Biotransformed soybean cream as a new nutraceutical for skin care: collagen stimulation in vitro and ex vivo.

Treatments that attenuate the effects of hypoestrogenism in menopausal women have been gaining visibility. This study investigated the skin response to a phytoestrogen-enriched cosmetic formulation created by incorporating a biotransformed soybean extract (BE) into a cream-like matrix. Collagen-I expression was analyzed both in vitro (fibroblast cells) and ex vivo (skin explants). The results revealed an increased amount of collagen-I both in fibroblasts and human skin when treated with BE and BE-incorporated cream. Also, this collagen-I overexpression was inhibited by PHTPP, indicating a dependence on estrogen hormone receptor beta (ERß) signaling. Moreover, BE was not harmful to skin microbiota, showing a promising nutricosmetic potential. Thus, this work presented a fully functional cream-like formulation that was shown to be safe and effectively increase collagen-I levels both in vitro and ex vivo.

Biotransformed soybean cream as a new nutraceutical for skin care: collagen stimulation in vitro and ex vivo

Treatments that attenuate the effects of hypoestrogenism in menopausal women have been gaining visibility. This study investigated the skin response to a phytoestrogen-enriched cosmetic formulation created by incorporating a biotransformed soybean extract (BE) into a cream-like matrix. Collagen-I expression was analyzed both in vitro (fibroblast cells) and ex vivo (skin explants). The results revealed an increased amount of collagen-I both in fibroblasts and human skin when treated with BE and BE-incorporated cream. Also, this collagen-I overexpression was inhibited by PHTPP, indicating a dependence on estrogen hormone receptor beta (ERβ) signaling. Moreover, BE was not harmful to skin microbiota, showing a promising nutricosmetic potential. Thus, this work presented a fully functional cream-like formulation that was shown to be safe and effectively increase collagen-I levels both in vitro and ex vivo.

Tertiary hospital sewage as reservoir of bacteria expressing MDR phenotype in Brazil / Esgoto de hospital terciário como reservatório de bactérias expressando o fenótipo MDR no Brasil

High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.

Altas doses de antibióticos utilizados em hospitais podem afetar a composição microbiana dos esgotos, selecionando bactérias resistentes. Nesse sentido, avaliamos o perfil de resistência a antibióticos e o fenótipo multirresistente de bactérias isoladas em esgoto de um hospital terciário no interior do estado de São Paulo, Brasil. Para o isolamento de bactérias, foram semeados 10 µL das amostras de esgoto em meios de cultura seletivos e os isolados foram identificados usando o sistema automatizado VITEK-2. O teste de sensibilidade aos antibióticos foi realizado por disco-difusão em ágar. Elevadas porcentagens de resistência foram encontradas para amoxicilina, ampicilina, ceftazidima, clindamicina, vancomicina e o fenótipo multirresistente (MDR) foi atribuído a 60,7% dos isolados. Nossos resultados mostram bactérias classificadas como prioridade crítica/alta pela Lista de Patógenos Prioritários da OMS (Enterococcus e Staphylococcus aureus resistentes à vancomicina e Enterobacteriaceae resistentes aos carbapenêmicos) no esgoto hospitalar. Sendo assim, implementação de tecnologias de desinfecção do esgoto hospitalar reduziriam a carga bacteriana no esgoto que chegará às estações de tratamento de esgoto urbanas, minimizando a contaminação dos ecossistemas hídricos receptores e a disseminação da resistência bacteriana no ambiente.

Tertiary hospital sewage as reservoir of bacteria expressing MDR phenotype in Brazil / Esgoto de hospital terciário como reservatório de bactérias expressando o fenótipo MDR no Brasil

High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.

Altas doses de antibióticos utilizados em hospitais podem afetar a composição microbiana dos esgotos, selecionando bactérias resistentes. Nesse sentido, avaliamos o perfil de resistência a antibióticos e o fenótipo multirresistente de bactérias isoladas em esgoto de um hospital terciário no interior do estado de São Paulo, Brasil. Para o isolamento de bactérias, foram semeados 10 µL das amostras de esgoto em meios de cultura seletivos e os isolados foram identificados usando o sistema automatizado VITEK-2. O teste de sensibilidade aos antibióticos foi realizado por disco-difusão em ágar. Elevadas porcentagens de resistência foram encontradas para amoxicilina, ampicilina, ceftazidima, clindamicina, vancomicina e o fenótipo multirresistente (MDR) foi atribuído a 60,7% dos isolados. Nossos resultados mostram bactérias classificadas como prioridade crítica/alta pela Lista de Patógenos Prioritários da OMS (Enterococcus e Staphylococcus aureus resistentes à vancomicina e Enterobacteriaceae resistentes aos carbapenêmicos) no esgoto hospitalar. Sendo assim, implementação de tecnologias de desinfecção do esgoto hospitalar reduziriam a carga bacteriana no esgoto que chegará às estações de tratamento de esgoto urbanas, minimizando a contaminação dos ecossistemas hídricos receptores e a disseminação da resistência bacteriana no ambiente.

Tertiary hospital sewage as reservoir of bacteria expressing MDR phenotype in Brazil

Abstract High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.

Resumo Altas doses de antibióticos utilizados em hospitais podem afetar a composição microbiana dos esgotos, selecionando bactérias resistentes. Nesse sentido, avaliamos o perfil de resistência a antibióticos e o fenótipo multirresistente de bactérias isoladas em esgoto de um hospital terciário no interior do estado de São Paulo, Brasil. Para o isolamento de bactérias, foram semeados 10 µL das amostras de esgoto em meios de cultura seletivos e os isolados foram identificados usando o sistema automatizado VITEK-2. O teste de sensibilidade aos antibióticos foi realizado por disco-difusão em ágar. Elevadas porcentagens de resistência foram encontradas para amoxicilina, ampicilina, ceftazidima, clindamicina, vancomicina e o fenótipo multirresistente (MDR) foi atribuído a 60,7% dos isolados. Nossos resultados mostram bactérias classificadas como prioridade crítica/alta pela Lista de Patógenos Prioritários da OMS (Enterococcus e Staphylococcus aureus resistentes à vancomicina e Enterobacteriaceae resistentes aos carbapenêmicos) no esgoto hospitalar. Sendo assim, implementação de tecnologias de desinfecção do esgoto hospitalar reduziriam a carga bacteriana no esgoto que chegará às estações de tratamento de esgoto urbanas, minimizando a contaminação dos ecossistemas hídricos receptores e a disseminação da resistência bacteriana no ambiente.

Tertiary hospital sewage as reservoir of bacteria expressing MDR phenotype in Brazil.

High doses of antibiotics used in hospitals can affect the microbial composition of sewers, selecting resistant bacteria. In this sense, we evaluated the antibiotic resistance profile and the multiresistant phenotype of bacteria isolated in sewage from a tertiary hospital in the interior São Paulo state, Brazil. For bacteria isolation, 10 µL of sewage samples were sown in selective culture media and the isolates were identified using VITEK-2 automatized system. The antibiotic sensitivity test was performed by disk diffusion. High percentages of resistance were found for amoxicillin, ampicillin, ceftazidime, clindamycin, vancomycin and the multidrug-resistant phenotype (MDR) was attributed to 60.7% of the isolates. Our results show bacteria classified as critical/high priority by WHO List of Priority Pathogens (Enterococcus and Staphylococcus aureus resistant to vancomycin and Enterobacteriaceae resistant to carbapenems) in hospital sewage. Therefore, the implementation of disinfection technologies for hospital sewage would reduce the bacterial load in the sewage that will reach urban wastewater treatment plants, minimizing superficial water contamination and bacterial resistance spread in the environment.

Double membrane based on lidocaine-coated polymyxin-alginate nanoparticles for wound healing: In vitro characterization and in vivo tissue repair.

The aim of this study was to develop and characterize a double layer biomembrane for dual drug delivery to be used for the treatment of wounds. The membrane was composed of chitosan, hydroxypropyl methylcellulose and lidocaine chloride (anesthetic drug) in the first layer, and of sodium alginate-polymyxin B sulphate (antibiotic) nanoparticles as the second layer. A product with excellent thickness (0.01-0.02 mm), adequate mechanical properties with respect to elasticity, stiffness, tension, and compatible pH for lesion application has been successfully obtained. The incorporation of the drugs was confirmed analysing the membrane cross-sections by scanning electron microscopy. A strong interaction between the drugs and the functional groups of respective polymers was confirmed by Fourier-Transform Infrared Spectroscopy, thermal analysis and X-ray diffraction. Microbiological assays showed a high antimicrobial activity when polymyxin B was present to act against the Staphylococcus aureus and Pseudomonas aeruginosa strains. Low cytotoxicity observed in a cell viability colorimetric assay and SEM analysis suggest biocompatibility between the developed biomembrane and the cell culture. The in vivo assay allowed visualizing the healing potential by calculating the wound retraction index and by histological analysis. Our results confirm the effectiveness of the developed innovative biomaterial for tissue repair and regeneration in an animal model.

Silver Nanoparticles-Composing Alginate/Gelatine Hydrogel Improves Wound Healing In Vivo.

Polymer hydrogels have been suggested as dressing materials for the treatment of cutaneous wounds and tissue revitalization. In this work, we report the development of a hydrogel composed of natural polymers (sodium alginate and gelatin) and silver nanoparticles (AgNPs) with recognized antimicrobial activity for healing cutaneous lesions. For the development of the hydrogel, different ratios of sodium alginate and gelatin have been tested, while different concentrations of AgNO3 precursor (1.0, 2.0, and 4.0 mM) were assayed for the production of AgNPs. The obtained AgNPs exhibited a characteristic peak between 430-450 nm in the ultraviolet-visible (UV-Vis) spectrum suggesting a spheroidal form, which was confirmed by Transmission Electron Microscopy (TEM). Fourier Transform Infra-red (FT-IR) analysis suggested the formation of strong intermolecular interactions as hydrogen bonds and electrostatic attractions between polymers, showing bands at 2920, 2852, 1500, and 1640 cm-1. Significant bactericidal activity was observed for the hydrogel, with a Minimum Inhibitory Concentration (MIC) of 0.50 µg/mL against Pseudomonas aeruginosa and 53.0 µg/mL against Staphylococcus aureus. AgNPs were shown to be non-cytotoxic against fibroblast cells. The in vivo studies in female Wister rats confirmed the capacity of the AgNP-loaded hydrogels to reduce the wound size compared to uncoated injuries promoting histological changes in the healing tissue over the time course of wound healing, as in earlier development and maturation of granulation tissue. The developed hydrogel with AgNPs has healing potential for clinical applications.

Chitosan/pvp-based mucoadhesive membranes as a promising delivery system of betamethasone-17-valerate for aphthous stomatitis.

Mucoadhesive membranes were proposed in this study as drug delivery system for betamethasone-17-valerate (BMV) in the treatment of recurrent aphthous stomatitis (RAS). The membranes were obtained by using the polymers chitosan (CHI) in both presence and absence of polyvinilpyrrolidone (PVP), following the solvent evaporation method. The presence of PVP in the membranes causes significant modifications in its thermal properties. Changes in the thermal events at 114 and 193 °C (related to BMV melting point), and losses in mass (39.38 and 30.68% for CH:PVP and CH:PVP-B, respectively), suggests the incorporation of BMV in these membranes. However, the morphological aspects of the membranes do not change after adding PVP and BMV. PVP causes changes in swelling ratios (>80%) of the membranes, and it is suggested that the reorganization of the polymer mesh was highlighted by the chemical interactions between the polymers leading to different percentages of BMV released ∼40% and ∼80% from CH-B and CH:PVP-B. BMV release profile follows Korsmeyer and Peppas model (n > 0.89) which suggests that the diffusion of the drug in the swollen matrix is driven by polymer relaxation. In addition, the membranes containing PVP (higher swelling ability) present high rates of tensile strength, and therefore, higher mucoadhesion. Moreover, given the results presented, the developed mucoadhesive membranes are a promising system to deliver BMV for the treatment of RAS.

Improving the therapeutic potential of endostatin by fusing it with the BAX BH3 death domain.

Endostatin (ES) inhibits angiogenesis, reducing tumor growth in animal models. However, it has low therapeutic effect in human clinical trials. BAX is a member of the BCL-2 family of proteins; its proapoptotic (BH3) domain interacts with other members of the family in the cytoplasm, to induce apoptosis. Here, we fused the BAX BH3 domain with murine ES, to enhance ES potency. Endothelial cells specifically internalize the fusion protein ES-BAX. The presence of the BAX domain enhances endothelial cell death by apoptosis by 1.8-fold and diminishes microvessel outgrowth in the rat aortic ring assay by 6.5-fold. Daily injections of 15 µg of ES-BAX/g in tumor-bearing mice reduce tumor weight by 86.9% as compared with ES-treated animals. Co-immunoprecipitation assays confirmed that ES-BAX interacts with members of the BCL-2 family. Also, ES interacts with BCL-2, BCL-XL, and BAK in endothelial cell lysates, suggesting a potential new mechanism for the apoptosis induction by ES. The superiority of the ES-BAX antiangiogenic effect indicates that this fusion protein could be a promising therapeutic alternative to treat cancer.

Detection of chromosomal bla(CTX-M-2) in diverse Escherichia coli isolates from healthy broiler chickens.

The rise of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in food-producing animals is a growing concern for public health. We investigated ESBL producers isolated from broiler chickens in Brazil and characterized 19 CTX-M-2-producing E. coli. The ISCR1 was detected upstream of the chromosome-located gene bla(CTX-M-2), associated with sul-1 type integron structure. CTX-M-2-producing E. coli exhibited different PFGE-types and phylogenetic groups, showing a non-clonal dissemination. The sequence types found (ST93, ST155 and ST2309) have been associated with humans and animals worldwide. Herein, we report the chromosomal location of bla(CTX-M-2) on E. coli, highlighting the risks of multidrug-resistant bacteria in food-producing animals.

Toll-like receptors 2, 3 and 4 and thymic stromal lymphopoietin expression in fatal asthma.

BACKGROUND: Airway inflammation in asthma involves innate immune responses. Toll-like receptors (TLRs) and thymic stromal lymphopoietin (TSLP) are thought to be involved in airway inflammation, but their expression in asthmatics' both large and small airways has not been investigated. OBJECTIVE: To analyse the expression of TLR2, TLR3, TLR4 and TSLP in large and small airways of asthmatics and compare their expression in smoking and non-smoking asthmatics; to investigate whether TLR expression is associated with eosinophilic or neutrophilic airway inflammation and with Mycoplasma pneumoniae and Chlamydophila pneumoniae infection. METHODS: Using immunohistochemistry and image analysis, we investigated TLR2, TLR3, TLR4 and TSLP expression in large and small airways of 24 victims of fatal asthma, FA, (13 non-smokers, 11 smokers) and nine deceased control subjects (DCtrl). TLRs were also measured in 18 mild asthmatics (MA) and 12 healthy controls (HCtrl). M. pneumoniae and C. pneumoniae in autopsy lung tissue were analysed using real-time polymerase chain reaction. Airway eosinophils and neutrophils were measured in all subjects. RESULTS: Fatal asthma patients had higher TLR2 in the epithelial and outer layers of large and small airways compared with DCtrls. Smoking asthmatics had lower TLR2 levels in the inner and outer layers of the small airways than non-smoking asthmatics. TSLP was increased in the epithelial and outer layers of the large airways of FA. FA patients had greater TLR3 expression in the outer layer of large airways and greater TLR4 expression in the outer layer of small airways. Eosinophilic airway inflammation was associated with TLR expression in the epithelium of FA. No bacterial DNA was detected in FA or DCtrls. MA and HCtrls had only a small difference in TLR3 expression. CONCLUSIONS AND CLINICAL RELEVANCE: Increased expression of TLR 2, 3 and 4 and TSLP in fatal asthma may contribute to the acute inflammation surrounding asthma deaths.

Determinants of beta-lactam resistance in meningitis-causing Enterobacteriaceae in Brazil.

This study analyzed resistance determinants in extended-spectrum beta-lactamase (ESBL)-producing enterobacteria and the epidemiology of 11 Escherichia coli isolates obtained from meningitis patients in a region of Brazil from 2000 to 2005. ESBL-encoding genes and their genetic environment were investigated by PCR and sequencing. The gene blaCTX-M-2 was identified in 3 different enterobacteria (E. coli, Serratia marcescens, and Proteus mirabilis) downstream of the insertion sequence ISCR1 (localized in class 1 integrons), but not as part of the resistance cassettes region. Multilocus sequence typing (MLST) was used to investigate genetic relationships between the 11 E. coli isolates in this study and strains associated with meningitis in the E. coli MLST database. MLST analysis indicated high genetic diversity among isolates, and no significant genetic relationship was identified with meningitis-causing E. coli in the database. The results in this report reinforce the need to be attentive to meningitis suspected to be due to ESBL-producing enterobacterial isolates, especially where ESBL epidemiology is well known.

Evaluation of environmental mycobacteria contamination in a specific pathogen free animal facility from a tropical country.

With the evidence showing the protection variability of bacille Calmette-Guérin, new potential vaccines for tuberculosis have been tested around the world. One of the general concerns in tuberculosis vaccine development is the possibility of priming the host immune system with prior exposure to environmental mycobacteria antigens, which can change the efficacy of subsequent vaccination. As there is a great homology between the species from Mycobacterium genera, the previous contact of experimental animals with environmental mycobacteria could sensitize the mice and, in this way, could influence subsequent vaccine research. The aim of our study was to investigate critical points in an animal facility to search for environmental mycobacteria that eventually could be in direct or indirect contact with the experimental animals. Samples were collected from surfaces of walls, floor, animal cages and shelves and analysed using the Ogawa-Kudoh decontamination method. Samples of drinking water, food and sawdust were collected for analysis by the NALC/NaOH decontamination method. Also, the samples were cultivated directly in broth medium, without any method for decontamination. After decontamination methods, we observed bacterial colony growth in 4.31% of the total of samples analysed. These samples were stained with Ziehl-Neelsen and we did not detect any acid-fast bacilli, suggesting that the animal facility analysed is free from contamination by environmental mycobacteria and is not a source of mycobacterial antigens. Furthermore, our study showed a new paradigm in tuberculosis vaccine development: concern about the animal facility environment in terms of immune system priming of experimental animals by nascent bacterial contaminants.