Negative Charge-Carrying Glycans Attached to Exosomes as Novel Liquid Biopsy Marker.

Prostate cancer (PCa) is the second most common cancer. In this paper, the isolation and properties of exosomes as potential novel liquid biopsy markers for early PCa liquid biopsy diagnosis are investigated using two prostate human cell lines, i.e., benign (control) cell line RWPE1 and carcinoma cell line 22Rv1. Exosomes produced by both cell lines are characterised by various methods including nanoparticle-tracking analysis, dynamic light scattering, scanning electron microscopy and atomic force microscopy. In addition, surface plasmon resonance (SPR) is used to study three different receptors on the exosomal surface (CD63, CD81 and prostate-specific membrane antigen-PMSA), implementing monoclonal antibodies and identifying the type of glycans present on the surface of exosomes using lectins (glycan-recognising proteins). Electrochemical analysis is used to understand the interfacial properties of exosomes. The results indicate that cancerous exosomes are smaller, are produced at higher concentrations, and exhibit more nega tive zeta potential than the control exosomes. The SPR experiments confirm that negatively charged α-2,3- and α-2,6-sialic acid-containing glycans are found in greater abundance on carcinoma exosomes, whereas bisecting and branched glycans are more abundant in the control exosomes. The SPR results also show that a sandwich antibody/exosomes/lectins configuration could be constructed for effective glycoprofiling of exosomes as a novel liquid biopsy marker.

MXene-based electrochemical devices applied for healthcare applications.

The initial part of the review provides an extensive overview about MXenes as novel and exciting 2D nanomaterials describing their basic physico-chemical features, methods of their synthesis, and possible interfacial modifications and techniques, which could be applied to the characterization of MXenes. Unique physico-chemical parameters of MXenes make them attractive for many practical applications, which are shortly discussed. Use of MXenes for healthcare applications is a hot scientific discipline which is discussed in detail. The article focuses on determination of low molecular weight analytes (metabolites), high molecular weight analytes (DNA/RNA and proteins), or even cells, exosomes, and viruses detected using electrochemical sensors and biosensors. Separate chapters are provided to show the potential of MXene-based devices for determination of cancer biomarkers and as wearable sensors and biosensors for monitoring of a wide range of human activities.

Graphitic Carbon Nitride Nanosheets Decorated with Zinc-Cadmium Sulfide for Type-II Heterojunctions for Photocatalytic Hydrogen Production.

In this study, we fabricated graphitic carbon nitride (g-C3N4) nanosheets with embedded ZnCdS nanoparticles to form a type II heterojunction using a facile synthesis approach, and we used them for photocatalytic H2 production. The morphologies, chemical structure, and optical properties of the obtained g-C3N4-ZnCdS samples were characterized by a battery of techniques, such as TEM, XRD, XPS, and UV-Vis DRS. The as-synthesized g-C3N4-ZnCdS photocatalyst exhibited the highest hydrogen production rate of 108.9 µmol·g-1·h-1 compared to the individual components (g-C3N4: 13.5 µmol·g-1·h-1, ZnCdS: 45.3 µmol·g-1·h-1). The improvement of its photocatalytic activity can mainly be attributed to the heterojunction formation and resulting synergistic effect, which provided more channels for charge carrier migration and reduced the recombination of photogenerated electrons and holes. Meanwhile, the g-C3N4-ZnCdS heterojunction catalyst also showed a higher stability over a number of repeated cycles. Our work provides insight into using g-C3N4 and metal sulfide in combination so as to develop low-cost, efficient, visible-light-active hydrogen production photocatalysts.

Complex treatment of oily polluted waters by modified melamine foams: from colloidal emulsions to a free oil removal.

This study deals with the efficient, low-cost, and scalable treatment of oily polluted waters including colloidal emulsions, oil-in-water mixtures, and free oil removal using melamine foams (MFs) modified by ferric chloride (FeCl3). Modified foams have superhydrophobic character due to the coordination of Fe3+ with free electron pairs on nitrogen and oxygen atoms within the melamine structure. The water contact angles (WCA) were 146° ± 2°, 148° ± 4°, 153° ± 2°, and 150° ± 4° for foams modified by the solutions with concentrations of 0.001 M, 0.005 M, 0.01 M, and 0.02 M, respectively. This modification enables the efficient treatment of various oil/water systems, including oil/water colloidal emulsions (99 vol% of the droplets have dimensions below 500 nm), oil-in-water mixtures up to 40 weight % of the oil component, and "free" oil removal as it was demonstrated in this study for the first time. The emulsions containing 100 ppm diesel oil (DO) were separated with 91.4% efficiency, and the mixtures containing 20 and 40 weight % DO were separated with 99.9% efficiency. Modified foams also quickly remove free DO from the water surface, absorbing 95 g/g DO, whereas water sorption was negligible. The separation of colloidal oil in water emulsions represents the key finding of this study as it indicates the applicability of the treated MFs for the treatment of emulsified industrial wastewater. The demulsification mechanism is based on multiple diffusion processes running at different time scales, including diffusion of the emulsion into the foam and diffusion of oil droplets within the foam, combined with parallel adsorption of oil droplets onto the solid skeleton of the foam. A multiplied usage of these foams for all these niche operations was also proven. The application of our current study with previous studies on modified MFs and polyurethane for water oil separation utilization is summarized in Table S1 ESI.

Amperometric Miniaturised Portable Enzymatic Nanobiosensor for the Ultrasensitive Analysis of a Prostate Cancer Biomarker.

Screen-printing technology is a game changer in many fields including electrochemical biosensing. Two-dimensional nanomaterial MXene Ti3C2Tx was integrated as a nanoplatform to immobilise enzyme sarcosine oxidase (SOx) onto the interface of screen-printed carbon electrodes (SPCEs). A miniaturised, portable, and cost-effective nanobiosensor was constructed using chitosan as a biocompatible glue for the ultrasensitive detection of prostate cancer biomarker sarcosine. The fabricated device was characterised with energy-dispersive X-ray spectroscopy (EDX), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Sarcosine was detected indirectly via the amperometric detection of H2O2 formed during enzymatic reaction. The nanobiosensor could detect sarcosine down to 7.0 nM with a maximal peak current output at 4.10 ± 0.35 × 10-5 A using only 100 µL of a sample per measurement. The assay run in 100 µL of an electrolyte showed the first linear calibration curve in a concentration window of up to 5 µM with a slope of 2.86 µA·µM-1, and the second linear calibration curve in the range of 5-50 µM with a slope of 0.32 ± 0.01 µA·µM-1 (R2 = 0.992). The device provided a high recovery index of 92.5% when measuring an analyte spiked into artificial urine, and could be used for detection of sarcosine in urine for at least a period of 5 weeks after the preparation.

How to choose proper magnetic particles for bioaffinity interactions? The case for immobilised glyconanoconjugate.

In this study, an assay for detection of the cancer biomarker Thomsen-nouvelle (Tn) antigen on the ELISA plates format was designed and developed. The effects of size and the interfacial density of the negative charge of magnetic beads (MBs) on the specific sensitivity of the bioaffinity interaction were studied. In particular, glyconanoconjugate, i.e. glycan Tn antigen conjugated to bovine serum albumin (BSA) was covalently immobilised on MBs for the bioaffinity detection of anti-Tn antibodies as cancer biomarkers. Six different MBs were used in the study, i.e. carboxy-modified MBs of 250 nm, 500 nm, 1000 nm and 2800 nm and epoxy-modified MBs of 2800 nm and 4500 nm. In order to evaluate which MBs are the best suited for detection of the analyte anti-Tn antibodies, sensitivities of detection (slopes from calibration curves) were calculated. Next, specific sensitivities were calculated for each type of MBs as a ratio of sensitivity of detection to the mass of MBs. From zeta potential ζ for each type of MBs, the interfacial charge density on MBs was calculated, expressed as the density of zeta potential ζd (ratio of zeta potential to surface area of MBs, i.e. ζd = Î¶/A). Then, we evaluated the effect of size and ζd on the specific sensitivity of detection of anti-Tn antibodies in order to understand the immobilisation process on nanoscale. We also identified an optimal value of ζd on MBs; this was essential to achieve highly sensitive detection of the analyte, which made it possible to attain limit of detection (LOD) of (0.31 ± 0.01) ng mL-1 or (2.10 ± 0.04) pM for analyte detection. In addition, the optimal assay configuration was highly selective and enabled reliable detection of the analyte in human serum with a recovery index in the range of 102-104%.

Redox features of hexaammineruthenium(III) on MXene modified interface: Three options for affinity biosensing.

In this article we describe construction of a bioreceptive interface for detection of a breast cancer biomarker carbohydrate antigen CA15-3. The conductive interface was patterned by a 2D nanomaterial MXene, to which a mixed layer containing sulfobetaine and carboxybetaine was electrochemically grafted through a diazonium moiety. Such a modified interface was then applied for covalent immobilisation of anti-CA15-3 antibody as a bioreceptive probe for detection of a breast cancer biomarker. Two different strategies were applied for final construction of an immunosensor i.e. an interface finally blocked by bovine serum albumin or an immunosensor without such modification. Finally, electrochemical reading was accomplished using a soluble redox probe Ru(NH3)63+ ion for detection of CA15-3 in a clinically relevant range up to 50 U mL-1. The results indicate that immunosensor based on non-blocked interface can be applied for biosensing using two modes of action: 1. differential pulse voltammetry (a plot of a peak current vs. analyte concentration) and 2. an electrochemical impedance spectroscopy (a plot of a charge transfer resistance vs. analyte concentration). The electrode blocked by bovine serum albumin (BSA) can be used by additional 3. mode of action: through detection of changes in the potential (a plot Epvs. c). Additionally, we reveal and explain that Ru(NH3)63+ is redox probe, which can be applied as interfacial molecular nanoscale ruler to distinguish negatively charged protein molecules present in the close proximity (≤ 6 nm) of the electrode (in our case adsorbed BSA molecules) from the negatively charged protein molecules at a larger distance (>12 nm) from the electrode (i.e. CA15-3 analyte).

Atom Transfer Radical Polymerization of Pyrrole-Bearing Methacrylate for Production of Carbonyl Iron Particles with Conducting Shell for Enhanced Electromagnetic Shielding.

The conducting polymer poly(2-(1H-pyrrole-1-yl)ethyl methacrylate (PPEMA) was synthesized by conventional atom transfer radical polymerization for the first time from free as well as surface-bonded alkyl bromide initiator. When grafted from the surface of carbonyl iron (CI) a substantial conducting shell on the magnetic core was obtained. Synthesis of the monomer as well as its polymer was confirmed using proton spectrum nuclear magnetic resonance (1H NMR). Polymers with various molar masses and low dispersity showed the variability of this approach, providing a system with a tailorable structure and brush-like morphology. Successful grafting from the CI surface was elucidate by transmission electron microscopy and Fourier-transform infrared spectroscopy. Very importantly, thanks to the targeted nanometer-scale shell thickness of the PPEMA coating, the magnetization properties of the particles were negligibly affected, as confirmed using vibration sample magnetometry. Smart elastomers (SE) consisting of bare CI or CI grafted with PPEMA chains (CI-PPEMA) and silicone elastomer were prepared and dynamic mechanical properties as well as interference shielding ones were investigated. It was found that short polymer chains grafted to the CI particles exhibited the plasticizing effect, which might be interesting from the magnetorheological point of view, and more interestingly, in comparison to the neat CI-based sample, it provided enhanced electromagnetic shielding of nearly 30 dB in thickness of 500 µm. Thus, SE containing the newly synthesized CI-PPEMA hybrid particles also exhibited considerably enhanced damping factor and proper mechanical performance, which make the material highly promising from various practical application points of view.

Engineering at Subatomic Scale: Achieving Selective Catalytic Pathways via Tuning of the Oxidation States in Functionalized Single-Atom Quantum Catalysts.

Regulating the catalytic pathways of single-atom sites in single atom catalysts (SACs) is an exciting debate at the moment, which has redirected the research towards understanding and modifying the single-atom catalytic sites through various strategies including altering the coordination environment of single atom for desirable outcomes as well as increasing their number. One useful aspect concerning the tunability of the catalytic pathways of SACs, which has been overlooked, is the oxidation state dynamics of the single atoms. In this study, iron single-atoms (FeSA) with variable oxidation states, dependent on the precursors, are harnessed inside a nitrogen-rich functionalized carbon quantum dots (CQDs) matrix via a facile one-step and low-temperature synthesis process. Dynamic electronic properties are imparted to the FeSAs by the simpler carbon dots matrix of CQDs in order to achieve the desired catalytic pathways of reactive oxygen species (ROS) generation in different environments, which are explored experimentally and theoretically for an in-depth understanding of the redox chemistry that drives the alternative catalytic pathways in FeSA@CQDs. These alternative and oxidation state-dependent catalytic pathways are employed for specific as well as cascade-like activities simulating natural enzymes as well as biomarkers for the detection of cancerous cells.

Fluorescent Zn(II)-Based Metal-Organic Framework: Interaction with Organic Solvents and CO2 and Methane Capture.

Adsorption of carbon dioxide (CO2), as well as many other kinds of small molecules, is of importance for industrial and sensing applications. Metal-organic framework (MOF)-based adsorbents are spotlighted for such applications. An essential for MOF adsorbent application is a simple and easy fabrication process, preferably from a cheap, sustainable, and environmentally friendly ligand. Herein, we fabricated a novel structural, thermally stable MOF with fluorescence properties, namely Zn [5-oxo-2,3-dihydro-5H-[1,3]-thiazolo [3,2-a]pyridine-3,7-dicarboxylic acid (TPDCA)] • dimethylformamide (DMF) •0.25 H2O (coded as QUF-001 MOF), in solvothermal conditions by using zinc nitrate as a source of metal ion and TPDCA as a ligand easy accessible from citric acid and cysteine. Single crystal X-ray diffraction analysis and microscopic examination revealed the two-dimensional character of the formed MOF. Upon treatment of QUF-001 with organic solvents (such as methanol, isopropanol, chloroform, dimethylformamide, tetrahydrofuran, hexane), interactions were observed and changes in fluorescence maxima as well as in the powder diffraction patterns were noticed, indicating the inclusion and intercalation of the solvents into the interlamellar space of the crystal structure of QUF-001. Furthermore, CO2 and CH4 molecule sorption properties for QUF-001 reached up to 1.6 mmol/g and 8.1 mmol/g, respectively, at 298 K and a pressure of 50 bars.

Glycan signatures for the identification of cisplatin-resistant testicular cancer cell lines: Specific glycoprofiling of human chorionic gonadotropin (hCG).

BACKGROUND: Testicular cancer (TC) is the most frequent type of cancer among young men aged between 15 and 34 years. TC is treated using cisplatin, but 3%-5% of TC patients fail to respond to cisplatin, with a very bad to fatal prognosis. Accordingly, it is most important to quickly and readily identify those TC patients who are resistant to cisplatin treatment. METHODS: This study seeks to investigate changes in the glycosylation associated with cisplatin resistance to TC cell lines. RESULTS: A specific glycoprofiling of human chorionic gonadotropin (hCG) was analysed in three TC cell lines and one cell line of female origin. A typical calibration curve for hCG glycoprofiling showed a dynamic range up to 50 ng/ml, with a limit of detection of 0.3 ng/ml and assay reproducibility represented by relative standard deviation of 3.0%. Changes in the glycan signatures on hCG were analysed in cisplatin-sensitive cell lines and in their cisplatin-resistant sub-lines using an enzyme-linked lectin assay (ELLA) protocol. An immobilised antibody was applied to a selective capture of hCG from a cytoplasmic fraction of cell lysates with final incubation using a lectin from a panel of 17 lectins. CONCLUSION: The results suggest that one particular lectin Dolichos biflorus agglutinin (DBA) can selectively discriminate sensitive TC cell lines from resistant TC cell lines. Moreover, there are additional lectins which can provide useful information about the strength of cisplatin resistance.

Amplified suspension magnetic bead-based assay for sensitive detection of anti-glycan antibodies as potential cancer biomarkers.

The development of a novel SUspension Magnetic-Bead-based Assay (SUMBA) for the detection of antibodies against aberrant glycans (AGA) as potential cancer biomarkers is presented here. The SUMBA method was extensively optimised by choosing proper commercially available AGA able to specifically, and with high affinity, recognise aberrant glycans, which were attached to the protein backbone working as a molecular scaffold (a glycoconjugate). The whole SUMBA was optimised using several analytical techniques such as Surface Plasmon Resonance and Energy Dispersive X-ray Spectroscopy. Additionally, the SUMBA method was extensively optimised for signal enhancement. With all steps optimised, we were able to detect AGA ultrasensitively with a limit of detection of 0.45 pM. Moreover, AGA could be detected in serum samples with a recovery index in the range of 98%-104%.

Identification of Whole-Serum Glycobiomarkers for Colorectal Carcinoma Using Reverse-Phase Lectin Microarray.

Colorectal cancer (CRC) is one of the most common types of cancer among men and women worldwide. Efforts are currently underway to find novel and more cancer-specific biomarkers that could be detected in a non-invasive way. The analysis of aberrant glycosylation of serum glycoproteins is a way to discover novel diagnostic and prognostic CRC biomarkers. The present study investigated a whole-serum glycome with a panel of 16 different lectins in search for age-independent and CRC-specific glycomarkers using receiver operating characteristic (ROC) curve analyses and glycan heat matrices. Glycosylation changes present in the whole serum were identified, which could lead to the discovery of novel biomarkers for CRC diagnostics. In particular, the change in the bisecting glycans (recognized by Phaseolus vulgaris erythroagglutinin) had the highest discrimination potential for CRC diagnostics in combination with human L selectin providing area under the ROC curve (AUC) of 0.989 (95% CI 0.950-1.000), specificity of 1.000, sensitivity of 0.900, and accuracy of 0.960. We also implemented novel tools for identification of lectins with strong discrimination power.

Breast cancer glycan biomarkers: their link to tumour cell metabolism and their perspectives in clinical practice.

INTRODUCTION: Breast cancer (BCa) is the most common cancer type diagnosed in women and 5th most common cause of deaths among all cancer deaths despite the fact that screening program is at place. This is why novel diagnostics approaches are needed in order to decrease number of BCa cases and disease mortality. AREAS COVERED: In this review paper, we aim to cover some basic aspects regarding cellular metabolism and signalling in BCa behind altered glycosylation. We also discuss novel exciting discoveries regarding glycan-based analysis, which can provide useful information for better understanding of the disease. The final part deals with clinical usefulness of glycan-based biomarkers and the clinical performance of such biomarkers is compared to already approved BCa biomarkers and diagnostic tools based on imaging. EXPERT OPINION: Recent discoveries suggest that glycan-based biomarkers offer high accuracy for possible BCa diagnostics in blood, but also for better monitoring and management of BCa patients. The review article was written using Web of Science search engine to include articles published between 2019 and 2021.

The Separation of Emulsified Water/Oil Mixtures through Adsorption on Plasma-Treated Polyethylene Powder.

This paper addresses the preparation and characterization of efficient adsorbents for tertiary treatment (oil content below 100 ppm) of oil/water emulsions. Powdered low-density polyethylene (LDPE) was modified by radio-frequency plasma discharge and then used as a medium for the treatment of emulsified diesel oil/water mixtures in the concentration range from 75 ppm to 200 ppm. Plasma treatment significantly increased the wettability of the LDPE powder, which resulted in enhanced sorption capability of the oil component from emulsions in comparison to untreated powder. Emulsions formed from distilled water and commercial diesel oil (DO) with concentrations below 200 ppm were used as a model of oily polluted water. The emulsions were prepared using ultrasonication without surfactant. The droplet size was directly proportional to sonication time and ranged from 135 nm to 185 nm. A sonication time of 20 min was found to be sufficient to prepare stable emulsions with an average droplet size of approximately 150 nm. The sorption tests were realized in a batch system. The effect of contact time and initial oil concentrations were studied under standard atmospheric conditions at a stirring speed of 340 rpm with an adsorbent particle size of 500 microns. The efficiency of the plasma-treated LDPE powder in oil removal was found to be dependent on the initial oil concentration. It decreased from 96.7% to 79.5% as the initial oil concentration increased from 75 ppm to 200 ppm. The amount of adsorbed oil increased with increasing contact time. The fastest adsorption was observed during the first 30 min of treatment. The adsorption kinetics for emulsified oils onto sorbent followed a pseudo-second-order kinetic model.

Ultrasensitive Ti3C2TX MXene/Chitosan Nanocomposite-Based Amperometric Biosensor for Detection of Potential Prostate Cancer Marker in Urine Samples.

Two-dimensional layered nanomaterial Ti3C2TX (a member of the MXene family) was used to immobilise enzyme sarcosine oxidase to fabricate a nanostructured biosensor. The device was applied for detection of sarcosine, a potential prostate cancer biomarker, in urine for the first time. The morphology and structures of MXene have been characterised by atomic force microscopy (AFM) and scanning electron microscopy (SEM). Electrochemical measurements, SEM and AFM analysis revealed that MXene interfaced with chitosan is an excellent support for enzyme immobilisation to fabricate a sensitive biosensor exhibiting a low detection limit of 18 nM and a linear range up to 7.8 µM. The proposed biosensing method also provides a short response time of 2 s and high recovery index of 102.6% for detection of sarcosine spiked into urine sample in a clinically relevant range.

Crystal Growth, Single Crystal Structure, and Biological Activity of Thiazolo-Pyridine Dicarboxylic Acid Derivatives.

Four novel TPDCA derivatives were prepared via a supersaturation method combining TPDCA with water, N-methyl-2-pyrrolidone (NMP), Na(PO2H2), and ammonia solution: 2(C9H7NO5S)H2O (1), (C9H7NO5S)C5H9NO (2), (C9H7NO5S)Na(PO2H2) (3), and (C9H5NO5S)(NH4)2(H2O) (4). Their crystal structures were determined by single-crystal X-ray diffraction. Compounds (1) and (2) crystallize in the monoclinic space groups P21 and P21/c, respectively, whereas compounds (3) and (4) crystallize in the triclinic space group P1̅. Weak and moderate hydrogen bonds were detected in the four compounds. In the biological tests, (1) and (3) exhibited significant antibacterial activity against Escherichia coli and Staphylococcus aureus; in addition, (1) was cytotoxic against leukemia HL-60 cells with the IC50 value of 158.5 ± 12.5 µM.

Analysis of serum glycome by lectin microarrays for prostate cancer patients - a search for aberrant glycoforms.

This is the first work focused on glycoprofiling of whole N- and O- glycome using lectins in an array format applied for analysis of serum samples from healthy individuals, benign prostate hyperplasia (BPH) patients, and prostate cancer (PCa) patients. Lectin microarray was prepared using traditional lectins with the incorporation of 2 recombinant bacterial lectins and 3 human lectins (17 lectins in total). Clinical validation of glycans as biomarkers was done in two studies: discrimination of healthy individuals with BPH patients vs. PCa patients (C vs. PCa) and discrimination of healthy individuals vs. BPH and PCa patients (H vs. PCond). Single lectins (17 lectins) and a combination of two lectins (136 binary lectin combinations) were applied in the clinical validation of glycan biomarkers providing 153 AUC values from ROC curves for both studies (C vs. PCa and H vs. PCond). Potential N- and O-glycans as biomarkers were identified and possible carriers of these glycans are shortly discussed.

Electrochemical Nanobiosensors for Detection of Breast Cancer Biomarkers.

This comprehensive review paper describes recent advances made in the field of electrochemical nanobiosensors for the detection of breast cancer (BC) biomarkers such as specific genes, microRNA, proteins, circulating tumor cells, BC cell lines, and exosomes or exosome-derived biomarkers. Besides the description of key functional characteristics of electrochemical nanobiosensors, the reader can find basic statistic information about BC incidence and mortality, breast pathology, and current clinically used BC biomarkers. The final part of the review is focused on challenges that need to be addressed in order to apply electrochemical nanobiosensors in a clinical practice.

A bioconjugated MoS2 based nanoplatform with increased binding efficiency to cancer cells.

We evaluate the application of surfactant-free liquid-phase exfoliated MoS2 nanosheets as a nanoplatform for a cancer detection and treatment system equipped with an antibody-antigen based recognition element. Employing antigen-antibody binding, we increased the probability of the endocytosis of MoS2 nanosheets into CAIX expressing cells by 30%. The nanosheets are functionalized with a specific antibody M75, which forms an antigen-antibody complex with CAIX. The bioconjugation of MoS2 nanosheets involves biocompatible components with low cytotoxicity, verified in the tested cell lines by fluorescence-based cell viability assay. The cellular internalization is quantified by flow cytometry, while the internalization is confirmed by label-free confocal Raman imaging. Raman measurements show increased lysosomal activity in the proximity of the internalized nanoplatforms.