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Modern imaging strategies are paramount to studying living systems such as cells, bacteria, and fungi and their response to pathogens, toxicants, and nanomaterials (NMs) as modulated by exposure and environmental factors. The need to understand the processes and mechanisms of damage, healing, and cell survivability of living systems continues to motivate the development of alternative imaging strategies. Of particular interest is the use of label-free techniques (microscopy procedures that do not require sample staining) that minimize interference of biological processes by foreign marking substances and reduce intense light exposure and potential photo-toxicity effects. This review focuses on the synergic capabilities of atomic force microscopy (AFM) as a well-developed and robust imaging strategy with demonstrated applications to unravel intimate details in biomedical applications, with the label-free, fast, and enduring Holotomographic Microscopy (HTM) strategy. HTM is a technique that combines holography and tomography using a low intensity continuous illumination laser to investigate (quantitatively and non-invasively) cells, microorganisms, and thin tissue by generating three-dimensional (3D) images and monitoring in real-time inner morphological changes. We first review the operating principles that form the basis for the complementary details provided by these techniques regarding the surface and internal information provided by HTM and AFM, which are essential and complimentary for the development of several biomedical areas studying the interaction mechanisms of NMs with living organisms. First, AFM can provide superb resolution on surface morphology and biomechanical characterization. Second, the quantitative phase capabilities of HTM enable superb modeling and quantification of the volume, surface area, protein content, and mass density of the main components of cells and microorganisms, including the morphology of cells in microbiological systems. These capabilities result from directly quantifying refractive index changes without requiring fluorescent markers or chemicals. As such, HTM is ideal for long-term monitoring of living organisms in conditions close to their natural settings. We present a case-based review of the principal uses of both techniques and their essential contributions to nanomedicine and nanotoxicology (study of the harmful effects of NMs in living organisms), emphasizing cancer and infectious disease control. The synergic impact of the sequential use of these complementary strategies provides a clear drive for adopting these techniques as interdependent fundamental tools.

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Nanomaterials such as titanium dioxide and magnetite are increasingly used in several fields, such as water remediation and agriculture. However, this has raised environmental concerns due to potential exposure to organisms like humans. Nanomaterials can cause adverse interactions depending on physicochemical characteristics, like size, morphology, and composition, when interacting with living beings. To ensure safe use and prevent the risk of exposure to nanomaterials, their biocompatibility must be assessed. In vitro cell cultures are beneficial for assessing nanomaterial-cell interactions due to their easy handling. The present study evaluated the biocompatibility of TiO2, Fe3O4, and TiO2/Fe3O4 nanomaterials thermally treated at 350 °C and 450 °C in erythrocytes and HepG2 cells. According to the hemolysis experiments, non-thermally treated NMs are toxic (>5% hemolysis), but their thermally treated counterparts do not present toxicity (<2%). This behavior indicates that the toxicity derives from some precursor (solvent or surfactant) used in the synthesis of the nanomaterials. All the thermally treated nanomaterials did not show hemolytic activity under different conditions, such as low-light exposure or the absence of blood plasma proteins. In contrast, non-thermally treated nanomaterials showed a high hemolytic behavior, which was reduced after the purification (washing and thermal treatment) of nanomaterials, indicating the presence of surfactant residue used during synthesis. An MTS cell viability assay shows that calcined nanomaterials do not reduce cell viability (>11%) during 24 h of exposure. On the other hand, a lactate dehydrogenase leakage assay resulted in a higher variability, indicating that several nanomaterials did not cause an increase in cell death as compared to the control. However, a holotomographic microscopy analysis reveals a high accumulation of nanomaterials in the cell structure at a low concentration (10 µg mL-1), altering cell morphology, which could lead to cell membrane damage and cell viability reduction.

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Fungal infections have become a significant public health concern due to their increasing recurrence and harmful effects on plants, animals, and humans. Opportunistic pathogens (among others from the genera Candida and Aspergillus) can be present in indoor air, becoming a risk for people with suppressed immune systems. Engineered nanomaterials are novel alternatives to traditional antifungal therapy. In this work, copper(I) iodide (CuI) and a copper-doped titanium dioxide-copper(I) iodide (TiO2-Cu2+/CuI) composite nanomaterials (NMs)-were synthesized and tested as antifungal agents. The materials were synthesized using sol-gel (TiO2-Cu2+) and co-precipitation (CuI) techniques. The resulting colloids were evaluated as antifungal agents against Candida parapsilosis and Aspergillus niger strains. The NMs were characterized by XRD, HRTEM, AFM, and DLS to evaluate their physicochemical properties. The NMs present a high size dispersion and different geometrical shapes of agglomerates. The antifungal capacity of the NMs by the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) was below 15 µg/mL against Candida parapsilosis and below 600 µg/mL against Aspergillus niger for both NMs. Holotomography microscopy showed that the NMs could penetrate cell membranes causing cell death through its rupture and reactive oxygen species (ROS) production. Cytotoxicity tests showed that NMs could be safe to use at low concentrations. The synthesized nanomaterials could be potential antifungal agents for biomedical or environmental applications.

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Fungal mycoses have become an important health and environmental concern due to the numerous deleterious side effects on the well-being of plants and humans. Antifungal therapy is limited, expensive, and unspecific (causes toxic effects), thus, more efficient alternatives need to be developed. In this work, Copper (I) Iodide (CuI) nanomaterials (NMs) were synthesized and fully characterized, aiming to develop efficient antifungal agents. The bioactivity of CuI NMs was evaluated using Sporothrix schenckii and Candida albicans as model organisms. CuI NMs were prepared as powders and as colloidal suspensions by a two-step reaction: first, the CuI2 controlled precipitation, followed by hydrazine reduction. Biopolymers (Arabic gum and chitosan) were used as surfactants to control the size of the CuI materials and to enhance its antifungal activity. The materials (powders and colloids) were characterized by SEM-EDX and AFM. The materials exhibit a hierarchical 3D shell morphology composed of ordered nanostructures. Excellent antifungal activity is shown by the NMs against pathogenic fungal strains, due to the simultaneous and multiple mechanisms of the composites to combat fungi. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of CuI-AG and CuI-Chitosan are below 50 µg/mL (with 5 h of exposition). Optical and Atomic Force Microscopy (AFM) analyses demonstrate the capability of the materials to disrupt biofilm formation. AFM also demonstrates the ability of the materials to adhere and penetrate fungal cells, followed by their lysis and death. Following the concept of safe by design, the biocompatibility of the materials was tested. The hemolytic activity of the materials was evaluated using red blood cells. Our results indicate that the materials show an excellent antifungal activity at lower doses of hemolytic disruption.

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Among the different properties of the hydrophobic semiconductor surfaces, self-cleaning promoted by solar illumination is probably one of the most attractive from the technological point of view. The use of sonochemistry for nanomaterials' synthesis has been recently employed for the associated shorter reaction times and efficient route for control over crystal growth and the management of the resulting material's photocatalytic properties. Moreover, the sol-gel method coupled to sonochemistry modifies the chemical environment, with reactive species such as •OH and H2O2, which yield a homogeneous synthesis. Therefore, in the following investigation, the sol-gel method was coupled to sonochemistry to synthesize a SiO2@TiO2 composite, for which the sonochemical amplitude of irradiation was varied to determine its effect on the morphology and mechanical and self-cleaning properties. SEM and AFM characterized the samples of SiO2@TiO2 composite, and while the micrographs indicate that a high ultrasonic energy results in an amorphous SiO2@TiO2 composite with a low rugosity, which was affected in the determination of the contact angle on the surface. On the other hand, FTIR analysis suggests a significant change in both SiO2-SiO and SiO2-TiO2 chemical bonds with changes in vibrations and frequency, corroborating an important influence of the sonochemical energy contribution to the hydrolysis process. Raman spectroscopy confirms the presence of an amorphous phase of silicon dioxide; however, the vibrations of TiO2 were not visible. The evaluation of hydrophobic and self-cleaning properties shows a maximum of ultrasonic energy needed to improve the contact angle and rhodamine B (RhB) removal.

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Safety on the use of magnetic nanomaterials (MNMs) has become an active topic of research given all the recent applications of these materials in various fields. It is known that the toxicity of MNMs depends on size, shape, and surface functionalization. In this study, we evaluate the biocompatibility with different aquatic organisms of engineered MNMs-CIT with excellent aqueous dispersion and long-term colloidal stability. Primary producers (the alga Pseudokirchneriella subcapitata), primary consumers (the rotifer Lecane papuana), and predators (the fish, Danio rerio) interacted with these materials in acute and sub-chronic toxicity tests. Our results indicate that P. subcaptita was the most sensitive taxon to MNMs-CIT. Inhibition of their population growth (IC50 = 22.84 mg L-1) elicited cell malformations and increased the content of photosynthetic pigments, likely due to inhibition of cell division (as demonstrated in AFM analysis). For L. papuana, the acute exposure to MNMs shows no significant mortality. However, adverse effects such as decreased rate of population and altered swimming patterns arise after chronic interaction with MNMs. For D. rerio organisms on early life stages, their exposure to MNMs results in delayed hatching of eggs, diminished survival of larvae, altered energy resources allocation (measured as the content of total carbohydrates, lipids, and protein), and increased glucose demand. As to our knowledge, this is the first study that includes three different trophic levels to assess the effect of MNMs in aquatic organisms; furthermore, we demonstrated that these MNMs pose hazards on aquatic food webs at low concentrations (few mgL-1).

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Nowadays, air pollution has become a global menace being responsible of a significant increase on the morbidity and mortality of human beings. In view of this, sustainable and efficient technologies for air purification are being sought. Air purification by photocatalytic treatment has received a lot of attention due to the unspecific and high oxidation capacity of the catalyst; however still some variables must be optimized to assure practical applications. In this work, visible light active TiO2-Cu2+@perlite and Ag@TiO2-Cu2+/perlite supported materials were fabricated. TiO2-Cu2+ (2 at. %) were synthesized using a sol-gel procedure followed of the impregnation of the support by immersion. For Ag@TiO2-Cu2+, silver deposition was conducted by chemical reduction using sodium citrate and sodium borohydride. The materials (powders and supported materials) were characterized by Scanning Electron Microscopy (SEM) to demonstrate their small size and adherence to the substrate. A prototype of a photocatalytic air purifier was built. The efficacy of the prototype was evaluated for the disinfection of indoor air (dentistry clinics). The photo-catalyst was activated using visible and UVA low-cost high-energy LEDs. The antibacterial activity of the air filter was evaluated. Ag@TiO2-Cu2+ exerts better air disinfection activity at lower doses in comparison to TiO2-Cu2+. Bacterial growth inhibitions up to 99% were achieved for both, Gram-negative and Gram-positive bacteria. The incorporation of Ag and Cu to TiO2 improves the antibacterial activity of the materials due to enhanced photocatalytic activity and the synergic activity of TiO2 and dopant elements (Ag, Cu) to inhibit microorganism's growth.

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Entamoeba histolytica is an invasive enteric protozoan, whose infections are associated to high morbidity and mortality rates. However, only less than 10% of infected patients develop invasive amebiasis. The ability of E. histolytica to adapt to the intestinal microenvironment could be determinant in triggering pathogenic behavior. Indeed, during chronic inflammation, the vagus nerve limits the immune response through the anti-inflammatory reflex, which includes acetylcholine (ACh) as one of the predominant neurotransmitters at the infection site. Consequently, the response of E. histolytica trophozoites to ACh could be implicated in the establishment of invasive disease. The aim of this study was to evaluate the effect of ACh on E. histolytica virulence. Methods include binding detection of ACh to plasma membrane, quantification of the relative expression of virulence factors by RT-PCR and western blot, evaluation of the effect of ACh in different cellular processes related to E. histolytica pathogenesis, and assessment of the capability of E. histolytica to migrate and form hepatic abscesses in hamsters. Results demonstrated that E. histolytica trophozoites bind ACh on their membrane and show a clear increase of the expression of virulence factors, that were upregulated upon stimulation with the neurotransmitter. ACh treatment increased the expression of L220, Gal/GalNAc lectin heavy subunit (170 kDa), amebapore C, cysteine proteinase 2 (ehcp-a2), and cysteine proteinase 5 (ehcp-a5). Moreover, erythrophagocytosis, cytotoxicity, and actin cytoskeleton remodeling were augmented after ACh treatment. Likewise, by assessing the formation of amebic liver abscess, we found that stimulated trophozoites to develop greater hamster hepatic lesions with multiple granulomas. In conclusion, ACh enhanced parasite pathogenicity by upregulating diverse virulence factors, thereby contributing to disease severity, and could be linked to the establishment of invasive amebiasis.

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The goal of this research is to find a more effective treatment for tequila vinasses (TVs) with potential industrial application in order to comply with the Mexican environmental regulations. TVs are characterized by their high content of solids, high values of biochemical oxygen demand (BODs), chemical oxygen demand (COD), low pH and intense colour; thus, disposal of untreated TVs severely impacts the environment. Physicochemical and biological treatments, and a combination of both, were probed on the remediation of TVs. The use of alginate for the physicochemical treatment of TVs reduced BOD5 and COD values by 70.6% and 14.2%, respectively. Twenty white-rot fungi (WRF) strains were tested in TV-based solid media. Pleurotus ostreatus 7992 and Trametes trogii 8154 were selected due to their ability to grow on TV-based solid media. Ligninolytic enzymes' production was observed in liquid cultures of both fungi. Using the selected WRF for TVs' bioremediation, both COD and BOD5 were reduced by 88.7% and 89.7%, respectively. Applying sequential physicochemical and biological treatments, BOD5 and COD were reduced by 91.6% and 93.1%, respectively. Results showed that alginate and selected WRF have potential for the industrial treatment of TVs.

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Fundamental research has been carried out to define optimal "green" synthesis conditions for the production of titania (TiO(2)) and silver (Ag) nanocomposites (TANCs) ranging from 12.7-22.8 nm in diameter. A bottom-up colloidal approach was employed to accurately control TANC monodispersity and composition. TANCs were found to be effective at inactivating Escherichia coli (E. coli) in water. The presence of Ag in the nanocomposites induced a decrease in TiO(2) band gap energy, which favoured valence to conduction band electron transfer and allowed for electron excitation using visible light. Aggregation of ultra-fine particles was prevented through the use of a long-chain polymer as evidenced by electrophoretic mobility studies. The TANCs catalyzed oxidation of bacterial membranes and cell death or disinfection. Theoretically, the TANC mode of E. coli disinfection is via water photolysis, which results in production of hydroxyl radicals and hydrogen peroxide. These interact with the outer membrane polysaccharides and lipids, leading to lipid peroxidation, membrane weakening and resulted in cell death. Our overarching goals were to optimize the variables involved in TANC "green" synthesis and to characterize its nanostructure. High resolution (HR) transmission and scanning electron microscopic (TEM and SEM) studies demonstrated that TANCs were highly crystalline and mono-dispersive. Elemental composition of Ag and Ti, as measured by X-ray energy dispersive (EDS) and X-ray photoelectron spectroscopy (XPS) confirmed sample purity. Ultraviolet-visible (UV-VIS) spectroscopy showed that the energy band-gap of Ag modified TiO(2) was in the visible range.

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The title compound, [Zn(3)(C(9)H(21)SiS)(6)] or [((i)Pr(3)SiS)Zn(mu-SSi(i)Pr(3))(2)Zn(mu-SSi(i)Pr(3))(2)Zn(SSi(i)Pr(3))], is the first structurally characterized homoleptic silanethiolate complex of zinc. A near-linear arrangement of three Zn(II) ions is observed, the metals at the ends being three-coordinate with one terminally bound silanethiolate ligand. The central Zn(II) ion is four-coordinate and tetrahedral, with two bridging silanethiolate ligands joining it to each of the two peripheral Zn(II) ions. The nonbonding intermetallic distances are 3.1344 (11) and 3.2288 (12) A, while the Zn...Zn...Zn angle is 172.34 (2) degrees. A trimetallic silanethiolate species of this type has not been previously identified by X-ray crystallography for any element.

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Silver nanoparticles (Ag-NPs) were synthesized using a facile green chemistry synthetic route. The reaction occurred at ambient temperature with four reducing agents introduced to obtain nanoscale Ag-NPs. The variables of the green synthetic route, such as acidity, concentration of starting materials, and molar ratio of reactants were optimized. Dispersing agents were employed to prevent Ag-NPs from aggregating. Advanced instrumentation techniques, such as X-ray powder diffraction (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), ultraviolet-visible spectroscopy (UV-vis), and phase analysis light scattering technique (ZetaPALS) were applied to characterize the morphology, particle size distribution, elemental composition, and electrokinetic behavior of the Ag-NPs. UV-vis spectra detected the characteristic plasmon at approximately 395-410 nm; and XRD results were indicative of face-centered cubic phase structure of Ag. These particles were found to be monodispersed and highly crystalline, displaying near-spherical appearance, with average particle size of 10.2 nm using citrate or 13.7 nm using ascorbic acid as reductants from particle size analysis by ZetaPALS, respectively. The rapid electrokinetic behavior of the Ag was evaluated using zetapotential (from -40 to -42 mV), which was highly dependant on nanoparticle acidity and particle size. The current research opens a new avenue for the green fabrication of nanomaterials (including variables optimization and aggregation prevention), and functionalization in the field of nanocatalysis, disinfection, and electronics.

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It is well known that exposure to chromium (Cr) can lead to nephrotoxicity. Quercetin is a flavonoid of interest because of its proposed health-promoting effects. The aim of this work was to elucidate the role of quercetin against the nephrotoxicity caused by Cr in rats. Quercetin may have positive effects in combating, or helping to prevent, nephrotoxicity. It was observed that a single dose of potassium dichromate resulted in both an increase of systemic peroxidation of lipids and a decrease of the renal clearance of para-aminohippuric acid and inulin. Our results show that treatment with quercetin protected and prevented against these damaging effects.

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