Conformational Behavior, Topographical Features, and Antioxidant Activity of Partly De-Esterified Arabinoxylans.

This study aimed to investigate the effect of arabinoxylans (AX) partial de-esterification with feruloyl esterase on the polysaccharide conformational behavior, topographical features, and antioxidant activity. After enzyme treatment, the ferulic acid (FA) content in AX was reduced from 7.30 to 5.48 µg FA/mg polysaccharide, and the molecule registered a small reduction in radius of gyration (RG), hydrodynamic radius (Rh), characteristic ratio (C∞), and persistence length (q). A slight decrease in α and a small increase in K constants in the Mark-Houwink-Sakurada equation for partially de-esterified AX (FAX) suggested a reduction in molecule structural rigidity and a more expanded coil conformation, respectively, in relation to AX. Fourier transform infrared spectroscopy spectra of AX and FAX presented a pattern characteristic for this polysaccharide. Atomic force microscopy topographic analysis of FAX showed a more regular surface without larger hollows in relation to AX. The antioxidant activity of FAX, compared to AX, was reduced by 30 and 41% using both 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS+) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) methods, respectively. These results suggest that feruloyl esterase treatment of AX could offer a strategy to tailor AX chains conformation, morphological features, and antioxidant activity, impacting the development of advanced biomaterials for biomedical and pharmaceutical applications.

Atomic force microscopy and Raman spectra profile of blood components associated with exposure to cigarette smoking.

Tobacco smoke contains several compounds with oxidant and pro-oxidant properties with the capability of producing structural changes in biomolecules, as well as cell damage. This work aimed to describe and analyse the effect of tobacco smoke on human blood components, red blood cell (RBC) membrane, haemoglobin (Hb) and blood plasma by Atomic Force Microscopy (AFM) and Raman spectroscopy. Our results indicate that tobacco induced RBC membrane nano-alterations characterized by diminished RBC diameter and increased nano-vesicles formation, and RBC fragility. The Raman spectra profile suggests modifications in chemical composition specifically found in peaks 1135 cm-1, 1156 cm-1, 1452 cm-1 and intensity relation of peaks 1195 cm-1 and 1210 cm-1 of blood plasma and by change of peaks 1338 cm-1, 1357 cm-1, 1549 cm-1 and 1605 cm-1 associated with the pyrrole ring of Hb. The relevance of these results lies in the identification of a profile of structural and chemical alterations that serves as a biomarker of physiological and pathological conditions in the human blood components induced by tobacco exposure using AFM and the Raman spectroscopy as tools for monitoring them.

A magnetic immunoconjugate nanoplatform for easy colorimetric detection of the NS1 protein of dengue virus in infected serum.

In this work, as a proof of principle, the design and performance evaluation of a simple, cheap and efficient colorimetric test for the detection of the NS1 protein of dengue virus, assisted by an immunoconjugate of magnetite (Fe3O4) nanoparticles coupled to anti-NS1 antibodies is reported. A monoclonal antibody against the NS1 antigen was covalently immobilized on the surface of superparamagnetic iron oxide nanoparticles (SPIONs ∼ 20 nm) and used for the immunodetection of this protein. When the magnetic immuno-nanoplatform is added into infected serum, it conjugates with the NS1 protein and can then be easily separated using an external magnetic field; then, the recovered immunoconjugate is transferred into a well containing a second immobilized NS1-antibody to form an ELISA-type system. When the NS1 protein is present, a color change to blue is induced by reaction with the Perls reagent, which is consistent with the formation of a SPION-antibody-NS1 antigen-antibody conjugate that confirms infection. No false positives were found when NS1 was not present or a different antibody and the NS1 protein were added into the system. The experimental findings could be extrapolated and scaled up to lead to future developments of simple, quick, and inexpensive, in situ biomolecular diagnostic tests for emergent viral infections.

Specific capture of glycosylated graphene oxide by an asialoglycoprotein receptor: a strategic approach for liver-targeting.

In this work, we report the evaluation of lactosylated graphene oxide (GO-AL) as a potential drug carrier targeted at an asialoglycoprotein receptor (ASGPR) from hepatic cancer cells. Structural-modification, safety evaluation, and functional analysis of GO-AL were performed. The structure and morphology of the composite were analyzed by scanning electron microscopy (SEM) and atomic force microscopy (AFM), while Raman and FTIR spectroscopy were used to track the chemical modification. For the safe application of GO-AL, an evaluation of the cytotoxic effect, hemolytic properties, and specific interactions of the glycoconjugate were also studied. SEM and AFM analysis of the GO showed graphene sheets with a layer size of 2-3 nm, though a few of them reached 4 nm. The Raman spectra presented characteristic peaks of graphene oxide at 1608 cm-1 and 1350 cm-1, corresponding to G and D bands, respectively. Besides, Si-O peaks for the APTES conjugates of GO were identified by FTIR spectroscopy. No cytotoxic or hemolytic effects were observed for GO samples, thus proving their biocompatibility. The interaction of Ricinus communis lectin confirmed that GO-AL has a biorecognition capability and an exposed galactose structure. This biorecognition capability was accompanied by the determination of the specific absorption of lactosylated GO by HepG2 cells mediated through the asialoglycoprotein receptor. The successful conjugation, hemolytic safety, and specific recognition described here for lactosylated GO indicate its promise as an efficient drug-delivery vehicle to hepatic tissue.

Effect of gamma irradiation doses in the structural and functional properties of mice splenic cells.

PURPOSE: Ionizing radiation is nowadays effectively used in cancer treatments. However, the effect of irradiation in immune-system cells is poorly understood and remains controversial. The aim of this work was to determine the effect of γ-irradiation in the structural and functional properties of mice splenic cells. MATERIALS AND METHODS: Structural traits of irradiated splenic cells were evaluated by Atomic Force Microscopy and Raman spectroscopy. Functional properties were measured by gene and protein expression by RT-qPCR and ELISA, respectively. The induced cytotoxic effect was evaluated by MTT assay and the phagocytic capability by flow cytometry. RESULTS: Membrane roughness and molecular composition of splenic adherent cells are not changed by irradiation doses exposure. An increase in transcription of pro-inflammatory cytokines was observed. While protein expression decreased in IL-2 dose-dependent, relevant differences were identified in the anti-inflammatory marker IL-10 at 27 Gy. An increase of cytotoxicity in irradiated cells at 7 Gy and 27 Gy doses was observed, while phagocytosis was slight increased at 7 Gy dose but not statistically significant. CONCLUSIONS: We have demonstrated that γ-irradiation affects the splenic cells and changes the cytokines profile toward a pro-inflammatory phenotype and a tendency to increase the cytotoxicity was found, which implies a stimulation of immune response induced by γ-irradiation.