Quantitative recovery of viable Lactobacillus paracasei CNCM I-1572 (L. casei DG®) after gastrointestinal passage in healthy adults

Probiotics are live microorganisms, and viability after transit through the gastrointestinal tract (GIT) is considered an inherent property of the health benefits of probiotics. The aim of the present study was to quantify the viable and total loads of Lactobacillus paracasei DG cells after passage through the GIT following the consumption of the probiotic product Enterolactis (L. casei DG\uae; L. paracasei CNCM I-1572; L. paracasei DG) from drinkable vials by healthy adults. We developed a novel method for discriminating and enumerating culturable L. paracasei DG cells based on the unique sticky, filamentous phenotype of this strain on MRS agar containing vancomycin and kanamycin. The identity of DG was also confirmed with strain-specific primers by colony PCR. This method was used for a recovery study of the DG strain to quantify viable cells in the fecal samples of 20 volunteers during a 1-week probiotic consumption period and a 1-week follow-up. We isolated L. paracasei DG from at least one fecal sample from all the volunteers. The highest concentration of viable DG cells [ranging from 3.6 to 6.7 log10colony-forming unit (CFU) per gram of feces] in the feces was observed between 4 and 8 days from the beginning of Enterolactis intake and for up to 5 days after cessation of intake. As expected, the total DG count determined by real-time quantitative PCR (qPCR) was mostly higher than the viable DG cells recovered. Viable count experiments, carried out by combining ad hoc culture-based discriminative conditions and strain-specific molecular biological protocols, unambiguously demonstrated that L. paracasei DG can survive gastrointestinal transit in healthy adults when ingested as Enterolactis in drinkable vials containing no less than one billion CFU at the end of shelf life

Pérola-da-terra - Eurhizococcus brasiliensis (Hempel) (Hemiptera: Margarodidae) na cultura da videira.

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Effect of oral consumption of capsules containing Lactobacillus paracasei LPC-S01 on the vaginal microbiota of healthy adult women: a randomized, placebo-controlled, double-blind crossover study

Oral consumption of probiotics is practical and can be an effective solution to preserve vaginal eubiosis. Here, we studied the ability of orally administered Lactobacillus paracasei LPC-S01 (DSM 26760) to affect the composition of the vaginal microbiota and colonize the vaginal mucosa in nondiseased adult women. A total of 40 volunteers took oral probiotic (24 billion CFU) or placebo capsules daily for 4 weeks, and after a 4-week washout, they switched to placebo or probiotic capsules according to the crossover design. A total of 23 volunteers completed the study according to the protocol. Before and after capsule ingestion, vaginal swabs were collected for qPCR quantification to detect L. paracasei LPC-S01 and for 16S rRNA gene sequencing. Vaginal swabs were grouped according to their bacterial taxonomic structure into nine community state types (CSTs), four of which were dominated by lactobacilli. Lactobacillus paracasei LPC-S01 was detected in the vagina of two participants. Statistical modeling (including linear mixed-effects model analysis) demonstrated that daily intake of probiotic capsules reduced the relative abundance of Gardnerella spp. Quantitative PCR with Gardnerella vaginalis primers confirmed this result. Considering the pathogenic nature of G. vaginalis, these results suggest a potential positive effect of this probiotic capsule on the vaginal microbial ecosystem

A dairy bacterium displays in vitro probiotic properties for the pharyngeal mucosa by antagonizing group A streptococci and modulating the immune response

The probiotic approach represents an alternative strategy in the prevention and treatment of infectious diseases, not only at the intestinal level but also at other sites of the body where the microbiota plays a role in the maintenance of physiological homeostasis. In this context, we evaluated in vitro the potential abilities of probiotic and dairy bacteria in controlling Streptococcus pyogenes infections at the pharyngeal level. Initially, we analyzed bacterial adhesion to FaDu hypopharyngeal carcinoma cells and the ability to antagonize S. pyogenes on FaDu cell layers and HaCat keratinocytes. Due to its promising adhesive and antagonistic features, we studied the dairy strain Lactobacillus helveticus MIMLh5, also through in vitro immunological experiments. First, we performed quantification of several cytokines and measurement of NF-\u3baB activation in FaDu cells. MIMLh5 efficiently reduced the induction of interleukin-6 (IL-6), IL-8, and tumor necrosis factor alpha (TNF-\u3b1), in a dose-dependent manner. After stimulation of cells with IL-1\u3b2, active NF-\u3baB was still markedly lowered. Nevertheless, we observed an increased secretion of IL-6, gamma interferon (IFN-\u3b3), and granulocyte-macrophage colony-stimulating factor (GM-CSF) under these conditions. These effects were associated with the ability of MIMLh5 to enhance the expression of the heat shock protein coding gene hsp70. In addition, MIMLh5 increased the GM-CSF/G-CSF ratio. This is compatible with a switch of the immune response toward a TH1 pathway, as supported by our observation that MIMLh5, once in contact with bone marrow-derived dendritic cells, triggered the secretion of TNF-\u3b1 and IL-2. In conclusion, we propose MIMLh5 as a potential probiotic bacterium for the human pharynx, with promising antagonistic and immunomodulatory properties

Surface Layer of Lactobacillus helveticus MIMLh5 Promotes Endocytosis by Dendritic Cells

Surface layers (S-layers) are proteinaceous arrays covering the cell walls of numerous bacteria. Their suggested properties, such as interactions with the host immune system, have been only poorly described. Here, we aimed to elucidate the role of the S-layer from the probiotic bacterial strain Lactobacillus helveticus MIMLh5 in the stimulation of murine bone-marrow-derived dendritic cells (DCs). MIMLh5 induced greater production of interferon beta (IFN-\u3b2), interleukin 10 (IL-10), and IL-12p70, compared to S-layer-depleted MIMLh5 (naked MIMLh5 [n-MIMLh5]), whereas the isolated S-layer was a poor immunostimulator. No differences in the production of tumor necrosis factor alpha (TNF-\u3b1) or IL-1\u3b2 were found. Inhibition of the mitogen-activated protein kinases JNK1/2, p38, and ERK1/2 modified IL-12p70 production similarly in MIMLh5 and n-MIMLh5, suggesting the induction of the same signaling pathways by the two bacterial preparations. Treatment of DCs with cytochalasin D to inhibit endocytosis before the addition of fluorescently labeled MIMLh5 cells led to a dramatic reduction in the proportion of fluorescence-positive DCs and decreased IL-12 production. Endocytosis and IL-12 production were only marginally affected by cytochalasin D pretreatment when fluorescently labeled n-MIMLh5 was used. Treatment of DCs with fluorescently labeled S-layer-coated polystyrene beads (Sl-beads) resulted in much greater uptake of beads, compared to noncoated beads. Prestimulation of DCs with cytochalasin D reduced the uptake of Sl-beads more than plain beads. These findings indicate that the S-layer plays a role in the endocytosis of MIMLh5 by DCs. In conclusion, this study provides evidence that the S-layer of L. helveticus MIMLh5 is involved in endocytosis of the bacterium, which is important for strong Th1-inducing cytokine production.IMPORTANCE Beneficial microbes may positively affect host physiology at various levels, e.g., by participating in immune system maturation and modulation, boosting defenses and dampening reactions, thus affecting the whole homeostasis. As a consequence, the use of probiotics is increasingly regarded as suitable for more extended applications for health maintenance, not only microbiota balancing. This implies a deep knowledge of the mechanisms and molecules involved in host-microbe interactions, for the final purpose of fine tuning the choice of a probiotic strain for a specific outcome. With this aim, studies targeted to the description of strain-related immunomodulatory effects and the identification of bacterial molecules responsible for specific responses are indispensable. This study provides new insights in the characterization of the food-origin probiotic bacterium L. helveticus MIMLh5 and its S-layer protein as a driver for the cross-talk with DCs

Methods for Collecting Milk from Mice

Mouse models offer unique opportunities to study mammary gland biology and lactation. Phenotypes within the mammary glands, especially those caused by genetic modification, often arise during lactation, and their study requires the collection of adequate volumes of milk. We describe two approaches for collecting milk from lactating mice. Both methods are inexpensive, are easy to use in the laboratory or classroom, are non-invasive, and yield adequate volumes of milk for subsequent analyses

On the Easy Use of Scientific Computing Services for Large Scale Linear Algebra and Parallel Decision Making with the P-Grade Portal

International audienceScientific research is becoming increasingly dependent on the large-scale analysis of data using distributed computing infrastructures (Grid, cloud, GPU, etc.). Scientific computing (Petitet et al. 1999) aims at constructing mathematical models and numerical solution techniques for solving problems arising in science and engineering. In this paper, we describe the services of an integrated portal based on the P-Grade (Parallel Grid Run-time and Application Development Environment) portal (http://www.p-grade.hu) that enables the solution of large-scale linear systems of equations using direct solvers, makes easier the use of parallel block iterative algorithm and provides an interface for parallel decision making algorithms. The ultimate goal is to develop a single sign on integrated multi-service environment providing an easy access to different kind of mathematical calculations and algorithms to be performed on hybrid distributed computing infrastructures combining the benefits of large clusters, Grid or cloud, when needed

Atrativos alimentares para o monitoramento de adultos da mosca-das-frutas sul-americana na cultura da ameixeira.

Nos últimos anos, falhas significativas no controle da mosca-das-frutas sul-americana Anastrephafraterculus (Wied., 1830) (Diptera: Tephritidae) tem sido observadas em pomares em diversas culturas. Uma das principais justificativas para esse problema é decorrente do sistema de monitoramento (baseado na utilização de suco de uva a 25% como atrativo). Este atrativo não tem sido confiável, principalmente no período de pré-colheita das frutas, onde em hipótese, as moscas não são atraídas pelas armadilhas em função da grande presença de voláteis emitidos pelas frutas maduras ou em decomposição.Resumo

Concentration analysis and cocompactness

Loss of compactness that occurs in may significant PDE settings can be expressed in a well-structured form of profile decomposition for sequences. Profile decompositions are formulated in relation to a triplet $(X,Y,D)$, where $X$ and $Y$ are Banach spaces, $X\hookrightarrow Y$, and $D$ is, typically, a set of surjective isometries on both $X$ and $Y$. A profile decomposition is a representation of a bounded sequence in $X$ as a sum of elementary concentrations of the form $g_kw$, $g_k\in D$, $w\in X$, and a remainder that vanishes in $Y$. A necessary requirement for $Y$ is, therefore, that any sequence in $X$ that develops no $D$-concentrations has a subsequence convergent in the norm of $Y$. An imbedding $X\hookrightarrow Y$ with this property is called $D$-cocompact, a property weaker than, but related to, compactness. We survey known cocompact imbeddings and their role in profile decompositions

Il percorso ideale nella gestione del paziente con osteoporosi severa:dalla valutazione del rischio all'approccio multidisciplinare

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