Obtaining more contaminant-resistant variants from a native Chlorella vulgaris strain.

Cildáñez stream (in Matanza-Riachuelo basin, Buenos Aires) is one of the most polluted watercourses of Argentina, containing a mixed contamination from agricultural and industrial wastes. The application of water bioremediation processes for this kind of effluent will require microorganisms with a high tolerance to contamination. In this sense, obtaining higher contaminant-resistant microalgae lines is widely desired. In this study, adaptive laboratory evolution (ALE) and random mutagenesis were used to obtain Chlorella vulgaris LMPA-40 strains adapted to grow in polluted water from the Cildáñez stream. The ALE process was performed by 22 successive subcultures under selective pressure (Cildáñez wastewater alone or with the addition of phenol or H2O2) while random mutagenesis was performed with UV-C radiation at 275nm. Not all the cell lines obtained after ALE could adapt enough to overcome the stress caused by the Cildáñez wastewater, indicating that the process is quite random and depends on the stressor used. The best results were obtained for the Cildáñez wastewater adapted cells (Cild 3 strain) that were more resistant than the original strain. The concentration of protein, Chlorophyll A, Chlorophyll B, and carotenoids in the Cild 3 ALE evolved strain was higher than that of the control strain. However, this strain exhibited half of the lipid content compared to the same control strain. Interestingly, these alterations and the acquired tolerance may be reversed over time during storage. These findings suggest that the acquisition of novel cell lines could not be permanent, a fact that must be considered for future trials.

The Thioredoxin Fold Protein (TFP2) from Extreme Acidophilic Leptospirillum sp. CF-1 Is a Chaperedoxin-like Protein That Prevents the Aggregation of Proteins under Oxidative Stress.

Extreme acidophilic bacteria like Leptospirillum sp. require an efficient enzyme system to counteract strong oxygen stress conditions in their natural habitat. The genome of Leptospirillum sp. CF-1 encodes the thioredoxin-fold protein TFP2, which exhibits a high structural similarity to the thioredoxin domain of E. coli CnoX. CnoX from Escherichia coli is a chaperedoxin that protects protein substrates from oxidative stress conditions using its holdase function and a subsequent transfer to foldase chaperones for refolding. Recombinantly produced and purified Leptospirillum sp. TFP2 possesses both thioredoxin and chaperone holdase activities in vitro. It can be reduced by thioredoxin reductase (TrxR). The tfp2 gene co-locates with genes for the chaperone foldase GroES/EL on the chromosome. The "tfp2 cluster" (ctpA-groES-groEL-hyp-tfp2-recN) was found between 1.9 and 8.8-fold transcriptionally up-regulated in response to 1 mM hydrogen peroxide (H2O2). Leptospirillum sp. tfp2 heterologously expressed in E. coli wild type and cnoX mutant strains lead to an increased tolerance of these E. coli strains to H2O2 and significantly reduced intracellular protein aggregates. Finally, a proteomic analysis of protein aggregates produced in E. coli upon exposition to oxidative stress with 4 mM H2O2, showed that Leptospirillum sp. tfp2 expression caused a significant decrease in the aggregation of 124 proteins belonging to fifteen different metabolic categories. These included several known substrates of DnaK and GroEL/ES. These findings demonstrate that Leptospirillum sp. TFP2 is a chaperedoxin-like protein, acting as a key player in the control of cellular proteostasis under highly oxidative conditions that prevail in extreme acidic environments.

Ozone and peroxone disinfection of Toxocara canis eggs in water.

Water pollution in developing countries continues to be a major health problem due to various anthropological activities that contribute to the spread of many parasitic diseases, including those caused by helminths. The aim of this study is to explore the ability of ozone and peroxone to disinfect drinking water contaminated samples with Toxocara canis eggs. The oxidants used were ozone and ozone-hydrogen peroxide combination. The treatment of Toxocara canis eggs was carried out in a 50 ml reactor with an operating volume of 10 ml. The pH conditions (5, 7 and 10) were varied for each treatment. The treatment effect was calculated by counting eggs and examining the condition of the larvae larval condition (whole, broken and hatched larvae) using an optical microscope. The experiment was carried out by exposing the eggs for 60 and 120 minutes to ozone and peroxone. The best results were obtained for helminths treated with the ozone/hydrogen peroxide combination at pH 10, with an inactivation of 79.2%. The synergistic effect of ozone combined with hydrogen peroxide allows higher helminth egg inactivation rates, demonstrating that advanced oxidation processes are a real alternative to apply in the inactivation of Toxocara canis eggs. The results obtained in this study show that the ozone and peroxone treatment could be a useful disinfection process to destroy or inactivate Toxocara canis eggs in processes commonly applied in water treatment.

NADPH oxidase 4-derived hydrogen peroxide counterbalances testosterone-induced endothelial dysfunction and migration.

High levels of testosterone (Testo) are associated with cardiovascular risk by increasing reactive oxygen species (ROS) formation. NADPH oxidases (NOX) are the major source of ROS in the vasculature of cardiovascular diseases. NOX4 is a unique isotype, which produces hydrogen peroxide (H2O2), and its participation in cardiovascular biology is controversial. So far, it is unclear whether NOX4 protects from Testo-induced endothelial injury. Thus, we hypothesized that supraphysiological levels of Testo induce endothelial NOX4 expression to attenuate endothelial injury. Human mesenteric vascular endothelial cells (HMECs) and human umbilical vein endothelial cells (HUVEC) were treated with Testo (10-7 M) with or without a NOX4 inhibitor [GLX351322 (10-4 M)] or NOX4 siRNA. In vivo, 10-wk-old C57Bl/6J male mice were treated with Testo (10 mg/kg) for 30 days to study endothelial function. Testo increased mRNA and protein levels of NOX4 in HMECs and HUVECs. Testo increased superoxide anion (O2-) and H2O2 production, which were abolished by NOX1 and NOX4 inhibition, respectively. Testo also attenuated bradykinin-induced NO production, which was further impaired by NOX4 inhibition. In vivo, Testo decreased H2O2 production in aortic segments and triggered endothelial dysfunction [decreased relaxation to acetylcholine (ACh)], which was further impaired by GLX351322 and by a superoxide dismutase and catalase mimetic (EUK134). Finally, Testo led to a dysregulated endothelial cell migration, which was exacerbated by GLX351322. These data indicate that supraphysiological levels of Testo increase the endothelial expression and activity of NOX4 to counterbalance the deleterious effects caused by Testo in endothelial function.NEW & NOTEWORTHY By inducing ROS formation, high levels of testosterone play a major role in the pathogenesis of cardiovascular disease. NOXs are the major sources of ROS in the vasculature of cardiovascular diseases. Herein, we describe a novel compensatory mechanism by showing that NOX4 is a protective oxidant enzyme and counterbalances the deleterious effects of testosterone in endothelial cells by modulating hydrogen peroxide formation.

In-office Bleaching Activated With Violet LED: Effect on Pulpal and Tooth Temperature and Pulp Viability.

OBJECTIVES: This study evaluated the influence of hydrogen peroxide (HP) with or without titanium dioxide nanotubes (TiO2) associated with violet LED (VL) regarding: a) the temperature change in the pulp chamber and facial surface; b) the decomposition of HP; and c) the cytotoxicity of the gels on pulp cells. METHODS AND MATERIALS: The experimental groups were: HP35 (35% HP/Whiteness HP, FGM); HP35+VL; HP35T (HP35+TiO2); HP35T+VL; HP7 (7.5% HP/White Class 7.5%, FGM); HP7+VL; HP7T (HP7+TiO2); and HP7T+VL. TiO2 was incorporated into the bleaching gels at 1%. Eighty bovine incisors were evaluated to determine temperature change in 8 experimental groups (n=10/group). A k-type thermocouple was used to evaluate the temperatures of the facial surface and in the pulp chamber, achieved by enabling endodontic access to the palatal surface, throughout the 30-minute session. HP decomposition (n=3) of gels was evaluated by using an automatic potentiometric titrator at the initial and 30-minute time points. Trans-enamel and trans-dentinal cell viability were assessed with a pulp chamber device as well as enamel and dentin discs (n=6), and the treatment extracts (culture medium + diffused components) were collected and applied to MDPC-23 odontoblast cells to evaluate cell viability according to the MTT test. RESULTS: A temperature increase in the pulp chamber was observed in the presence of VL at 30 minutes (p<0.05) (Mann-Whitney test). Also at 30 minutes, HP35 showed greater decomposition in the presence of VL rather than in its absence (p<0.05) (mixed linear models and the Tukey-Kramer test). HP7 provided greater cell viability than the groups treated with HP35 (p<0.05) (generalized linear models test). Cell viability was significantly lower for HP7 in the presence of VL (p<0.05). CONCLUSION: Pulpal temperature increased with VL (maximum of 1.9°C), but did not exceed the critical limit to cause pulp damage. Less concentrated HP resulted in higher cell viability, even when associated with VL.

Toward Enhanced Antioxidant and Protective Potential: Conjugation of Corn Cob Xylan with Gallic Acid as a Novel Approach.

Maize ranks as the second most widely produced crop globally, yielding approximately 1.2 billion tons, with corn cob being its primary byproduct, constituting 18 kg per 100 kg of corn. Agricultural corn production generates bioactive polysaccharide-rich byproducts, including xylan (Xyl). In this study, we used the redox method to modify corn cob xylan with gallic acid, aiming to enhance its antioxidant and protective capacity against oxidative stress. The conjugation process resulted in a new molecule termed conjugated xylan-gallic acid (Xyl-GA), exhibiting notable improvements in various antioxidant parameters, including total antioxidant capacity (1.4-fold increase), reducing power (1.2-fold increase), hydroxyl radical scavenging (1.6-fold increase), and cupric chelation (27.5-fold increase) when compared with unmodified Xyl. At a concentration of 1 mg/mL, Xyl-GA demonstrated no cytotoxicity, significantly increased fibroblast cell viability (approximately 80%), and effectively mitigated intracellular ROS levels (reduced by 100%) following oxidative damage induced by H2O2. Furthermore, Xyl-GA exhibited non-toxicity toward zebrafish embryos, offered protection against H2O2-induced stress, and reduced the rate of cells undergoing apoptosis resulting from H2O2 exposure. In conclusion, our findings suggest that Xyl-GA possesses potential therapeutic value in addressing oxidative stress-related disturbances. Further investigations are warranted to elucidate the molecular structure of this novel compound and establish correlations with its pharmacological activities.

Role of induced nitric oxide synthases in orofacial nociception/discomfort after dental tooth bleaching with hydrogen peroxide.

OBJECTIVE: To evaluate the role of induced nitric oxide synthase (iNOS) in nociception/orofacial discomfort in rats submitted to tooth whitening with hydrogen peroxide (H2O2). DESIGN: Wistar rats were divided into three groups (n = 24/group): a sham group not submitted to whitening treatment, a saline group submitted to whitening treatment, and a test group submitted to whitening treatment and blockade of iNOS with aminoguanidine 50 mg/kg/day. After 24 and 48 h, and 7 days, the animals were euthanized to collect trigeminal ganglia and maxillae to histomorphometric analysis (size of neuronal bodies and percentage of pulp area filled by vessels) and behavior/nociception (Grimace scales, scratching and biting counting, weight loss and nociception assay). ANOVA-1- or - 2-way tests were used (p < 0.05, GraphPadPrism 5.0). RESULTS: The aminoguanidine-treated group showed a reduction in nociceptive threshold in the masseteric region (p < 0.001), Grimace scale scores (p < 0.001), number of scratching (p = 0.011) and body mass loss (p = 0.007). After 24 and 48 h of tooth bleaching, the saline group showed a significant increase in the mean area of the blood vessels (p = 0.020) and iNOS immunostaining in odontoblasts (p = 0.002) and non-odontoblasts cells (p = 0.025). Aminoguanidine reversed both increases. Tooth bleaching reduced the mean area of neuronal bodies, and aminoguanidine significantly reversed it (p = 0.019), but an increase in GFAP immunostaining in neuronal bodies did not reduce after seven-days or after aminoguanidine treatment (p = 0.003). CONCLUSION: iNOS blockage by aminoguanidine plays an important role in nociception and orofacial discomfort by control of inflammation in dental pulp after tooth bleaching with hydrogen peroxide (H2O2) 35%.

Exploring the potential anti-thyroid activity of Acetyl-L-carnitine: Lactoperoxidase inhibition profile, iodine complexation and scavenging power against H2O2. Experimental and theoretical studies.

L-Acetylcarnitine (ALC), a versatile compound, has demonstrated beneficial effects in depression, Alzheimer's disease, cognitive impairment, and other conditions. This study focuses on its antithyroid activity. The precursor molecule, L-carnitine, inhibited the uptake of triiodothyronine (T3) and thyroxine (T4), and it is possible that ALC may reduce the iodination process of T3 and T4. Currently, antithyroid drugs are used to control the excessive production of thyroid hormones (TH) through various mechanisms: (i) forming electron donor-acceptor complexes with molecular iodine, (ii) eliminating hydrogen peroxide, and (iii) inhibiting the enzyme thyroid peroxidase. To understand the pharmacological properties of ALC, we investigated its plausible mechanisms of action. ALC demonstrated the ability to capture iodine (Kc = 8.07 ± 0.32 x 105 M-1), inhibit the enzyme lactoperoxidase (LPO) (IC50 = 17.60 ± 0.76 µM), and scavenge H2O2 (39.82 ± 0.67 mM). A comprehensive physicochemical characterization of ALC was performed using FTIR, Raman, and UV-Vis spectroscopy, along with theoretical DFT calculations. The inhibition process was assessed through fluorescence spectroscopy and vibrational analysis. Docking and molecular dynamics simulations were carried out to predict the binding mode of ALC to LPO and to gain a better understanding into the inhibition process. Furthermore, albumin binding experiments were also conducted. These findings highlight the potential of ALC as a therapeutic agent, providing valuable insights for further investigating its role in the treatment of thyroid disorders.

Dithiothreitol reduces oxidative stress and necrosis caused by ultraviolet A radiation in L929 fibroblasts.

Ultraviolet A (UVA) radiation, present in sunlight, can induce cell redox imbalance leading to cellular damage and even cell death, compromising skin health. Here, we evaluated the in vitro antioxidant and photochemoprotective effect of dithiothreitol (DTT). DTT neutralized the free radicals 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS·+), 2,2-diphenyl-1-picrylhydrazyl (DPPH·), and superoxide anion (O2·-) in in vitro assays, as well as the ferric ion (Fe3+) in the ferric reducing antioxidant power (FRAP) assay. We also evaluated the effect of DTT pre-treatment in L929 dermal fibroblasts and DTT (50 and 100 µM) led to greater cell viability following UVA-irradiation compared to cells that were untreated. Furthermore, the pre-treatment of cells with DTT prevented the increase of intracellular reactive oxygen species (ROS) production, including hydrogen peroxide (H2O2), lipid peroxidation, and DNA condensation, as well as the decrease in mitochondrial membrane potential (Δψm), that occurred following irradiation in untreated cells. The endogenous antioxidant system of cells was also improved in irradiated cells that were DTT pre-treated compared to the untreated cells, as the activity of the superoxide dismutase (SOD) and catalase (CAT) enzymes remained as high as non-irradiated cells, while the activity levels were depleted in the untreated irradiated cells. Furthermore, DTT reduced necrosis in UVA-irradiated fibroblasts. Together, these results showed that DTT may have promising use in the prevention of skin photoaging and photodamage induced by UVA, as it provided photochemoprotection against the harmful effects of this radiation, reducing oxidative stress and cell death, due mainly to its antioxidant capacity.

Catalysis-based approaches with biopolymers and violet LED to improve in-office dental bleaching.

This in vitro experimental investigation aimed to evaluate the impact of the combined application of a nanofiber scaffold (NS), a polymeric catalyst primer (PCP) containing 10 mg/mL of heme peroxidase enzyme, and violet LED (LEDv) on the esthetic efficacy (EE), trans-amelodentinal cytotoxicity (TC), and procedural duration of conventional in-office bleaching therapy. To achieve this, 96 standardized enamel/dentin discs were individually placed in artificial pulp chambers. A 35% hydrogen peroxide (H2O2) bleaching gel was administered for 45, 30, or 15 min to the enamel, either previously coated with NS + PCP or left uncoated, followed by irradiation with LEDv for 15 min or no irradiation. The established groups were as follows: G1, negative control (no treatment); G2, 35% H2O2/45 min; G3, NS + PCP + LEDv; G4, NS + PCP + 35%H2O2/45 min + LEDv; G5, NS + PCP + 35%H2O2/30 min + LEDv; and G6, NS + PCP + 35%H2O2/15 min + LEDv. Extracts (culture medium + gel components diffused through the discs) were collected and applied to odontoblast-like MDPC-23 cells. EE (ΔE00 and ΔWI) and TC were assessed using ANOVA/Tukey analysis (p < 0.05). The EE analysis revealed no statistical differences between G6 and G2 (p > 0.05). Cells in G6 exhibited higher viability and lower oxidative stress compared to other bleached groups (p < 0.05). In conclusion, employing NS + PCP + LEDv to catalyze a 35%H2O2 bleaching gel applied for 15 min to the enamel resulted in successful esthetic improvements and reduced the cytotoxicity commonly linked with traditional in-office bleaching procedures.

Effect of Three Chemical Agents on Stain Removal from Dentin Caries Lesions Treated with Silver Diamine Fluoride.

INTRODUCTION: This in vitro study aimed to evaluate the effect of three different chemical agents on stain removal and mineral uptake of artificial dentin caries (ADC) lesions treated with silver diamine fluoride (SDF). METHODS: Baseline L*a*b* values were determined in polished human permanent dentin blocks, and ADC lesions were induced with an acid gel for 1 week. Samples were assigned to four groups; in three groups, half of each sample received SDF (30% SDF for 3 min), while the other half received SDF followed by a bleaching treatment protocol (garlic extract, bentonite, or 35% hydrogen peroxide). The fourth group had one SDF-treated half and one half without SDF. Color changes (ΔE) were measured by spectrometry, and transversal microradiography was used to quantify integrated mineral loss (ΔZ) 24 h after treatment (SDF or SDF + bleaching). A two-way mixed ANOVA was applied to thirty percent. RESULTS: SDF application increased mineral uptake by ADC (p = 0.001). The type of chemical agent evaluated (p < 0.0001), time (p = 0.01), and their interaction (p < 0.0001) bleached the ADC treated with SDF. However, 35% hydrogen peroxide was the only compound with a bleaching effect (p < 0.001), without returning to baseline color. None of the compounds altered the mineral uptake effect of SDF (p = 0.30). CONCLUSION: This in vitro study showed mineral uptake effect in ACD within 24 h after SDF application and the ability of hydrogen peroxide to partially remove (reduction of 24%) the staining caused by SDF without affecting its mineral uptake effect.

Antioxidant effects of LEDT in dystrophic muscle cells: involvement of PGC-1α and UCP-3 pathways.

PURPOSE: Reactive oxygen species and mitochondrial dysfunction play a crucial role in the pathophysiology of Duchenne muscular dystrophy (DMD). The light-emitting diode therapy (LEDT) showed beneficial effects on the dystrophic muscles. However, the mechanisms of this therapy influence the molecular pathways in the dystrophic muscles, particularly related to antioxidant effects, which still needs to be elucidated. The current study provides muscle cell-specific insights into the effect of LEDT, 48 h post-irradiation, on oxidative stress and mitochondrial parameters in the dystrophic primary muscle cells in culture. METHODS: Dystrophic primary muscle cells were submitted to LEDT, at multiple wavelengths (420 nm, 470 nm, 660 nm and 850 nm), 0.5 J dose, and evaluated after 48 h based on oxidative stress markers, antioxidant enzymatic system and biogenesis, and functional mitochondrial parameters. RESULTS: The mdx muscle cells treated with LEDT showed a significant reduction of H2O2 production and 4-HNE, catalase, SOD-2, and GR levels. Upregulation of UCP3 was observed with all wavelengths while upregulation of PGC-1α and a slight upregulation of electron transport chain complexes III and V was only observed following 850 nm LEDT. In addition, the mitochondrial membrane potential and mitochondrial mass mostly tended to be increased following LEDT, while parameters like O2·- production tended to be decreased. CONCLUSION: The data shown here highlight the potential of LEDT as a therapeutic agent for DMD through its antioxidant action by modulating PGC-1α and UCP3 levels.

In vitro evaluation of the effect of different bleaching varnishes: Hydrogen peroxide penetration into the pulp chamber and color change.

OBJECTIVE: To evaluate the penetration of hydrogen peroxide (HP) into the pulp chamber and the color change of different bleaching varnishes in low concentrations used for at-home bleaching. MATERIALS AND METHODS: Ninety healthy premolars were used, randomly distributed into nine groups (n = 10) according to bleaching varnish (PL, PolaLuminate; VS, VivaStyle Paint On Plus; CA, Cavex Bite&White whitening pen and; AW AlignerWhite) and time (10 and 30 min), and a control group (no bleaching). The penetration of HP was evaluated by UV-Vis spectroscopy. To evaluate the color change (ΔEab , ΔE00 , ΔWID ) a digital spectrophotometer was used (α = 0.05). RESULTS: The AW group in 10 min and the control group showed similar and lower HP penetration in the pulp chamber when compared to the other groups (p = 0.003). Increasing the application time to 30 minutes elevated the amount of HP inside the pulp chamber for all groups (p = 0.003), except for PL (p > 0.05). When applied for 30 min all bleaching varnishes showed higher color change (ΔWID ) when compared to 10 min (p = 0.04). CONCLUSIONS: For all bleaching varnishes evaluated, PolaLuminate applied for 30 min showed lower penetration into the pulp chamber and higher bleaching effects. CLINICAL SIGNIFICANCE: The use of bleaching varnishes seems promising for teeth bleaching, but it varies according to user product and protocol.

Elevated extracellular inorganic phosphate inhibits ecto-phosphatase activity in breast cancer cells: Regulation by hydrogen peroxide.

For cells to obtain inorganic phosphate, ectoenzymes in the plasma membrane, which contain a catalytic site facing the extracellular environment, hydrolyze phosphorylated molecules. In this study, we show that increased Pi levels in the extracellular environment promote a decrease in ecto-phosphatase activity, which is associated with Pi-induced oxidative stress. High levels of Pi inhibit ecto-phosphatase because Pi generates H2 O2 . Ecto-phosphatase activity is inhibited by H2 O2 , and this inhibition is selective for phospho-tyrosine hydrolysis. Additionally, it is shown that the mechanism of inhibition of ecto-phosphatase activity involves lipid peroxidation. In addition, the inhibition of ecto-phosphatase activity by H2 O2 is irreversible. These findings have new implications for understanding ecto-phosphatase regulation in the tumor microenvironment. H2 O2 stimulated by high Pi inhibits ecto-phosphatase activity to prevent excessive accumulation of extracellular Pi, functioning as a regulatory mechanism of Pi variations in the tumor microenvironment.

Exploring the antioxidant activity of Fe(III), Mn(III)Mn(II), and Cu(II) compounds in Saccharomyces cerevisiae and Galleria mellonella models of study.

Reactive oxygen species (ROS) are closely related to oxidative stress, aging, and the onset of human diseases. To mitigate ROS-induced damages, extensive research has focused on examining the antioxidative attributes of various synthetic/natural substances. Coordination compounds serving as synthetic antioxidants have emerged as a promising approach to attenuate ROS toxicity. Herein, we investigated the antioxidant potential of a series of Fe(III) (1), Mn(III)Mn(II) (2) and Cu(II) (3) coordination compounds synthesized with the ligand N-(2-hydroxybenzyl)-N-(2-pyridylmethyl)[(3-chloro)(2-hydroxy)]-propylamine in Saccharomyces cerevisiae exposed to oxidative stress. We also assessed the antioxidant potential of these complexes in the alternative model of study, Galleria mellonella. DPPH analysis indicated that these complexes presented moderate antioxidant activity. However, treating Saccharomyces cerevisiae with 1, 2 and 3 increased the tolerance against oxidative stress and extended yeast lifespan. The treatment of yeast cells with these complexes decreased lipid peroxidation and catalase activity in stressed cells, whilst no change in SOD activity was observed. Moreover, these complexes induced the Hsp104 expression. In G. mellonella, complex administration extended larval survival under H2O2 stress and did not affect the insect's life cycle. Our results suggest that the antioxidant potential exhibited by these complexes could be further explored to mitigate various oxidative stress-related disorders.

Exosome Liberation by Human Neutrophils under L-Amino Acid Oxidase of Calloselasma rhodostoma Venom Action.

L-Amino acid oxidase (LAAO) is an enzyme found in snake venom that has multifaceted effects, including the generation of hydrogen peroxide (H2O2) during oxidative reactions, leading to various biological and pharmacological outcomes such as apoptosis, cytotoxicity, modulation of platelet aggregation, hemorrhage, and neutrophil activation. Human neutrophils respond to LAAO by enhancing chemotaxis, and phagocytosis, and releasing reactive oxygen species (ROS) and pro-inflammatory mediators. Exosomes cellular nanovesicles play vital roles in intercellular communication, including immune responses. This study investigates the impact of Calloselasma rhodostoma snake venom-derived LAAO (Cr-LAAO) on human neutrophil exosome release, including activation patterns, exosome formation, and content. Neutrophils isolated from healthy donors were stimulated with Cr-LAAO (100 µg/mL) for 3 h, followed by exosome isolation and analysis. Results show that Cr-LAAO induces the release of exosomes with distinct protein content compared to the negative control. Proteomic analysis reveals proteins related to the regulation of immune responses and blood coagulation. This study uncovers Cr-LAAO's ability to activate human neutrophils, leading to exosome release and facilitating intercellular communication, offering insights into potential therapeutic approaches for inflammatory and immunological disorders.

Chemical, morphological and microhardness analysis of coronary dentin submitted to internal bleaching with hydrogen peroxide and violet LED.

BACKGROUND: Violet LED has been used for internal bleaching, however its implications on coronary dentin composition are unclear. The present study aims to evaluate the effect of bleaching with violet LED, either associated with 35 % hydrogen peroxide or not, on microhardness, chemical composition, and morphological characteristics of coronal dentin. METHODS: Thirty maxillary canines were selected to obtain 30 blocks of coronal dentin, distributed in 3 groups (n = 10): 35 % hydrogen peroxide (HP); violet LED (LED); HP 35 % + LED, (HP+LED). The chemical analysis was performed by FTIR and the morphological evaluation of the dentin structure by confocal laser scanning microscopy before (T0) and after treatment (T1). The microhardness analysis was performed by microdurometer after bleaching. The data were submitted to repeated measures ANOVA test (P> 0.05). RESULTS: The intensity of the inorganic peaks decreased after bleaching for all groups (P = 0.003). There was an increase in the organic peak intensity after bleaching with HP, a decrease for LED, while HP+LED did not change the intensity (P = 0.044). Moreover, the inorganic/organic ratio decreased for HP (P = 0.022), while for LED and HP+LED there was no significant changes (P>0.05). HP and HP+LED showed lower microhardness values compared to LED (P< 0.05). Regarding morphological changes, an increase in the perimeter of the dentinal tubules was found for all groups, with the smallest increase being observed for LED. CONCLUSION: HP bleaching decreased the chemical stability and microhardness of the coronal dentin, while the violet LED treatments had no significant impact on dentin stability. In all groups, there was an increase in exposure of the dentinal tubules after bleaching, which was less pronounced with the violet LED bleaching.

Influence of bleaching gels formulated with nano-sized sodium trimetaphosphate and fluoride on the physicochemical, mechanical, and morphological properties of dental enamel.

OBJECTIVES: To evaluate in vitro the effects of sodium fluoride (F) and nano-sized sodium trimetaphosphate (TMPnano) added to a 35% hydrogen peroxide (H2O2) bleaching gel on the color alteration, enamel mechanical and morphological properties, and H2O2 transamelodentinal diffusion. MATERIALS AND METHODS: Bovine enamel/dentin discs (n = 180) were divided according to the bleaching gel: 35% H2O2 (HP); 35% H2O2 + 0.1% F (HP/F); 35% H2O2 + 1% TMPnano (HP/TMPnano); 35% H2O2 + 0.1% F + 1% TMPnano (HP/F/TMPnano) and 35% H2O2 + 2% calcium gluconate (HP/Ca). The gels were applied 3 times by 40 min; once each 7-day. The Commission Internationale de l'Eclairage (CIE) L*a*b* total color alteration (ΔE), color alteration by CIEDE2000 (ΔE00), whitening index (ΔWID), surface (SH) and cross-sectional hardness (ΔKHN), surface roughness (Ra), and transamelodentinal diffusion were determined. Enamel surfaces were evaluated by Scanning Electron Microscopy (SEM) and X-ray Dispersive Energy (EDX). Data were submitted to ANOVA, followed by the Student-Newman-Keuls test (p <0.05). RESULTS: ΔE, ΔE00, and ΔWID were similar among the gels that promoted a bleaching effect after treatment (p <0.001). Mineral loss (SH and ΔKHN), Ra, and H2O2 diffusion were lower for HP/F/TMPnano; the HP and HP/Ca groups presented the highest values (p <0.001). For SEM/EDX, surface changes were observed in all bleached groups, but less intense with TMPnano. CONCLUSIONS: Gels containing F/TMPnano do not interfere with the bleaching effect and reduce enamel demineralization, roughness, H2O2 diffusion, and morphological changes. CLINICAL RELEVANCE: Whitening gels containing F/TMPnano can be used as a new strategy to increase safety and maintain clinical performance.

Solar heterogeneous photo-Fenton for complete inactivation of Escherichia coli and Salmonella typhimurium in secondary-treated wastewater effluent.

In this work, complete elimination of Escherichia coli and Salmonella typhimurium was achieved in 120 min using a heterogeneous photo-Fenton process under sunlight at pH 6.5 in distilled water. A face-centered composite central design 22 with one categoric factor and three replicates at the central point was used to evaluate the effect of iron (III) oxide concentration (0.8-3.4 mg L-1), H2O2 (2-10 mg L-1), and the type of iron oxide phase (maghemite and hematite) on the inactivation of both bacteria. The results showed that the amount of catalyst, H2O2 concentration and their interaction were significant factors (p < 0.05) in the elimination of the microorganisms. Thus, under the best conditions (3.4 mg L-1 of iron (III) oxide and 10 mg L-1 of H2O2) in the experimental ranges, complete inactivation of E. coli and S. typhimurium was achieved (6-log reduction) in 120 min using the photo-Fenton treatment with both iron-oxide phases. Furthermore, the photocatalytic elimination of both bacteria by the photo-Fenton process using hematite and maghemite in secondary-treated wastewater effluent was performed obtaining slower inactivation rates (1.2-5.9 times) than in distilled water due to the matrix effect of the effluent from a wastewater treatment plant. Nevertheless, the process continued to be effective in the effluent, achieving complete bacterial elimination in 150 min using the hematite phase. Additionally, the SEM images of the bacterial cells showed that the heterogeneous photo-Fenton treatment generated permanent and irreversible cell damage, resulting in complete cell death.

The role of Vav3 expression for inflammation and cell death during experimental myocardial infarction.

OBJECTIVES: Myocardial Infarction (MI) is the leading cause of chronic heart failure. Previous studies have suggested that Vav3, a receptor protein tyrosine kinase signal transducer, is associated with a variety of cellular signaling processes such as cell morphology regulation and cell transformation with oncogenic activity. However, the mechanism of Vav3-mediated MI development requires further investigation. METHOD: Here, The authors established an MI rat model by ligating the anterior descending branch of the left coronary artery, and an MI cell model by treating cardiomyocytes with H2O2. Microarray analysis was conducted to identify genes with differential expression in heart tissues relevant to MI occurrence and development. Vav3 was thus selected for further investigation. RESULTS: Vav3 downregulation was observed in MI heart tissue and H2O2-treated cardiomyocytes. Administration of Lentiviral Vav3 (LV-VAV3) in MI rats upregulated Vav3 expression in MI heart tissue. Restoration of Vav3 expression reduced infarct area and ameliorated cardiac function in MI rats. Cardiac inflammation, apoptosis, and upregulation of NFκB signal in heart tissue of MI animals were assessed using ELISA, TUNEL staining, real-time PCR, and WB. Vav3 overexpression reduced cardiac inflammation and apoptosis and inhibited NFκB expression and activation. Betulinic Acid (BA) was then used to re-activate NFκB in Vav3-overexpressed and H2O2-induced cardiomyocytes. The expression of P50 and P65, as well as nuclear P65, was significantly increased by BA exposure. CONCLUSIONS: Vav3 might serve as a target to reduce ischemia damage by suppressing the inflammation and apoptosis of cardiomyocytes.