A asma alérgica é uma doença pulmonar de inflamação crônica caracterizada por uma resposta imune do tipo Th2 e uma das principais abordagens terapêuticas para o seu tratamento no futuro, poderia ser a imunoterapia baseada na modulação da resposta imune Th2 para um perfil Th1 e anti-inflamatório. Nosso grupo já demonstrou a eficácia de uma imunoterapia, baseada em um plasmídeo de DNA contendo o gene hsp65 de M. Leprae (DNAHsp65) em modelo murino de asma alérgica. No entanto, apesar dos excelentes resultados, o grupo está procurando outras alternativas imunoterapêuticas, usando a Hsp65 micobacteriana, para futura utilização clinica na área de saúde humana e veterinária. Efeitos benéficos na prevenção e tratamento de doenças alérgicas vêm sendo obtidos com o uso de probióticos. Nossa hipótese é de que uma nova formulação terapêutica, como o probiótico Lactococcus lactis expressando Hsp65 micobacteriana (LLHsp65), apresentaria vantagens significativas no desenvolvimento de pesquisas translacionais. Diante disso, este estudo teve como objetivo avaliar se o probiótico LLHsp65 apresenta atividades imunoterapêuticas no modelo de asma experimental induzida por ovalbumina (OVA). Em diferentes grupos experimentais, 5x109 CFU de LLHsp65 ou de L. lactis selvagem (LLSELV), foram administrados por via oral durante 10 dias consecutivos aos camundongos BALB/c previamente sensibilizados e desafiados com OVA. Em seguida, investigamos os efeitos do tratamento na inflamação alérgica, modulação do padrão de citocinas, produção de anticorpos específicos, hiper-responsividade das vias aéreas e inflamação pulmonar. Nossos resultados demonstraram que o tratamento oral com LLhsp65 modula a resposta imune alérgica de padrão Th2 para o perfil Th1 com aumento dos níveis de IFN-?, IL-12, TNF-?, IL-10, IL-6 e IL- 17, e com a redução das citocinas IL-4, IL-5 e IL-13. Os níveis de IgE e IgG1 anti-OVA no soro também foram significativamente diminuídos. Como consequência desses resultados, também observamos diminuição significativa do infiltrado de eosinófilos no lavado broncoalveolar, na hiper-responsividade nas vias aéreas e a atenuação da inflamação e produção de muco no tecido pulmonar, quando comparados com o grupo controle (OVA/SAL). Por conseguinte, nossos resultados demonstraram que a administração oral de LLHsp65 proporciona uma melhora significativa do processo inflamatório alérgico induzido por OVA, sendo portanto, uma estratégia terapêutica promissora para o desenvolvimento de pesquisa translacional no tratamento de asma alérgica. / Allergic asthma is a chronic inflammatory lung disease characterized by a Th2-type immune response and one of the main therapeutic approaches to its treatment in the future could be immunotherapy based on the modulation of the Th2 immune response to a Th1 and anti-inflammatory profile. Our group has already demonstrated the efficacy of an immunotherapy based on a DNA plasmid carrying the mycobacterial hsp65 gene (DNAHsp65) in murine model of allergic asthma. However, despite the excellent results, our group is looking for other immunotherapeutic alternatives, using mycobacterial Hsp65, for future clinical use in the area of human and veterinary health. Beneficial effects in the prevention and treatment of allergic diseases have been obtained with the use of probiotics. Our hypothesis is that a new therapeutic formulation, such as the probiotic Lactococcus lactis expressing mycobacterial Hsp65 (LLHsp65), would present significant advantages in the development of translational research. The objective of this study was to evaluate whether the probiotic LLHsp65 has immunotherapeutic activities in the ovalbumin-induced asthma model. In different experimental groups, 5x109 CFU of LLHsp65 or control L. lactis (ctLL) were administered orally for 10 consecutive days to BALB/c mice previously sensitized and challenged with OVA. We then investigated the effects of treatment on allergic inflammation, modulation of the cytokine pattern, production allergen-specific antibodies, airway hyperresponsiveness and pulmonary inflammation. Our results demonstrated that oral treatment with LLhsp65 modulates the allergic Th2 immune response to Th1 profile with increased levels of IFN-?, IL-12, TNF-?, IL-10, IL-6 and IL-17 and with the reduction of IL-4, IL-5 and IL-13 cytokines. Serum anti-OVA IgE and IgG1 levels were also significantly decreased. Correspondingly with these results, we also observed a significant decrease in eosinophil infiltrate in bronchoalveolar lavage, airway hyper responsiveness and attenuation of inflammation and mucus production in lung tissue when compared to the control group (OVA/SAL). Therefore, our results demonstrated that oral administration of LLHsp65 provides a significant improvement of the OVA-induced allergic inflammatory process and is therefore a promising therapeutic strategy for the development of translational research in the treatment of allergic asthma.

Orientador: Ana Lia Anbinder / Banca: Maria Aparecida Neves Jardini / Banca: Lucilene Fernandes Ricardo / Resumo: A doença periodontal (DP) é caracterizada pela destruição dos tecidos de proteção e suporte dentário, osso alveolar, ligamento periodontal, cemento e gengiva, e é a principal causa de perda dentária na população adulta, um grande problema socioeconômico e de saúde pública. Com o aumento da resistência antibiótica, existe interesse no uso de outras substâncias para o combate de doenças infecciosas, como os probióticos. O uso de probióticos na prevenção e tratamento de doença periodontal em animais tem apresentado bons resultados. O estudo dos efeitos do Lactobacillus reuteri viável e inativado (paraprobiótico) pode esclarecer mecanismos de ação e possibilitar o uso comercial de outras preparações, dado que os paraprobióticos teriam produção facilitada. O objetivo deste trabalho foi analisar os efeitos de L. reuteri, viável e inativado, em doença periodontal induzida em ratos. Foram utilizados 32 ratos machos, divididos em 4 grupos: (1) Grupo Controle: com saúde periodontal (n=8); (2) Grupo LIP: periodontite induzida (n=8); (3) Grupo LIP/PRO: periodontite induzida e probiótico (n=8); e (4) Grupo LIP/PARAPRO: periodontite induzida e paraprobiótico (n=8). A indução da doença periodontal foi realizada com a inserção de fio de algodão ao redor dos primeiros molares inferiores e foi realizado o tratamento com L. reuteri viável ou inativado pelo calor, via oral, diariamente, 30 dias antes da ligadura e 14 dias após, quando ocorreu a eutanásia. As hemimandíbulas direitas foram removid... (Resumo completo, clicar acesso eletrônico abaixo) / Periodontal disease (PD) is characterized by the destruction of dental protection and support tissues, alveolar bone, periodontal ligament, cementum and gingiva, and is the major cause of tooth loss in the adult population, a major socioeconomic and public health problem. With the increase of antibiotic resistance, there is interest in the use of other substances to combat infectious diseases, such as probiotics. The use of probiotics in the prevention and treatment of periodontal disease in animals has shown good results. The study of the effects of viable and inactivated Lactobacillus reuteri (paraprobiotic) can clarify mechanisms of action and enable the commercial use of other preparations, since paraprobiotics would have facilitated production. The aim of this study was to evaluate the effects of viable and inactivated L. reuteri on induced periodontal disease in rats. Thirty-two male rats were divided into four groups: (1) Control group: with periodontal health (n = 8); (2) LIP group: induced periodontitis (n = 8); (3) LIP/PRO group: induced periodontitis and probiotic (n = 8); and (4) LIP/PARAPRO group: periodontitis induced and paraprobiotic (n = 8). Induction of periodontal disease was performed with the insertion of cotton thread around the lower first molars and treatment was performed with viable or heatinactivated L. reuteri daily, 30 days prior to ligation and 14 days later, when euthanasia occurred. The right hemimandible were removed and submitted to radiographic analysis for quantification of the bone support. A computerized microtomography was performed to evaluate the alveolar microarchitecture in the first molar region. The use of paraprobiotic led to a higher percentage of weight gain when compared to other treatments. In the radiographic and microtomographic analyzes, the efficacy of the DP induction was verified, with a significant difference... (Complete abstract click electronic access below) / Mestre

BACKGROUND: In a 2006 WHO report, diarrheal diseases ranked second among conditions afflicting children. Pediatric acute diarrhea, although most often the result of a gastrointestinal infection, can also occur as a result of antibiotic exposure. This is often referred to as antibiotic-associated diarrhea (AAD). Previous research suggests that probiotics may be effective in the treatment or prevention of various types of PAD. METHODS: The first study involved a systematic review and meta-analysis of RCTs involving probiotics as an adjunct to antibiotics for preventing AAD in children. The second study was a systematic review of definitions and primary outcome measures employed in RCTs of PAD. The third study used a modified Delphi consensus procedure to develop a new instrument for evaluating the severity of PAD. The study involved steering committee discussions (phase 1) and two electronic surveys (phase 2 and 3) of leading experts in measurement and clinical gastroenterology. RESULTS: The per protocol meta-analysis of ten RCTs significantly favored probiotics to prevent the incidence of diarrhea (NNT = 10). However, this effect did not withstand ITT analysis and among included trials there was considerable inconsistency regarding definitions for the reviews primary outcome measure, the incidence of diarrhea. Study two identified 121 RCTs that reported 62 unique definitions of diarrhea, 64 unique definitions of diarrhea resolution and 62 unique primary outcome measures. Thirty-one trials used grading systems to support outcome evaluation. However, none of the trials (or their citations) reported evidence of their validation. In study three experts agreed on the inclusion of five attributes containing 13 items. Attributes proposed for the IPADDS include: Diarrhea Frequency and Duration, Vomiting Frequency and Duration, Fever, Restrictions in Normal Daily Activities and Dehydration. CONCLUSION: It is premature to draw a valid conclusion about the efficacy of probiotics for pediatric AAD. Definitions of diarrhea and primary outcome measures in RCTs of PAD are heterogeneous and lack evidence of validity. The third study represents content validity evidence for IPADDS. A numerical scoring system needs to be added and further empirical evidence of reliability and validity are required. / Experimental Medicine

Probiotics can influence intestinal responses and mucosal immunity either directly or indirectly through transient modulation of the endogenous microenvironment or the immune system. During space travel, astronauts experience various physiological stresses including putting them at risk for infections or inappropriate immune responses. Macrophages and monocytes are a key cell type involved in innate immunity. The effects of dairy milk or soy milk base fermented with S. thermophilus ST5 in combination with either B. longum R0175 or L. helveticus R0052 on the cell line U937 and all-trans retinoic acid differentiated U937 were examined under normal gravity and simulated microgravity conditions, in order to screen for effects on monocytes and macrophages. Soy and milk ferments demonstrated the ability to modulate certain aspects of the innate immune system, both in normal gravity and in simulated microgravity. These probiotics affected U937 cells differently depending on differentiation stage (monocyte or macrophage) and whether or not the cells were tested in regular gravity or in simulated microgravity conditions. These results provide insight into effects on this aspect of innate immunity and may provide guidelines to potential in vivo administration. / UOIT

Groupers are an important mariculture fish in Taiwan and Southeast Asian countries. The rapidly growing orange spotted grouper (Epinephelus coioides) has experienced relatively severe bacterial disease problems. The proliferation of pathogens in fish can be suppressed by commensal microbiota. In this context, probiotic seem to offer an attractive alternative. Bacillus subtilisis a probiotic bacteriumthat is administered in diet to suppress proliferation of pathogens. In the present study, E.coioideswere fed for 6 months with diets containing B.subtilis at 0 (control), 0.1 % and 1 %. Percent weight gain and feed efficiency of the 0.1 and 1 % groups were significantlybetter than the control group. The innate cellular response, respiratory burst of the fish fed the 1 % and 0.1 % diet was significantly higher compared to the control group on 10 or 20 days after feeding, and even moresignificanton 30 days.ProbioticBacillus subtilis increased the fish¡¦s intestinal microbial diversity as measured by visible band number and Shannon diversity indexin DGGE analysis. Probiotic Bacillus subtilis also stimulated the population of bacterial species likePaenibacillussp,Lactobacillus oenistrain 59 b, and Methilacidophiluminfernorumstrain V4 that beneficial for Epinephelus coioides. The best dose of probiotic Bacillus subtilis based on growth performances, innate cellular responses and profile of microbiota in fish intestines is 0.1 %, which showed equal efficacy as the 1% diet.

A dextran sulphate (DS) model of ulcerative colitis was used to examine the effects of macronutrient restriction with (MR+PRO) and without (MR) probiotic supplementation (VSL#3RTM) on protein metabolism and growth. MR and MR+PRO decreased weight and chest circumference gain, but had no effect on linear growth of piglets. MR decreased the protein fractional synthesis rate (FSR) of liver, masseter, longissimus dorsi, colon, as well as plasma albumin, measured by stable isotope tracer L-[ring-2H 5]phenylalanine. MR+PRO increased the FSR of hepatic proteins by greater than 70% and increased both the FSR and absolute synthesis rate (ASR) of the total plasma protein pool, albumin and fibrinogen. Over 11 days, MR+PRO piglets showed a smaller decrease in plasma total protein concentrations than WN piglets, and maintained higher albumin levels than either WN or MR piglets. These findings highlight the importance of adequate nutrition during gastrointestinal inflammation and show that when food intake is decreased, probiotics stimulate liver protein synthesis and increase the acute phase protein response, similar to the well-nourished state.

The aim of this work is to collect, analyze and summarize information about prebiotics and probiotics using in producible dairy products in Lithuania, about features and quality of their consumption, production and labeling marking. Prebiotics and probiotics are mostly inserting into dairy products because it results their vitality. In process of this work was done written consumers quiz, sensual analysis of dairy products with prebiotics and probiotics, analysis of requests of labeling marking of those products according valid legislation. On purpose to find out if consumers have enough information about prebiotics and probiotics was preparing the questionnaire. During the poll it was aimed if consumers use products with prebiotics and probiotics. 50 Lithuanian residents were investigated. The data of the poll showed that consumers know about prebiotics and probiotics but not large amount of residents use those products. It is the reason of the deficit of information.-According valid legislation was done sensual analysis of yogurts: were evaluated sensual characteristics of yogurts and sensual descriptive- profiled analysis. Was formed the group of 5 estimators to valuate following characteristics: design, consistency, savour and smell. And was found out the average. During the sensual descriptive- profiled analysis consumers excluded 7 characteristics: sweetness, acidity, consistency, homogeneity, intensity of smell, intensity of savour, residual taste. The average of... [to full text]

1. Probiotinio Bactocell® preparato poveikyje bandymo pabaigoje paukščių svoris buvo 1% didesnis nei I kontrolinės grupės, o lesalų sąnaudos 1 kg priesvorio gauti buvo 4% mažesnės lyginant su I kontroline grupe; 2. Probiotinio Bactocell® + prebiotinio Agrimos® priedų poveikyje bandymo pabaigoje viščiukų broilerių svoris buvo 2% didesnis lyginant su I kontroline grupe, o lesalų sąnaudos 1 kg priesvorio gauti 3% mažesnės nei I kontrolinės grupės; 3. Preparatai Bactocell® ir Agrimos® paukščių išsaugojimui esminės įtakos neturėjo; 4. Preparatai Bactocell® ir Agrimos® liaukinio skrandžio (pars glandularis ventriculi), raumeninio skrandžio (pars muscularis ventriculi), dvylikapirštės žarnos (duodenum), aklųjų maišų (caecum), plonųjų žarnų paskutinio segmento (intestinum tenue), storosios žarnos (intestinum crassum) himusų pH esminės įtakos neturėjo; 5. Ženklus sausųjų medžiagų sumažėjimas iki 5,54% himusuose liaukinio skrandžio (pars glandularis ventriculi), raumeninio skrandžio (pars muscularis ventriculi), dvylikapirštės žarnos (duodenum), aklųjų maišų (caecum), plonųjų žarnų paskutinio segmento (intestinum tenue), storosios žarnos (intestinum crassum) nustatytas, naudojant preparatus Bactocell® + Agrimos® ; 6. Preparatų probiotinio Bactocell® ir probiotinio Bactocell® + prebiotinio Agrimos® poveikyje padidėjo– tiek bendra, tiek ir atskirų riebalų rūgščių koncentracija; 7. Naudojant probiotinį Bactocell® + prebiotinį Agrimos® preparatus amoniako kiekis viščiukų broilerių... [toliau žr. visą tekstą] / The objectives of this work is the analysis of effect of probiotics Bactocell® and prebiotics Agrimos® on process digestive and on productivity of broiler chickens. 800 items of Cobb500 broiler chickens from 1 to 35 days of age were taken, for this research to be made. The birds were divided into 4 treatment groups. They were kept on deep litter, watered from stationary drinking - bowls. They were fed and allocated according to the recommendations of suppliers of broiler cross. Grown and slaughtered broilers were analyzed by evaluation of activity of fermentation, amount of dry substances, pH- level within the alimentary canal. Also, index of growth intensity, feed expenditures were registered. In addition, characteristics of meat of chicken legs and breast were examined on the background of sensual perception. After the research was undertaken, it was determined that after using probiotics Bactocell® and prebiotics Agrimos® the feed expenditure decreased by 1-2 %, whereas, the daily weight increased by 2 – 4 %. level. The influence on the pH- level within the alimentary canal wasn't significant; the level of ammonia and dry substances tended to decrease. Whereas, activity of fermentation was registered as increased. It was determined, the probiotics Bactocell® and prebiotics Agrimoss® treated seed meal hasn't any significant effect for the sensual perception of the meat.

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Probiotics are a live microbial supplement that reside within the intestinal tract and are considered normal flora. The Balb/c mouse model was used to determine if the elimination of probiotics, general LAB species, by antibiotics plays a role in the breakdown of oral tolerance leading to the generation of an immune response to oral antigens. A mouse model was developed for in vivo research regarding probiotic populations and the effect on the induction of oral tolerance. The Balb/c mouse was used to determine if the mouse model had a colonized intestinal tract with probiotics followed by a reduction of probiotics that was done with orally administered antibiotics. After the reduction of probiotics, mice were fed oral antigen, ovalbumin, to determine that an immune response was not shown with oral antigen alone. After the mouse model was set up, mice were then fed oral antigen and then stimulated with immunizations to study the induction of oral tolerance and the possible effect of the absence of probiotics. The results indicated that mice with reduced probiotics and fed with oral antigen alone do not show an immune response. In contrast, mice fed with oral antigen followed by immunization indicate a higher OVA-specific serum IgG. This is evidence that correlates with clinical findings in disease states such as Crohn's Disease and Irritable Bowel Syndrome (IBD). / Department of Biology