RESUMO
The present work aimed to produce a cupuassu juice (Theobroma grandiflorum) fermented by the probiotic bacterium Lactiplantibacillus plantarum Lp62 and to analyze its antioxidant potential, antimicrobial activity, and resistance to biological barriers. The fermented beverage showed an increase in the content of phenolics, flavonoids, and antioxidant potential. The culture showed antagonistic activity against pathogens, but this result was not observed when the juice was tested. The probiotic strain remained viable under refrigeration, even in an acidified environment, and survived simulated gastrointestinal transit in vitro. L. plantarum Lp62 showed 30% adherence to HT-29 intestinal cells and proved to be safe in terms of antibiotic resistance and production of virulence factors. Fermentation increased the functional characteristics of cupuassu juice. This drink proved to be a good vehicle for the delivery of the probiotic bacteria L. plantarum Lp62.
Assuntos
Sucos de Frutas e Vegetais , Lactobacillaceae , Malvaceae , Probióticos , Humanos , Farmacorresistência Bacteriana , Fermentação , Sucos de Frutas e Vegetais/análise , Sucos de Frutas e Vegetais/microbiologia , Células HT29 , Lactobacillaceae/efeitos dos fármacos , Lactobacillaceae/metabolismoRESUMO
Escherichia coli is one of the main pathogens that impacts swine production. Given the need for methods for its control, the in vitro effect of lactic acid bacteria (LAB) and their metabolites against E. coli F4 was evaluated through cell culture and microbiological analysis. The strains Limosilactobacillus fermentum 5.2, Lactiplantibacillus plantarum 6.2, and L. plantarum 7.1 were selected. To evaluate the action of their metabolites, lyophilized cell-free supernatants (CFS) were used. The effect of CFS was evaluated in HT-29 intestinal lineage cells; in inhibiting the growth of the pathogen in agar; and in inhibiting the formation of biofilms. The bioprotective activity of LAB was evaluated via their potential for autoaggregation and coaggregation with E. coli. The CFS did not show cytotoxicity at lower concentrations, except for L. fermentum 5.2 CFS, which is responsible for cell proliferation at doses lower than 10 mg ml-1. The CFS were also not able to inhibit the growth of E. coli F4 in agar; however, the CFS of L. plantarum 7.1 resulted in a significant decrease in biofilm formation at a dose of 40 mg ml-1. Regarding LAB, their direct use showed great potential for autoaggregation and coaggregation in vitro, thus suggesting possible effectiveness in animal organisms, preventing E. coli fixation and proliferation. New in vitro tests are needed to evaluate lower doses of CFS to control biofilms and confirm the bioprotective potential of LAB, and in vivo tests to assess the effect of LAB and their metabolites interacting with animal physiology.
Assuntos
Escherichia coli Enterotoxigênica , Lactobacillales , Limosilactobacillus fermentum , Ágar , Animais , SuínosRESUMO
Traditionally, probiotic microorganisms are isolated from human and animal intestinal microbiota. However, the demand for diversification of biofunctional products has driven the search for new sources of probiotic candidates, such as fermented foods and vegetables. The present study found that strains isolated from the fermentation of fine cocoa from southern Bahia have biotechnological potential for use as a probiotic, since they showed capacity for self-aggregation and co-aggregation, antimicrobial activity against intestinal pathogens and resistance to gastrointestinal transits. Scores of importance for each property were established in order to more accurately assess the probiotic potential of the strains. The tests carried out contemplate the criteria previously established for the selection of probiotic candidates.
Assuntos
Lactobacillus , Probióticos , Animais , Humanos , Fermentação , Intestinos , Probióticos/farmacologiaRESUMO
BACKGROUND: Probiotics are important tools in therapies against vaginal infections and can assist traditional antibiotic therapies in restoring healthy microbiota. Recent research has shown that microorganisms belonging to the genus Lactobacillus have probiotic potential. Thus, this study evaluated the potential in vitro probiotic properties of three strains of Lactiplantibacillus plantarum, isolated during the fermentation of high-quality cocoa, against Gardnerella vaginalis and Neisseria gonorrhoeae. Strains were evaluated for their physiological, safety, and antimicrobial characteristics. RESULTS: The hydrophobicity of L. plantarum strains varied from 26.67 to 91.67%, and their autoaggregation varied from 18.10 to 30.64%. The co-aggregation of L. plantarum strains with G. vaginalis ranged from 14.73 to 16.31%, and from 29.14 to 45.76% with N. gonorrhoeae. All L. plantarum strains could moderately or strongly produce biofilms. L. plantarum strains did not show haemolytic activity and were generally sensitive to the tested antimicrobials. All lactobacillus strains were tolerant to heat and pH resistance tests. All three strains of L. plantarum showed antimicrobial activity against the tested pathogens. The coincubation of L. plantarum strains with pathogens showed that the culture pH remained below 4.5 after 24 h. All cell-free culture supernatants (CFCS) demonstrated activity against the two pathogens tested, and all L. plantarum strains produced hydrogen peroxide. CFCS characterisation in conjunction with gas chromatography revealed that organic acids, especially lactic acid, were responsible for the antimicrobial activity against the pathogens evaluated. CONCLUSION: The three strains of L. plantarum presented significant probiotic characteristics against the two pathogens of clinical importance. In vitro screening identified strong probiotic candidates for in vivo studies for the treatment of vaginal infections.
Assuntos
Antibiose/fisiologia , Cacau/microbiologia , Alimentos Fermentados/microbiologia , Gardnerella vaginalis/fisiologia , Lactobacillus plantarum/fisiologia , Neisseria gonorrhoeae/fisiologia , Probióticos , Fermentação , Humanos , Lactobacillus plantarum/isolamento & purificaçãoRESUMO
PURPOSE: Lactobacillus plantarum Lp62 is a lactic acid bacteria strain that has been isolated from cocoa beans and exhibited probiotic potential. The influence of oral administration of L. plantarum Lp62 on the growth of rat's pups; on yield, cytokines and milk composition was studied. METHODS: Lactobacillus plantarum Lp62 is a lactic acid bacteria strain that has been isolated from cocoa beans. It was administered daily by gavage to Wistar rats (n = 8), from the 7th day before delivery and for 20 days during lactation, in a concentration of 1.44 × 109 CFU/rat. The dam and pups were weighed and milk was collected at 12th and 19th day for determination of protein, triglycerides, cholesterol and lactose by colorimetric assays. TGF-ß1 milk levels were analyzed by ELISA. The mammary glands of rats were removed for histological analysis. To detect statistical differences between the groups, tests of mean differences at a significance level of 5% was performed. RESULTS: Supplementation with L. plantarum L62 resulted in significant higher weight of pups (p < 0.05), with similar weight on dams (p > 0.05). The milk yield was not altered by L. plantarum treatment, but the levels of protein, triglycerides and cholesterol were increased (p < 0.05), with no difference in lactose concentration (p > 0.05). Levels of TGF-ß1 were higher in the milk of L. plantarum treatment (p < 0.05). CONCLUSIONS: The treatment of dams at the end of pregnancy and lactation with L. plantarum Lp62 increased nutritional content of milk, probably contributing to the higher weight of the pups. The higher levels of TGF-ß1 in the milk, could promote immune benefits to the pups. Further studies in this field are needed to prove the potential use of L. plantarum Lp62 as a probiotic.
Assuntos
Peso Corporal/efeitos dos fármacos , Lactação/efeitos dos fármacos , Lactobacillus plantarum , Fator de Crescimento Transformador beta1/efeitos dos fármacos , Administração Oral , Animais , Animais Recém-Nascidos , Animais Lactentes , Feminino , Lactação/metabolismo , Modelos Animais , Gravidez , Prenhez , Ratos , Ratos Wistar , Fator de Crescimento Transformador beta1/metabolismoRESUMO
Neutrophils respond differently to violations of the body's physiological barriers during infections. Extracellular traps comprise one of the mechanisms used by these cells to reduce the spread of pathogens to neighboring tissues, as well as ensure a high concentration of antimicrobial agents at the site of infection. To date, this innate defense mechanism has not been previously demonstrated in neutrophils of cats exposed to Toxoplasma gondii. The aim of this study was to characterize the in vitro release of neutrophil extracellular traps (NETs) when neutrophils isolated from cats were exposed to T. gondii. First, cellular viability was tested at different time points after parasite exposure. The production of reactive oxygen species (ROS) and lactate dehydrogenase and the amount of extracellular DNA were quantified. In addition, the number of parasites associated with neutrophils was determined, and the observed NETs formed were microscopically characterized. Results showed that (i) in culture, neutrophils isolated from cats presented diminished cellular viability after 4â¯h of incubation, and when neutrophils were incubated with T. gondii, they displayed cytotoxic effects after 3â¯h of interaction; (ii) neutrophils were able to release structures composed of DNA and histones, characterized as NETs under optical, immunofluorescence, and electron scanning microscopy, when stimulated with T. gondii; (iii) only 11.4% of neutrophils were able to discharge NETs during 3â¯h of incubation; however, it was observed through extracellular quantification of DNA that this small number of cells were able to display different behavior compared to a negative control (no parasite) group; (iv) significant differences in ROS production were observed in neutrophils exposed to T. gondii. In conclusion, our results showed that neutrophils isolated from cats exposed to T. gondii release structures composed of DNA and histones, similar to what has already been described in other neutrophil species infected with the parasite.
Assuntos
Armadilhas Extracelulares/metabolismo , Neutrófilos/parasitologia , Toxoplasma/imunologia , Animais , Gatos , Sobrevivência Celular , Chlorocebus aethiops , DNA/análise , Formazans/metabolismo , L-Lactato Desidrogenase/metabolismo , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Neutrófilos/imunologia , Neutrófilos/ultraestrutura , Espécies Reativas de Oxigênio/metabolismo , Superóxidos/análise , Sais de Tetrazólio/metabolismo , Células VeroRESUMO
BACKGROUND: Some sexually transmitted infectious agents, such as Chlamydia trachomatis and Herpes simplex, cause local inflammation, and could contribute to Human Papillomavirus (HPV) and cervical lesion progression. Thus, the aim of this study was to determine any association between the presence of microorganisms of gynecological importance, sexual behavior, clinical and demographical variables to the development and progress of cervical lesions. METHODS: One hundred and thirty-two women between 14 and 78 years and living at Vitória da Conquista, Bahia, Brazil, were included (62 individuals with cervical lesions and 70 without lesions). They answered a questionnaire to provide data for a socioeconomic and sexual activity profile. Samples of cervical swabs were collected and analyzed by PCR to detect genital microorganisms and HPV. Quantitative PCR was used to detect and quantify Ureaplasma urealyticum and Ureaplasma parvum. Univariate and multiple logistic regression were performed to measure the association with the cervical lesions, and an odds ratio (OR) with 95% confidence intervals (95%CI) were calculated. The Mann-Whitney U test was also used to compare the microorganism load in the case and control groups. The significance level was 5% in all hypotheses tested. RESULTS: Cervical lesions were associated with: women in a stable sexual relationship (OR = 14.21, 95%CI = 3.67-55.018), positive PCR for HPV (OR = 16.81, 95%CI = 4.19-67.42), Trichomonas vaginalis (OR = 8.566, 95%CI = 2.04-35.94) and Gardnerella vaginalis (OR = 6.13, 95%CI = 1.53-24.61), adjusted by age and qPCR for U. parvum. U. parvum load showed a statistical difference between the case and control groups (p-value = 0.002). CONCLUSION: Variables such as stable relationship, HPV, T. vaginalis, G. vaginalis were associated with cervical lesions in epidemiological studies. U. parvum load was higher in woman with cervical lesions compared with women without lesions. Additional studies are needed to better understand the role of these factors in cervical lesion development.
Assuntos
Infecções por Papillomavirus/diagnóstico , Infecções Sexualmente Transmissíveis/diagnóstico , Doenças do Colo do Útero/diagnóstico , Adolescente , Adulto , Idoso , Brasil , Colo do Útero/microbiologia , Colo do Útero/virologia , Coinfecção/diagnóstico , Coinfecção/microbiologia , Coinfecção/virologia , DNA Bacteriano/isolamento & purificação , DNA Bacteriano/metabolismo , DNA Viral/isolamento & purificação , DNA Viral/metabolismo , Feminino , Gardnerella vaginalis/genética , Gardnerella vaginalis/isolamento & purificação , Humanos , Modelos Logísticos , Pessoa de Meia-Idade , Razão de Chances , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/transmissão , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real , Infecções Sexualmente Transmissíveis/microbiologia , Infecções Sexualmente Transmissíveis/transmissão , Infecções Sexualmente Transmissíveis/virologia , Inquéritos e Questionários , Trichomonas vaginalis/genética , Trichomonas vaginalis/isolamento & purificação , Ureaplasma/genética , Ureaplasma/isolamento & purificação , Ureaplasma urealyticum/genética , Ureaplasma urealyticum/isolamento & purificação , Doenças do Colo do Útero/microbiologia , Doenças do Colo do Útero/virologia , Adulto JovemRESUMO
BACKGROUND: Biofilm production represents an important virulence and pathogenesis factor for Staphylococcus aureus. The formation of biofilms on medical devices is a major concern in hospital environments, as they can become a constant source of infection. Probiotic bacteria, such as Lactobacillus fermentum and L. plantarum, have been found to inhibit biofilm formation; however little is known about the underlying mechanism. In this study, we tested the activity of supernatants produced by L. fermentum TCUESC01 and L. plantarum TCUESC02, isolated during the fermentation of fine cocoa, against S. aureus CCMB262 biofilm production. We measured inhibition of biofilm formation in vitro and analyzed biofilm structure by confocal and electronic microscopy. Additionally, we quantified the expression of S. aureus genes icaA and icaR involved in the synthesis of the biofilm matrix by real-time PCR. RESULTS: Both Lactobacillus supernatants inhibited S. aureus growth. However, only L. fermentum TCUESC01 significantly reduced the thickness of the biofilm, from 14 µm to 2.83 µm (at 18 mgâmL-1, 90 % of the minimum inhibitory concentration, MIC), 3.12 µm (at 14 mgâmL-1, 70 % of the MIC), and 5.21 µm (at 10 mgâmL-1, 50 % of the MIC). Additionally, L. fermentum TCUESC01 supernatant modulated the expression of icaA and icaR. CONCLUSIONS: L. fermentum TCUESC01 reduces the formation of S. aureus biofilm under subinhibitory conditions. Inhibition of biofilm production probably depends on modulation of the ica operon.
Assuntos
Biofilmes/crescimento & desenvolvimento , Chocolate/microbiologia , Lactobacillus/fisiologia , Staphylococcus aureus/fisiologia , Antibiose , Meios de Cultura , Fermentação , Lactobacillus/isolamento & purificação , Limosilactobacillus fermentum/fisiologia , Lactobacillus plantarum/fisiologia , Testes de Sensibilidade Microbiana , Microscopia Confocal , Microscopia Eletrônica de Varredura , Fenótipo , Poliestirenos , ProbióticosRESUMO
Ants cause a series of accidents involving humans. Such accidents generate different reactions in the body, ranging from a mild irritation at the bite site to anaphylactic shock, and these reactions depend on the mechanism of action of the venom. The study of animal venom is a science known as venomics. Through venomics, the composition of the venom of several ant species has already been characterized and their biological activities described. Thus, the aim of this study was to evaluate the protein composition and biological activities (hemolytic and immunostimulatory) of the venom of Neoponera villosa (N. villosa), an ant widely distributed in South America. The protein composition was evaluated by proteomic techniques, such as two-dimensional electrophoresis. To assess the biological activity, hemolysis assay was carried out and cytokines were quantified after exposure of macrophages to the venom. The venom of N. villosa has a profile composed of 145 proteins, including structural and metabolic components (e.g., tubulin and ATPase), allergenic and immunomodulatory proteins (arginine kinase and heat shock proteins (HSPs)), protective proteins of venom (superoxide dismutase (SOD) and catalase) and tissue degradation proteins (hyaluronidase and phospholipase A2). The venom was able to induce hemolysis in human erythrocytes and also induced release of both pro-inflammatory cytokines, as the anti-inflammatory cytokine release by murine macrophages. These results allow better understanding of the composition and complexity of N. villosa venom in the human body, as well as the possible mechanisms of action after the bite.
Assuntos
Venenos de Formiga/farmacologia , Hemolíticos/farmacologia , Proteoma , Animais , Venenos de Formiga/química , Venenos de Formiga/isolamento & purificação , Formigas , Brasil , Células Cultivadas , Citocinas/metabolismo , Eritrócitos/efeitos dos fármacos , Humanos , Macrófagos/efeitos dos fármacos , CamundongosRESUMO
Background: People in low-income countries, especially those with low socio-economic conditions, are likelier to test positive for SARS-CoV-2. The unequal conditions of public health systems also increase the infection rate and make early identification and treatment of at-risk patients difficult. Here, we aimed to characterize the epidemiological profile of COVID-19 patients in intensive care and identify laboratory and clinical markers associated with death. Materials and methods: We conducted an observational, descriptive, and cross-sectional study in a reference hospital for COVID-19 treatment in the Southern Region of Bahia State, in Brazil, to evaluate the epidemiological, clinical, and laboratory characteristics of COVID-19 patients admitted to the intensive care unit (ICU). Additionally, we used the area under the curve (AUC) to classify survivors and non-survivors and a multivariate logistic regression analysis to assess factors associated with death. Data was collected from the hospital databases between April 2020 and July 2021. Results: The use of bladder catheters (OR 79.30; p < 0.0001) and central venous catheters (OR, 45.12; p < 0.0001) were the main factors associated with death in ICU COVID-19 patients. Additionally, the number of non-survivors increased with age (p < 0.0001) and prolonged ICU stay (p < 0.0001). Besides, SAPS3 presents a higher sensibility (77.9%) and specificity (63.1%) to discriminate between survivors and non-survivor with an AUC of 0.79 (p < 0.0001). Conclusion: We suggest that multi-laboratory parameters can predict patient prognosis and guide healthcare teams toward more assertive clinical management, better resource allocation, and improved survival of COVID-19 patients admitted to the ICU.
Assuntos
COVID-19 , Humanos , COVID-19/epidemiologia , Brasil/epidemiologia , SARS-CoV-2 , Tratamento Farmacológico da COVID-19 , Estudos Transversais , Unidades de Terapia Intensiva , HospitaisRESUMO
This work reports the distribution of an oral dose of Salmonella enterica serovar Enteritidis (SE) in C57Bl/6-Bcgr mice, to study its pathogenesis in a latent carrier animal. Mice orally inoculated with a high dose of SE developed a latent infection characterized by the absence of clinical symptoms in which the cecum is functioning as a "strategic site" of SE proliferation, releasing bacteria into feces intermittently over the 4-week study. A sequence of disruptions occurred in the small intestine at 1 day postinculation (PI). The microvilli exhibited different degrees of degeneration, which were reversible as the cells became vacuolated. From 2 days PI, SE was detected in the mononuclear phagocytic system, and an exponential growth of the remaining bacteria in tissues was observed until 4 days PI. The production of interferon gamma from 3 days PI is restricting the SE growth, and a plateau phase was observed from 4 to 15 days PI. A recurrence of the bacterial growth in tissue occurred from 15 to 28 days PI, especially in the cecum. Increasing our knowledge about the host-pathogen interaction of adapted pathogens with the ability to develop latency is essential for the development of an efficient strategy for Salmonella control.
Assuntos
Portador Sadio/microbiologia , Ceco/microbiologia , Interações Hospedeiro-Patógeno , Salmonelose Animal/microbiologia , Salmonella enteritidis/patogenicidade , Animais , Derrame de Bactérias , Fezes/microbiologia , Interferon gama/imunologia , Interleucina-12/imunologia , Intestino Delgado/microbiologia , Intestino Delgado/patologia , Intestino Delgado/ultraestrutura , Camundongos , Camundongos Endogâmicos C57BL , Sistema Fagocitário Mononuclear/imunologia , Salmonelose Animal/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
COVID-19 is a lethal disease caused by the pandemic SARS-CoV-2, which continues to be a public health threat. COVID-19 is principally a respiratory disease and is often associated with sputum retention and cytokine storm, for which there are limited therapeutic options. In this regard, we evaluated the use of BromAc®, a combination of Bromelain and Acetylcysteine (NAC). Both drugs present mucolytic effect and have been studied to treat COVID-19. Therefore, we sought to examine the mucolytic and anti-inflammatory effect of BromAc® in tracheal aspirate samples from critically ill COVID-19 patients requiring mechanical ventilation. METHOD: Tracheal aspirate samples from COVID-19 patients were collected following next of kin consent and mucolysis, rheometry and cytokine analysis using Luminex kit was performed. RESULTS: BromAc® displayed a robust mucolytic effect in a dose dependent manner on COVID-19 sputum ex vivo. BromAc® showed anti-inflammatory activity, reducing the action of cytokine storm, chemokines including MIP-1alpha, CXCL8, MIP-1b, MCP-1 and IP-10, and regulatory cytokines IL-5, IL-10, IL-13 IL-1Ra and total reduction for IL-9 compared to NAC alone and control. BromAc® acted on IL-6, demonstrating a reduction in G-CSF and VEGF-D at concentrations of 125 and 250 µg. CONCLUSION: These results indicate robust mucolytic and anti-inflammatory effect of BromAc® ex vivo in tracheal aspirates from critically ill COVID-19 patients, indicating its potential to be further assessed as pharmacological treatment for COVID-19.
Assuntos
Acetilcisteína/farmacologia , Bromelaínas/farmacologia , COVID-19/patologia , Quimiocinas/efeitos dos fármacos , Citocinas/efeitos dos fármacos , Escarro/citologia , Acetilcisteína/administração & dosagem , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Bromelaínas/administração & dosagem , Síndrome da Liberação de Citocina/patologia , Relação Dose-Resposta a Droga , Regulação para Baixo , Combinação de Medicamentos , Expectorantes/farmacologia , Feminino , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Pessoa de Meia-Idade , Respiração Artificial , Reologia , SARS-CoV-2 , Traqueia/patologia , Adulto JovemRESUMO
BACKGROUND: Fungi of the genus Paracoccidioides are the etiological agents of paracoccidioidomycosis, a highly prevalent mycosis in Latin America. Infection in humans occurs by the inhalation of conidia, which later revert to the form of yeast. In this context, macrophages are positioned as an important line of defense, assisting in the recognition and presentation of antigens, as well as producing reactive oxygen species that inhibit fungal spreading. AIMS: The objective of this study was to identify differentially expressed proteins during the interaction between Paracoccidioides lutzii Pb01 strain and human U937 monocytes. METHODS: Two-dimensional electrophoresis, combined with mass spectrometry, was used to evaluate the differential proteomic profiles of the fungus P. lutzii (Pb01) interacting with U937 monocytes. RESULTS: It was possible to identify 25 proteins differentially expressed by Pb01 alone and after interacting with U937 monocytes. Most of these proteins are directly associated with fungal metabolism for energy generation, such as glyceraldehyde-3-phosphate dehydrogenase, and intracellular adaptation to monocytes. Antioxidant proteins involved in the response to oxidative stress, such as peroxiredoxin, cytochrome, and peroxidase, were expressed in greater quantity in the interaction with monocytes, suggesting their association with survival mechanisms inside phagocytic cells. We also identified 12 proteins differentially expressed in monocytes before and after the interaction with the fungus; proteins involved in the reorganization of the cytoskeleton, such as vimentin, and proteins involved in the response to oxidative stress, such as glioxalase 1, were identified. CONCLUSIONS: The results of this proteomic study of a P. lutzii isolate are novel, mimicking in vitro what occurs in human infections. In addition, the proteins identified may aid to understand fungal-monocyte interactions and the pathogenesis of paracoccidioidomycosis.
Assuntos
Paracoccidioides , Paracoccidioidomicose , Proteínas Fúngicas/genética , Humanos , Monócitos , ProteômicaRESUMO
Traditional therapy for malignant neoplasms involving surgical procedures, radiotherapy and chemotherapy aims to kill neoplastic cells, but also affects normal cells. Therefore, exogenous proteases are the target of studies in cancer therapy, as they have been shown to be effective in suppressing tumors and reducing metastases. Pluronic F127 (F127) is a copolymer of amphiphilic blocks that has shown significant potential for drug administration, as it is capable of incorporating hydrophobic drugs and self-assembling in micrometers of nanometric size. This study investigated the effects of immobilization of the alkaline protease PR4A3 with pluronic F127 micelles on the enzyme-induced cytotoxicity. Protease immobilization was demonstrated through UV-visible and circular dichroism (CD) spectroscopies, as the enzyme interacts with the polymeric micelle of Pluronic F127 without changing its secondary structure. In addition, the immobilized form of the enzyme showed greater bioavailability after passing through the simulated gastrointestinal transit. Cell viability was assessed using the tetrazoic methylthiazole (MTT) assay. The results open perspectives for new research and development for PR4A3 in the treatment of colorectal carcinoma.
Assuntos
Adenocarcinoma/patologia , Proteínas de Bactérias/metabolismo , Neoplasias Colorretais/patologia , Endopeptidases/metabolismo , Enzimas Imobilizadas/metabolismo , Micelas , Poloxâmero/química , Disponibilidade Biológica , Morte Celular , Linhagem Celular Tumoral , Dicroísmo Circular , Trânsito Gastrointestinal , Humanos , Hidrólise , Proteólise , Espectrofotometria UltravioletaRESUMO
Staphylococcus aureus is a Gram-positive bacterium that is considered an important human pathogen. Due to its virulence and ability to acquire mechanisms of resistance to antibiotics, the clinical severity of S. aureus infection is driven by inflammatory responses to the bacteria. Thus, the present study aimed to investigate the modulating role of citral in inflammation caused by S. aureus infection. For this, we used an isolate obtained from a nasal swab sample of a healthy child attending a day-care centre in Vitória da Conquista, Bahia, Brazil. The role of citral in modulating immunological factors against S. aureus infection was evaluated by isolating and cultivating human peripheral blood mononuclear cells. The monocytes were treated with 4%, 2%, and 1% citral before and after inoculation with S. aureus. The cells were analysed by immunophenotyping of monocyte cell surface molecules (CD54, CD282, CD80, HLA-DR, and CD86) and cytokine dosage (IL-1ß, IL-6, IL-10, IL-12p70, IL-23, IFN-γ, TGF-ß, and TNF-α), and evaluated for the expression of 84 genes related to innate and adaptive immune system responses. GraphPad Prism software and variables with P values < 0.05, were used for statistical analysis. Our data demonstrated citral's action on the expression of surface markers involved in recognition, presentation, and migration, such as CD14, CD54, and CD80, in global negative regulation of inflammation with inhibitory effects on NF-κB, JNK/p38, and IFN pathways. Consequently, IL-1ß, IL-6, IL-12p70, IL-23, IFN-γ, and TNF-α cytokine expression was reduced in groups treated with citral and groups treated with citral at 4%, 2%, and 1% and infected, and levels of anti-inflammatory cytokines such as IL-10 were increased. Furthermore, citral could be used as a supporting anti-inflammatory agent against infections caused by S. aureus. There are no data correlating citral, S. aureus, and the markers analysed here; thus, our study addresses this gap in the literature.
Assuntos
Monoterpenos Acíclicos/farmacologia , Fatores Imunológicos/farmacologia , Monócitos/efeitos dos fármacos , Infecções Estafilocócicas/imunologia , Staphylococcus aureus/imunologia , Adulto , Brasil , Células Cultivadas , Criança , Humanos , Inflamação/complicações , Inflamação/tratamento farmacológico , Inflamação/imunologia , Masculino , Monócitos/imunologia , Infecções Estafilocócicas/complicações , Infecções Estafilocócicas/tratamento farmacológico , Staphylococcus aureus/efeitos dos fármacos , Adulto JovemRESUMO
Microorganisms native to mangroves are expected to contain enzymes capable of hydrolyzing different carbon sources. However, most of these microorganisms aren't cultivable; hence, alternative techniques as metagenomics are tools for studying and obtaining some of the natural genomes, genes and enzymes of biotechnological interest. The ß-glucanase was produced using a metagenomic clone of mangrove sediments and detected by functional screening on carboxymethylcellulose substrate. The enzyme was purified by cation exchange chromatography. The peptides detected by mass spectrometry showed 20% identity with the polypeptide deduced from the genomic fragment sequenced. The ORF identified as BglfosD9 possessed 729â¯bp and the encoded protein showed predicted MW and pI of 28kD and 6.8, respectively. The enzyme was active in a wide range of pH (5-10) with optimum pH at 8, had relative activity greater than 50% at all temperatures tested (5-90⯰C), was stable at temperatures of 5, 50 and 90⯰C and showed excellent relative activity at high NaCl concentrations. This ß-glucanase also showed high relative activity in the presence of SDS and it could hydrolyze ß-glucan, CMC and Avicel as substrates. These findings support the idea of a new thermostable and active enzyme at basic pH from metagenomic library of mangrove sediment.
Assuntos
Bacillus , Sedimentos Geológicos/microbiologia , Glicosídeo Hidrolases , Áreas Alagadas , Bacillus/enzimologia , Bacillus/genética , Carboximetilcelulose Sódica/química , Clonagem Molecular , Estabilidade Enzimática , Glicosídeo Hidrolases/química , Glicosídeo Hidrolases/genética , Temperatura Alta , Metagenoma , SalinidadeRESUMO
In the present work we aimed to demonstrate the influence of inoculum starter in support high quality fermentation. Cocoa fermentations were performed in wooden boxes and eight yeasts strains were used in separated fermentations of fine cocoa, type Scavina, as starter inoculum. Temperature, pH, titirable acidity, reducing sugar and free amino acids were evaluated during or after fermentation. The influence of starters yeasts on the decrease of acidity, sugar concentration and free amino acids was significant. The strains Candida parapsilosis, Torulaspora delbrueckii and Pichia kluyveri showed greater changes in the reducing sugar and free amino acids in fermented cocoa beans. These results indicate the ability of yeast used as inoculum starter to modify the end condition and further enhance the quality of fine cocoa beans.
Assuntos
Cacau , Microbiologia de Alimentos/métodos , Leveduras , Aminoácidos/análise , Aminoácidos/metabolismo , Cacau/química , Cacau/metabolismo , Candida parapsilosis/genética , Candida parapsilosis/metabolismo , Chocolate , Fermentação , Concentração de Íons de Hidrogênio , Pichia/genética , Pichia/metabolismo , Sementes/química , Sementes/microbiologia , Temperatura , Torulaspora/genética , Torulaspora/metabolismo , Leveduras/genética , Leveduras/metabolismoRESUMO
Bacteria in the genera Mycoplasma and Ureaplasma do not have cell walls and therefore interact with host cells through lipid-associated membrane proteins (LAMP). These lipoproteins are important for both surface adhesion and modulation of host immune responses. Mycoplasma and Ureaplasma have been implicated in cases of bacterial vaginosis (BV), which can cause infertility, abortion, and premature delivery. In contrast, bacteria of the genus Lactobacillus, which are present in the vaginal microbiota of healthy women, are thought to inhibit local colonization by pathogenic microorganisms. The aim of the present study was to evaluate the in vitro interactions between lipoproteins of Mycoplasma and Ureaplasma species and vaginal lineage (HMVII) cells and to study the effect of Lactobacillus isolates from cocoa fermentation on these interactions. The tested Lactobacillus strains showed some important probiotic characteristics, with autoaggregation percentages of 28.55% and 31.82% for L. fermentum FA4 and L. plantarum PA3 strains, respectively, and percent adhesion values of 31.66 and 41.65%, respectively. The two strains were hydrophobic, with moderate to high hydrophobicity values, 65.33% and 71.12% for L. fermentum FA4 and L. plantarum PA3 in toluene. Both strains secreted acids into the culture medium with pH=4.32 and pH=4.33, respectively, and showed antibiotics susceptibility profiles similar to those of other lactobacilli. The strains were also able to inhibit the death of vaginal epithelial cells after incubation with U. parvum LAMP from 41.03% to 2.43% (L. fermentum FA4) and 0.43% (L. plantarum PA3) and also managed to significantly decrease the rate of cell death caused by the interaction with LAMP of M. hominis from 34.29% to 14.06% (L. fermentum FA4) and 14.61% (L. plantarum PA3), thus demonstrating their potential for maintaining a healthy vaginal environment.
Assuntos
Cacau/microbiologia , Células Epiteliais/microbiologia , Lactobacillus plantarum/crescimento & desenvolvimento , Limosilactobacillus fermentum/crescimento & desenvolvimento , Microbiota/fisiologia , Vagina/microbiologia , Linhagem Celular , Feminino , Humanos , Limosilactobacillus fermentum/isolamento & purificação , Lactobacillus plantarum/isolamento & purificaçãoRESUMO
OBJECTIVE: The objective of this study is to evaluate the inflammatory response caused by endovascular stents in the treatment of aortic aneurysms. METHODS: Twenty-five patients underwent endovascular stent treatment from March through December 2005. The evolution of mediators (sedimentation velocity, C reactive protein, interleukin-6, interleukin-8, tumor necrosis factor-alpha, intercellular adhesion molecule-1, l-selectin), inflammatory cells (leukocytes, lymphocytes, platelets), serum creatinine and body temperature within preoperative period and in the following postoperative periods--1, 6, 24 and 48 h, 7 days, 1-3 months, was analyzed. In order to achieve statistic significance, Friedman test and Wilcoxon test were used, with index of significance of 5% (p<0.05). RESULTS: Peak values of sedimentation velocity, C reactive protein and interleukin-6 were observed at 7 days (p<0.0001), 48 h (p<0.0001) and 24h (p<0.0001), respectively. Tumor necrosis factor-alpha and interleukin-8 did not show statistically significant variability during the entire follow-up. In terms of intercellular adhesion molecule-1 and l-selectin, their expressive values were found in late phase of follow-up, although without statistical significance. Elevation of leukocytes count occurred in premature phase of follow-up (p<0.0001), while lymphocyte and platelet count occurred in a late phase of follow-up (p<0.0001). Serum levels of creatinine did not show significant variability during follow-up. The period between 24 and 48 h corresponded to major frequency for fever (p<0.0001). CONCLUSION: Individual mediators analysis and inflammatory cells demonstrated variability of their values during postoperative follow-up. This could help in the analysis of the inflammatory response evolution caused by endovascular stent treatment for aortic aneurysms in premature and late phases after implantation of the vascular prosthesis.
Assuntos
Aneurisma Aórtico/cirurgia , Implante de Prótese Vascular/efeitos adversos , Mediadores da Inflamação/sangue , Inflamação/etiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Aneurisma da Aorta Abdominal/cirurgia , Aneurisma da Aorta Torácica/cirurgia , Feminino , Seguimentos , Humanos , Molécula 1 de Adesão Intercelular/sangue , Interleucina-6/sangue , Interleucina-8/sangue , Selectina L/sangue , Masculino , Pessoa de Meia-Idade , Stents/efeitos adversos , Fator de Necrose Tumoral alfa/sangueRESUMO
The use of intestinal probiotic bacteria is very common in the food industry and has been the focus of the majority of research in this field. Yet in recent years, research on extraintestinal microorganisms has greatly increased due to their well-known potential as probiotics. Thus, we studied a strain of Lactobacillus fermentum (TCUESC01) extracted from fermenting cocoa. First, we examined the impact of pH on the growth of this strain and studied its survival under conditions similar to those of the human gastrointestinal tract. L. fermentum TCUESC01 demonstrated resistance to conditions mimicking the human stomach and intestines and grew well between pH 5 and pH 7. Next, we subjected L. fermentum TCUESC01 to storage at 4°C in a milk solution and found that it survived well for 28 days. Lastly, we measured the susceptibility of this strain to numerous antibiotics and its tendency to autoaggregate. L. fermentum TCUESC01 showed significant autoaggregation, as well as susceptibility to the majority of antibiotics tested. Overall, our findings support the potential use of this extraintestinal bacterium as a dietary probiotic.