Preliminary evaluation of probiotic properties of Lactobacillus strains isolated from Sardinian dairy products

Twenty-three Lactobacillus strains of dairy origin were evaluated for some functional properties relevant to their use as probiotics. A preliminary subtractive screening based on the abilities to inhibit the growth of microbial pathogens and hydrolyze conjugated bile salts was applied, and six strains were selected for further characterization including survival under gastrointestinal environmental conditions, adhesion to gut epithelial tissue, enzymatic activity, and some safety properties. All selected strains maintained elevated cell numbers under conditions simulating passage through the human gastrointestinal tract, well comparable to the values obtained for the probiotic strain Lactobacillus rhamnosus GG, and were able to adhere to Caco-2 cells to various extents (from 3 to 20%). All strains exhibited high aminopeptidase, and absent or very low proteolytic and strong β-galactosidase activities; none was found to be haemolytic or to produce biogenic amines and all were susceptible to tetracycline, chloramphenicol, erythromycin, ampicillin, and amoxicillin/clavulanic acid. Our results indicate that the Lactobacillus strains analyzed could be considered appropriate probiotic candidates, due to resistance to GIT simulated conditions, antimicrobial activity, adhesion to Caco-2 cell-line, and absence of undesirable properties. They could be used as adjunct cultures for contributing to the quality and health related functional properties of dairy products

Estimation of traffic matrices for LRD traffic

The estimation of traffic matrices in a communications network on the basis of a set of traffic measurements on the network links is a well known problem, for which a number of solutions have been proposed when the traffic does not show dependence over time, as in the case of the Poisson process. However, extensive measurements campaigns conducted on IP networks have shown that the traffic exhibits long range dependence. Here two methods are proposed for the estimation of traffic matrices in the case of long range dependence, their asymptotic properties are studied, and their relative merits are compared

Is Mandevilla (Apocynaceae, Mesechiteae) monophyletic? Evidence from five plastid DNA loci and morphology

In order to test the monophyly of Mandevilla Lindl., the largest genus in tribe Mesechiteea (Apocynaceae, Apocynoideae). and its affinities to other genera, in the tribe. maximum parsimony analysis was conducted on a data set comprising DNA sequence, from five plastid loci (rp/16. rps16, and trnK introns: trnS(GEE)-trnG(LPC) intergenic spacer; and matK gene), as well as morphological data for 65 taxa of Mesechiteae (48, Mandevilla) and nine taxa from other tribes of the subfamily. Mandevilla, as circumscribed by Pichon, was found to be monophyletic. whereas Woodson's circumscription proved to be polyphyletic. Thus defined, Mandevilla form, it strongly supported clade that call be divided into six clades of species groups. Most of the infrageneric taxa of Mandevilla proposed by Woodson and Pichon are polyphyletic. Many of the diagnostic characters previously used to define taxonomic groups are shown to have arisen multiple times, rendering them unsuitable lot classificatory purposes. The similar growth form and tubular flowers of Macrosiphonia Mull. Arg. and Telosiphonia (Woodson) Henr, two geographically disjunct segregates, represent the most extreme case of parallel evolution within Mandevilla, with their striking similarities most likely correlated to colonization of open. dry habitats and pollination by hawkmoths.93456559

Characterization of Italian honeys (Marche Region) on the basis of their mineral content and some typical quality parameters

<p>Abstract</p> <p>Background</p> <p>The characterization of three types of Marche (Italy) honeys (Acacia, Multifloral, Honeydew) was carried out on the basis of the their quality parameters (pH, sugar content, humidity) and mineral content (Na, K, Ca, Mg, Cu, Fe, and Mn). Pattern recognition methods such as principal components analysis (PCA) and linear discriminant analysis (LDA) were performed in order to classify honey samples whose botanical origins were different, and identify the most discriminant parameters. Lastly, using ANOVA and correlations for all parameters, significant differences between diverse types of honey were examined.</p> <p>Results</p> <p>Most of the samples' water content showed good maturity (98%) whilst pH values were in the range 3.50 – 4.21 confirming the good quality of the honeys analysed. Potassium was quantitatively the most relevant mineral (mean = 643 ppm), accounting for 79% of the total mineral content. The Ca, Na and Mg contents account for 14, 3 and 3% of the total mineral content respectively, while other minerals (Cu, Mn, Fe) were present at very low levels. PCA explained 75% or more of the variance with the first two PC variables. The variables with higher discrimination power according to the multivariate statistical procedure were Mg and pH. On the other hand, all samples of acacia and honeydew, and more than 90% of samples of multifloral type have been correctly classified using the LDA. ANOVA shows significant differences between diverse floral origins for all variables except sugar, moisture and Fe.</p> <p>Conclusion</p> <p>In general, the analytical results obtained for the Marche honeys indicate the products' high quality. The determination of physicochemical parameters and mineral content in combination with modern statistical techniques can be a useful tool for honey classification.</p

Frequency of educational computer use as a longitudinal predictor of educational outcomes in young people with specific language impairment

Computer use draws on linguistic abilities. Using this medium thus presents challenges for young people with Specific Language Impairment (SLI) and raises questions of whether computer-based tasks are appropriate for them. We consider theoretical arguments predicting impaired performance and negative outcomes relative to peers without SLI versus the possibility of positive gains. We examine the relationship between frequency of computer use (for leisure and educational purposes) and educational achievement; in particular examination performance at the end of compulsory education and level of educational progress two years later. Participants were 49 young people with SLI and 56 typically developing (TD) young people. At around age 17, the two groups did not differ in frequency of educational computer use or leisure computer use. There were no associations between computer use and educational outcomes in the TD group. In the SLI group, after PIQ was controlled for, educational computer use at around 17 years of age contributed substantially to the prediction of educational progress at 19 years. The findings suggest that educational uses of computers are conducive to educational progress in young people with SLI

Strain-controlled criticality governs the nonlinear mechanics of fibre networks

Disordered fibrous networks are ubiquitous in nature as major structural components of living cells and tissues. The mechanical stability of networks generally depends on the degree of connectivity: only when the average number of connections between nodes exceeds the isostatic threshold are networks stable (Maxwell, J. C., Philosophical Magazine 27, 294 (1864)). Upon increasing the connectivity through this point, such networks undergo a mechanical phase transition from a floppy to a rigid phase. However, even sub-isostatic networks become rigid when subjected to sufficiently large deformations. To study this strain-controlled transition, we perform a combination of computational modeling of fibre networks and experiments on networks of type I collagen fibers, which are crucial for the integrity of biological tissues. We show theoretically that the development of rigidity is characterized by a strain-controlled continuous phase transition with signatures of criticality. Our experiments demonstrate mechanical properties consistent with our model, including the predicted critical exponents. We show that the nonlinear mechanics of collagen networks can be quantitatively captured by the predictions of scaling theory for the strain-controlled critical behavior over a wide range of network concentrations and strains up to failure of the material

Actin and myosin contribute to mammalian mitochondrial DNA maintenance.

Mitochondrial DNA maintenance and segregation are dependent on the actin cytoskeleton in budding yeast. We found two cytoskeletal proteins among six proteins tightly associated with rat liver mitochondrial DNA: non-muscle myosin heavy chain IIA and β-actin. In human cells, transient gene silencing of MYH9 (encoding non-muscle myosin heavy chain IIA), or the closely related MYH10 gene (encoding non-muscle myosin heavy chain IIB), altered the topology and increased the copy number of mitochondrial DNA; and the latter effect was enhanced when both genes were targeted simultaneously. In contrast, genetic ablation of non-muscle myosin IIB was associated with a 60% decrease in mitochondrial DNA copy number in mouse embryonic fibroblasts, compared to control cells. Gene silencing of β-actin also affected mitochondrial DNA copy number and organization. Protease-protection experiments and iodixanol gradient analysis suggest some β-actin and non-muscle myosin heavy chain IIA reside within human mitochondria and confirm that they are associated with mitochondrial DNA. Collectively, these results strongly implicate the actomyosin cytoskeleton in mammalian mitochondrial DNA maintenance.Medical Research Council; the European Union; the Intramural Research Program of the Eunice Kennedy Shriver National Institute of Child Health and Human Development and National Heart; Lung and Blood Institute; National Institutes of Health and grants [CMRPG360491-2, 380651, NSC 97-2321-B-182A-002-MY2] from the Chang Gung Memorial Hospital, Lin-Kou, Taiwan (to C.C.M.). Funding for open access charge: Medical Research Council

PO-299 In vivo shRNA screening to identify quiescence-related genes required for AML growth

Introduction AML is hierarchically organised with at the apex Leukaemia Stem Cells (LSCs), a rare cell population able to initiate and sustain the tumour growth. LSCs share many functional properties with normal Hematopoietic Stem Cells (HSCs) including self-renewal capacity and quiescence. Quiescent LSCs can survive to radiation and chemotherapy acting as a reservoir for leukaemia relapse, the major cause of death for AML patients. Therefore, LSCs quiescence is critical for leukaemia maintenance and few evidences suggest that quiescence regulation in pre-leukemic phase plays a pivotal role for leukemogenic process as well. Material and methods We analysed the transcriptional deregulations induced by the expression of different leukemic oncogenes in HSCs and we examined the contribution of representative quiescence related genes in AML growth by in vivo RNA interference screening. Results and discussions The transcriptional profile of oncogene-expressing HSCs is enriched in a quiescent stem cell gene signature, compared to normal HSCs. Therefore, we hypothesised that enhancement of the quiescent phenotype in HSCs could be a shared mechanism for leukaemia development and maintenance. The in vivo shRNA screening allowed the identification of genes whose silencing in AML blasts was sufficient to significantly decrease in vitro self-renewal and delay leukaemia growth in vivo . Conclusion We identified quiescence-related genes, commonly deregulated by leukemic oncogenes at pre-leukemic level, which may offer new therapeutic targets in a wide group of AML patients

PO-272 Leukemia-associated NPM mutations promote quiescence of hematopoietic stem cells and prevent their functional exhaustion upon oncogene-induced hyper-proliferation

Introduction Acute Myeloid Leukaemia (AML) is a heterogeneous and multi-step disease. The serial acquisition of mutations and the environmental pressure allow one or more clones to expand and contribute to the disease. In particular, 6% of AMLs are characterised by an initial mutation in the DNMT3a gene, followed by mutations in NPM (NPMc) and FLT3 loci (FLT3-ITD). We previously shown that NPMc can drive AML development in mouse model and highly cooperates with FLT3-ITD. Moreover, it has been reported that normal Hematopoietic Stem Cells (HSCs) of elderly people may bear some somatic early AML mutations and this correlate with an increased risk of hematologic diseases suggesting that mutations can shape pre-leukemic HSCs to be more prone to the acquisition of further mutations giving rise to Leukaemia Initiating Cells (LIC). While the ability of FLT3-ITD to drive HSC compartment exhaustion has been already shown, the impact of NPMc on HSCs remains unclear. Material and methods Taking advantage of the extended pre-leukemic phase of our inducible NPMc mouse model, we elucidate the role of NPMc in HSCs by functional and transcriptional analysis. Moreover, to investigate the basis of NPMc and FLT3-ITD cooperation we generate mice carrying both the conditional NPMc transgene and the FLT3-ITD constitutive mutation and, before AML onset, we analyse double mutant HSCs behaviour. Results and discussions We have found that NPMc expression lead to the expansion of the HSC compartment through the enforcement of a stem-cell transcriptional program that increases self-renewal by promoting quiescence. We then investigated how the NPMc dependent quiescence program is linked to its oncogenic function. The expression of NPMc +in the FLT3-ITD background prevents the HSCs exhaustion imposed by FLT3-ITD and restores their repopulating capacity. Accordingly, gene expression analysis revealed a strong dominance of NPMc +with the restoration of the same transcriptional program observed in the NPMc HSCs. These data strongly suggest that NPMc imposes a HSC-specific program that, in combination with the oncogenic signal provided by FLT3-ITD, allows the selection of the LIC and the occurrence of AML. Conclusion In conclusion, enforcement of quiescence might be a critical function for the maintenance of the transformed clone during both the pre-leukemic and the leukemic phase. As consequence, interfering with quiescence key determinants may eradicate the reservoir of quiescent cells responsible for disease recurrence