Quantum Dots for Cancer-Related miRNA Monitoring.

Quantum dots (QDs) possess exceptional optoelectronic properties that enable their use in the most diverse applications, namely, in the medical field. The prevalence of cancer has increased and has been considered the major cause of death worldwide. Thus, there has been a great demand for new methodologies for diagnosing and monitoring cancer in cells to provide an earlier prognosis of the disease and contribute to the effectiveness of treatment. Several molecules in the human body can be considered relevant as cancer markers. Studies published over recent years have revealed that micro ribonucleic acids (miRNAs) play a crucial role in this pathology, since they are responsible for some physiological processes of the cell cycle and, most important, they are overexpressed in cancer cells. Thus, the analytical sensing of miRNA has gained importance to provide monitoring during cancer treatment, allowing the evaluation of the disease's evolution. Recent methodologies based on nanochemistry use fluorescent quantum dots for sensing of the miRNA. Combining the unique characteristics of QDs, namely, their fluorescence capacity, and the fact that miRNA presents an aberrant expression in cancer cells, the researchers created diverse strategies for miRNA monitoring. This review aims to present an overview of the recent use of QDs as biosensors in miRNA detection, also highlighting some tutorial descriptions of the synthesis methods of QDs, possible surface modification, and functionalization approaches.

Minimizing the Silver Free Ion Content in Starch Coated Silver Nanoparticle Suspensions with Exchange Cationic Resins.

This work describes the optimization of a methodology for the reduction of silver ions from silver nanoparticle suspensions obtained from low-yield laboratory procedures. The laboratory synthesis of silver nanoparticles following a bottom-up approach starting from silver nitrate, originates silver ions that were not reduced to their fundamental state for nanoparticles creation at the end of the process. However, it is well known that silver ions can easily influence chemical assays due to their chemical reactivity properties and can limit biological assays since they interfere with several biological processes, namely intracellular ones, leading to the death of living cells or organisms. As such, the presence of silver ions is highly undesirable when conducting biological assays to evaluate the influence of silver nanoparticles. We report the development of an easy, low-cost, and rapid methodology that is based on cation exchange resins to minimize the silver ion content in a raw suspension of silver nanoparticles while preserving the integrity of the nanomaterials. This procedure preserves the physical-chemical properties of the nanoparticles, thus allowing the purified nanoparticulate systems to be biologically tested. Different types of cationic resins were tested, and the developed methodology was optimized by changing several parameters. A reduction from 92% to 10% of free silver/total silver ratio was achieved when using the Bio-Rad 50W-X8 100-200 mesh resin and a contact time of 15 min. Filtration by vacuum was used to separate the used resin from the nanoparticles suspension, allowing it to be further reused, as well as the purified AgNPs suspension.

Formulation of Nano/Micro-Carriers Loaded with an Enriched Extract of Coffee Silverskin: Physicochemical Properties, In Vitro Release Mechanism and In Silico Molecular Modeling.

Designing strategies for an effective transformation of food waste into high-value products is a priority to address environmental sustainability concerns. Coffee silverskin is the major by-product of the coffee roasting industry, being rich in compounds with health benefits. Such composition gives it the potential to be transformed into high-value products. In this study, coffee silverskin extracts were enriched, regarding caffeine and chlorogenic acid contents, by adsorbent column chromatography. The compounds content increased 3.08- and 2.75-fold, respectively, compared to the original extract. The enriched fractions were loaded into nano-phytosomes or cholesterol-incorporated nano-phytosomes (first coating layers) to improve the physiochemical properties and permeation rate. These nano-lipid carriers were also subjected to a secondary coating with different natural polymers to improve protection and stability against degradation. In parallel, and for comparison, different natural polymers were also used as first coating layers. The produced particles were evaluated regarding product yield, encapsulation efficiency, loading capacity, particle size, surface charge, and in vitro release simulating gastrointestinal conditions. All samples exhibited anionic surface charge. FTIR and molecular docking confirmed interactions between the phytoconstituents and lipid bilayers. The best docking score was observed for 5-caffeoylquinic acid (chlorogenic acid) exhibiting a stronger hydrogen binding to the lipid bilayer. Among several kinetic models tested, the particle release mechanism fitted well with the First-order, Korsmeyer-Peppas, and Higuchi models. Moreover, most of the formulated particles followed the diffusion-Fick law and anomalous transport.

From Impure to Purified Silver Nanoparticles: Advances and Timeline in Separation Methods.

AgNPs have exceptional characteristics that depend on their size and shape. Over the past years, there has been an exponential increase in applications of nanoparticles (NPs), especially the silver ones (AgNPs), in several areas, such as, for example, electronics; environmental, pharmaceutical, and toxicological applications; theragnostics; and medical treatments, among others. This growing use has led to a greater exposure of humans to AgNPs and a higher risk to human health and the environment. This risk becomes more aggravated when the AgNPs are used without purification or separation from the synthesis medium, in which the hazardous synthesis precursors remain unseparated from the NPs and constitute a severe risk for unnecessary environmental contamination. This review examines the situation of the available separation methods of AgNPs from crude suspensions or real samples. Different separation techniques are reviewed, and relevant data are discussed, with a focus on the sustainability and efficiency of AgNPs separation methods.

You Don't Learn That in School: An Updated Practical Guide to Carbon Quantum Dots.

Carbon quantum dots (CQDs) have started to emerge as candidates for application in cell imaging, biosensing, and targeted drug delivery, amongst other research fields, due to their unique properties. Those applications are possible as the CQDs exhibit tunable fluorescence, biocompatibility, and a versatile surface. This review aims to summarize the recent development in the field of CQDs research, namely the latest synthesis progress concerning materials/methods, surface modifications, characterization methods, and purification techniques. Furthermore, this work will systematically explore the several applications CQDs have been subjected to, such as bioimaging, fluorescence sensing, and cancer/gene therapy. Finally, we will briefly discuss in the concluding section the present and future challenges, as well as future perspectives and views regarding the emerging paradigm that is the CQDs research field.

Silver Nanoparticles as Carriers of Anticancer Drugs for Efficient Target Treatment of Cancer Cells.

Since the last decade, nanotechnology has evolved rapidly and has been applied in several areas, such as medicine, pharmaceutical, microelectronics, aerospace, food industries, among others. The use of nanoparticles as drug carriers has been explored and presents several advantages, such as controlled and targeted release of loaded or coupled drugs, and the improvement of the drug's bioavailability, in addition to others. However, they also have some limitations, related to their in vivo toxicity, which affects all organs including the healthy ones, and overall improvement in the disease treatment, which can be unnoticeable or minimal. Silver nanoparticles have been increasingly investigated due to their peculiar physical, chemical, and optical properties, which allows them to cover several applications, namely in the transport of drugs to a specific target in the body. Given the limitations of conventional cancer chemotherapy, which include low bioavailability and the consequent use of high doses that cause adverse effects, strategies that overcome these difficulties are extremely important. This review embraces an overview and presentation about silver nanoparticles used as anticancer drug carrier systems and focuses a discussion on the state of the art of silver nanoparticles exploited for transport of anticancer drugs and their influence on antitumor effects.

Starch-Capped AgNPs' as Potential Cytotoxic Agents against Prostate Cancer Cells.

One of the major therapeutic approaches of prostate cancer (PC) is androgen deprivation therapy (ADT), but patients develop resistance within 2-3 years, making the development of new therapeutic approaches of great importance. Silver nanoparticles (AgNPs) synthesized through green approaches have been studied as anticancer agents because of their physical-chemical properties. This study explored the cytotoxic capacity of starch-capped AgNPs, synthesized through green methods, in LNCaP and in PC-3 cells, a hormonal-sensitive and hormone-resistant PC cell line, respectively. These AgNPs were synthesized in a microwave pressurized synthesizer and characterized by ultraviolet-visible (UV-Vis) spectroscopy, transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). Their cytotoxicity was assessed regarding their ability to alter morphological aspect (optical microscopy), induce damage in cytoplasmic membrane (Trypan Blue Assay), mitochondria (WST-1 assay), cellular proliferation (BrdU assay), and cell cycle (Propidium iodide and flow-cytometry). AgNPs showed surface plasmon resonance (SPR) of approximately 408 nm and average size of 3 nm. The starch-capped AgNPs successfully induced damage in cytoplasmic membrane and mitochondria, at concentrations equal and above 20 ppm. These damages lead to cell cycle arrest in G0/G1 and G2/M, blockage of proliferation and death in LNCaP and PC-3 cells, respectively. This data shows these AgNPs' potential as anticancer agents for the different stages of PC.

Cytotoxic Effect of Silver Nanoparticles Synthesized by Green Methods in Cancer.

Cancer is a major public health problem, but despite the several treatment approaches available, patients develop resistance in short time periods, making overcoming resistance or finding more efficient treatments an imperative challenge. Silver nanoparticles (AgNPs) have been described as an alternative option due to their physicochemical properties. The scope of this review was to systematize the available scientific information concerning these characteristics in AgNPs synthesized according to green chemistry's recommendations as well as their cytotoxicity in different cancer models. This is the first paper analyzing, correlating, and summarizing AgNPs' main parameters that modulate their cellular effect, including size, shape, capping, and surface plasmon resonance profile, dose range, and exposure time. It highlights the strong dependence of AgNPs' cytotoxic effects on their characteristics and tumor model, making evident the strong need of standardization and full characterization. AgNPs' application in oncology research is a new, open, and promising field and needs additional studies.

Urtica spp.: Phenolic composition, safety, antioxidant and anti-inflammatory activities.

Urtica dioica and other less studied Urtica species (Urticaceae) are often used as a food ingredient. Fifteen hydroxycinnamic acid derivatives and sixteen flavonoids, flavone and flavonol-type glycosides were identified in hydroalcoholic extracts from aerial parts of Urtica dioica L., Urtica urens L. and Urtica membranacea using HPLC-PDA-ESI/MSn. Among them, the 4-caffeoyl-5-p-coumaroylquinic acid and three statin-like 3-hydroxy-3-methylglutaroyl flavone derivatives were identified for the first time in Urtica urens and U. membranacea respectively. Urtica membranacea showed the higher content of flavonoids, mainly luteolin and apigenin C-glycosides, which are almost absent in the other species studied. In vitro, Urtica dioica exhibited greater antioxidant activity but Urtica urens exhibited stronger anti-inflammatory potential. Interestingly, statin-like compounds detected in Urtica membranacea have been associated with hypocholesterolemic activity making this plant interesting for future investigations. None of the extracts were cytotoxic to macrophages and hepatocytes in bioactive concentrations (200 and 350µg/mL), suggesting their safety use in food applications.

Antioxidant capacity automatic assay based on inline photogenerated radical species from L-glutathione-capped CdTe quantum dots.

This work aimed at the development of a methodology implemented in an automatic flow system for determination of the antioxidant capacity in food samples, based on the luminol oxidation by inline photogenerated radical species from cadmium telluride nanoparticles capped with L-glutathione. Radical species were generated inline by a high-power visible light obtained by Light Emitting Diodes (LEDs) assembled in a multipumping flow system (MPFS). The use of visible light instead of UV radiation allowed the development of a new methodology for antioxidant capacity determination, more environment friendly and to circumvent the risk for UV photo-induced degradation of sample antioxidant compounds. Additionally, the formation of superoxide radical species was theoretically predicted considering the variation of the redox potential with the size of CdTe QDs and the values of redox potential of the oxidizing and oxidable species present in the irradiated medium. The obtained results of trolox equivalent antioxidant capacity (TEAC) from the analysis of commercial beverages were compared with the results of ABTS and DPPH batch assays through Spearman's-Rho correlation coefficients and no correlation was found (for ABTS: ρ=0.2, p<0.6 and for DPPH: ρ=0.5, p<0.1) since the mechanism of action of the proposed methodology was based on the scavenging capacity of ROS species rather than the reduction of a colored oxidant. An analytical linear response range between 0.0001 and 0.005mmol L(-1) of trolox and a limit of detection of 0.00005mmol L(-1) was found. The QDs based MPFS methodology allowed a determination rate of about 79h(-1), a total waste generation of 20.5mL h(-1) and the consumption of 0.100mg h(-1) of QDs and 2.1mg h(-1) of luminol.

Immobilization of Distinctly Capped CdTe Quantum Dots onto Porous Aminated Solid Supports.

Immobilization of quantum dots (QDs) onto solid supports could improve their applicability in the development of sensing platforms and solid-phase reactors by allowing the implementation of reusable surfaces and the execution of repetitive procedures. As the reactivity of QDs relies mostly on their surface chemistry, immobilization could also limit the disruption of solution stability that could prevent stable measurements. Herein, distinct strategies to immobilize QDs onto porous aminated supports, such as physical adsorption and the establishment of chemical linking, were evaluated. This work explores the influence of QD capping and size, concentration, pH, and contact time between the support and the QDs. Maximum QD retention was obtained for physical adsorption assays. Freundlich and Langmuir isotherms were used to analyze the equilibrium data. Gibbs free energy, enthalpy, and entropy were calculated and the stability of immobilized QDs was confirmed.

Competitive metal-ligand binding between CdTe quantum dots and EDTA for free Ca2+ determination.

In this work, a fluorometric approach for the selective determination of calcium by using CdTe nanocrystals as chemosensors, was developed. The quantum dots interacted not with the metal, but with a ligand that also bonded the metal. The fluorescence response was modulated by the extension of the competitive metal-ligand binding, and therefore the amount of free ligand. CdTe quantum dots (QDs) with different capping layers were evaluated, as the QDs surface chemistry and capping nature affected recognition, thus the magnitude of the ensuing fluorescence quenching. The developed procedure was automated by using a multipumping flow system. Upon optimization, thioglycolic acid (TGA) and EDTA were selected as capping and ligand, respectively, providing a linear working range for calcium concentrations between 0.80-3.20 mg L(-1), and a detection limit of 0.66 mg L(-1). A quenching mechanism relying on nanocrystal destabilization upon detachment of surface Cd by the ligand was proposed.

Fluorescence enhancement of CdTe MPA-capped quantum dots by glutathione for hydrogen peroxide determination.

The manipulation of the surface chemistry of semiconductor nanocrystals has been exploited to implement distinct sensing strategies in many analytical applications. In this work, reduced glutathione (GSH) was added at reaction time, as an electron-donor ligand, to markedly increase the quantum yield and the emission efficiency of MPA-capped CdTe quantum dots. The developed approach was employed in the implementation of an automated flow methodology for hydrogen peroxide determination, as this can oxidize GSH preventing its surface passivating effect and producing a manifest fluorescence quenching. After optimization, linear working calibration curve for hydrogen peroxide concentrations between 0.0025% and 0.040% were obtained (n=6), with a correlation coefficient of 0.9975. The detection limit was approximately 0.0012%. The developed approach was employed in the determination of H2O2 in contact lens preservation solutions and the obtained results complied with those furnished by the reference method, with relative deviations comprised between -1.18 and 4.81%.

Application of quantum dots as analytical tools in automated chemical analysis: a review.

Colloidal semiconductor nanocrystals or quantum dots (QDs) are one of the most relevant developments in the fast-growing world of nanotechnology. Initially proposed as luminescent biological labels, they are finding new important fields of application in analytical chemistry, where their photoluminescent properties have been exploited in environmental monitoring, pharmaceutical and clinical analysis and food quality control. Despite the enormous variety of applications that have been developed, the automation of QDs-based analytical methodologies by resorting to automation tools such as continuous flow analysis and related techniques, which would allow to take advantage of particular features of the nanocrystals such as the versatile surface chemistry and ligand binding ability, the aptitude to generate reactive species, the possibility of encapsulation in different materials while retaining native luminescence providing the means for the implementation of renewable chemosensors or even the utilisation of more drastic and even stability impairing reaction conditions, is hitherto very limited. In this review, we provide insights into the analytical potential of quantum dots focusing on prospects of their utilisation in automated flow-based and flow-related approaches and the future outlook of QDs applications in chemical analysis.

Photoactivation by visible light of CdTe quantum dots for inline generation of reactive oxygen species in an automated multipumping flow system.

Quantum dots (QD) are semiconductor nanocrystals able to generate free radical species upon exposure to an electromagnetic radiation, usually in the ultraviolet wavelength range. In this work, CdTe QD were used as highly reactive oxygen species (ROS) generators for the control of pharmaceutical formulations containing epinephrine. The developed approach was based on the chemiluminometric monitoring of the quenching effect of epinephrine on the oxidation of luminol by the produced ROS. Due to the relatively low energy band-gap of this chalcogenide a high power visible light emitting diode (LED) lamp was used as photoirradiation element and assembled in a laboratory-made photocatalytic unit. Owing to the very short lifetime of ROS and to ensure both reproducible generation and time-controlled reaction implementation and development, all reactional processes were implemented inline by using an automated multipumping micro-flow system. A linear working range for epinephrine concentration of up to 2.28×10(-6) mol L(-1) (r=0.9953; n=5) was verified. The determination rate was about 79 determinations per hour and the detection limit was about 8.69×10(-8) mol L(-1). The results obtained in the analysis of epinephrine pharmaceutical formulations by using the proposed methodology were in good agreement with those furnished by the reference procedure, with relative deviations lower than 4.80%.

Exploiting adsorption and desorption at solid-liquid interface for the fluorometric monitoring of glibenclamide in adulterated drinks.

Nowadays, the use of a drug to modify a person's behavior with criminal intentions has become a growing public concern. In fact, stealthy drink spiking with certain drugs can cause the incapacitation of a potential victim of assault and in extreme cases can even lead to death. Belonging to the group of drugs used to commit drug-facilitated crimes is glibenclamide, which not only exhibits high sedation secondary effects but when subject to an overdose intake can lead to intense hypoglycemic episodes that could end with death. Suicide attempts and homicides through overdose with glibenclamide have already been reported. In this work and for the first time, it was developed a new methodology for detection of glibenclamide in spiked liquid samples (teas) by fluorometry (λ(ex)=300 nm; λ(em)=404 nm). The novel methodology was also implemented in a miniaturized and portable automatic flow system based in the concept of multipumping with an in-line pre-separation unit. The separation of the drug from the liquid samples was achieved through adsorption of the drug into activated charcoal packed within a mini column followed by elution with a solution composed by ethanol, hydrochloric acid and the surfactant CTAB (70%, 1.0 mol L(-1), 0.01 mol L(-1), respectively). The results allowed to obtain a linear working range for glibenclamide concentrations of up to 50 mg L(-1) (r=0.9999) and the detection limit was about 0.81 mg L(-1) of glibenclamide.

Chemiluminometric evaluation of melatonin and selected melatonin precursors' interaction with reactive oxygen and nitrogen species.

Melatonin is a hormone, a derivative of tryptophan, that possesses a potent scavenging capacity for the most reactive and dangerous free radicals, being an important protection against oxidative stress. In this work, an automated flow-based procedure for assessment of melatonin, tryptophan, and 5-hydroxytryptophan scavenging capacity was developed. The presented methodology involved a multi-pumping flow system and exploited the ability of selected compounds to inhibit the chemiluminescence reaction of luminol with hydrogen peroxide, hydroxyl radical, and peroxynitrite anion. The system was based on the use of several solenoid actuated micro-pumps as the only active components of the flow manifold. This enabled the reproducible insertion and efficient mixing of very low volumes of sample and reagents as well as the transportation of the sample zone toward detection for monitoring the chemiluminometric response. Furthermore, the high versatility of the proposed multi-pumping flow system allowed the implementation of distinct reactions for the in-line generation of the different reactive species assayed without requiring physical reconfiguration. The results obtained demonstrated that 5-hydroxytryptophan is the most potent scavenger, followed by melatonin and tryptophan. The developed multi-pumping flow system exhibited good measurement precision (relative standard deviations typically <2%, n=10), low operational costs, and low reagent consumption.

Automatic miniaturized fluorometric flow system for chemical and toxicological control of glibenclamide.

In this work, and for the first time, it was developed an automatic and fast screening miniaturized flow system for the toxicological control of glibenclamide in beverages, with application in forensic laboratory investigations, and also, for the chemical control of commercially available pharmaceutical formulations. The automatic system exploited the multipumping flow (MPFS) concept and allowed the implementation of a new glibenclamide determination method based on the fluorometric monitoring of the drug in acidic medium (λ(ex)=301 nm; λ(em)=404 nm), in the presence of an anionic surfactant (SDS), promoting an organized micellar medium to enhance the fluorometric measurements. The developed approach assured good recoveries in the analysis of five spiked alcoholic beverages. Additionally, a good agreement was verified when comparing the results obtained in the determination of glibenclamide in five commercial pharmaceutical formulations by the proposed method and by the pharmacopoeia reference procedure.

Automated determination of diazepam in spiked alcoholic beverages associated with drug-facilitated crimes.

In this work, a multipumping flow system (MPFS) coupled to a photodegradation unit was developed, for the first time, for the determination of diazepam (a benzodiazepine) in spiked alcoholic beverages by fluorimetry. The main features of MPFS such as, high portability, versatility and straightforward automation and control combined with the efficiency and simplicity of photodegradation and the selectivity and sensitivity of fluorimetric detection makes the developed analytical methodology an attractive tool and a valuable contribution for the prevention of drug-facilitated crimes (DFC). Drug-facilitated crimes involve the unauthorized administration of strong central nervous system depressant drugs, which have the capability of preventing victims from resist to the action of the perpetrator or fighting off. Most often, the drugs identified as being used in DFC are surreptitiously placed in drinks served to potential victims in entertainment places, like night clubs. Five commercial alcoholic beverages (Eristoff, Smirnoff, Bacardi, Dry Gin and Brazilian Cachaça 51) spiked with diazepam were analyzed by the proposed methodology, and the results revealed good agreement with those obtained through a HPLC comparison procedure. Relative deviations comprised between -1.97 and 2.05% were achieved, and additionally, the application of a paired t-test, revealed the absence of any statistical difference for a confidence level of 95% (n=5). The detection limit was approximately 2.02 mg L(-1).

Diazepam fluorimetric monitoring upon photo-degradation in an automatic miniaturized flow system.

The present work describes the fully integration in line of a photo-degradation unit, comprising a low pressure UV lamp, in a Multipumping Flow System (MPFS), for the fluorimetric chemical control of commercially available pharmaceutical formulations containing diazepam. The utilization of an organized micellar medium provided enhanced fluorescence emission. The results allowed to obtain a linear working range for diazepam concentrations of up to 40 mg L(-1) (r = 0.9998) and the detection limit was about 0.97 mg L(-1). The results obtained by the miniaturized and automatic flow system were in agreement with those furnished by the reference procedure, with relative deviations comprised between -2.09% and 2.13%.