The intestinal anti-inflammatory effect of dersalazine sodium is related to a down-regulation in IL-17 production in experimental models of rodent colitis.

BACKGROUND AND PURPOSE: Dersalazine sodium (DS) is a new chemical entity formed by combining, through an azo bond, a potent platelet activating factor (PAF) antagonist (UR-12715) with 5-aminosalicylic acid (5-ASA). DS has been demonstrated to have anti-inflammatory effects on trinitrobenzene sulphonic acid (TNBS)-induced colitis in rats and recently in UC patients in phase II PoC. There is Increasing evidence that Th17 cells have an important role in the pathogenesis of inflammatory bowel disease (IBD). The aim of this study was to further characterize the anti-inflammatory effects of DS. EXPERIMENTAL APPROACH: Effect of DS (10 or 30 mg·kg(-1) b.i.d.) on TNBS-induced colitis in rats was studied after 2 and 7 days with special focus on inflammatory mediators. Additionally, its anti-inflammatory properties were analysed in two different models of dextran sodium sulphate (DSS)-induced colitis, BALB/c and C57BL/6 mice, the latter being dependent on IL-17. KEY RESULTS: DS, when administered for 7 days, showed intestinal anti-inflammatory effects in TNBS-induced colitis; these effects were observed both macroscopically and through the profile of inflammatory mediators (TNF, IL-1ß, IL-6 and IL-17). Although the 2 day treatment with DS did not induce intestinal anti-inflammatory effects, it was sufficient to reduce the enhanced IL-17 expression. DS showed beneficial effects on DSS-induced colitis in C57BL/6 mice and reduced colonic pro-inflammatory cytokines IL-1ß, IL-6 and IL-17. In contrast, it did not exert intestinal anti-inflammatory effects on DSS-induced colitis in BALB/c mice. CONCLUSIONS AND IMPLICATIONS: DS exerts intestinal anti-inflammatory activity in different rodent models of colitis through down-regulation of IL-17 expression.

Oral administration of Lactobacillus strains isolated from breast milk as an alternative for the treatment of infectious mastitis during lactation.

In this study, 20 women with staphylococcal mastitis were randomly divided in two groups. Those in the probiotic group daily ingested 10 log(10) CFU of Lactobacillus salivarius CECT5713 and the same quantity of Lactobacillus gasseri CECT5714 for 4 weeks, while those in the control one only ingested the excipient. Both lactobacillus strains were originally isolated from breast milk. On day 0, the mean staphylococcal counts in the probiotic and control groups were similar (4.74 and 4.81 log(10) CFU/ml, respectively), but lactobacilli could not be detected. On day 30, the mean staphylococcal count in the probiotic group (2.96 log(10) CFU/ml) was lower than that of the control group (4.79 log(10) CFU/ml). L. salivarius CECT5713 and L. gasseri CECT5714 were isolated from the milk samples of 6 of the 10 women of the probiotic group. At day 14, no clinical signs of mastitis were observed in the women assigned to the probiotic group, but mastitis persisted throughout the study period in the control group women. In conclusion, L. salivarius CECT5713 and L. gasseri CECT5714 appear to be an efficient alternative for the treatment of lactational infectious mastitis during lactation.

Efectos beneficiosos de los probióticos de la leche materna / Beneficial effects of the probiotics of breast milk

La leche materna es el mejor alimento para los bebés durante sus fases de rápido desarrollo, puesto que no sólo aporta todos los nutrientes necesarios, sino que además contiene importantes factores funcionales implicados en el desarrollo y la maduración del sistema inmunitario neonatal, así como en la protección frente a infecciones. Entre estos factores cabe incluir también las bacterias comensales de la leche materna. Este trabajo pretende mostrar los posibles efectos beneficiosos ejercidos por las bacterias presentes en la leche materna, así como de cepas probióticas aisladas de dicha fuente. Entre ellos, cabe resaltar los efectos antimicrobianos, antiinflamatorios y/o moduladores de la respuesta inmunitaria, tanto en modelos de experimentación animal como en estudios clínicos .La demostración de la existencia de bacterias en la leche materna y los efectos beneficiosos potencialmente ejercidos por éstas en el lactante ofrecen nuevas ideas para la sustentación de las propuestas dirigidas a la inclusión de determinadas cepas probióticas en las fórmulas infantiles(AU)

Breast milk is the best food for the neonate because it provides a unique combination of proteins, carbohydrates, lipids, minerals and vitamins that ensures the correct growth and development of infants. In addition, it also contains bioactive compounds responsible for a wide range of beneficial effects, such as the promotion of immune system maturation and protection against infections. Among these bioactive agents, probiotic bacteria have recently been isolated from human milk. The present report reviews the beneficial effects of these bacteria both in animal models and in clinical trials. The promotion of immune system maturation and defence against infections, as well as the anti-inflammatory properties, are among the major health effects of these bacteria. The isolation of probiotic bacteria with beneficial effects for the host provides scientific support for the supplementation of infant formula with these bacteria, in order to advance toward the main target of these formula, to mimic the functional effects observed in breastfed infants(AU)

A comparative study of the preventative effects exerted by three probiotics, Bifidobacterium lactis, Lactobacillus casei and Lactobacillus acidophilus, in the TNBS model of rat colitis.

AIMS: The intestinal anti-inflammatory effects of three probiotics with immunomodulatory properties, Lactobacillus casei, Lactobacillus acidophilus and Bifidobacterium lactis, were evaluated and compared in the trinitrobenzenesulphonic acid (TNBS) model of rat colitis. METHODS AND RESULTS: Colitis was induced in rats by intracolonic administration of 10 mg of TNBS dissolved in 0.25 ml of 50% ethanol. Each probiotic was administered orally (5x10(8) CFU suspended in 0.5 ml of skimmed milk) for 3 weeks, starting 2 weeks before the administration of TNBS. Colonic damage was evaluated histologically and biochemically 1 week after TNBS instillation. The results obtained revealed that all probiotics assayed showed intestinal anti-inflammatory effects, macroscopically evidenced by a significant reduction in the colonic weight/length ratio. Only B. lactis showed a lower incidence of diarrhoea in comparison with untreated rats. Biochemically, all probiotics restored colonic glutathione levels, depleted as a consequence of the oxidative stress of the inflammatory process. Bifidobacterium lactis treatment reduced colonic tumour necrosis factor (TNF)-alpha production, and inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) expression; L. acidophilus administration reduced colonic leukotriene B4 production and iNOS expression and L. casei intake was associated with a decrease in colonic COX-2 expression. CONCLUSION: The three probiotics assayed have shown intestinal anti-inflammatory activity in the TNBS model of rat colitis, although each probiotic shows its own anti-inflammatory profile. SIGNIFICANCE AND IMPACT OF THE STUDY: These probiotics could be considered as potential adjuvants in the treatment of inflammatory bowel disease, although more studies are required in order to demonstrate their efficacy in humans.

Efectos beneficiosos en niños sanos del consumo de un producto lácteo que contiene dos cepas probióticas. Lactobacillus coryniformis CECT5711 y lactobacillus gasseri CECT5714 / Beneficial effects os consumption of a dairy product containing two probiotic strains, lactobacillus coryniformis CECT5711 and lactobacillus gasseru CECT5714 in healthy children

Objetivo o antecedente: En las últimas décadas la manipulación de la microbiota intestinal mediante el uso de probióticos ha adquirido un gran interés en el tratamiento y prevención de determinadas patologías infantiles. Además actualmente existen multitud de estudios que demuestran que las bacterias probióticas podrían tener un importante papel en el desarrollo del sistema inmunitario. Estudios recientes sugieren que dos cepas probióticas, Lactobacillus coryniformis CECT5711 y Lactobacillus gasseri CECT5714 mejoran la función intestinal de adultos sanos y potencian la respuesta inmunitaria. Dado que son muy pocos los estudios que analizan el papel de los probióticos en niños sanos, principales consumidores de estos productos, el objetivo del presente trabajo fue analizar los efectos de la administración conjunta de estas dos cepas probióticas en un producto lácteo fermentado en niños sanos. Intervenciones: Se reclutaron 30 niños de entre 3 y 12 años sin patología gastrointestinal conocida. Además de su dieta habitual, durante las 3 primeras semanas los niños recibieron 200 ml de un yogurt convencional que contenía Lactobacillus bulgaricus y Streptococcus thermophilus. A continuación este yogurt se sustituyó por 80 ml de un producto probiótico (Max Defensas® Puleva FOOD S.L.) que contenía la misma cantidad de Streptococcus thermophilus y donde el L. bulgaricus fue substituido por la misma cantidad de una mezcla de las bacterias probióticas objeto del estudio: L. coryniformis CECT5711 y L. gasseri CECT5714. Se tomaron muestras de heces y de saliva al comienzo del estudio, a las 3 semanas y al finalizar el estudio. Durante estas 6 semanas los niños no tomaron otro yogurt o probiótico que no fueran los citados anteriormente. Se analizó la microbiotafecal de los niños, la toxicidad de las aguas fecales y la capacidad de éstas para inhibir la adhesión de Salmonella cholerasusis ssp. cholerasuis a mucinas intestinales. Finalmente se determinó la concentración de IgA en heces y en saliva. Resultados: El consumo del producto probiótico objeto del estudio fue bien tolerado por todos los niños. Se observó un aumento del número de lactobacilos en las heces tras 3 semanas de consumo del probiótico. Además la toxicidad de las aguas fecales fue significativamente inferior tras el consumo del probiótico (P < 0,05). La inhibición de la adhesión de S. cholerasuis a mucinas intestinales fue significativamente (P < 0,05) mayor con las aguas fecales de los niños tras el consumo del probiótico en comparación con los homogeneizados iniciales y los obtenidos tras el consumo del yogurt convencional. Por último, el consumo del probiótico aumentó significativamente la concentración de IgA en las heces y en la saliva (P < 0,05). Conclusiones: La administración de un producto probiótico que contiene L. coryniformis CECT5711 y L. gasserii CECT5714 mejora la flora intestinal de niños sanos, favoreciendo la defensa frente a agresiones e infecciones gastrointestinales por inhibición de la adhesión de patógenos y potenciación de la respuesta inmunitaria

Objective: In the last decades there has been an increasing interest in the manipulation of intestinal microbiota with probiotics for the prevention and treatment of certain paediatric diseases. In addition, it has been suggested that probiotics could play a role in the development of immune system. Recent studies suggest that the administration of two probiotic strains, Lactobacillus coryniformis CECT5711 and Lactobacillus gasseri CECT5714 improves intestinal function of healthy adults and enhances the immune response. Since there are few studies reporting the use of probiotic in children, the main consumers of these products, the aim of the present study was to analyze the effects of the administration of the mentioned probiotic strains in healthy children. Interventions: 30 children (age range 3-12) with no gastrointestinal pathology were included in the study. In addition to their usual diet, during the first 3 weeks they received 200 ml of a conventional yogurt containing Lactobacillus bulgaricus and Streptococcus thermophilus. During the following three weeks this yogurt was substi-tuted for 80 ml of a probiotic product (Max Defensas®, Puleva Food S.L.) containing the same amounts of Streptococcus thermophilus and the L. bulgaricus was substituted by a mixture of the target probiotic strains: L. coryniformis CECT5711 and L. gasseri CECT5714. Samples of faeces and saliva were taken at the beginning of the protocol, at week 3 and at the end of the study. Intestinal microbiota, faecal citotoxicity and the inhibition of Salmonella cholerasusis ssp. cholerasuis adhesion to intestinal mucins by the faeces were analyzed. Finally, IgA concentration was determined in the faecal and saliva samples. Results: Tolerance of the probiotic product was good in all the children included in the study. An increase in faecal lactobacilli counts was shown at the end of the experimental protocol (P < 0,05). In addition citotoxicity of faecal samples was significantly (p < 0.05) reduced after probiotic consumption. The inhibition of S. cholerasuis adhesion to intestinal mucins was significantly higher (P < 0.05) for faecal waters from children in week 6 compared to samples form week 0 and 3. Probiotic con-sumption was also shown to increase IgA concentration in faeces and saliva (P < 0.05). Conclusions: The consumption of a probiotic product containing L. coryniformis CECT5711 and L. gasseri CECT5714 improves intestinal flora of healthy children, enhancing the defence against gastrointestinal aggressions and infections both by inhibiting pathogen adhesion to intestinal mucins and enhancing the immune function

Safety assessment of the human milk-isolated probiotic Lactobacillus salivarius CECT5713.

The potential probiotic bacteria Lactobacillus salivarius CECT5713 has recently been isolated from human milk and characterized. The objective of the present study was to evaluate the oral toxicity of this potential probiotic bacteria in mice. With this aim, 50 Balb/C mice were divided in 5 groups (n = 10). Three of these groups were treated orally with different doses of L. salivarius CECT5713: 5 x 10(8), 2 x 10(9), or 10(10) cfu/mouse per d for 28 d. One additional group was administered the vehicle alone and was used as a control. The last group were injected intraperitoneally with 10(8) cfu/mouse in a single dose and killed 2 (n = 5) and 5 (n = 5) d after intraperitoneal injection. Food intake, body weight, bacterial translocation, serum alpha-amyloid protein, and different biochemical parameters were analyzed. Oral administration of L. salivarius CECT5713 to mice had no adverse effects on mouse body weight or food intake. No bacteremia was shown and there was no treatment-associated bacterial translocation to the liver or spleen. Intraperitoneal administration caused a significant bacterial translocation to the liver and spleen, but not to the blood. However, this translocation was not related to illness or death at either d 2 or d 5, although an increase in plasma serum alpha-amyloid protein was observed at d 2. These results suggest that the strain L. salivarius CECT5713 is nonpathogenic for mice, even in doses 10,000 times higher (expressed per kilograms of body weight) than those normally consumed by humans. Thus, this strain is likely to be safe for human consumption.

[Beneficial effects of consumption of a dairy product containing two probiotic strains, Lactobacillus coryniformis CECT5711 and Lactobacillus gasseri CECT5714 in healthy children]. / Efectos beneficiosos en niños sanos del consumo de un producto lácteo que contiene dos cepas probióticas. Lactobacillus coryniformis CECT5711 y Lactobacillus gasseri CECT5714.

OBJECTIVE: In the last decades there has been an increasing interest in the manipulation of intestinal microbiota with probiotics for the prevention and treatment of certain paediatric diseases. In addition, it has been suggested that probiotics could play a role in the development of immune system. Recent studies suggest that the administration of two probiotic strains, Lactobacillus coryniformis CECT5711 and Lactobacillus gasseri CECT5714 improves intestinal function of healthy adults and enhances the immune response. Since there are few studies reporting the use of probiotic in children, the main consumers of these products, the aim of the present study was to analyze the effects of the administration of the mentioned probiotic strains in healthy children. INTERVENTIONS: 30 children (age range 3-12) with no gastrointestinal pathology were included in the study. In addition to their usual diet, during the first 3 weeks they received 200 ml of a conventional yogurt containing Lactobacillus bulgaricus and Streptococcus thermophilus. During the following three weeks this yogurt was substi-tuted for 80 ml of a probiotic product (Max Defensas, Puleva Food S.L.) containing the same amounts of Streptococcus thermophilus and the L. bulgaricus was substituted by a mixture of the target probiotic strains: L. coryniformis CECT5711 and L. gasseri CECT5714. Samples of faeces and saliva were taken at the beginning of the protocol, at week 3 and at the end of the study. Intestinal microbiota, faecal citotoxicity and the inhibition of Salmonella cholerasusis ssp. cholerasuis adhesion to intestinal mucins by the faeces were analyzed. Finally, IgA concentration was determined in the faecal and saliva samples. RESULTS: Tolerance of the probiotic product was good in all the children included in the study. An increase in faecal lactobacilli counts was shown at the end of the experimental protocol (P < 0,05). In addition citotoxicity of faecal samples was significantly (p < 0.05) reduced after probiotic consumption. The inhibition of S. cholerasuis adhesion to intestinal mucins was significantly higher (P < 0.05) for faecal waters from children in week 6 compared to samples form week 0 and 3. Probiotic consumption was also shown to increase IgA concentration in faeces and saliva (P < 0.05). CONCLUSIONS: The consumption of a probiotic product containing L. coryniformis CECT5711 and L. gasseri CECT5714 improves intestinal flora of healthy children, enhancing the defence against gastrointestinal aggressions and infections both by inhibiting pathogen adhesion to intestinal mucins and enhancing the immune function.

Safety assessment of two probiotic strains, Lactobacillus coryniformis CECT5711 and Lactobacillus gasseri CECT5714.

AIMS: The object of the present study was to evaluate the oral toxicity of the recently isolated probiotic bacteria Lactobacillus coryniformis CECT5711 and Lactobacillus gasseri CECT5714. METHODS AND RESULTS: Enzymatic activity and antibiotic resistance profile were evaluated in vitro. Then, the oral toxicity was analysed by an in vivo experiment using 20 Balb/C mice, which were orally treated with CECT5711 or CECT5714 (10(10) CFU mouse(-1) day(-1)) during 30 days. Results showed that CECT5711 and CECT5714 have no deleterious enzymatic activities and present intrinsic antibiotic resistance profile. Administration of both strains to mice had no adverse effects on body weight or food intake. No bacteraemia was present in liver or spleen and there was no treatment-associated bacterial translocation to these tissues. Liver glutathione content as well as plasma malondialdehide concentration were not statistically different in probiotic-treated mice when compared with control mice. Probiotic treatment did not cause changes in the biochemical and haematological parameters analysed. CONCLUSIONS: These results suggest that strains CECT5711 and CECT5714 are nonpathogenic and likely to be safe for human consumption. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reveals the oral safety of two new lactobacilli strains that are aimed to be used as probiotics in food and pharmaceutical applications.

Two Lactobacillus strains, isolated from breast milk, differently modulate the immune response.

AIMS: The ability of two different Lactobacillus strains (Lactobacillus salivarius CECT5713 and Lactobacillus fermentum CECT5716), isolated from human breast milk, to modulate the immune response was examined. METHODS AND RESULTS: In rodent bone-marrow-derived macrophages (BMDM), the presence of Lact. fermentum CECT5716 induced pro-inflammatory cytokines, in contrast to the activation of IL-10 induced by Lact. salivarius CECT5713. Although both strains reduced the lipopolysaccharide (LPS)-induced inflammatory response in BMDM, the effect of Lact. salivarius CECT5713 was more efficient, probably because of the production of higher amounts of IL-10 cytokine. In vivo assays in mice showed similar results; the consumption of Lact. fermentum CECT5716 enhanced the production of Th1 cytokines by spleen cells and increased the IgA concentration in faeces. However, the consumption of Lact. salivarius CECT5713 induced IL-10 production by spleen cells. CONCLUSION: Therefore, in general, the effect of Lact. fermentum CECT5716 is immunostimulatory in contrast to the anti-inflammatory effect of Lact. salivarius CECT5713. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study show that two Lactobacillus strains isolated from breast milk can exert different and even opposing effects on immune response demonstrating the specificity of each strain.

Antimicrobial potential of four Lactobacillus strains isolated from breast milk.

AIMS: The antimicrobial potential of four lactobacilli (Lactobacillus salivarius CECT5713, Lactobacillus gasseri CECT5714, L. gasseri CECT5715 and Lactobacillus fermentum CECT5716), isolated from fresh human breast milk, was evaluated in this study and compared with Lactobacillus coryniformis CECT5711, a reuterin-producing strain isolated from an artisan goat's cheese. METHODS AND RESULTS: Agar diffusion tests, competitive adhesion assays and mucin expression assays were carried out in order to value the antibacterial properties of the lactobacilli strains. The antibacterial capability of the strains was tested in vivo by using a murine infection model with Salmonella choleraesuis. The results revealed that all the strains studied, displayed antibacterial properties against pathogenic bacteria. However, the antibacterial potential varied among the lactobacilli tested and, in fact, L. salivarius CECT5713 showed not only the best in vitro antibacterial activity, but also the highest protective effect against a Salmonella strain in the murine infection model. CONCLUSION: The four breast-milk lactobacilli, and particularly L. salivarius CECT5713, possess potent antibacterial activities that result in a higher protection against S. choleraesuis CECT4155 in a mouse infection model. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest that lactobacilli from breast milk could contribute to an anti-infective protection in neonates and would be excellent candidates for the development of infant probiotic products.

Lactobacillus salivarius CECT 5713, a potential probiotic strain isolated from infant feces and breast milk of a mother-child pair.

In this study, Lactobacillus salivarius CECT 5713 was originally isolated from feces of a one-month-old breast-fed infant. Since it has been suggested that the gut microbiota of breast-fed infants reflects that of the maternal breast milk, we investigated if this specific strain was present in breast milk of the respective mother. RAPD and PFGE analysis revealed the presence of the strain L. salivarius CECT 5713 in this biological fluid. To our knowledge, this is the first report of a L. salivarius strain isolated from breast milk. L. salivarius CECT 5713 produced l-lactate, acetate and hydrogen peroxide, which may be responsible for its antimicrobial activity against most of the indicator organisms used in this study; in addition, this strain showed a high survival rate after exposition to conditions simulating those found in the gastrointestinal tract. Finally, it was strongly adhesive to Caco-2 and HT-29 cells did not produce biogenic amines and were unable to degrade gastric mucin in vitro.

Intestinal anti-inflammatory activity of combined quercitrin and dietary olive oil supplemented with fish oil, rich in EPA and DHA (n-3) polyunsaturated fatty acids, in rats with DSS-induced colitis.

BACKGROUND AND AIMS: Previous studies have described the intestinal anti-inflammatory effects exerted by the bioflavonoid quercitrin (QR) and by an n-3 polyunsaturated fatty acids (PUFA)-enriched diet in experimental models of rat colitis. The aim of the present study was to test if the combination of both treatments would result in an improvement in the intestinal anti-inflammatory effect achieved separately. METHODS: Colitis was induced in female Wistar rats by incorporating dextran sodium sulfate (DSS) in drinking water at 5% (w/v) for 5 days and at 2% (w/v) for the following 10 days. Five groups of rats (n=10) were used: two of them received an olive-oil-based diet with fish oil, rich in n-3 PUFA (FO diet) for 2 weeks before colitis induction and until the end of the experiment, and one of those also was administered daily QR (1mg/kg, PO), starting when DSS concentration was changed. DSS colitis was induced in other two groups fed with standard rat diet, one of them being administered QR as before. A non-colitic group fed standard diet was also included. After that period, the rats were sacrificed and colonic damage was assessed both histologically and biochemically. RESULTS: The concurrent administration of FO diet and QR exhibited an intestinal anti-inflammatory effect, as evidenced by a significant improvement of all biochemical parameters of colonic inflammation assayed in comparison with non-treated colitic rats. Thus, both colonic myeloperoxidase (MPO) and alkaline phosphatase (AP) activities were significantly reduced compared with untreated colitic rats. In addition, a complete restoration of colonic glutathione content, which was depleted as a consequence of the colonic insult, was obtained in rats treated with QR plus FO diet; this content was even higher than that obtained when colitic rats were treated with FO diet alone. When compared with the control colitic group, the combined treatment was also associated with a lower colonic nitric oxide synthase and cyclooxygenase-2 expression as well as with a significant reduction in different colonic proinflammatory mediators assayed, i.e. leukotriene B(4), tumor necrosis factor alpha and interleukin 1beta, showing a significantly greater inhibitory effect of the latter in comparison with rats receiving FO diet without the flavonoid. CONCLUSIONS: These results support the potential synergism between the administration of the flavonoid and the incorporation of olive oil and n-3 PUFA to the diet for the treatment of these intestinal inflammatory disorders.

Characterization of a reuterin-producing Lactobacillus coryniformis strain isolated from a goat's milk cheese.

Lactobacillus coryniformis CECT 5711, a strain isolated from a goat's milk cheese, displayed a broad-spectrum antimicrobial activity; as a consequence, its ability to produce the antagonistic compounds associated to lactic acid bacteria, including bacteriocins, hydrogen peroxide, lactic acid, acetic acid, and reuterin (3-hydroxypropionaldehyde, 3-HPA) was investigated. Production of bacteriocins or hydrogen peroxide by this strain could not be detected. However, in addition to lactic acid and acetic acid, it produced reuterin and cobalamin, a cofactor required for conversion of glycerol to 3-HPA through a glycerol dehydratase. The gene encoding a glycerol dehydratase subunit was detected by PCR and the corresponding amplicon was sequenced. This strain showed a high survival after exposition to conditions simulating those existing in the gastrointestinal tract as well as a notable ability to adhere to intestinal cells, which suggests that its reuterin-producing ability may be used for the host benefit. In addition, the strain showed a strong beta-galactosidase activity. Production of biogenic amines and degradation of mucin could not be detected.

The balance between caseins and whey proteins in cow's milk determines its allergenicity.

Cow's milk allergy is quite common in the first years of human life. Protein composition plays an important role in this pathology, particularly the casein/whey protein ratio. It is known that milks from different species have different sensitization capacities although their protein sources are quite similar. Thus, the objective of this work was to compare the allergenicity of native cow's milk and milk with a modified ratio of casein and whey proteins in a murine model of atopy. Twenty-four Balb/c mice were orally sensitized to native cow's milk or modified cow's milk with a casein/whey protein ratio of 40:60. During the sensitization period, the number of mice suffering from diarrhea was significantly higher in the native cow's milk-sensitized group than in the modified milk-sensitized group. Once mice were killed, plasma histamine levels were shown to be significantly higher in native cow's milk-sensitized mice. In addition, cow's milk proteins induced a higher lymphocyte sensitization in the native milk-sensitized mice, with a significant increase in the specific proliferation ratio of these cells. These results suggest that the balance between caseins and whey proteins plays an important role in the sensitization capacity of cow's milk, and its modification might be a way to reduce the allergenicity of cow's milk.

La expresión de IL-10 interviene en la regulación de la respuesta inflamatoria por los ácidos grasos omega 3 / IL-10 expression is involved in the regulation of the immune response by omega 3 fatty acids

Objetivo o antecedente: Los ácidos grasos poliinsaturados son importantes para el organismo humano debido a su implicación en numerosas funciones biológicas. Las dietas occidentales se caracterizan por ser excesivamente ricas en ácidos grasos w-6 y pobres en ácidos grasos w-3.Los ácidos grasos de la serie w-3 son necesarios para el normal crecimiento y desarrollo del individuo así como para la regulación de la respuesta inmunológica. El objetivo de este estudio es analizar el efecto de una dieta enriquecida en ácidos grasos w-3 frente a un proceso inflamatorio así como el estudio de los mecanismos implicados en dicho efecto. Intervenciones: Para ello, ratones Balb/c fueron alimentados durante un mes con una dieta cuya fuente lipídica era 100 por ciento aceite de girasol (control), o con la misma dieta en la que el 12 por ciento de la grasa era aceite de pescado y el resto aceite de girasol (W-3). Doce horas antes de su sacrificio se indujo en una de las orejas de cada animal una dermatitis de contacto que cursó con inflamación y edema. Como agente inflamatorio se utilizó 2,4 dinitrofluorobenceno. Tras el sacrificio se tomaron diversas muestras y se analizaron. Resultados: La inflamación, medida como peso y contenido de agua de las orejas, disminuyó significativamente en los ratones alimentados con w-3. La medida de la infiltración leucocitaria y los parámetros de oxidación revelaron también la mejora en el proceso inflamatorio de dichos ratones. Para explicar estos hechos se analizó la expresión de diversas citocinas, observándose un incremento de IL-10 y una disminución de citocinas tanto Th1 como Th2.Conclusiones: Los ácidos grasos w-3 poseen un efecto inmunomodulador al actuar como antiinflamatorios y antialérgicos, al tiempo que aumentan algunas defensas del organismo. La citocina reguladora IL-10 podría ser la responsable del efecto antiinflamatorio ejercido por los ácidos grasos w-3 (AU)

Introduction: Polyunsaturated fatty acids play a key role in a huge number of biological functions. Western diets are highly rich in w-6 fatty acids. However the content of w-3 fatty acids is not suitable in those diets, despite of their importance in normal development of the human body and regulation of immune response. 
The aim of this work is to examine the effect of w-3 fatty acids enriched diet in the regulation of inflammatory response. Material and methods: Balb/c mice were fed either w-6 fatty acids rich diet (100% sunflower oil) or w-3 fatty acids fortified diet (12% fish oil plus 88% sunflower oil) during 28 days. Twelve hours prior to sacrifice, the mice were treated with 2,4-ninitro-1-fluorobezene on the left ear to induce the inflammatory reaction. Afterwards the mice were sacrificed and the different samples collected were analized. Results: Ear inflammation of mice fed the w-3 diet was significantly lower. Leukocyte infiltration and oxidative stress were also lower in those mice. To explain these results, cytokine expression and plasma eicosanoid concentration were measured. An increase in IL-10 levels and a down regulation of Th1 and Th2 responses were observed in mice fed the w-3 diet. Conclusion: Not only n-3 fatty acids exerts an antiinflammatory and an antialergical role but also they enhance some of the organism defenses. Our data suggest that w-3 fatty acids down regulate the inflammatory response by enhancing IL10 expression (AU)

[Il-10 expression is involved in the regulation of the immune response by omega 3 fatty acids]. / La expresión de IL-10 interviene en la regulación de la respuesta inflamatoria por los acidos grasos omega 3.

INTRODUCTION: Polyunsaturated fatty acids play a key role in a huge number of biological functions. Western diets are highly rich in w-6 fatty acids. However the content of w-3 fatty acids is not suitable in those diets, despite of their importance in normal development of the human body and regulation of immune response. The aim of this work is to examine the effect of w-3 fatty acids enriched diet in the regulation of inflammatory response. MATERIAL AND METHODS: Balb/c mice were fed either w-6 fatty acids rich diet (100% sunflower oil) or w-3 fatty acids fortified diet (12% fish oil plus 88% sunflower oil) during 28 days. Twelve hours prior to sacrifice, the mice were treated with 2,4-ninitro-1-fluorobezene on the left ear to induce the inflammatory reaction. Afterwards the mice were sacrificed and the different samples collected were analized. RESULTS: Ear inflammation of mice fed the w-3 diet was significantly lower. Leukocyte infiltration and oxidative stress were also lower in those mice. To explain these results, cytokine expression and plasma eicosanoid concentration were measured. An increase in IL-10 levels and a down regulation of Th1 and Th2 responses were observed in mice fed the w-3 diet. CONCLUSION: Not only n-3 fatty acids exerts an antiinflammatory and an antialergical role but also they enhance some of the organism defenses. Our data suggest that w-3 fatty acids downregulate the inflammatory response by enhancing IL10 expression.

Treatment with anti-interferon-gamma monoclonal antibodies modifies experimental autoimmune encephalomyelitis in interferon-gamma receptor knockout mice.

The role of interferon-gamma (IFN-gamma) in the pathogenesis of multiple sclerosis and experimental autoimmune encephalomyelitis (EAE) is still controversial. We have studied the function of IFN-gamma and its receptor in the EAE model using two different IFN-gamma receptor knockout (IFN-gamma R(-/-)) mouse types: C57Bl/6x129Sv, with a disruption of the IFN-gamma receptor cytoplasmic domain, and 129Sv, homozygous for a disrupted IFN-gamma receptor gene. Mice were immunized with peptide 40-55 from rat myelin oligodendrocyte glycoprotein. A subgroup of mice was treated with anti-IFN-gamma monoclonal antibodies (mAb) on day 8 postimmunization. Clinical scoring and both histological and immunohistochemical studies were undertaken for all groups. We hereby show that treatment with anti-IFN-gamma mAb worsened the disease course of 129Sv wild-type mice. However, it decreased the mean daily score in IFN-gamma R(-/-) 129Sv and the incidence of the disease down to 50% in C57Bl/6x129Sv IFN-gamma R(-/-) mice. Moreover, after anti-IFN-gamma mAb treatment, oxidative stress levels, metallothionein I and II antioxidant protein expression, and apoptoticneuronal death were increased in wild-type mice while decreased in IFN-gamma R(-/-) mice. These results suggest a putative alternative mechanism of action of this cytokine that works independent of its receptor.

Macrophages require different nucleoside transport systems for proliferation and activation.

To evaluate the mechanisms involved in macrophage proliferation and activation, we studied the regulation of the nucleoside transport systems. In murine bone marrow-derived macrophages, the nucleosides required for DNA and RNA synthesis are recruited from the extracellular medium. M-CSF induced macrophage proliferation and DNA and RNA synthesis, whereas interferon gamma (IFN-gamma) led to activation, blocked proliferation, and induced only RNA synthesis. Macrophages express at least the concentrative systems N1 and N2 (CNT2 and CNT1 genes, respectively) and the equilibrative systems es and ei (ENT1 and ENT2 genes, respectively). Incubation with M-CSF only up-regulated the equilibrative system es. Inhibition of this transport system blocked M-CSF-dependent proliferation. Treatment with IFN-gamma only induced the concentrative N1 and N2 systems. IFN-gamma also down-regulated the increased expression of the es equilibrative system induced by M-CSF. Thus, macrophage proliferation and activation require selective regulation of nucleoside transporters and may respond to specific requirements for DNA and RNA synthesis. This report also shows that the nucleoside transporters are critical for macrophage proliferation and activation.

Decorin inhibits macrophage colony-stimulating factor proliferation of macrophages and enhances cell survival through induction of p27(Kip1) and p21(Waf1).

Decorin is a small proteoglycan that is ubiquitous in the extracellular matrix of mammalian tissues. It has been extensively demonstrated that decorin inhibits tumor cell growth; however, no data have been reported on the effects of decorin in normal cells. Using nontransformed macrophages from bone marrow, results of this study showed that decorin inhibits macrophage colony-stimulating factor (M-CSF)-dependent proliferation by inducing blockage at the G(1) phase of the cell cycle without affecting cell viability. In addition, decorin rescues macrophages from the induction of apoptosis after growth factor withdrawal. Decorin induces the expression of the cdk inhibitors p21(Waf1) and p27(Kip1). Using macrophages from mice where these genes have been disrupted, inhibition of proliferation mediated by decorin is related to p27(Kip1) expression, whereas p21(Waf1) expression is necessary to protect macrophages from apoptosis. Decorin also inhibits M-CSF-dependent expression of MKP-1 and extends the kinetics of ERK activity, which is characteristic when macrophages become activated instead of proliferating. The effect of decorin on macrophages is not due to its interaction with epidermal growth factor or interferon-gamma receptors. Furthermore, decorin increases macrophage adhesion to the extracellular matrix, and this may be partially responsible for the expression of p27(Kip1) and the modification of ERK activity, but not for the increased cell survival.

Lipopolysaccharide-induced apoptosis of macrophages determines the up-regulation of concentrative nucleoside transporters Cnt1 and Cnt2 through tumor necrosis factor-alpha-dependent and -independent mechanisms.

In murine bone marrow macrophages, lipopolysaccharide (LPS) induces apoptosis through the autocrine production of tumor necrosis factor-alpha (TNF-alpha), as demonstrated by the fact that macrophages from TNF-alpha receptor I knock-out mice did not undergo early apoptosis. In these conditions LPS up-regulated the two concentrative high affinity nucleoside transporters here shown to be expressed in murine bone marrow macrophages, concentrative nucleoside transporter (CNT) 1 and 2, in a rapid manner that is nevertheless consistent with the de novo synthesis of carrier proteins. This effect was not dependent on the presence of macrophage colony-stimulating factor, although LPS blocked the macrophage colony-stimulating factor-mediated up-regulation of the equilibrative nucleoside transport system es. TNF-alpha mimicked the regulatory response of nucleoside transporters triggered by LPS, but macrophages isolated from TNF-alpha receptor I knock-out mice similarly up-regulated nucleoside transport after LPS treatment. Although NO is produced by macrophages after LPS treatment, NO is not involved in these regulatory responses because LPS up-regulated CNT1 and CNT2 transport activity and expression in macrophages from inducible nitric oxide synthase and cationic amino acid transporter (CAT) 2 knock-out mice, both of which lack inducible nitric oxide synthesis. These data indicate that the early proapoptotic responses of macrophages, involving the up-regulation of CNT transporters, follow redundant regulatory pathways in which TNF-alpha-dependent- and -independent mechanisms are involved. These observations also support a role for CNT transporters in determining extracellular nucleoside availability and modulating macrophage apoptosis.