Assessing the reinforcement effect by response surface methodology of holocellulose from spent coffee grounds on biopolymeric films as food packaging materials.

The pollution caused by petroleum-derived plastic materials has become a major environmental problem that has encouraged the development of new compostable and environmentally friendly materials for food packaging based on biomodified polymers with household residues. This study aims to design, synthesize, and characterize a biobased polymeric microstructure film from polyvinyl alcohol and chitosan reinforced with holocellulose from spent coffee grounds for food-sustainable packaging. Chemical isolation with a chlorite-based solution was performed to obtain the reinforced holocellulose from the spent coffee ground, and the solvent casting method was used to obtain the films to study. Physicochemical and microscopic characterizations were conducted to identify and select the best formulations using a simplex-centroid design analysis. The response surface methodology results indicate that the new packaging material obtained with equal amounts of polymers and reinforced material (1:1:1) possesses the appropriate barrier properties and microstructural character to prevent water attack and hydrophobic behavior and thus could be used as an alternative for food packaging materials.

Ecotoxicological evaluation of chitosan biopolymer films particles in adult zebrafish (Danio rerio): A comparative study with polystyrene microplastics.

To mitigate the environmental impact of microplastics (MPs), the scientific community has innovated sustainable and biodegradable polymers as viable alternatives to traditional plastics. Chitosan, the deacetylated form of chitin, stands as one of the most thoroughly investigated biopolymers and has garnered significant interest due to its versatile applications in both medical and cosmetic fields. Nevertheless, there is still a knowledge gap regarding the impact that chitosan biopolymer films (CBPF) may generate in aquatic organisms. In light of the foregoing, this study aimed to assess and compare the potential effects of CBPF on the gastrointestinal tract, gills, brain, and liver of Danio rerio against those induced by MPs. The findings revealed that both CBPF and MPs induced changes in the levels of oxidative stress biomarkers across all organs. However, it is essential to note that our star plots illustrate a tendency for CBPF to activate antioxidant enzymes and for MPs to produce oxidative damage. Regarding gene expression, our findings indicate that MPs led to an up-regulation in the expression of genes associated with apoptotic response (p53, casp3, cas9, bax, and bcl2) in all fish organs. Meanwhile, CBPF produced the same effect in genes related to antioxidant response (nrf1 and nrf2). Overall, our histological observations substantiated these effects, revealing the presence of plastic particles and tissue alterations in the gills and gastrointestinal tract of fish subjected to MPs. From these results, it can be concluded that CBPF does not represent a risk to fish after long exposure.

The Role of ZO-2 in Modulating JAM-A and γ-Actin Junctional Recruitment, Apical Membrane and Tight Junction Tension, and Cell Response to Substrate Stiffness and Topography.

This work analyzes the role of the tight junction (TJ) protein ZO-2 on mechanosensation. We found that the lack of ZO-2 reduced apical membrane rigidity measured with atomic force microscopy, inhibited the association of γ-actin and JAM-A to the cell border, and instead facilitated p114RhoGEF and afadin accumulation at the junction, leading to an enhanced mechanical tension at the TJ measured by FRET, with a ZO-1 tension probe, and increased tricellular TJ tension. Simultaneously, adherens junction tension measured with an E-cadherin probe was unaltered. The stability of JAM-A and ZO-2 binding was assessed by a collaborative in silico study. The absence of ZO-2 also impacted the cell response to the substrate, as monolayers plated in 20 kPa hydrogels developed holes not seen in parental cultures and displayed a retarded elongation and formation of cell aggregates. The absence of ZO-2 was sufficient to induce YAP and Snail nuclear accumulation in cells cultured over glass, but when ZO-2 KD cells were plated in nanostructured ridge arrays, they displayed an increased abundance of nuclear Snail and conspicuous internalization of claudin-4. These results indicate that the absence of ZO-2 also impairs the response of cells to substrate stiffness and exacerbates transformation triggered by substrate topography.

Comparison of the SMLM technique and the MSSR algorithm in confocal microscopy for super-resolved imaging of cellulose fibres.

Nowadays, the use of super-resolution microscopy (SRM) is increasing globally due to its potential application in several fields of life sciences. However, a detailed and comprehensive guide is necessary for understanding a single-frame image's resolution limit. This study was performed to provide information about the structural organisation of isolated cellulose fibres from garlic and agave wastes through fluorophore-based techniques and image analysis algorithms. Confocal microscopy provided overall information on the cellulose fibres' microstructure, while techniques such as total internal reflection fluorescence microscopy facilitated the study of the plant fibres' surface structures at a sub-micrometric scale. Furthermore, SIM and single-molecule localisation microscopy (SMLM) using the PALM reconstruction wizard can resolve the network of cellulose fibres at the nanometric level. In contrast, the mean shift super-resolution (MSSR) algorithm successfully determined nanometric structures from confocal microscopy images. Atomic force microscopy was used as a microscopy technique for measuring the size of the fibres. Similar fibre sizes to those evaluated with SIM and SMLM were found using the MSSR algorithm and AFM. However, the MSSR algorithm must be cautiously applied because the selection of thresholding parameters still depends on human visual perception. Therefore, this contribution provides a comparative study of SRM techniques and MSSR algorithm using cellulose fibres as reference material to evaluate the performance of a mathematical algorithm for image processing of bioimages at a nanometric scale. In addition, this work could act as a simple guide for improving the lateral resolution of single-frame fluorescence bioimages when SRM facilities are unavailable.

Polystyrene microplastics mitigate the embryotoxic damage of metformin and guanylurea in Danio rerio.

Microplastics (MPs) alone may endanger the health and fitness of aquatic species through different mechanisms. However, the harmful effects of these when mixed with other emerging contaminants require additional research. Herein, we aimed to determine whether a mixture of MPs with metformin (MET) or guanylurea (GUA) might induce embryotoxicity and oxidative stress in Danio rerio. Upon exposure to mixtures, our results showed MPs reduced the mortality rate of MET and GUA in embryos. Moreover, the severity and the rate of malformations were also decreased in all mixtures with MPs. Concerning oxidative stress, our findings indicated MET, GUA, MPs, and the mixtures increased the levels of lipoperoxidation, hydroperoxide content, and protein carbonyl content in D. rerio larvae. However, the oxidative damage induced in all mixtures was lower than that produced by both drugs alone. Thus, it is likely that the accumulation of MPs avoided the entrance of MET and GUA into the embryos. Once the embryo hatched, MPs did only remain accumulated in the yolk sac of larvae and did not translocate to other organs. Our risk assessment analysis confirmed that MPs shrunk the damage produced by MET and GUA. In a nutshell, MPs mitigate the embryotoxic damage of metformin and guanylurea in D. rerio by blocking their entrance.

Characterization of Structural Changes of Casein Micelles at Different PH Using Microscopy and Spectroscopy Techniques.

This study aimed to evaluate the influence of pH changes on morphometric parameters of casein micelles and a general overview of their conformational structure through microscopy techniques, Raman spectroscopy and multivariate analysis. It was found that casein micelles morphology and protein secondary structure depend strongly upon pH. The changes of arithmetic average roughness (Ra), size, and shape of casein micelles at different pH are properly characterized by atomic force and cryo-transmission electron microscopy. Morphometric changes of casein micelles were correlated correctly with folding and unfolding of casein molecules as evaluated by Raman spectroscopy when the pH was varied. The novelty of this contribution consists in demonstrating that there is a close structure-functionality relationship between the morphometric parameters of proteins and their secondary structure. Knowledge about casein micelles can help improve their use of its diverse applications.

Bioaccumulation and oxidative stress caused by aluminium nanoparticles and the integrated biomarker responses in the common carp (Cyprinus carpio).

The use of nanoparticles (NPs) in various industries has experienced significant growth due to the advantages they offer, so the increase in their use has generated the continuous discharge of these products in numerous water bodies, which can affect the organisms that inhabit them. Previous studies have shown that Al is capable of producing oxidative stress in aquatic organisms; however, so far the impact of AlNP on hydrobionts is limited. Therefore, the objective of this work was to determine the oxidative stress produced by AlNP in liver, gill and blood of Cyprinus carpio, as well as their bioconcentration factor (BCF) in various tissues. For this purpose, the organisms were exposed to 50 µg L-1 AlNP for 12-96 h. Subsequently, the tissues were obtained and the activity of antioxidant enzymes, oxidative damage to lipids and proteins were determined, and the BCF was calculated for liver, brain, gill and muscle. The results showed alterations in the activity of antioxidant enzymes and increased levels of lipoperoxidation, hydroperoxides and oxidized proteins. When establishing the integrated biomarker response, it was observed that the liver is the most affected organ and these effects are related to the Al content in the tissue. Finally, it was observed that muscle and gills presented a higher BCF, compared to brain and liver. These findings show that AlNP are capable of generating oxidative stress in carp, affecting tissue function and accumulating, which represents an important risk for the health of fish such as common carp.

Electrospun Mats Based on PVA/NaDDBS/CNx Nanocomposite for Electrochemical Sensing.

This study presents a nanocomposite developed with PVA, multiwall carbon nanotubes (CNTs) doped with nitrogen, and NaDDBS, which change the electrical properties of the polymer and its viscosity to be used in electrospinning process for obtaining mats of nano/macro fibers. The proposed nanocomposite was characterized using Fourier transform-infrared and Raman spectroscopy techniques, confirming the presence of the CNxs immersed in the polymer. High-resolution transmission electron microscopy was used to obtain the micrographs that showed the characteristic interplanar distances of the multiwall CNT in the polymeric matrix, with values of 3.63 Å. Finally, the CNx mats were exposed to various aqueous solutions in a potentiostat to demonstrate the effectiveness of the nanofibers for electrochemical analysis. The CNx-induced changes in the electrical properties of the polymer were identified using cyclic voltammograms, while the electrochemical analysis revealed supercapacitor behavior.

Effects of TiO2 Nanoparticles Incorporation into Cells of Tomato Roots.

In this study, tomato plants were grown in vitro with and without incorporation of TiO2 nanoparticles in Murashige and Skoog (MS) growth medium. The aim of this study was to describe the morphological (area and roundness cell) and mechanical (Young's Modulus) change in the different tissue of tomato root, epidermis (Ep), parenchyma (Pa), and vascular bundles (Vb), when the whole plant was exposed to TiO2 nanoparticles (TiO2 NPs). light microscopy (LM), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM), wavelength dispersive X-ray fluorescence (WDXRF) techniques were used to identify changes into the root cells when TiO2 NPs were incorporated. TiO2 NPs incorporation produces changes in the area, roundness, and Young's Modulus of the tomato root. When tomato root is exposed to TiO2 NPs, the Ep and Vb area size decreases from 260.92 µm2 to 160.71 µm2 and, 103.08 µm2 to 52.13 µm2, respectively, compared with the control area, while in Pa tissue the area size was increased considerably from 337.72 mm2 to 892.96 mm2. Cellular roundness was evident in tomato root that was exposed to TiO2 NPs in the Ep (0.49 to 0.67), Pa (0.63 to 0.79), and Vb (0.76 to 0.71) area zones. Young's Modulus in Pa zone showed a rigid mechanical behavior when tomato root is exposed to TiO2 NPs (0.48 to 4.98 MPa control and TiO2 NPs, respectively). Meanwhile, Ep and Vb were softer than the control sample (13.9 to 1.06 MPa and 6.37 to 4.41 MPa respectively). This means that the Pa zone was stiffer than Ep and Vb when the root is exposed to TiO2 NPs. Furthermore, TiO2 NPs were internalized in the root tissue of tomato, accumulating mainly in the cell wall and intercellular spaces, with a wide distribution throughout the tissue, as seen in TEM.

Effect of ball milling on cellulose nanoparticles structure obtained from garlic and agave waste.

The aim of this study was to obtain cellulose and cellulose nanoparticles (CNP) from garlic and agave wastes, as well as elucidating its structure at different scales using microscopy and spectroscopy techniques. Cellulose is isolated by using a sequential extractive process and monitored by CLSM and SEM, while CNP are produced in a high-energy planetary mill. FTIR and XRD confirmed the presence of cellulose type I and CI and Dhkl was used to evaluate the size of CNP. The corresponding crystalline structure, d-spacing and angles obtained from crystalline regions of CNP were estimated by TEM and computational simulation. It is shown that the triclinic phase is predominant in G, and a monoclinic conformation in CNP for A. The novelty of this contribution is to demonstrate that the crystalline structure of CNP, extracted from different agro-food wastes, depends on its initial microstructural arrangement (laminar or fibrillar).

Study of cellular architecture and micromechanical properties of cuajilote fruits (Parmentiera edulis D.C.) using different microscopy techniques.

The cuajilote (Parmentiera edulis D.C.) tree produces fibrous fruits with a high content of lignocellulosic compounds. However, this fruit and their fibers have been scarcely studied. For this reason, an integral study of their cellular architecture, physicochemical, micromechanical, and structural properties in two maturity stages were carried out. Physicochemical tests, light, confocal and electron microscopy, microindentation, and X-ray diffraction were used for the characterization of fruit and their fibers. Chemical analysis showed that the unripe fruits have the highest cellulose content (42.17%), but in ripe fruit the cellulose content decreases (32.76%) while lignin content increases from 35.26 to 40.79%, caused by the lignification of the sclerenchyma fibers. Microstructural and micromechanical studies in the different regions of the fruit provided relevant information about its cellular architecture, distribution of lignocellulosic compounds and its role in the micromechanical properties of their fibers. The thickening cell wall of sclerenchyma fibers was caused by the cellular lignification of the ripe fruits. According to the physicochemical and structural studies, cuajilote fibers are comparable to other fibers obtained from crops rich in lignocellulosic compounds. The current study provided new knowledge about the cellular architecture of fruit and criteria for selecting the ripening stage adequate for the extraction of cellulose or lignin. Furthermore, information regarding the micromechanical properties of their fibers and which structural arrangement could be more convenient for mechanical reinforcement of biodegradable materials was obtained.

Detection of Brucella abortus by a platform functionalized with protein A and specific antibodies IgG.

Oriented immobilization of antibodies on a sensor surface is critical for enhancing both the antigen-binding capacity and the sensitivity of immunosensors. In this study, we describe a strategy to adsorb immunoglobulin G (IgG) anti-Brucella antibodies onto a silicon surface, oriented by protein A obtained from Staphylococcus aureus (SpA). X-ray photoelectron spectroscopy and atomic force microscopy were used to characterize topographically, morphologically, and chemical changes of the sensor functionalization. The activity of the biosensor was assessed by confocal microscopy, scanning electronic microscopy, and bacteria capture assays (BCA). According to the BCA, the efficiency of Brucella abortus detection with the SpA-IgG anti Brucella biosensor was three-fold higher than that of the random orientated IgG anti Brucella biosensor. The limit of detection was 1 × 106 CFU/ml. These data show that the orientation of antibodies immobilization is crucial to developing immunosensors for bacterial antigen detection as Brucella spp and improve its sensibility level. Functionalization with protein A increases Brucella detection by an antibody-coated surface. Functionalized silicon surface for Brucella detection was characterized by atomic force microscopy, X-ray photoelectron spectroscopy and confocal microscopy.

Design of Nano-Structured Micro-Thermoelectric Generator: Load Resistance and Inflections in the Efficiency.

In recent years the interest for the harvest of energy with micro thermoelectric generators ( µ TEG) has increased, due to its advantages compared to technologies that use fossil fuels. There are three ways to improve the performance of the device, by modifying its structure, type of material and operation control. In this study, the role of the load resistance R L on the performance of a µ TEG with nanostructured materials is investigated. The interaction of the load resistance with the thermoelements exhibits interesting features, arising from the coupling of the temperature-dependent electrical and thermal transport properties at different temperature ranges and the architecture of nanostructured thermoelectric materials. This coupling results in inflections on the efficiency, i.e., maximum and minimum values of the efficiency at higher temperatures, 600-900 K. We show the explicit dependence of the performance of the µ TEG in terms of the load resistance and discuss the underlying physics. The unusual features of the efficiency of nanostructured thermoelectric materials are a result of the behavior of the power factor and the nonequilibrium properties of the system. We also analyze the effect of the geometric shape of the thermoelements on the device. We determine the performance of the µ TEG, evaluating the generation power and its efficiency. The results show that the efficiency of the device can decrease or increase depending on the value of R L , while the power decreases with an increase of the load resistance.

Morphometric study of adipocytes on breast cancer by means of photonic microscopy and image analysis.

Worldwide, breast cancer (BrCa) is currently the leading cause of deaths associated to malignant lesions in adult women. Given that some studies have mentioned that peritumoral adipocytes may contribute to breast carcinogenesis, present work sought to quantitative evaluate the morphometry of these cells in a group of adult women. Three thousand six hundred sixty four breast adipocytes, that came from biopsies of a group of adult females with different types of breast carcinomas (ductal, lobular, and mixed) and one with normal tissues, were evaluated through an image analysis (IA) process regarding six morphometric descriptors: area (A), perimeter (P), Feret diameter (FD ), aspect ratio (AR), roundness factor (RF), and fractal dimension of cellular contour (FDC ). Data showed that the adipocytes of the normal tissues group were bigger (A: 3398 ± 2331 µm2 , P: 239 ± 83 µm, and FD : 79.9 ± 24.5 µm) than those from BrCa samples (A: 2860 ± 1933 µm2 , P: 214 ± 66 µm, and FD : 73.2 ± 22.5 µm), and presented a more irregular contour (FDC of 1.370 ± 0.037 for normal group and of 1.335 ± 0.049 for the oncologic one). Moreover, it could be accounted that adipocytes of mixed carcinomas were largest (FD : 75.1 ± 22.4 µm) than those of lobular lesions (FD : 61.6 ± 22.6 µm), while the adipocytes of ductal carcinomas were the most oval (AR: 1.421 ± 0.524) and roughest (FDC : 1.324 ± 0.050) cells. IA results suggest that BrCa lesions can be categorized through a quantitative morphometric evaluation of peritumoral adipocytes. These findings could let the development of an analytical tool to help the Pathologist to enhance the accuracy of the oncologic diagnose.

Microstructural characterization of chitosan and alginate films by microscopy techniques and texture image analysis.

The aim of this work is to characterize the microstructure of chitosan and alginate edible films by microscopy techniques and texture image analysis. Edible films were obtained by solution casting and solvent evaporation. The microscopy techniques used in this work were: light, environmental scanning electron and atomic force microscopy. Textural features and fractal dimension were extracted from the images. Entropy and fractal dimension were more useful to evaluate the complexity and roughness of films. The highest values of entropy and fractal dimension corresponded to alginate/chitosan, followed of alginate and chitosan films. An entropy/fractal dimension ratio, proposed here, was useful to characterize the degree of image complexity and roughness of edible films at different magnifications. It was possible to postulate that microscopy techniques combined with texture image analysis are efficient tools to quantitatively evaluate the surface morphology of edible films made of chitosan and alginate.