RESUMO
The term postbiotics has acquired great interest in recent years. Numerous studies have shown a potential beneficial effect of its use in many inflammatory pathologies. However, it has not been much explored in ocular inflammatory diseases. The aims of this study were to develop and characterise an ophthalmic formulation with a postbiotic of Lactiplantibacillus plantarum CRL 759, and to evaluate its anti-inflammatory actions on murine macrophage stimulated with lipopolysaccharides (LPS) in vitro. First, we evaluated the ability of L. plantarum CRL 759 to generate a supernatant with anti-inflammatory property using different buffers. Then, we studied the stability at different temperatures and storage times of the generated postbiotic. In vitro assays showed that incubation of L. plantarum CRL 759 in modified phosphate buffer according to Sorensen (called POF-759), generated a supernatant that significantly reduced the production of interleukin-6, tumour necrosis factor-α, and nitric oxide by RAW 264.7 cells stimulated with LPS. Furthermore, POF-759 maintained its anti-inflammatory activity at room temperature, 4 and -20 °C, up to 30 days of storage. From the studies reported here, a postbiotic product with anti-inflammatory properties and optimal characteristics for the formulation of eye drops was obtained.
Assuntos
Lactobacillus plantarum , Probióticos , Camundongos , Animais , Lipopolissacarídeos , Macrófagos , Anti-Inflamatórios/farmacologiaRESUMO
Anti-inflammatory effect of soluble secreted compounds of probiotic bacteria was widely demonstrated as therapy for different inflammatory diseases, but was not investigated in inflammatory eye disorders. The aim of this study was to determine whether Lactiplantibacillus plantarum CRL759 cell-free supernatant reduced inflammatory parameters and clinical signs in ocular inflammations. First, we evaluated the effect of L. plantarum CRL759 supernatant in vitro on human retinal cell line, ARPE-19 cells, stimulated with lipopolysaccharide (LPS). Then, we investigated in vivo its capacity to decrease inflammation by local administration on the eyes of mice with endotoxin induced inflammation. In vitro assays demonstrated that L. plantarum CRL759 supernatant reduced the production of interleukin (IL)-6, IL-8, nitric oxide and thiobarbituric acid reactive substances in LPS-stimulated ARPE-19 cells. Our in vivo data proved that L. plantarum supernatant significantly reduced the clinical score of endotoxin treated mice and diminished levels of tumour necrosis factor alpha, interferon gamma and protein concentration in aqueous humour. Histological examination showed reduction of infiltrating inflammatory cells in the posterior segment of the eyes. As far as we know, this is the first report showing that Lactobacillus spp. supernatant administered as drops reduces some parameters of ocular inflammation. This promising strategy is safe and could alleviate symptoms and signs of ocular inflammation in people that are refractories to the conventional therapies.
Assuntos
Oftalmopatias/tratamento farmacológico , Oftalmopatias/imunologia , Probióticos/administração & dosagem , Animais , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Oftalmopatias/etiologia , Oftalmopatias/genética , Feminino , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-8/genética , Interleucina-8/imunologia , Lactobacillus plantarum/fisiologia , Lipopolissacarídeos/efeitos adversos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Soluções Oftálmicas/administração & dosagem , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Haemonchus contortus is a blood-sucking parasite causing the presence of faecal occult blood (FOB). The objective was to study three different FOB tests in order to have a new indicator of H. contortus infection in sheep that could be included in the genetic evaluation system as an alternative selection criterion to faecal worm egg count (FEC). A total of 29 Corriedale lambs were experimentally infected with 10.000 larvae of H. contortus. Stool samples were recorded for FEC and FOB tests (Hexagon, Hematest(®) and Multistix(®)), blood for packed cell volume (PCV), haemoglobin, white and red blood cell count (RBC), and FAMACHA(©) for scoring anaemia. At the end of the experiment lambs were slaughtered to worm burden count. Field infection was achieved in 309 Merino lambs under natural parasite challenge. FEC data were normalized through logarithmic transformation (LnFEC). Pearson correlation was estimated to examine the relationship between all traits. The three tests were able to detect the presence of FOB at day 11. FEC, PCV and RBC decreased to sub-normal values from day 18. FAMACHA(©) score 3 was considered to be indicative of anaemia. Most of the correlations were of high magnitude, with the exception of Multistix(®) test that was moderately correlated with haematological parameters, LnFEC and FEC. In field infection, most samples were negative to FOB tests and the correlations were lower than those calculated under experimental infection. In conclusion, FOB tests were able to detect haemonchosis earlier than FEC under high experimental parasite challenge. However, they were not able to detect FOB under natural mixed parasite challenge. FAMACHA(©) and PCV demonstrated to be good indicators of Haemonchosis, having moderate to high correlations with FEC.
RESUMO
Acetyl-salicylic acid (ASA) is a nonsteroidal antiinflammatory/analgesic drug, which may cause gastritis or stomach ulcers if intensively employed. Exopolysaccharide (EPS)-producing lactic acid bacteria have been claimed to induce immunostimulatory/antiulcer effects in the host. This study investigated the potential preventive effect of fermented milks (FM) with EPS-producing Streptococcus thermophilus strains (CRL 1190 and CRL 804) on an in vivo model of chronic gastritis. Fermented milks (2 EPS(+) and 1 EPS(-), separately) were fed to BALB/c mice for 7 d before inducing gastritis with ASA (400 mg/kg of body weight per day for 10 d; gastritis group, n = 5). Appropriate control groups (ASA administered but not given FM, n = 5; and ASA not administered but given FM) were included in this study. Gastric inflammatory activity was evaluated through the stomach's histology and the number of IFNgamma(+) and IL-10(+) cytokine-producing cells in the gastric mucosa. Only mice preventively treated with the EPS-producing Strep. thermophilus CRL 1190 FM and later administered ASA did not develop gastritis, showing a conserved gastric mucosa structure similar to those of healthy mice. A marked decrease of IFNgamma(+)- and increase of IL-10(+)-producing cells compared with the gastritis group mice were observed. Purified EPS from Strep. thermophilus CRL 1190 resuspended in autoclaved milk was also effective for gastritis prevention. The EPS-protein interaction might be responsible for the observed gastroprotective effect; such interactions may be affected by industrial manufacturing conditions. The results indicate that the FM with Strep. thermophilus CRL 1190 or its EPS could be used in novel functional foods for preventing chronic gastritis.
Assuntos
Produtos Fermentados do Leite , Gastrite/prevenção & controle , Streptococcus thermophilus/fisiologia , Animais , Aspirina/farmacologia , Peso Corporal , Doença Crônica , Produtos Fermentados do Leite/microbiologia , Inibidores de Ciclo-Oxigenase/farmacologia , Mucosa Gástrica/química , Mucosa Gástrica/efeitos dos fármacos , Mucosa Gástrica/patologia , Gastrite/induzido quimicamente , Concentração de Íons de Hidrogênio , Camundongos , Camundongos Endogâmicos BALB C , Polissacarídeos Bacterianos/farmacologiaRESUMO
Phosphatidylinositol-specific phospholipase C (PI-PLC) activity was investigated in 25 different lactic acid bacteria (LAB) strains belonging to the genera Lactobacillus, Weisella, and Enterococcus. PI-PLC activity was detected in 44% of the strains studied in culture medium without carbon source. From the PI-PLC positive strains, Lactobacillus rhamnosus ATCC 7469 was selected for translocation studies. Healthy mice were orally administered with a daily dose of 2.0 x 10(9) of viable L. rhamnosus suspension. Viable bacteria were detected in liver and spleen of mice fed with LAB for 7 days. Bacterial colonies isolated from liver were biochemically characterized, and further subjected to randomly amplified polymorphic DNA. Amplification patterns of five strains displayed identical profiles to L. rhamnosus. PI-PLC activity was determined in the strains recovered from liver.
Assuntos
Translocação Bacteriana , Lactobacillus/enzimologia , Lactobacillus/fisiologia , Probióticos , Fosfolipases Tipo C/metabolismo , Animais , Humanos , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Fígado/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Fosfatidilinositol Diacilglicerol-Liase , Fosfoinositídeo Fosfolipase C , Técnica de Amplificação ao Acaso de DNA Polimórfico , Baço/microbiologiaRESUMO
OBJECTIVE: To demonstrate the utility of fine needle aspiration biopsy in the diagnosis of prostatic tumors, basically carcinoma of the prostate. METHODS/RESULTS: The clinical records of 497 patients who underwent fine needle aspiration biopsy were reviewed. Age, skin color, clinical diagnosis by DRE, transrectal core biopsy, correlation between clinical and histological diagnosis, distribution according to type of adenocarcinoma, correlation between cytological and histological diagnosis, and complications were analyzed. Of the 497 patients, 162 had histological confirmation. Of these 162 patients, the Galen and Gambino statistical method was utilized in 132 which represented the true negatives and positives. Sensitivity was 86%, specificity 96%, positive predictive value 91%, negative predictive value 93%, and efficiency was 92%. The complication rate was low. CONCLUSIONS: Fine needle aspiration biopsy is a safe and efficient diagnostic method in prostatic tumors and causes minimal discomfort to the patient.