Spectroscopic Identification of Bacteria Resistance to Antibiotics by Means of Absorption of Specific Biochemical Groups and Special Machine Learning Algorithm.

FTIR (Fourier transform infrared spectroscopy) is one analytical technique of the absorption of infrared radiation. FTIR can also be used as a tool to characterize profiles of biomolecules in bacterial cells, which can be useful in differentiating different bacteria. Considering that different bacterial species have different molecular compositions, it will then result in unique FTIR spectra for each species and even bacterial strains. Having this important tool, here, we have developed a methodology aimed at refining the analysis and classification of the FTIR absorption spectra obtained from samples of Staphylococcus aureus, with the implementation of machine learning algorithms. In the first stage, the system conforming to four specified species groups, Control, Amoxicillin induced (AMO), Gentamicin induced (GEN), and Erythromycin induced (ERY), was analyzed. Then, in the second stage, five hidden samples were identified and correctly classified as with/without resistance to induced antibiotics. The total analyses were performed in three windows, Carbohydrates, Fatty Acids, and Proteins, of five hundred spectra. The protocol for acquiring the spectral data from the antibiotic-resistant bacteria via FTIR spectroscopy developed by Soares et al. was implemented here due to demonstrating high accuracy and sensitivity. The present study focuses on the prediction of antibiotic-induced samples through the implementation of the hierarchical cluster analysis (HCA), principal component analysis (PCA) algorithm, and calculation of confusion matrices (CMs) applied to the FTIR absorption spectra data. The data analysis process developed here has the main objective of obtaining knowledge about the intrinsic behavior of S. aureus samples within the analysis regions of the FTIR absorption spectra. The results yielded values with 0.7 to 1 accuracy and high values of sensitivity and specificity for the species identification in the CM calculations. Such results provide important information on antibiotic resistance in samples of S. aureus bacteria for potential application in the detection of antibiotic resistance in clinical use.

Multifunctional bilayer membranes composed of poly(lactic acid), beta-chitin whiskers and silver nanoparticles for wound dressing applications.

Nanomaterial-based wound dressings have been extensively studied for the treatment of both minor and life-threatening tissue injuries. These wound dressings must possess several crucial characteristics, such as tissue compatibility, non-toxicity, appropriate biodegradability to facilitate wound healing, effective antibacterial activity to prevent infection, and adequate physical and mechanical strength to withstand repetitive dynamic forces that could potentially disrupt the healing process. Nevertheless, the development of nanostructured wound dressings that incorporate various functional micro- and nanomaterials in distinct architectures, each serving specific purposes, presents significant challenges. In this study, we successfully developed a novel multifunctional wound dressing based on poly(lactic acid) (PLA) fibrous membranes produced by solution-blow spinning (SBS) and electrospinning. The PLA-based membranes underwent surface modifications aimed at tailoring their properties for utilization as effective wound dressing platforms. Initially, beta-chitin whiskers were deposited onto the membrane surface through filtration, imparting hydrophilic character. Afterward, silver nanoparticles (AgNPs) were incorporated onto the beta-chitin layer using a spray deposition method, resulting in platforms with antimicrobial properties against both Staphylococcus aureus and Escherichia coli. Cytotoxicity studies demonstrated the biocompatibility of the membranes with the neonatal human dermal fibroblast (HDFn) cell line. Moreover, bilayer membranes exhibited a high surface area and porosity (> 80%), remarkable stability in aqueous media, and favorable mechanical properties, making them promising candidates for application as multifunctional wound dressings.

Structural basis of antibacterial photodynamic action of curcumin against S. aureus.

Antimicrobial photodynamic therapy (aPDT) is an alternative tool to commercial antibiotics for the inactivation of pathogenic bacteria (e.g., S. aureus). However, there is still a lack of understanding of the molecular modeling of the photosensitizers and their mechanism of action through oxidative pathways. Herein, a combined experimental and computational evaluation of curcumin as a photosensitizer against S. aureus was performed. The radical forms of keto-enol tautomers and the energies of curcumin's frontier molecular orbitals were evaluated by density functional theory (DFT) to point out the photodynamic action as well as the photobleaching process. Furthermore, the electronic transitions of curcumin keto-enol tautomers were undertaken to predict the transitions as a photosensitizer during the antibacterial photodynamic process. Moreover, molecular docking was used to evaluate the binding affinity with the S. aureus tyrosyl-tRNA synthetase as the proposed a target for curcumin. In this regard, the molecular orbital energies show that the curcumin enol form has a character of 4.5% more basic than the keto form - the enol form is a more promising electron donor than its tautomer. Curcumin is a strong electrophile, with the enol form being 4.6% more electrophilic than its keto form. In addition, the regions susceptible to nucleophilic attack and photobleaching were evaluated by the Fukui function. Regarding the docking analysis, the model suggested that four hydrogen bonds contribute to the binding energy of curcumin's interaction with the ligand binding site of S. aureus tyrosyl-tRNA synthetase. Finally, residues Tyr36, Asp40, and Asp177 contact curcumin and may contribute to orienting the curcumin in the active area. Moreover, curcumin presented a photoinactivation of 4.5 log unit corroborating the necessity of the combined action of curcumin, light, and O2 to promote the photooxidation damage of S. aureus. These computational and experimental data suggest insights regarding the mechanism of action of curcumin as a photosensitizer to inactivate S. aureus bacteria.

Polyurethane films formation from microcrystalline cellulose as a polyol and cellulose nanocrystals as additive: Reactions favored by the low viscosity of the source of isocyanate groups used.

To simultaneously form films while synthesizing solvent-free and catalyst-free bio-based polyurethanes, hexamethylene diisocyanate trimer was selected as an isocyanate group source to produce a low-viscosity reaction medium for dispersing high contents of microcrystalline cellulose (MCC, polyol) and cellulose nanocrystals (CNC). Castor oil was used as an additional polyol source. Up to 80 % of the MCC was dispersed, producing a film exhibiting the highest Tg (72 °C), tensile strength (18 MPa), and Young's modulus (522.4 MPa). 12.5 % (30 % MCC) and 7.5 % (50 % MCC) of CNC dispersed in the reaction medium formed films stiffer than their counterparts. All the films exhibited transparency and high crystallinity. The contact angle/zeta potential (ζ) indicated hydrophobic film surfaces. At pH 7.4, ζ suggested that the films interacted with physiological fluids favorably. The films were non-cytotoxic, and the composites exhibited cell growth compared with the control. The reported results, as far as it is known, are unprecedented.

Environmentally Safe Photodynamic Control of Aedes aegypti Using Sunlight-Activated Synthetic Curcumin: Photodegradation, Aquatic Ecotoxicity, and Field Trial.

This study reports curcumin as an efficient photolarvicide against Aedes aegypti larvae under natural light illumination. Larval mortality and pupal formation were monitored daily for 21 days under simulated field conditions. In a sucrose-containing formulation, a lethal time 50 (LT50) of 3 days was found using curcumin at 4.6 mg L-1. This formulation promoted no larval toxicity in the absence of illumination, and sucrose alone did not induce larval phototoxicity. The photodegradation byproducts (intermediates) of curcumin were determined and the photodegradation mechanisms proposed. Intermediates with m/z 194, 278, and 370 were found and characterized using LC-MS. The ecotoxicity of the byproducts on non-target organisms (Daphnia, fish, and green algae) indicates that the intermediates do not exhibit any destructive potential for aquatic organisms. The results of photodegradation and ecotoxicity suggest that curcumin is environmentally safe for non-target organisms and, therefore, can be considered for population control of Ae. aegypti.

Formulations of curcumin and d-mannitol as a photolarvicide against Aedes aegypti larvae: Sublethal photolarvicidal action, toxicity, residual evaluation, and small-scale field trial.

Dengue, Zika, chikungunya, and yellow fever are arboviruses transmitted by Aedes aegypti mosquito. In this regard, a number of techniques have emerged aiming to combat its proliferation. Elimination of Aedes aegypti larvae by photodynamic action has been reported as an efficient approach. In this regard, this study was aimed at synthetize and characterize formulations with different proportions (w/w) of the plant-based photolarvicidal curcumin and d-mannitol (CCD 1-4) and their evaluation on sublethal photolarvicidal efficiency, photodegradation profile,solubility, internalization, elimination time, persistence in simulated field, growth of microorganisms in water and the toxicity using an animal models (Zebrafish). CCD 3 (curcumin:d-mannitol 50:50 w/w) showed the best efficacy (LC50-24h = 0.01 mg/L), and also presented the shortest internalization and longest elimination time, 60 min and 8 days, respectively. This formulation caused an extrusion into the intestine and peritrophic membrane. Moreover, CCD 3 showed a photodegradation of 50% (in 24 h) under white fluorescent lamps. In a small-scale field trial, CCD 3 had a residual time of 14 days and abnormal microbial growth was not observed. Finally, CCD 3 did not present any toxicity in Zebrafish, after exposition for 24 h at 100 mg/L. Overall, these results raise the possibility of reducing virus transmission through the controlled photoinactivation of Aedes aegypti larvae using a non-toxic plant-based formulated photolarvicide.

Bacterial Photoinactivation Using PLGA Electrospun Scaffolds.

The use of ultraviolet (UV) and blue irradiation to sterilize surfaces is well established, but commercial applications would be enhanced if the light source is replaced with ambient light. In this paper, it is shown that nanofibers can be explored as an alternative methodology to UV and blue irradiation for bacterial inactivation. It is demonstrated that this is indeed possible using spun nanofibers of poly[lactic-co-(glycolic acid)] (PLGA). This work shows that PLGA spun scaffolds can promote photoinactivation of Staphylococcus aureus and Escherichia coli bacteria with ambient light or with laser irradiation at 630 nm. With the optimized scaffold composition of PLGA85:15 nanofibers, the minimum intensity required to kill the bacteria is much lower than in antimicrobial blue light applications. The enhanced effect introduced by PLGA scaffolds is due to their nanofiber structures since PLGA spun nanofibers were able to inactivate both S. aureus and E. coli bacteria, but cast films had no effect. These findings pave the way for an entirely different method to sterilize surfaces, which is less costly and environmentally friendly than current procedures. In addition, the scaffolds could also be used in cancer treatment with fewer side effects since photosensitizers are not required.

Photosensitizing nanoclays for efficient cell uptake and in vitro photodynamic therapy.

In this study a straightforward strategy to deliver photosensitizing molecules into cancer cells by using Laponite RD® (LAP) nanodisks as carrier is presented. We report the application of LAP functionalized with a highly hydrophobic Silicon phthalocyanine photosensitizer (SiPc) for efficient cell uptake and photodynamic therapy (PDT) of MCF-7 breast cancer cells in vitro. This inorganic-organic hybrid nanomaterial caused a threefold increase in intracellular ROS levels and 99.7% of cancer cell inactivation after light treatment (630 nm; fluency of 108 J/cm²). With its theranostic capabilities (simultaneous fluorescence and singlet oxygen generation), uptake into cancer cells was monitored to control the photo-inactivation. The functionalized nanodisks already proved to be an efficient photo-inactivator of pathogenic Gram-(+) bacteria and were previously reported as LS10. By extending the use of LS10 to PDT of cancer cells, it is an example of a photosensitizer functionalized nanoclay with multipurpose drug characteristics, demonstrating an intriguing potential for future phototherapeutic studies and applications.

Hydrogel from all in all lignocellulosic sisal fibers macromolecular components.

The heterogeneous structure of lignocellulosic biomass makes it difficult to dissolve its main components (cellulose, hemicelluloses, and lignin) by solvent action with the aim of further applying the mixture of the biological macromolecules generated in the solvent medium. In the present study, the dissolution efficiency (DE) of lignocellulosic sisal fibers in the lithium chloride/dimethylacetamide solvent system (LiCl/DMAc) was evaluated for further application in the formation of hydrogels. Catalytic amounts of trifluoroacetic acid (TFA) were used in some experiments, which increased the DE from 40% to 90%. The regeneration of the solutions, either previously filtered or not, led to hydrogels based on sisal lignocellulosic biomass. In brief, the properties of the hydrogels were influenced by the content of the lignocellulosic components in the hydrogels, present both in the dissolved fraction and in the incorporated undissolved fraction (when nonfiltered solutions were used). Hydrogels presented water absorption up to 7479% and resorption content in the lyophilized hydrogel up to 2133%. Extracts obtained from preselected hydrogels exhibited cell viability up to 127% compared to the control group when in contact with fibroblast cultures, exhibiting their noncytotoxic properties. This attribute increased the range of possible applications of these hydrogels, ranging from agriculture to biocompatible materials.

Graphene Oxide Theranostic Effect: Conjugation of Photothermal and Photodynamic Therapies Based on an in vivo Demonstration.

INTRODUCTION: Cancer is the second leading cause of death globally and is responsible, where about 1 in 6 deaths in the world. Therefore, there is a need to develop effective antitumor agents that are targeted only to the specific site of the tumor to improve the efficiency of cancer diagnosis and treatment and, consequently, limit the unwanted systemic side effects currently obtained by the use of chemotherapeutic agents. In this context, due to its unique physical and chemical properties of graphene oxide (GO), it has attracted interest in biomedicine for cancer therapy. METHODS: In this study, we report the in vivo application of nanocomposites based on Graphene Oxide (nc-GO) with surface modified with PEG-folic acid, Rhodamine B and Indocyanine Green. In addition to displaying red fluorescence spectra Rhodamine B as the fluorescent label), in vivo experiments were performed using nc-GO to apply Photodynamic Therapy (PDT) and Photothermal Therapy (PTT) in the treatment of Ehrlich tumors in mice using NIR light (808 nm 1.8 W/cm2). RESULTS: This study based on fluorescence images was performed in the tumor in order to obtain the highest concentration of nc-GO in the tumor as a function of time (time after intraperitoneal injection). The time obtained was used for the efficient treatment of the tumor by PDT/PTT. DISCUSSION: The current study shows an example of successful using nc-GO nanocomposites as a theranostic nanomedicine to perform simultaneously in vivo fluorescence diagnostic as well as combined PDT-PTT effects for cancer treatments.

Features of third generation photosensitizers used in anticancer photodynamic therapy: Review.

Cancer remains a main public health issue and the second cause of mortality worldwide. Photodynamic therapy is a clinically approved therapeutic option. Effective photodynamic therapy induces cancer damage and death through a multifactorial manner including reactive oxygen species-mediated damage and killing, vasculature damage, and immune defense activation. Anticancer efficiency depends on the improvement of photosensitizers drugs used in photodynamic therapy, their selectivity, enhanced photoproduction of reactive species, absorption at near-infrared spectrum, and drug-delivery strategies. Both experimental and clinical studies using first- and second-generation photosensitizers had pointed out the need for developing improved photosensitizers for photodynamic applications and achieving better therapeutic outcomes. Bioconjugation and encapsulation with targeting moieties appear as a main strategies for the development of photosensitizers from their precursors. Factors influencing cellular biodistribution and uptake are briefly discussed, as well as their roles as cancer diagnostic and therapeutic (theranostics) agents. The two-photon photodynamic approach using third-generation photosensitizers is present as an attempt in treating deeper tumors. Although significant advances had been made over the last decade, the development of next-generation photosensitizers is still mainly in the developmental stage.

Avoiding ventilator-associated pneumonia: Curcumin-functionalized endotracheal tube and photodynamic action.

Hospital-acquired infections are a global health problem that threatens patients' treatment in intensive care units, causing thousands of deaths and a considerable increase in hospitalization costs. The endotracheal tube (ETT) is a medical device placed in the patient's trachea to assist breathing and delivering oxygen into the lungs. However, bacterial biofilms forming at the surface of the ETT and the development of multidrug-resistant bacteria are considered the primary causes of ventilator-associated pneumonia (VAP), a severe hospital-acquired infection for significant mortality. Under these circumstances, there has been a need to administrate antibiotics together. Although necessary, it has led to a rapid increase in bacterial resistance to antibiotics. Therefore, it becomes necessary to develop alternatives to prevent and combat these bacterial infections. One possibility is to turn the ETT itself into a bactericide. Some examples reported in the literature present drawbacks. To overcome those issues, we have designed a photosensitizer-containing ETT to be used in photodynamic inactivation (PDI) to avoid bacteria biofilm formation and prevent VAP occurrence during tracheal intubation. This work describes ETT's functionalization with curcumin photosensitizer, as well as its evaluation in PDI against Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli A significant photoinactivation (up to 95%) against Gram-negative and Gram-positive bacteria was observed when curcumin-functionalized endotracheal (ETT-curc) was used. These remarkable results demonstrate this strategy's potential to combat hospital-acquired infections and contribute to fighting antimicrobial resistance.

Antibacterial Photodynamic Inactivation of Antibiotic-Resistant Bacteria and Biofilms with Nanomolar Photosensitizer Concentrations.

Gram-negative bacteria and bacteria in biofilms are very difficult to eradicate and are the most antibiotic-resistant bacteria. Therapeutic alternatives less susceptible to mechanisms of resistance are urgently needed to respond to an alarming increase of resistant nosocomial infections. Antibacterial photodynamic inactivation (PDI) generates oxidative stress that triggers multiple cell death mechanisms that are more difficult to counteract by bacteria. We explore PDI of multidrug-resistant bacterial strains collected from patients and show how positive charge distribution in the photosensitizer drug impacts the efficacy of inactivation. We demonstrate the relevance of size for drug diffusion in biofilms. The designed meso-imidazolyl porphyrins of small size with positive charges surrounding the macrocycle enabled the inactivation of bacteria in biofilms by 6.9 log units at 5 nM photosensitizer concentration and 5 J cm-2, which offers new opportunities to treat biofilm infections.

Core-sheath nanostructured chitosan-based nonwovens as a potential drug delivery system for periodontitis treatment.

Core-sheath nanofibers were successfully prepared via coaxial electrospinning by using chitosan with well-defined structural characteristics as the shell layer and poly (vinyl alcohol) (PVA) containing tetracycline hydrochloride (TH) as the core layer. The effects of the average degree of deacetylation (DD‾) of chitosan and the post-electrospinning genipin crosslinking on physicochemical and biological properties of resulting nonwovens were evaluated. Defect-free and geometrically uniform nanofibers with diameters predominantly in the range of 100-300 nm were prepared, and transmission electron microscopy (TEM) revealed the core-sheath structures and its preservation after crosslinking. The mechanical properties, as well as the stability of nonwovens in aqueous medium, were greatly improved by genipin-crosslinking, which enabled a sustained release of TH over 14 days. Results also revealed that the release profile of TH in the presence of lysozyme was affected by the composition of the shell layer, as the TH release rate increases with decreasing of DD‾. Further in vitro antimicrobial activity demonstrated that the cross-linked nonwovens containing TH showed strong activity against bacterial strains associated with periodontal disease. Additionally, the nonwovens did not demonstrate cytotoxic toward fibroblast (HDFn) cells, hence showing their potential for applications as a novel drug delivery platform for periodontitis treatment.

Field cancerization treatment using topical photodynamic therapy: A comparison between two aminolevulinate derivatives.

The objective of this study was to evaluate and compare the clinical response to PDT (Photodynamic Therapy) in field cancerization using two aminolevulinate derivatives. Forty patients with multiple actinic keratosis (AK) on forearms and hands scattered received two sessions of ALA and MAL-PDT at 630 nm (36 J/cm2). The AK clearance rate was 72 % for both drugs with a significant decrease in AK observed clinically (p < 00,001). Clinical improvement in field cancerization using two aminolevulinate derivatives in PDT is proven with no significant difference in the efficacy of drugs.

Visible-Light-Mediated Photodynamic Water Disinfection @ Bimetallic-Doped Hybrid Clay Nanocomposites.

This study reports a new class of photocatalytic hybrid clay nanocomposites prepared from low-cost sources (kaolinite clay and Carica papaya seeds) doped with Zn and Cu salts via a solvothermal process. X-ray diffraction analysis suggests that Cu-doping and Cu/Zn-doping introduce new phases into the crystalline structure of Kaolinite clay, which is linked to the reduced band gap of kaolinite from typically between 4.9 and 8.2 eV to 2.69 eV for Cu-doped and 1.5 eV for Cu/Zn hybrid clay nanocomposites (Nisar, J.; Århammar, C.; Jämstorp, E.; Ahuja, R. Phys. Rev. B 2011, 84, 075120). In the presence of solar light irradiation, Cu- and Cu/Zn-doped nanocomposites facilitate the electron-hole pair separation. This promotes the generation of singlet oxygen which in turn improves the water disinfection efficiencies of these novel nanocomposite materials. The nanocomposite materials were further characterized using high-resolution scanning electron microscopy, fluorimetry, thermogravimetric analysis, and Raman spectroscopy. The breakthrough times of the nanocomposites for a fixed bed mode of disinfection of water contaminated with 2.32 × 107 cfu/mL E. coli ATCC 25922 under solar light irradiation are 25 h for Zn-doped, 30 h for Cu-doped, and 35 h for Cu/Zn-doped nanocomposites. In the presence of multidrug and multimetal resistant strains of E. coli, the breakthrough time decreases significantly. Zn-only doped nanocomposites are not photocatalytically active. In the absence of light, the nanocomposites are still effective in decontaminating water, although less efficient than under solar light irradiation. Electrostatic interaction, metal toxicity, and release of singlet oxygen (only in the Cu-doped and Cu/Zn-doped nanocomposites) are the three disinfection mechanisms by which these nanocomposites disinfect water. A regrowth study indicates the absence of any living E. coli cells in treated water even after 4 days. These data and the long hydraulic times (under gravity) exhibited by these nanocomposites during photodisinfection of water indicate an unusually high potential of these nanocomposites as efficient, affordable, and sustainable point-of-use systems for the disinfection of water in developing countries.

Luminescent Mesoporous Silica Nanohybrid Based on Drug Derivative Terbium Complex.

Mesoporous silica nanoparticles prepared by organic template-driven synthesis have been successfully explored as carriers of the drug-derivate green luminescent complex of terbium (III) with the nonsteroidal anti-inflammatory drug ketoprofen. The terbium (III) complex was synthesized by reacting ketoprofen sodium salt with terbium (III) chloride, which was further adsorbed onto the surface of mesoporous nanoparticles with a mean particle size of 47 ± 4 nm and pore size of 11 nm. The incorporation of the complex into mesoporous silica nanoparticles was tracked by the decrease in the surface area and pore size of the nanoparticles, and successfully demonstrated by substantial changes in the adsorption isotherms and thermal and vibrational spectroscopy results. The cytotoxicity assay and confocal microscopy have shown that the novel luminescent nanohybrid presents high cell viability and the characteristic terbium (III) emission can be assessed through two-photon excitation, which paves the way for bioimaging applications in nanomedicine.

Nebulization as a tool for photosensitizer delivery to the respiratory tract.

To this day, any photosensitizers for the photodynamic treatment of pulmonary illnesses have been administered intravenously. There is, however, an intrinsic difficulty in reaching the target cells or bacteria in the respiratory system. Nebulization could overcome distribution problems and alleviate side effects by delivering the photosensitizers directly to the lungs. In this study, we evaluated the viability of three photosensitizers (indocyanine green, the chlorine Photodithazine, and the porphyrin Photogem) was evaluated comparatively in a jet nebulizer. Quantitative analysis was performed by looking at the droplet size, extent of nebulization, output over time and stability of the solutions. All of the tested photosensitizers were found to be adequately nebulized. We also demonstrated the delivery of indocyanine green to the pulmonary tract and its activation with infrared light in a murine model using extracorporeal detection of fluorescence. This was an important step toward clinical implementation of the extracorporeally illuminated photodynamic inactivation of pneumonia, recently demonstrated in vivo by this research group.

Biofilm Destruction on Endotracheal Tubes by Photodynamic Inactivation.

BACKGROUND: Hospital infections are a public health problem that can occur with the use of catheters and endotracheal tubes (ETT). Pathogenic microorganisms may adhere to surfaces of these materials forming a biofilm and produce an extracellular polymer matrix that promotes resistance of microorganisms to factors such as pH, temperature and drugs. The conventional treatment is being made by antibiotics, which has serious adverse effects in immunocompromised patients. Photodynamic therapy (PDT) is an alternative for microbial inactivation noninvasive without the stimulus of microbial resistance. PDT combines light and a photosensitive molecule for produce reactive oxygen species leading to bacterial death. OBJECTIVE: The objective of this study was to determine the efficacy of a PDT protocol in bacterial inactivation of biofilm ETT. METHOD: The photosensitizer (PS) used was curcumin and the light source LED at 450nm. A statistical experimental design was used for optimization of antimicrobial PDT. RESULTS: The highest microbial inactivation was observed with 70% biofilm reduction in conditions 1.25 mg/mL curcumin, 2 h of PS incubation and 50 J/cm2. CONCLUSION: This study described the photodynamic death of bacteria forming a biofilm on ETT. Parameters optimization was important for clinical application of this system.

Mangifera indica L. extract (Vimang®) reduces plasma and liver cholesterol and leucocyte oxidative stress in hypercholesterolemic LDL receptor deficient mice.

Cardiovascular diseases are major causes of death worldwide. Beyond the classical cholesterol risk factor, other conditions such as oxidative stress are well documented to promote atherosclerosis. The Mangifera indica L. extract (Vimang®) was reported to present antioxidant and hypocholesterolemic properties. Thus, here we evaluate the effects of Vimang treatment on risk factors of the atherosclerosis prone model of familial hypercholesterolemia, the LDL receptor knockout mice. Mice were treated with Vimang during 2 weeks and were fed a cholesterol-enriched diet during the second week. The Vimang treated mice presented significantly reduced levels of plasma (15%) and liver (20%) cholesterol, increased plasma total antioxidant capacity (10%) and decreased reactive oxygen species (ROS) production by spleen mononuclear cells (50%), P < 0.05 for all. In spite of these benefits, the average size of aortic atherosclerotic lesions stablished in this short experimental period did not change significantly in Vimang treated mice. Therefore, in this study we demonstrated that Vimang has protective effects on systemic and tissue-specific risk factors, but it is not sufficient to promote a reduction in the initial steps of atherosclerosis development. In addition, we disclosed a new antioxidant target of Vimang, the spleen mononuclear cells that might be relevant for more advanced stages of atherosclerosis.