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Bio-oil derived from the pyrolysis of lignocellulosic biomass residues cannot be used directly as biofuel due to the high content of oxygenated compounds. As an alternative, bio-oil must undergo a deoxygenation process, such as catalytic hydrodeoxygenation (HDO). In this sense, this work studied the effect of different concentrations of Ce3+ and Ce4+ in layered double hydroxides (LDHs) in order to obtain mixed oxide catalysts containing NiAlCe (NiO-NiAl2O4-CeO2) with low cost and high performance for the hydrodeoxygenation of anisole as a model bio-oil compound. Mixed oxides were obtained from the thermal decomposition of layered double hydroxides (LDHs) by using terephthalic acid as compensation anion, with molar ratio: Ni2+/(Al3++ M) = 1.0, where M = Ce3+; Ce4+ or Ce3+-Ce4+; and Al/Ce ratios of 9 and 1. Characterization analyses confirmed the formation of LDHs for all materials, although at lower Al/Ce ratio it is observed a loss of crystallinity, due to a greater repulsion and distortion of the layer structure caused by the incorporation, in greater amount, of cations with a high ionic radius. The increase in the content of cerium in mixed oxides also led to an increase in the acidity of the catalysts, in addition to a reduction in the surface area, justified by the pore blockage by CeO2 on the catalyst surface. The greater structural and thermal stability was evidenced in the LDHs derived from Ce4+, as well as a greater dispersion of the NiO phase in the corresponding mixed oxides, and therefore presenting greater anisole conversion. The obtained data indicated that the presence of Ce4+ ions on the catalyst surface was decisive in the conversion of anisole to cyclohexane, the main deoxygenated product, obtained via the direct deoxygenation and hydrogenation mechanism.

A simple and fast aqueous synthesis of silver selenide quantum dots is proposed with the aid of multivariate optimization.

Candida albicans is the main cause of superficial candidiasis. While the antifungals available are defied by biofilm formation and resistance emergence, antimicrobial photodynamic inactivation (aPDI) arises as an alternative antifungal therapy. The tetracationic metalloporphyrin Zn(II) meso-tetrakis(N-n-hexylpyridinium-2-yl)porphyrin (ZnTnHex-2-PyP4+) has high photoefficiency and improved cellular interactions. We investigated the ZnTnHex-2-PyP4+ as a photosensitizer (PS) to photoinactivate yeasts and biofilms of C. albicans strains (ATCC 10231 and ATCC 90028) using a blue light-emitting diode. The photoinactivation of yeasts was evaluated by quantifying the colony forming units. The aPDI of ATCC 90028 biofilms was assessed by the MTT assay, propidium iodide (PI) labeling, and scanning electron microscopy. Mammalian cytotoxicity was investigated in Vero cells using MTT assay. The aPDI (4.3 J/cm2) promoted eradication of yeasts at 0.8 and 1.5 µM of PS for ATCC 10231 and ATCC 90028, respectively. At 0.8 µM and same light dose, aPDI-treated biofilms showed intense PI labeling, about 89% decrease in the cell viability, and structural alterations with reduced hyphae. No considerable toxicity was observed in mammalian cells. Our results introduce the ZnTnHex-2-PyP4+ as a promising PS to photoinactivate both yeasts and biofilms of C. albicans, stimulating studies with other Candida species and resistant isolates.

Non-stochiometric copper selenide nanocrystals are p-type semiconductors, which present localized surface plasmon resonances (LSPRs) in the near-infrared region, which are proportional to the Cu(I) vacancies. Here, an easy, low-cost, and environmentally friendly methodology for the one-pot aqueous colloidal synthesis of non-stoichiometric copper selenide nanocrystals (NCs) is described. The developed methodology consists in the use of CuCl and Na2O3Se as precursors of copper and selenium, respectively, stabilized by mercaptosuccinic acid (MSA). The best conditions for the non-stoichiometric NCs regarding Cu/stabilizer and Cu/Se molar ratios, and the initial pH, that provided the prominent plasmonic, were those with a Cu/Se molar ratio of 1 : 2 and pH 5.0, and both molar ratios of Cu/MSA used (1 : 6 and 1 : 9) with stirring for 60 min, at 90 °C, and under N2 flux. On the other hand, the nominal stoichiometric condition (Cu2−XSe, X = 0) presented a very weak LSPR behavior, suggesting lower copper vacancy defects, as expected. The obtained plasmonic NCs were characterized optically by electron absorption spectroscopy and structurally by X-ray diffractometry, transmission electron microscopy (TEM), Raman, and infrared spectroscopy (FTIR). For the best synthetic condition, the NCs produced presented a maximum absorption band at 1124 nm. TEM images for non-stoichiometric copper selenide systems showed highly crystalline and mostly spheroid particles, and a rather large size dispersion ranging from 2 to 13 nm, with a mean size of d = 8.6 ± 2.0 nm. The crystalline structure determined by the X-ray diffractometry profile for these as-prepared non-stoichiometric nanoparticles matches those observed for the Klockmannite hexagonal structure for CuSe (X = 1) structures. Furthermore, the use of the synthesized Cu2−XSe-MSA NCs in surface-enhanced infrared absorption (SEIRA) spectroscopy promoted an effective enhancement of infrared signatures of molecules in a very low concentration, as shown in our preliminary results with standard dye molecules.

We report the development of a new nanostructured electrochemical immunosensing platform for the detection of the Zika virus envelope protein (EP-ZIKV). For this, quantum dots (QDs) were explored in combination with screen-printed carbon electrodes (SPCEs) functionalized with a conductor polymeric film, formed from 2-(1H-pyrrol-1-yl)ethanamine (Py

In this work, the optical activity of Ag prismatic nanoplatelets was tuned to the methylene molecules absorption region and we applied these conjugated systems to enhance the photodynamic action against Staphylococcus aureus.

Quantum dots have generated great interest because of their optical properties, both to life sciences and electronics applications. However, possible risks to the environment associated with these nanoparticles are still under investigation. The present study aimed to evaluate the toxicity of suspensions of cadmium telluride (CdTe) quantum dots to Biomphalaria glabrata mollusks, a very sensitive aquatic environmental bioindicator for physical and chemical agents. Toxicity was examined by using embryos and adult mollusks as well as hemocytes. The distribution of cadmium in the organs of adults was also assessed. Effects of the stabilizing agent of the quantum dots were also evaluated. Animals were exposed to suspensions of quantum dots for 24 h, at concentrations varying from 1.2 to 20 nM for embryos and from 50 to 400 nM for adult mollusks. Results showed that suspensions of quantum dots induced malformations and mortality in embryos and mortality in adults, depending on the concentration applied. In the cytotoxicity study, hemocyte apoptosis was observed in adults exposed to the highest concentration of quantum dots applied as well as to the stabilizing agent. Cell binucleation and micronucleus frequencies were not significative. Bioaccumulation evaluation revealed that quantum dots targeted the digestive gland (hepatopancreas). Taken together, outcomes suggested that specific nano-effects related directly not only to composition but also to the aggregation of quantum dots may be mediating the observed toxicity. Thus B. glabrata was determined to be a very sensitive species for interpreting possible nano-effects in aquatic environments. Environ Toxicol Chem 2019;38:2128-2136. © 2019 SETAC.

Biflorin (6,9-dimethyl-3-(4-methylpent-3-en-1-yl) benzo[de]chromene-7,8-dione) is a promising substance that has been increasingly studied in the past decades due to its diverse pharmacological properties (i.e. antitumor, antioxidant, antiinflamatory, antimicrobial activity etc.). Aiming the comprehension of its antitumoral activity we investigated the cell proliferation and cytotoxicity habilities of biflorin against mice splenocytes Balb/c. Biflorin was able to stimulate mice splenocytes Balb/c in 48 h of incubation at a concentration of 20.2 μM. Its immunostimulation promoted the production of cytokines such as: TNF-α, IFN-γ, IL-2, IL-6 and IL-17, inducing the immune profile toward a Th1 response. Moreover, an original method which led to an excellent yield with less processing time compared to the methods described in the literature was developed to obtain biflorin, from sawdust of Capraria biflora L., Scrophulariaceae. This method shows a great potential of increasing the production of this pharmacological active compound. Keywords: Biflorin, Original method, Biological activity, Immunomodulatory effect

Markers are regularly used to detect fraud and adulteration in fuel as well as to track products in a distribution chain. Herein, we tested the use of CdSe semiconductor fluorescent nanocrystals, quantum dots, as a new class of fuel markers. Quantum dots show bright fluorescence and they can be excited by light sources covering a broad spectral range, from ultraviolet up to the near infrared region. This class of markers presents great advantages over conventional ones, such as high detectability potential, fluorescence emission in multicolor wavelengths, high photostability, and easy preparation procedures. In this work, we prepared CdSe quantum dots by colloidal controlled growth in an organic medium to be used for evaluation of diesel oil. The detection method was either by electronic absorption or fluorescence spectroscopies. The nanomarkers could be detected down to 1 mg/kg level in diesel fuel. The overall results indicated that CdSe quantum dots have the potential for use as a new class of fuel nanomarkers.

Magnetic resonance imaging (MRI) is a powerful non-invasive diagnostic tool that enables distinguishing healthy from pathological tissues, with high anatomical detail. Nevertheless, MRI is quite limited in the investigation of molecular/cellular biochemical events, which can be reached by fluorescence-based techniques. Thus, we developed bimodal nanosystems consisting in hydrophilic quantum dots (QDs) directly conjugated to Gd(III)-DO3A monoamide chelates, a Gd(III)-DOTA derivative, allowing for the combination of the advantages of both MRI and fluorescence-based tools. These nanoparticulate systems can also improve MRI contrast, by increasing the local concentration of paramagnetic chelates. Transmetallation assays, optical characterization, and relaxometric analyses, showed that the developed bimodal nanoprobes have great chemical stability, bright fluorescence, and high relaxivities. Moreover, fluorescence correlation spectroscopy (FCS) analysis allowed us to distinguish nanosystems containing different amounts of chelates/QD. Also, inductively coupled plasma optical emission spectrometry (ICP – OES) indicated a conjugation yield higher than 75%. Our nanosystems showed effective longitudinal relaxivities per QD and per paramagnetic ion, at least 5 times [per Gd(III)] and 100 times (per QD) higher than the r1 for Gd(III)-DOTA chelates, suitable for T1-weighted imaging. Additionally, the bimodal nanoparticles presented negligible cytotoxicity, and efficiently labeled HeLa cells as shown by fluorescence. Thus, the developed nanosystems show potential as strategic probes for fluorescence analyses and MRI, being useful for investigating a variety of biological processes.

Background Site-specific multimodal nanoplatforms with fluorescent-magnetic properties have great potential for biological sciences. For this reason, we developed a multimodal nanoprobe (BNPs-Tf), by covalently conjugating an optical-magnetically active bimodal nanosystem, based on quantum dots and iron oxide nanoparticles, with the human holo-transferrin (Tf). Methods The Tf bioconjugation efficiency was evaluated by the fluorescence microplate assay (FMA) and the amount of Tf immobilized on BNPs was quantified by fluorescence spectroscopy. Moreover, relaxometric and fluorescent properties of the BNPs-Tf were evaluated, as well as its ability to label specifically HeLa cells. Cytotoxicity was also performed by Alamar Blue assay. Results The FMA confirmed an efficient bioconjugation and the fluorescence spectroscopy analysis indicated that 98% of Tf was immobilized on BNPs. BNPs-Tf also presented a bright fluorescence and a transversal/longitudinal relaxivities ratio (r2/r1) of 65. Importantly, the developed BNPs-Tf were able to label, efficiently and specifically, the Tf receptors in HeLa cells, as shown by fluorescence and magnetic resonance imaging assays. Moreover, this multimodal system did not cause noteworthy cytotoxicity. Conclusions The prepared BNPs-Tf hold great promise as an effective and specific multimodal, highly fluorescent-magnetic, nanoplatform for fluorescence analyses and T2-weighted images. General significance This study developed an attractive and versatile multimodal nanoplatform that has potential to be applied in a variety of in vitro and in vivo studies, addressing biological processes, diagnostic, and therapeutics. Moreover, this work opens new possibilities for designing other efficient multimodal nanosystems, considering other biomolecules in their composition able to provide them important functional properties.

Short chain sodium polyphosphates – (NaPO3)9 – PolyP – were added to 2-in-1 pediatric parenteral nutrition (PN) formulations as new alternative phosphate sources. Solutions of PolyP and Ca2+ in the presence of Mg2+, glucose, amino acids and trace elements were analyzed by electronic absorption and 31P NMR spectroscopies, as well as by pH measurements. Additionally, to verify the biocompatibility of short chain PolyP solutions, red blood cell (RBCs) units were evaluated by analyzing hemoglobin, hematocrit, and hemolysis. The blood clotting action of the PolyP was determined through Prothrombin time. We observed that the short chain PolyP anions did not interfere in the quality control parameters for RBCs units or in the blood extrinsic coagulation pathway. The spectral changes observed in the 31P NMR confirm the association of Mg2+ and Ca2+ with PolyP in the admixtures. Compatibility diagrams showed that the PolyP-Ca/Mg systems reflect a gel behavior based on the PolyP chains interaction with the incoming cations maintaining a supramolecular equilibrium until a limiting concentration. We observed that the greater the PolyP concentration, the greater the amount of Ca2+ that could be added to the admixture without phase separation and that the increased complexity of species (glucose, amino acid, trace elements and Mg2+) in the admixture favored a still higher final Ca2+ concentration. The overall colloidal behavior of short chain PolyPs shows that these polyanions function not only as alternative and safe phosphate sources in PN formulations but also as polyanions that increase the admixture stability against phase separation by efficiently sequestering metal cations present.

The development of multimodal nanoprobes has been growing in recent years. Among these novel nanostructures are bimodal systems based on quantum dots (QDs) and low molecular weight Gd3+ chelates, prepared for magnetic resonance imaging (MRI) and optical analyses. MRI is a technique used worldwide that provides anatomic resolution and allows distinguishing of physiological differences at tissue and organ level. On the other hand, optical techniques are very sensitive and allow events to be followed at the cellular or molecular level. Thus, the association of these two techniques has the potential to achieve a more complete comprehension of biological processes. In this review, we present state-of-the-art research concerning the development of potential multimodal optical/paramagnetic nanoprobes based on Gd3+ chelates and QDs, highlighting their preparation strategies and overall properties.

Mastitis is a bacterial infection that affects all lactating mammals, and in dairy cattle, it leads to a reduction in their milk production and, in worse cases, it may lead to animal death. One viable therapeutic modality for overcoming bacterial resistance can be photodynamic inactivation (PDI), a therapeutic modality for bacterial infection treatment. One of the main factors that can lead to an efficient PDI process is the association of metallic nanoparticles in the close vicinity of photosensitizers, which has shown promising results due to localized surface plasmon resonance phenomena. In this work, methylene blue (MB) molecules were associated with Ag prismatic nanoplatelets (AgNPrs) to use as PDI photosensitizer against Staphylococcus aureus isolated from bubaline mastitis. The optical plasmonic activity of AgNPrs was tuned to the MB absorption region (600-700 nm) by inducing their growth into prismatic shapes by a seed-mediated procedure, using poly (sodium 4-styrene sulfonate) as the surfactant. A simulation on the plasmonic properties of the nanoprisms, applying particle size within the dimensions determined by TEM image analysis (d = 32 ± 6 nm), showed a 30 % increase of the incident field on the prismatic tips. Photodynamic results showed that the electrostatic AgNPr-MB conjugates promoted enhancement (ca. 15 %) of the reactive oxygen species production. Besides, PDI mediated by AgNPrs-MB led to the complete inactivation of the mastitis S. aureus strain after 6 min inactivation, in contrast to PDI mediated by MB, which reduced less than a 0.5 bacterial log. Thus, the results show this plasmonic enhanced photodynamic tool's potential to be applied in the inactivation of multi-resistant bacterial strains.

Photodynamic inactivation (PDI) is emerging as a promising alternative for cutaneous leishmaniasis (CL). The chemotherapy currently used presents adverse effects and cases of drug resistance have been reported. ZnTnHex-2-PyPZnTnHex-2-PyPZnTnHex-2-PyPZnTnHex-2-PyPThis report highlighted promising photodynamic effects mediated by ZnTnHex-2-PyP

Quantum dots (QDs) have attracted considerable attention as fluorescent probes for life sciences. The advantages of using QDs in fluorescence-based studies include high brilliance, a narrow emission band allowing multicolor labeling, a chemically active surface for conjugation, and especially, high photostability. Despite these advantageous features, the size of the QDs prevents their free transport across the plasma membrane, limiting their use for specific labeling of intracellular structures. Over the years, various methods have been evaluated to overcome this issue to explore the full potential of the QDs. Thus, in this review, we focused our attention on physical and biochemical QD delivery methods—electroporation, microinjection, cell-penetrating peptides, molecular coatings, and liposomes—discussing the benefits and drawbacks of each strategy, as well as presenting recent studies in the field. We hope that this review can be a useful reference source for researches that already work or intend to work in this area. Graphic Abstract: Strategies for the intracellular delivery of quantum dots discussed in this review (electroporation, microinjection, cell-penetrating peptides, molecular coatings, and liposomes).[Figure not available: see fulltext.]

Zika virus (ZIKV) infection has been declared a public health emergency due to its associations with severe disorders in newborns and adults. The detection of ZIKV and its associated biomolecules is challenging and has been stimulating the search for new methods that can offer greater sensitivity, specificity, and practicality, among other advantages. Quantum dots (QDs), fluorescent semiconductor nanocrystals, can be useful in the development of innovative (bio)detection assays for Zika. QDs have high brightness, resistance to photodegradation, and versatility for conjugations with biological molecules or other nanoparticles, which can also enhance/complement the QDs properties. Thus the first part of this chapter covers a brief introduction to the characteristics, advantages, and potentialities of QDs associated with biosensing. Then, in the second part, studies involving QD-based assays for the detection of the ZIKV ribonucleic acid (RNA) or nonstructural NS1 protein, as well as anti-ZIKV envelope E protein immunoglobulin G (IgG) antibodies, are described.

The development of multimodal nanoprobes has been growing in recent years. Among these novel nanostructures are bimodal systems based on quantum dots (QDs) and low molecular weight Gd

Quantum dots (QDs) have attracted considerable attention as fluorescent probes for life sciences. The advantages of using QDs in fluorescence-based studies include high brilliance, a narrow emission band allowing multicolor labeling, a chemically active surface for conjugation, and especially, high photostability. Despite these advantageous features, the size of the QDs prevents their free transport across the plasma membrane, limiting their use for specific labeling of intracellular structures. Over the years, various methods have been evaluated to overcome this issue to explore the full potential of the QDs. Thus, in this review, we focused our attention on physical and biochemical QD delivery methods-electroporation, microinjection, cell-penetrating peptides, molecular coatings, and liposomes-discussing the benefits and drawbacks of each strategy, as well as presenting recent studies in the field. We hope that this review can be a useful reference source for researches that already work or intend to work in this area. Strategies for the intracellular delivery of quantum dots discussed in this review (electroporation, microinjection, cell-penetrating peptides, molecular coatings, and liposomes).

Enhancement of hydrophilicity and functionalization of CdTe QDs (Quantum Dots) via surface modifications have made them suitable to be used as specific probes for cell imaging. Applications for targeting cell surfaces have been widely demonstrated in vitro but their use in animal models is not trivial. Here, we reported the interaction of mercaptosuccinic-coated (MSA) CdTe QDs with the epidermis of living and Carnoy-fixed zebrafish embryos. QDs concentrate along adherent junctions and reveal the characteristic pattern of actin microridges at the apical surface of the enveloping layer. In our study, labeling with anionic QDs is attained within few minutes at submicromolar concentrations in whole mounted Carnoy-fixed zebrafish embryos, providing a faster approach compared with immunodetection or standard Phalloidin staining of actin for visualization by fluorescence microscopy. Fil: Sigot, Valeria. Universidad Nacional de Entre Ríos. Instituto de Investigación y Desarrollo en Bioingeniería y Bioinformática - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación y Desarrollo en Bioingeniería y Bioinformática; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina Fil: Cabral Filho, Paulo E.. Universidade Federal de Pernambuco; Brasil Fil: Sampedro, Maria Florencia. Universidad Nacional de Entre Ríos. Instituto de Investigación y Desarrollo en Bioingeniería y Bioinformática - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación y Desarrollo en Bioingeniería y Bioinformática; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina Fil: Santos, Beate S.. Universidade Federal de Pernambuco; Brasil Fil: Fontes, Adriana. Universidade Federal de Pernambuco; Brasil

The use of Quantum Dots (QDs) as fluorescent probes for understanding biological functions has emerged as an advantageous alternative over application of conventional fluorescent dyes. Intracellular delivery of QDs is currently a specific field of research. When QDs are tracking a specific target in live cells, they are mostly applied for extracellular membrane labeling. In order to study intracellular molecules and structures it is necessary to deliver free QDs into the cell cytosol. In this work, we adapted the freeze and thaw method to encapsulate water dispersed carboxyl-coated CdTe QDs into liposomes of different compositions, including cationic liposomes with fusogenic properties. We showed that labeled liposomes were able to fuse with live human stem cells and red blood cells in an endocytic-independent way. We followed the interactions of liposomes containing QDs with the cells. The results were minutely discussed and showed that QDs were delivered, but they were not freely diffused in the cytosol of those cells. We believe that this approach has the potential to be applied as a general route for encapsulation and delivery of any membrane-impermeant material into living cells.

The increasing applications of quantum dots (QDs) as optic tools in life science have stimulated researchers to evaluate the effects of these nanoprobes in cell viability using a variety of methods, especially colorimetric ones. One of the most applied tests is the MTT assay. In comparison to MTT, for example, the resazurin-based method has the main advantage of not evaluating the cells directly, thus eliminating false-positive results that may arise from the overlap of the absorbances of the QD with the colorimetric compound. Therefore, herein, we describe the resazurin assay as an alternative, simple, quick, sensitivity, reproducible, and nontoxic test to evaluate the in vitro cell viability after QD exposure. Moreover, this test presents an additional advantage; the cells remain viable for complementary experimental procedures, such as cell migration or adhesion.

Sialic acids (SAs) modulate essential physiological and pathological conditions, including cell-cell communication, immune response, neurological disorders, and cancer. Besides, SAs confer negative charges to cell membranes, also contributing to hemorheology. Phenylboronic acids, called as mimetic lectins, have been highlighted to study SA profiles. The association of these interesting molecules with the optical properties of quantum dots (QDs) can provide a deeper/complementary understanding of mechanisms involving SAs. Herein, we explored the thiol affinity to the QD surface to develop a simple, fast and direct attachment procedure to functionalize these nanocrystals with 3-mercaptophenylboronic acids (MPBAs). The functionalization was confirmed by fluorescence correlation spectroscopy and inductively coupled plasma spectrometry. The conjugate specificity/efficiency was proved in experiments using red blood cells (RBCs). A labeling >90% was found for RBCs incubated with conjugates, which reduced to 17% after neuraminidase pretreatment. Moreover, QDs-MPBA conjugates were applied in a comparative study using acute (KG-1) and chronic (K562) myelogenous leukemia cell lines. Results indicated that KG-1 membranes have a greater level of SA, with 100% of cells labeled and a median of fluorescence intensity of ca. 2.5-fold higher when compared to K562 (94%). Therefore, this novel QDs-MPBA conjugate can be considered a promising nanoplatform to evaluate SA contents in a variety of biological systems.

Folic acid content was evaluated in food preparations containing wheat and corn flour submitted to baking, deep-frying, and steaming. Commercially fortified flours showed the absence of folic acid. Flours with laboratory folic acid fortification showed 487 and 474 μg of folic acid in 100 g of wheat and corn flours, respectively. In the corn flour preparations, the cake had the highest retention (99%) when compared to couscous (97%). Besides, the cake showed higher retention when compared to the wheat flour preparations due to the interactions of the folic acid with the hydrophobic amino acids of the Zein, a protein found in corn. In wheat flour preparations, vitamin retention was 87%, 80% and 57% in bread, cake, and White sauce respectively. These findings relate to the change of the physicochemical properties of food components that occurs during mixing and cooking of the ingredients.

The increasing applications of quantum dots (QDs) as optic tools in life science have stimulated researchers to evaluate the effects of these nanoprobes in cell viability using a variety of methods, especially colorimetric ones. One of the most applied tests is the MTT assay. In comparison to MTT, for example, the resazurin-based method has the main advantage of not evaluating the cells directly, thus eliminating false-positive results that may arise from the overlap of the absorbances of the QD with the colorimetric compound. Therefore, herein, we describe the resazurin assay as an alternative, simple, quick, sensitivity, reproducible, and nontoxic test to evaluate the in vitro cell viability after QD exposure. Moreover, this test presents an additional advantage; the cells remain viable for complementary experimental procedures, such as cell migration or adhesion.

Quantum Dots (QDs) are fluorescent semiconductor nanocrystals with unique optical properties, which are being highly applied in opto-electronic devices and biological sciences. Aiming at biological applications we developed highly blue fluorescent Cysteamine coated ZnSe QDs. They were prepared in aqueous medium at near physiological pH by a fast precipitation process followed by hydrothermal growth. The characteristic excitonic emission output was enhanced by decreasing their surface defects via an UV photoactivation procedure. We tested their in vitro cytotoxicity towards macrophage cells and probed their functionality by labeling two different cell types: Candida albicans and red blood cells by electrostatic interaction with the negative cell membranes. The overall results show that these positively charged QDs can be applied as new low toxic and promising platforms for biolabeling purposes.

The optical properties of quantum dots (QDs) make them useful tools for biology, especially when combined with biomolecules such as lectins. QDs conjugated to lectins can be used as nanoprobes for carbohydrate expression analysis, which can provide valuable information about glycosylation changes related to cancer and pathogenicity of microorganisms, for example. In this study, we evaluated the best strategy to conjugate Cramoll lectin to QDs and used the fluorescent labeling of Candida albicans cells as a proof-of-concept. Cramoll is a mannose/glucose-binding lectin with unique biological properties such as immunomodulatory, antiparasitic, and antitumor activities. We probed covalent coupling and adsorption as conjugation strategies at different pH values. QDs conjugated to Cramoll at pH7.0 showed the best labeling efficiency in the fluorescence microscopy analysis. Moreover, QD-Cramoll conjugates remained brightly fluorescent and preserved identical biological activity according to hemagglutination assays. Flow cytometry revealed that approximately 17% of C. albicans cells were labeled after incubation with covalent conjugates, while approximately 92% of cells were labeled by adsorption conjugates (both at pH7.0). Inhibition assays confirmed QD-Cramoll specificity, which reduced the labeling to at most 3%. Therefore, the conjugates obtained by adsorption (pH7.0) proved to be promising and versatile fluorescent tools for glycobiology.

Over the years, microorganisms have developed several resistance mechanisms against standard treatments, thus limiting the effect of drugs and rendering ineffective therapies. Considering the growing number of resistant pathogens and adverse effects of conventional therapies, new antimicrobial technologies able to provide more effective, rapid, and safer treatments to inactivate pathogens, with unlikely chances of inducing resistance, are needed. In this regard, antimicrobial photodynamic inactivation (aPDI) has emerged as an alternative modality of treatment. In particular, Zn(II) porphyrins (ZnPs) hold great potential as photosensitizers (PSs) for aPDI and have been attracting increasing attention. The chemical structure of ZnPs can be tailored to produce PSs with improved chemical stability and photophysical properties, also modulating their amphiphilic and ionic characters, bioavailability, and (sub)cellular distribution. Thus, in this review, we provide a detailed report of studies published in about the last 10 years (2010–2021) focusing on aPDI mediated by ZnPs over a variety of pathogens, including bacteria, fungi, viruses, and protozoa. Fundamentals of aPDI, and porphyrin and its derivatives, especially ZnPs, are also included herein. We hope that this review can guide and be a reference for future studies related to aPDI mediated by ZnPs, and encourages more detailed studies on ZnP photophysical and photochemical properties, aiming to improve the fight against infectious diseases.

Background Photodynamic inactivation (PDI) is emerging as a promising alternative for cutaneous leishmaniasis (CL). The chemotherapy currently used presents adverse effects and cases of drug resistance have been reported. ZnTnHex-2-PyP4+ is a porphyrin with a high potential as a photosensitizer (PS) for PDI, due to its photophysical properties, structural stability, and cationic/amphiphilic character that can enhance interaction with cells. This study aimed to investigate the photodynamic effects mediated by ZnTnHex-2-PyP4+ on Leishmania parasites. Methods ZnTnHex-2-PyP4+ stability was evaluated using accelerated solvolysis conditions. The photodynamic action on promastigotes was assessed by (i) viability assays, (ii) mitochondrial membrane potential evaluation, and (iii) morphological analysis. The PS-promastigote interaction was studied. PDI on amastigotes and the cytotoxicity on macrophages were also analyzed. Results ZnTnHex-2-PyP4+, under submicromolar concentration, led to immediate inactivation of more than 95% of promastigotes. PDI promoted intense mitochondrial depolarization, loss of the fusiform shape, and plasma membrane wrinkling in promastigotes. Fluorescence microscopy revealed a punctate PS labeling in the parasite cytoplasm. PDI also led to reductions of ca. 64% in the number of amastigotes/macrophage and 70% in the infection index after a single treatment session. No noteworthy toxicity was observed on mammalian cells. Conclusions ZnTnHex-2-PyP4+ is stable against demetallation and more efficient as PS than the ethyl analogue ZnTE-2-PyP4+, indicating readiness for evaluation in in vivo studies as an alternative approach to CL. General significance This report highlighted promising photodynamic effects mediated by ZnTnHex-2-PyP4+ on Leishmania parasites, opening up perspectives for applications in CL pre-clinical assays and PDI of other microorganisms.

During carcinogenesis, changes in the glycosylation can modulate many biological processes. Thus, the interest in exploring and understanding the roles of carbohydrates as cancer biomarkers has been increasing. Lectins have been applied as useful tools in glycobiology, especially when associated with fluorescent reporters. Therefore, to take advantage of the physicochemical properties of quantum dots (QDs), herein, we conjugated Cramoll, a lectin that recognizes glucose/mannose residues, with those nanoparticles. We applied the conjugates to investigate the glycocode of normal, fibroadenoma (FB), and invasive ductal carcinoma (IDC) human breast tissues. Additionally, we proposed a method to quantitatively evaluate the tissue labeling intensity by a fluorescence microplate assay (FMA). Conjugates showed intense fluorescence and specificity. The lectin activity and secondary structure were also preserved after the conjugation with QDs. Moreover, fluorescence images showed that ductal cells of normal and FB tissues were preferentially labeled by conjugates, whereas both cells and stroma were strongly labeled in IDC. FMA showed in a quantitative, practical, and sensitive way that the level of exposed glucose/mannose residues increased accordingly to the sample malignancy degree. In conclusion, QDs-Cramoll conjugates can be considered effective, specific, and versatile probes to evaluate glycan profiles in normal and transformed tissues, by fluorescence microscopy as well as FMA quantification. Furthermore, FMA showed to be a potential method that can be applied with other fluorescent conjugates.

Quantum dots (QDs) are semiconductor nanocrystals, which present unique photophysical properties, enabling their application as new fluorescent platforms for biomedical sciences. Colloidal QDs are end-capped with organic or inorganic compounds, not only to prevent their agglomeration but also to provide reaction sites for the attachment of targeting (bio)molecules, nanoparticles or other interfaces, for specific biological purposes. The (bio)conjugation can involve non-covalent or covalent interactions, which can be accomplished through different strategies. The final assembly needs to maintain its chemical and optical stability and biochemical functionality. Although a relative good comprehension of the experimental procedures has been established, the bioconjugation process is still a challenge. The present manuscript aims to review the main (bio)conjugation strategies successfully applied to QDs, describing the steps necessary to prepare stable targeting fluorescent nanoplatforms, as well as some usual methods used to evaluate and optimize this process.

Crack cocaine and tobacco are usually used combined. Many crack cocaine users utilize tobacco ashes to facilitate the drug burning process. Crack cocaine pyrolysis products potentiate cocaine stimulant effects, and tobacco use acts as trigger to crack cocaine use. There are many discussions on how nicotine acts as gateway to other drugs use, but multiple drugs use seems to facilitate the use of other drugs. This multiple use operates in many different mechanisms at central nervous system, as tobacco and crack cocaine simultaneous use. Nicotine and cocaine act, mainly, in mechanisms mediated by dopamine and acetylcholine in reward pathway in a synergic way. Besides, nicotine has an important role in cocaine craving decrease. A better understanding of these interactions can facilitate the development of new treatment strategies for users who combine these two drugs.

In this study we evaluated the absolute fluorescence quantum yield ( Φ ) of hydrophilic CdTe QDs in function of different pHs, modified from the alkaline to acid, by using two different chemicals compounds, the mercaptosuccinic acid (MSA-the stabilizing agent of the QDs synthesis) or hydrochloric acid (HCl). The pH control of QDs suspensions is essential for the use of fluorescent nanoparticles in biological systems. We used the eclipsing thermal lens spectroscopy technique to determine the absolute fluorescence quantum yield values. The results showed variations on the Φ values as a function of the pH, which allowed a better understanding of QDs emission characteristics, establishing parameters for their use in biomedical applications such as optical images of biological systems, immunoassays, flow cytometry, biosensors and others.

In this study, we present the optimization of the optical properties of Mn2+ doped ZnSe QDs (also referred to as d-dots) coated with thioglycolic acid in aqueous medium. The nanoparticles were characterized by ionic coupled plasma, electron paramagnetic resonance, transmission electron microscopy and X-ray diffractometry. By applying a controlled dose of UV irradiation, we obtained efficient orange emitting d-dots (4T1 6A1 transition centered at 580 nm). The results point out to a doping fraction in the nanoparticles smaller than 1% (0.89%), and that the Mn2+ ions are preferentially located close to the particle's surface. The UV photoactivation procedure has a definite influence on the emission intensity and on the colloidal stability of the particles. Photoactivated Mn2+ d-dots were conjugated to Concanavalin A and labeled specifically Candida albicans yeast cells, by targeting carbohydrate residues depicting the successful use of these QDs as fluorescent probes. (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Background Overexpression of transferrin receptors (TfRs), which are responsible for the intracellular uptake of ferric transferrin (Tf), has been described in various cancers. Although molecular biology methods allow the identification of different types of receptors in cancer cells, they do not provide features about TfRs internalization, quantification and distribution on cell surface. This information can, however, be accessed by fluorescence techniques. In this work, the quantum dots (QDs)' unique properties were explored to strengthen our understanding of TfRs in cancer cells. Methods QDs were conjugated to Tf by covalent coupling and QDs-(Tf) bioconjugates were applied to quantify and evaluate the distribution of TfRs in two human glioblastoma cells lines, U87 and DBTRG-05MG, and also in HeLa cells by using flow cytometry and confocal microscopy. Results HeLa and DBTRG-05MG cells showed practically the same TfR labeling profile by QDs-(Tf), while U87 cells were less labeled by bioconjugates. Furthermore, inhibition studies demonstrated that QDs-(Tf) were able to label cells with high specificity. Conclusions HeLa and DBTRG-05MG cells presented a similar and a higher amount of TfR than U87 cells. Moreover, DBTRG-05MG cells are more efficient in recycling the TfR than the other two cells types. General significance This is the first study about TfRs in human glioblastoma cells using QDs. This new fluorescent tool can contribute to our understanding of the cancer cell biology and can help in the development of new therapies targeting these receptors.

Folic acid (FA) regulates metabolic activities essential to the human body. FA receptor (FR) overexpression has been reported for many cancers, but there are still few or conflicting data about FRs in breast cancer cells. Quantum dots (QDs) have arisen as tools to elucidate aspects on FRs, due to their unique physicochemical properties. Herein, QDs conjugated to FA were explored to study the internalization and recycling of FRs in breast cancer cells, using HeLa as an out-group control. QDs were covalently conjugated to FA under different conditions. The best conjugate was applied to study FRs in HeLa, MCF7, MDA-MB231, and T47D cells applying confocal microscopy and flow cytometry analyses. The conjugation efficiency and specificity were evaluated, respectively, using fluorescence correlation spectroscopy (FCS) and saturation assays. FCS confirmed the effectiveness of the conjugation. HeLa and T47D had/internalized a higher amount of FRs (95% and 90% of labeling, respectively) than MDA-MB231 cells (68%). MCF7 cells seem to have very low functional FRs (3%). Saturation assays proved the specificity of QD-FA conjugates and suggested that FR recycling rate is low in the majority of cells studied, except for T47D. QD-FA conjugates were successfully developed. Therapies targeting FRs may be more effective for HeLa, T47D, and MDA-MB231.

We developed a biosensing platform by electropolymerizing 2-(1H-Pyrrol-1-yl)ethanamine on glassy carbon (GC) surfaces, followed by the immobilization of carboxyl-coated CdTe QDs onto the films. The immobilization of QDs was studied by applying two procedures: adsorption (electrostatic interactions) and covalent (using crosslinking agents). At a later stage, the covalent immobilization of IgG antibodies was also tested in platforms prepared with and without QDs. The formation of the polymeric film was confirmed by scanning electron microscopy. Furthermore, all surface modifications were monitored using cyclic voltammetry and electrochemical impedance spectroscopy, and K3[Fe(CN)6]/K4[Fe(CN)6] as the redox probe. The covalent crosslinking showed reproducibility and effective QD immobilization. Our study also showed that the QD coating by carboxylic groups leads to a decrease in the charge transfer process. The nanometric size of QDs and their active surfaces highly increased the functional platform area, enabling a high number of available sites for IgG antibodies. Indeed, in the absence of QDs, the platform was not able to detect the anti-IgG molecules. These results were supported by saturation curves constructed from differential pulse voltammetry. Thus, the use of carboxyl-coated CdTe QDs combined with 2-(1H-Pyrrol-1-yl)ethanamine polymer favored the development of a promising and sensitive biosensing electrochemical platform.