Search

The present study investigated potential modulatory effects of low doses of nano-sized titanium dioxide (TiO 2 ) on intestinal cells in vivo and in vitro . After short-term exposure to TiO 2 nanoparticles in rats, histopathological analysis of intestinal tissues indicated a gender-specific effect with increased length of intestinal villi in male rats only. Moreover the intestinal tissue showed nanoparticle deposition as revealed by ICP-MS determination of titanium. Increased serum testosterone levels were also detected. Considering the male-specific effects detected in vivo , the TiO 2 nanoparticle interaction with intestinal cells was further characterized in vitro and the modulating effect of testosterone and a hormone-induced growth factor, namely Insulin-like Growth Factor 1 (IGF-1), was also assessed. Cytotoxicity assays and analysis of Reactive Oxygen Species (ROS) production showed neither cellular alteration nor oxidative stress for nanoparticles at low concentrations, even though they were able to penetrate intestinal cells, as revealed by electron microscopy. Cell treatments with nanoparticles in association with testosterone or IGF-1 showed increased cell proliferation, compared to nanoparticles or testosterone/IGF-1 alone. Since long-term intake of TiO 2 nanoparticles at low doses is a relevant scenario for human exposure, attention should be given to the potential modulating activity of this nanomaterial on cell proliferation. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is the agent of listeriosis, a food-borne disease. It represents a serious problem for the food industry because of its environmental persistence mainly due to its ability to form biofilm on a variety of surfaces. Microrganisms attached on the surfaces are a potential source of contamination for environment and animals and humans. Titanium dioxide nanoparticles (TiO2 NPs) are used in food industry in a variety of products and it was reported that daily exposure to these nanomaterials is very high. Anti-listerial activity of TiO2 NPs was investigated only with UV-irradiated nanomaterials, based on generation of reactive oxigen species (ROS) with antibacterial effect after UV exposure. Since both Listeria monocytogenes and TiO2 NPs are veicolated with foods, this study explores the interaction between Listeria monocytogenes and non UV-irradiated TiO2 NPs, with special focus on biofilm formation and intestinal cell interaction. Scanning electron microscopy and quantitative measurements of biofilm mass indicate that NPs influence both production and structural architecture of listerial biofilm. Moreover, TiO2 NPs show to interfere with bacterial interaction to intestinal cells. Increased biofilm production due to TiO2 NPs exposure may favour bacterial survival in environment and its transmission to animal and human hosts. [ABSTRACT FROM AUTHOR]

Exosomes are among the most puzzling vehicles of intercellular communication, but several crucial aspects of their biogenesis remain elusive, primarily due to the difficulty in purifying vesicles with similar sizes and densities. Here we report an effective methodology for labelling small extracellular vesicles (sEV) using Bodipy FL C16, a fluorescent palmitic acid analogue. In this study, we present compelling evidence that the fluorescent sEV population derived from Bodipy C16‐labelled cells represents a discrete subpopulation of small exosomes following an intracellular pathway. Rapid cellular uptake and metabolism of Bodipy C16 resulted in the incorporation of fluorescent phospholipids into intracellular organelles specifically excluding the plasma membrane and ultimately becoming part of the exosomal membrane. Importantly, our fluorescence labelling method facilitated accurate quantification and characterization of exosomes, overcoming the limitations of nonspecific dye incorporation into heterogeneous vesicle populations. The characterization of Bodipy‐labelled exosomes reveals their enrichment in tetraspanin markers, particularly CD63 and CD81, and in minor proportion CD9. Moreover, we employed nanoFACS sorting and electron microscopy to confirm the exosomal nature of Bodipy‐labelled vesicles. This innovative metabolic labelling approach, based on the fate of a fatty acid, offers new avenues for investigating exosome biogenesis and functional properties in various physiological and pathological contexts. [ABSTRACT FROM AUTHOR]

Caveolin‐1 (Cav‐1) is a fundamental constituent of caveolae, whose functionality and structure are strictly dependent on cholesterol. In this work the U18666A inhibitor was used to study the role of cholesterol transport in the endosomal degradative‐secretory system in a metastatic human melanoma cell line (WM266‐4). We found that U18666A induces a shift of Cav‐1 from the plasma membrane to the endolysosomal compartment, which is involved, through Multi Vesicular Bodies (MVBs), in the formation and release of small extracellular vesicles (sEVs). Moreover, this inhibitor induces an increase in the production of sEVs with chemical–physical characteristics similar to control sEVs but with a different protein composition (lower expression of Cav‐1 and increase of LC3II) and reduced transfer capacity on target cells. Furthermore, we determined that U18666A affects mitochondrial function and also cancer cell aggressive features, such as migration and invasion. Taken together, these results indicate that the blockage of cholesterol transport, determining the internalization of Cav‐1, may modify sEVs secretory pathways through an increased fusion between autophagosomes and MVBs to form amphisome, which in turn fuses with the plasma membrane releasing a heterogeneous population of sEVs to maintain homeostasis and ensure correct cellular functionality. [ABSTRACT FROM AUTHOR]

Increasing interest is being addressed to the development of a reliable, reproducible and relevant in vitro model of intestinal barrier, mainly for engineered nanomaterials hazard and risk assessment, in order to meet regulatory and scientific demands. Starting from the consolidated Caco-2 cell model, widely used for determining translocation of drugs and chemicals, the establishment of an advanced intestinal barrier model with different level of complexity is important for overcoming Caco-2 monoculture limitations. For this purpose, a tri-culture model, consisting of two human intestinal epithelial cells (Caco-2 and HT29-MTX) and a human lymphocyte B cell (Raji B), was developed by several research groups to mimic the in vivo intestinal epithelium, furnishing appropriate tools for nanotoxicological studies. However, tri-culture model shows high levels of variability in ENM uptake/translocation studies. With the aim of implementing the standardization and optimization of this tri-culture for ENM translocation studies, the present paper intends to identify and discuss such relevant parameters involved in model establishment as: tri-culture condition set-up, barrier integrity evaluation, mucus characterization, M-cell induction. SiO2 fluorescent nanoparticles were used to compare the different models. Although a low level of SiO2 translocation is reported for all the different culture conditions. a relevant role of mucus and M-cells in NPs uptake/translocation has been highlighted. [ABSTRACT FROM AUTHOR]

The analytical issues connected with chromium determination in foodstuffs by quadrupole inductively coupled plasma–mass spectrometry (Q-ICP-MS) were addressed, including signal stability, spectral interferences and the use of mathematical correction equations. The analytical performance was compared to that of electrothermal atomisation–atomic absorption spectrometry (ETA-AAS), selected as reference method. Five food certified reference materials (CRMs), including two meat-based CRMs not previously characterized for their Cr content, were included in the study. The use of ultrasonic nebulization (UN) and the adoption of 53Cr as analytical mass allowed precise and accurate results to be obtained by Q-ICP-MS, with lower detection limits than ETA-AAS. [Copyright &y& Elsevier]

The features of Graphene Oxide (GO) could produce unexpected interactions with ecosystem, triggering negative effects on biota. In this work we evaluated the effects of two different GO samples (chemically synthesized GOC and electrochemically synthesized GOE) on Lemna gibba L., after incubation with different concentrations over a period of 21 days. The stress conditions were evidenced by the measurements of frond number, fresh weight, dry weight, dry/fresh weight ratio, and inhibition of growth rate. Furthermore, a morphological characterization was also carried out to verify the mechanical effects on L. gibba plants of these two GO. Results demonstrated that the smallest GOE nano sheets were very well dispersed on L. gibba fronds than GOC. This effect could be explained considering the lower GOE functionalization degree and, as a result, a less electrostatic adhesion among nanosheets. The presence of electrically charged functionalities mainly provides electrostatic interactions with plants causing growth inhibition as shown by GOC. [ABSTRACT FROM AUTHOR]

In this work, the authors prepared and characterized two different graphene oxides: one chemically synthesized (GO sample) and the other one electrochemically synthesized (GO(LiCl)). Both samples were fully characterized with atomic force microscopy (AFM), Raman and Fourier transform infrared (FTIR) spectroscopies, X-ray photo electron spectroscopy (XPS), thermal analysis (TG/DTA), and Z-potential. The antibacterial properties of both graphene oxides were studied using Gram-negative Escherichia coli ATCC 25922 and Gram-positive Staphylococcus aureus ATCC 25923 by spectrophotometer and viable cell count as indirect and direct methods, respectively. Results demonstrated that the GO(LiCl) exhibited a significant antibacterial activity compared to GO that showed a bacteriostatic effect on both pathogens. Electron microscopy analysis confirmed the antibacterial effects of both graphene oxides toward the pathogens, especially working at 80 μg/mL, for 24 h. Additional studies were also performed and both GO samples were not cytotoxic at 2 μg/mL toward neuroblastoma cells. Moreover, 2 μg of GO was suitable to carry the minimum effective dose (5.74 ng/mL) of kinase inhibitor S29 (1-(2-chloro-2-(4-chlorophenyl)ethyl)-N-(4-fluorobenzyl)-1H-pyrazolo[3,4-d] pyrimidin-4-amine), providing negligible side effects related to the S29 treatment (this latter being specifically active on the neuroblastoma cell lines (SK-N-BE(2))). [ABSTRACT FROM AUTHOR]

Introduction: Recently, we introduced a new deposition method, based on Ion Plating Plasma Assisted technology, to coat titanium implants with a thin but hard nanostructured layer composed of titanium carbide and titanium oxides, clustered around graphitic carbon. The nanostructured layer has a double effect: protects the bulk titanium against the harsh conditions of biological tissues and in the same time has a stimulating action on osteoblasts. Results: The aim of this work is to describe the biological effects of this layer on osteoblasts cultured in vitro. We demonstrate that the nanostructured layer causes an overexpression of many early genes correlated to proteins involved in bone turnover and an increase in the number of surface receptors for α3β1 integrin, talin, paxillin. Analyses at single-cell level, by scanning electron microscopy, atomic force microscopy, and single cell force spectroscopy, show how the proliferation, adhesion and spreading of cells cultured on coated titanium samples are higher than on uncoated titanium ones. Finally, the chemistry of the layer induces a better formation of blood clots and a higher number of adhered platelets, compared to the uncoated cases, and these are useful features to improve the speed of implant osseointegration. Conclusion: In summary, the nanostructured TiC film, due to its physical and chemical properties, can be used to protect the implants and to improve their acceptance by the bone. [ABSTRACT FROM AUTHOR]

Abstract: It has been suggested that the rarity of human listeriosis due to Listeria ivanovii reflects not only host tropism factors but also the rare occurrence of this species in the environment, compared with Listeria monocytogenes. In the present study we evaluate the effects on the reference strain L. ivanovii ATCC 19119 behaviour of two combined stresses, low iron availability and acid environment, that bacteria can encounter in the passage from saprophytic life to the host. In these conditions, L. ivanovii evidenced a different behaviour compared to L. monocytogenes exposed to similar conditions. L. ivanovii was not able to mount an acid tolerance response (ATR) even if, upon entry into the stationary phase in iron-loaded medium, growth phase-dependent acid resistance (AR) was evidenced. Moreover, bacteria grown in iron excess and acidic pH showed the higher invasion value in Caco-2 cells, even though it was not able to efficiently multiply. On the contrary, low iron and acidic conditions improved invasion ability in amniotic WISH cells. [Copyright &y& Elsevier]

The Gram-negative bacterium Pseudomonas aeruginosa is one of the most dreaded pathogens in the hospital setting, and represents a prototype of multi-drug resistant “superbug” for which effective therapeutic options are very limited. The identification and characterization of new cellular functions that are essential for P. aeruginosa viability and/or virulence could drive the development of anti-Pseudomonas compounds with novel mechanisms of action. In this study we investigated whether TolB, the periplasmic component of the Tol-Pal trans-envelope protein complex of Gram-negative bacteria, represents a potential drug target in P. aeruginosa. By combining conditional mutagenesis with the analysis of specific pathogenicity-related phenotypes, we demonstrated that TolB is essential for P. aeruginosa growth, both in laboratory and clinical strains, and that TolB-depleted P. aeruginosa cells are strongly defective in cell-envelope integrity, resistance to human serum and several antibiotics, as well as in the ability to cause infection and persist in an insect model of P. aeruginosa infection. The essentiality of TolB for P. aeruginosa growth, resistance and pathogenicity highlights the potential of TolB as a novel molecular target for anti-P. aeruginosa drug discovery. [ABSTRACT FROM AUTHOR]

Simple Summary: Early detection of cutaneous melanoma is the key to increasing survival and proper therapeutic adjustment, especially in stages II–IV. We investigated whether the fatty acid (FA) and protein compositions of small extracellular vesicles (sEV) expressing CD81, derived from the plasma of stage 0–I, II and III–IV melanoma patients, could reflect disease stage. Results showed a higher content of FA and differences in C18:0/C18:1 ratio, a marker of cell membrane fluidity, that distinguished patients' CD81sEV from those of healthy donors (HD). By proteomic analysis (identifier PXD024434) we identified significant increases in CD14, PON1, PON3 and APOA5 in stage II CD81sEV compared to HD. In stage III–IV, CD81sEV' RAP1B expression was decreased. These stage-related signatures may support the potential of sEV to provide information for early diagnosis, prediction of metastatic behavior, treatment and follow-up of melanoma patients. The early detection of cutaneous melanoma, a potentially lethal cancer with rising incidence, is fundamental to increasing survival and therapeutic adjustment. In stages II–IV especially, additional indications for adjuvant therapy purposes after resection and for treatment of metastatic patients are urgently needed. We investigated whether the fatty acid (FA) and protein compositions of small extracellular vesicles (sEV) derived from the plasma of stage 0–I, II and III–IV melanoma patients (n = 38) could reflect disease stage. The subpopulation of sEV expressing CD81 EV marker (CD81sEV) was captured by an ad hoc immune affinity technique from plasma depleted of large EV. Biological macromolecules were investigated by gas chromatography and mass spectrometry in CD81sEV. A higher content of FA was detectable in patients with respect to healthy donors (HD). Moreover, a higher C18:0/C18:1 ratio, as a marker of cell membrane fluidity, distinguished early (stage 0–I) from late (III–IV) stages' CD81sEV. Proteomics detected increases in CD14, PON1, PON3 and APOA5 exclusively in stage II CD81sEV, and RAP1B was decreased in stage III–IV CD81sEV, in comparison to HD. Our results suggest that stage dependent alterations in CD81sEV' FA and protein composition may occur early after disease onset, strengthening the potential of circulating sEV as a source of discriminatory information for early diagnosis, prediction of metastatic behavior and following up of melanoma patients. [ABSTRACT FROM AUTHOR]

• An Electrogenic and hydrocarbonoclastic biofilm as response to oil spill • Biofilm microbes are involved in electrogenic and hydrocarbonoclastic activity • Biofilm was connected to bacterial filaments to transfer electrons at distance • Electrogenic filamentation was associated to "myelin like" expression pattern • A vertical and spontaneous Liquid Microbial Fuel Cell (L-MFC) was created The oil-water interface formed during an oil spill represents a challenging environment for pelagic communities living in aquatic ecosystems. At this anoxic barrier, we report the formation of a microbial hydrocarbonoclastic biofilm capable of electron transfer along the water column. This biofilm generated a membrane of surface-active compounds that allowed the spontaneous separation of electrical charges, causing the establishment of an anodic and a cathodic region and, as a result, the spontaneous creation of a liquid microbial fuel cell. Such floating biofilm was connected to the water column underneath by floating filaments that could contribute to oxygen reduction at distance. The filaments revealed an unusual lipid content induced by anoxic conditions, with prominent ultrastructural features similar to myelin found in oligodendrocytes of the vertebrate nervous system. Furthermore, these filaments showed an interesting cross-reactivity towards different epitopes of the myelin basic protein (MBP) and Claudin 11 (O 4) of human oligodendrocytes. The presence of a network of filaments similar to myelin suggests the probable existence of evolutionary connections between very distant organisms. Collectively these results suggest a possible mechanism for how lake microbial communities can adapt to oil spills while offering an interesting starting point for technological developments of liquid microbial fuel cells related to the study of hydrocarbon-water interfaces. The data that support the findings of this study are openly available in figshare at https://figshare.com/s/72bc73ae14011dc7920d. [Display omitted] [ABSTRACT FROM AUTHOR]