals, the inheritance of traits needs to be explored. Additionally, for the inheritance of a trait like scent, which remains a mystery, it is crucial to know the success of parental traits in transmitting them to the next generation. Understanding this allows for accurate parental selection, ensuring sustainability in meeting market demand and providing convenience to breeders. The aim of this study was to assess the success of cross-combinations between scented old garden roses and hybrid tea roses used in cut roses in transferring their existing traits, with the objective of achieving scented cut roses. The evaluated traits included recurrent blooming, flower stem length, flower diameter, petal number, scent, and bud length of both parents and progenies. The inheritance of these traits was evaluated through theoretical evaluations, including calculating heterosis and heterobeltiosis and determining narrow-sense heritability. The combinations and examined traits were assessed using a hierarchical clustering heat map. The results of this study indicated that flower stem length, flower diameter, petal number, and bud length traits had a moderate degree of narrow-sense heritability, suggesting the influence of non-additive genes on these traits. This study observed a low success rate in obtaining progenies with scent in cross combinations between cut roses and old garden roses, indicating the challenges in obtaining scented genotypes. The discrepancy between the observed phenotypic rates and the expected phenotypic and genotypic rates, according to Punnett squares, suggests that the examined traits could be controlled by polygenic genes. The progenies were observed to exhibit a greater resemblance to old garden roses than hybrid tea roses and did not meet the commercial quality standards for cut flowers. The significant negative heterosis observed in 65.12% (petal number) and 99.61% (flower diameter) of the progenies provides strong evidence of resemblance to old garden roses. Considering these findings, it is recommended to consider old garden roses as parents, taking into account their suitability for other breeding objectives.
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The success of rose breeding programs is low due to poor seed sets and germination rates. Determining fertile parents and cross combinations that show high compatibility could increase the effectiveness of breeding programs. In this study, three rose varieties belonging to Rosa × hybrida (Jumilia, First Red and Magnum), and two old garden rose species (Black Rose and Cabbage Rose) with known ploidy levels were reciprocally crossbred under controlled conditions to determine the successful crosses by checking fertility. The pollen germination rate (PG), crossability rate (CR), seed number per fruit (SNpF), seed production efficiency (SPE), seed germination rate (SGR), fruit weight (FW), seed weight (SW) and stigma number (SiN), etc. were recorded. Comprehensive fertility index value was calculated. Principal component analysis (PCA), correlation matrix, and hierarchical heat map were used to evaluate the data. The findings showed that old garden roses had more viable pollen than hybrid tea roses. The crossing success improved as pollen fertility increased. Also, female parent fertility improved crossing success just as much as pollen fertility. Although the pollen fertility and stigma numbers were low, some combinations had higher CR and SPE. The maximum SPE (from 8.67% to 19.46%) was determined in combinations where Black Rose was the female parent despite the lower stigma number and low pollen fertility. The highest CR was recorded in Black Rose × First Red (94.36%). All combinations in which Black Rose was used as the female parent had a more stable CR. The SNpF of combinations where hybrid rose varieties were female parents and old garden roses were pollen parents was higher than other combinations where hybrid rose varieties were both female and pollen parents. The SPE in intraspecific crosses was lower than that obtained from interspecific crosses. Moreover, the SGR decreased in combinations that produced heavier seeds. The results suggested that SPE is a more accurate parameter than SNpF in demonstrating combination success in breeding programs. Black Rose × First Red, Black Rose × Jumilia, Black Rose × Magnum and Black Rose × Cabbage Rose combinations can be used successfully as the PCA and heat map showed. Black Rose showed better performance as both seed and pollen parents according to the comprehensive fertility index. From the correlation matrix, it is understood that the number of stigmas cannot be an important criterion in parent selection. Old garden roses can be used as parents to increase the success of breeding programs. However, it is necessary to reveal how successful they are in transferring desired characteristics such as scent, petal number, and color.
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Rosa , Rosa/genética , Melhoramento Vegetal , Hibridização Genética , Fertilidade/genética , CháRESUMO
The emerging field of liquid biopsy stands at the forefront of novel diagnostic strategies for cancer and other diseases. Liquid biopsy allows minimally invasive molecular characterization of cancers for diagnosis, patient stratification to therapy, and longitudinal monitoring. Liquid biopsy strategies include detection and monitoring of circulating tumor cells, cell-free DNA, and extracellular vesicles. In this review, we address the current understanding and the role of existing liquid-biopsy-based modalities in cancer diagnostics and monitoring. We specifically focus on the technical and clinical challenges associated with liquid biopsy and biomarker development being addressed by the Liquid Biopsy Consortium, established through the National Cancer Institute. The Liquid Biopsy Consortium has developed new methods/assays and validated existing methods/technologies to capture and characterize tumor-derived circulating cargo, as well as addressed existing challenges and provided recommendations for advancing biomarker assays.
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Ácidos Nucleicos Livres , Vesículas Extracelulares , Células Neoplásicas Circulantes , Humanos , Biópsia Líquida , Ácidos Nucleicos Livres/genética , Biomarcadores , Células Neoplásicas Circulantes/patologiaRESUMO
Surface chemistry critically affects the diagnostic performance of biosensors. An ideal sensor surface should be resistant to nonspecific protein adsorption, yet be conducive to analytical responses. Here a new polymeric material, zwitterionic polypyrrole (ZiPPy), is reported to produce optimal surface condition for biosensing electrodes. ZiPPy combines two unique advantages: the zwitterionic function that efficiently hydrates electrode surface, hindering nonspecific binding of hydrophobic proteins; and the pyrrole backbone, which enables rapid (<7 min), controlled deposition of ZiPPy through electropolymerization. ZiPPy-coated electrodes show lower electrochemical impedance and less nonspecific protein adsorption (low fouling), outperforming bare and polypyrrole-coated electrodes. Moreover, affinity ligands for target biomarkers can be immobilized together with ZiPPy in a single-step electropolymerization. ZiPPy-coated electrodes are developed with specificity for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The prepared sensor detects SARS-CoV-2 antibodies in human saliva down to 50 ng mL-1 , without the need for sample purification or secondary labeling.
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Anticorpos Antivirais/análise , Técnicas Biossensoriais/métodos , COVID-19/diagnóstico , Polímeros/química , Pirróis/química , Técnicas Biossensoriais/instrumentação , COVID-19/virologia , Técnicas Eletroquímicas , Eletrodos , Galvanoplastia , Ouro/química , Humanos , Limite de Detecção , SARS-CoV-2/imunologia , SARS-CoV-2/isolamento & purificação , Saliva/metabolismo , Propriedades de SuperfícieRESUMO
Detecting protein markers in extracellular vesicles (EVs) is becoming a useful tool for basic research and clinical diagnoses. Most EV protein assays, however, require lengthy processes-conjugating affinity ligands onto sensing substrates and affixing EVs with additional labels to maximize signal generation. Here, we present an iPEX (impedance profiling of extracellular vesicles) system, an all-electrical strategy toward fast, multiplexed EV profiling. iPEX adopts one-step electropolymerization to rapidly functionalize sensor electrodes with antibodies; it then detects EV proteins in a label-free manner through impedance spectroscopy. The approach streamlines the entire EV assay, from sensor preparation to signal measurements. We achieved (i) fast immobilization of antibodies (<3 min) per electrode; (ii) high sensitivity (500 EVs/mL) without secondary labeling; and (iii) parallel detection (quadruple) in a single chip. A potential clinical utility was demonstrated by directly analyzing plasma samples from glioblastoma multiforme patients.
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Organs-on-chips have emerged as viable platforms for drug screening and personalized medicine. While a wide variety of human organ-on-a-chip models have been developed, rarely have there been reports on the inclusion of sensors, which are critical in continually measuring the microenvironmental parameters and the dynamic responses of the microtissues to pharmaceutical compounds over extended periods of time. In addition, automation capacity is strongly desired for chronological monitoring. To overcome this major hurdle, in this protocol we detail the fabrication of electrochemical affinity-based biosensors and their integration with microfluidic chips to achieve in-line microelectrode functionalization, biomarker detection and sensor regeneration, allowing continual, in situ and noninvasive quantification of soluble biomarkers on organ-on-a-chip platforms. This platform is almost universal and can be applied to in-line detection of a majority of biomarkers, can be connected with existing organ-on-a-chip devices and can be multiplexed for simultaneous measurement of multiple biomarkers. Specifically, this protocol begins with fabrication of the electrochemically competent microelectrodes and the associated microfluidic devices (~3 d). The integration of electrochemical biosensors with the chips and their further combination with the rest of the platform takes ~3 h. The functionalization and regeneration of the microelectrodes are subsequently described, which require ~7 h in total. One cycle of sampling and detection of up to three biomarkers accounts for ~1 h.
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Técnicas Biossensoriais/instrumentação , Reutilização de Equipamento , Dispositivos Lab-On-A-Chip , Biomarcadores/metabolismo , Eletroquímica , Microeletrodos , Fatores de TempoRESUMO
Geobacillus sp. D413 and Geobacillus toebii E134 are aerobic, non-pathogenic, endospore-forming, obligately thermophilic bacilli. Gram-positive thermophilic bacilli can produce heat-resistant spores. The bacteria are indicator organisms for assessing the manufacturing process's hygiene and are capable of forming biofilms on surfaces used in industrial sectors. The present study aimed to determine the biofilm-forming properties of Geobacillus isolates and how to eliminate this formation with sanitation agents. According to the results, extracellular DNA (eDNA) was interestingly not affected by the DNase I, RNase A, and proteinase K. However, the genomic DNA (gDNA) was degraded by only DNase I. It seemed that the eDNA had resistance to DNase I when purified. It is considered that the enzymes could not reach the target eDNA. Moreover, the eDNA resistance may result from the conserved folded structure of eDNA after purification. Another assumption is that the eDNA might be protected by other extracellular polymeric substances (EPS) and/or extracellular membrane vesicles (EVs) structures. On the contrary, DNase I reduced unpurified eDNA (mature biofilms). Biofilm formation on surfaces used in industrial areas was investigated in this work: the D413 and E134 isolates adhered to all surfaces. Various sanitation agents could control biofilms of Geobacillus isolates. The best results were provided by nisin for D413 (80%) and α-amylase for E134 (98%). This paper suggests that sanitation agents could be a solution to control biofilm structures of thermophilic bacilli.
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Antibacterianos/farmacologia , Bacillaceae/crescimento & desenvolvimento , Biofilmes/crescimento & desenvolvimento , Geobacillus/crescimento & desenvolvimento , Bacillaceae/efeitos dos fármacos , Biofilmes/efeitos dos fármacos , DNA Bacteriano/química , DNA Bacteriano/metabolismo , Desoxirribonuclease I/metabolismo , Endopeptidase K/metabolismo , Geobacillus/efeitos dos fármacos , Nisina/farmacologia , Conformação de Ácido Nucleico , Ribonuclease Pancreático/metabolismo , alfa-Amilases/farmacologiaRESUMO
Rapid spread of coronavirus disease 2019 (COVID-19) is ravaging the globe. Since its first report in December 2019, COVID-19 cases have exploded to over 14 million as of July 2020, claiming more than 600,000 lives. Implementing fast and widespread diagnostic tests is paramount to contain COVID-19, given the current lack of an effective therapeutic or vaccine. This review focuses on a broad description of currently available diagnostic tests to detect either the virus (SARS-CoV-2) or virus-induced immune responses. We specifically explain the working mechanisms of these tests and compare their analytical performance. These analyses will assist in selecting most effective tests for a given application, for example, epidemiology or global pandemic research, population screening, hospital-based testing, home-based and point-of-care testing, and therapeutic trials. Finally, we lay out the shortcomings of certain tests and future needs.
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Schizophrenia treatment may see a paradigm shift due to development of new atypical antipsychotic drugs (APDs), with better tolerability due to more selective dopamine (DA) receptor blockade. Monitoring of these APD candidates in biological fluids is of great importance to reduce the development cost, to clarify the mechanism of action and ultimately to support the demonstration of efficacy of these molecules. Electrochemical approaches have attracted great attention for monitoring DA and APD levels but none of the methods developed so far aimed to screen APD candidates. Herein, by this work, we propose for the first time an electrochemical ligand-binding approach for antipsychotic drug screening where competitive binding of a novel APD and DA to a dopamine D3 receptor (D3R) was investigated by looking at electrochemical signals of DA and drug before and after D3R interaction. D3R peptide was incubated with DA and/or drug first and then changes in electrochemical oxidation signals of free DA and the drug was measured by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). Circular Dichroism spectroscopy was used to investigate the secondary structure of the peptide upon binding with either drug and/or DA.
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Antipsicóticos/farmacologia , Técnicas Biossensoriais/métodos , Dopamina/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Técnicas Eletroquímicas/métodos , Receptores de Dopamina D3/metabolismo , Humanos , Modelos Moleculares , Estrutura Secundária de Proteína/efeitos dos fármacos , Receptores de Dopamina D3/química , Esquizofrenia/tratamento farmacológicoRESUMO
As being the most extensively studied, non-coding, evolutionary conserved, post-transcriptional gene regulators of genome, microRNAs (miRNAs) have taken great attention among various disciplines due to their important roles in biological processes and link with cancer. Due to their diagnostic value, there have been many conventional methods used in detection of miRNAs including northern blotting, quantitative real time PCR (qRT-PCR) and microarray technology besides novel techniques based on various nanotechnology approaches and molecular biology tools including miRNA biosensors. The aim of this review is to explain the importance of miRNAs in biomedical field with an emphasis on early cancer diagnosis by overviewing both research based and commercially available miRNA detection methods in the last decade considering their strengths and weakness with an emphasis on miRNA biosensors.
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Técnicas Biossensoriais/métodos , MicroRNAs/isolamento & purificação , Nanotecnologia/métodos , Técnicas Biossensoriais/tendências , Regulação da Expressão Gênica/genética , Humanos , MicroRNAs/genética , Nanotecnologia/tendênciasRESUMO
Nanoscale extracellular vesicles (EVs) including exosomes (50-150 nm membrane particles) have emerged as promising cancer biomarkers due to the carried genetic information about the parental cells. However the sensitive detection of these vesicles remains a challenge. Here we present a label-free electrochemical sensor to measure the EVs secretion levels of hypoxic and normoxic MCF-7 cells. The sensor design includes two consecutive steps; i) Au electrode surface functionalization for anti-CD81 Antibody and ii) EVs capture. The label-free detection of EVs was done via Differential Pulse Voltammetry (DPV) and Electrochemical Impedance Spectroscopy (EIS). The working linear range for the sensor was 102-109 EVs/ml with an LOD 77 EVs/mL and 379 EVs/ml for EIS and DPV based detection. A blood-abundant protein, RhD was used for the selectivity test. In order to assess the performance of the biosensor, the level of EVs secretion by the human breast cancer MCF-7 cell line was compared with enzyme-linked immunosorbent assays (ELISA) and Nanoparticle Tracking Analysis (NTA). Designed label-free electrochemical sensors utilized for quantification of EVs secretion enhancement due to CoCl2-induced hypoxia and 1.23 fold increase with respect to normoxic conditions was found.