1,721,078 research outputs found
Fluorescent in situ hybridization - bacterial detection and identification
No full textFluorescent in situ hybridization (FISH) with ribosomal RNA (rRNA) targeted fluorescent oligonucleotide probes is a culture-independent technique for the detection and identification of bacteria. The cells remain morphologically intact allowing the visualization and enumeration of cells. Briefly, bacterial cells are first fixed and permeabilized if necessary, hybridized with specific probes under stringent conditions, and detected either by epifluorescent microscope or by flow cytometry (FCM). The technique is suitable for species-specific detection of pathogens and for the analysis of complex microbial populations. Multiplexed detection can be achieved by selecting unique fluorochromes for different specific probes. Fluorescent in situ hybridization provides information about the presence, number, morphology, and spatial distribution of bacteria. It can be performed in a few hours and therefore holds great promise for medical microbiology. This review summarizes methodological aspects and diagnostic applications of FISH
Modulation of Gut Microbiota for Health by Current and Next-Generation Probiotics
Full text linkThe human gut microbiota is a complex ecosystem and has an essential role in maintaining intestinal and systemic health. Microbiota dysbiosis is associated with a number of intestinal and systemic conditions and its modulation for human health is of great interest. Gut microbiota is a source of novel health-promoting bacteria, often termed as next-generation probiotics in order to distinguish them from traditional probiotics. The previous lessons learned with traditional probiotics can help the development of next-generation probiotics that target specific health issues and needs
Molecular identification and characterisation of bifidobacteria and lactobacilli in the human gastrointestinal tract
No full textBifidobacteria and lactobacilli are considered to be members of the beneficial microbiota in the human gastrointestinal (GI) tract. The present study describes the development and validation of new molecular methods for the detection and analysis of bifidobacteria and lactobacilli and the application of new techniques to study Bifidobacterium and Lactobacillus populations in the human intestine. A method based on genus-specific PCR of 16S rDNA and denaturing gradient gel electrophoresis (DGGE) was developed and validated for profiling Bifidobacterium populations in human faeces. The PCR-DGGE method is a qualitative tool for assessing species/strain composition of complex communities by a single PCR reaction and subsequent resolution of the amplification products by DGGE in a sequence-dependent manner. The approach greatly facilitates the monitoring of faecal samples from large numbers of subjects to reveal bifidobacterial diversity and shifts occurring in it. The identification of DGGE fragments can be done by subsequent cloning and sequencing of the PCR products. Genotypic methods were developed and evaluated for the identification and characterisation of Lactobacillus casei -group lactobacilli ( L. casei, L. paracasei, L. rhamnosus , and L. zeae ). L. rhamnosus species-specific PCR was developed and validated. The discriminatory power of the three fingerprinting techniques, pulsed field gel electrophoresis (PFGE), randomly amplified polymorphic DNA (RAPD) and ribotyping, was compared. All three techniques were highly effective in differentiating strains below the species level and they can be placed in the following order with respect to their discriminatory power: PFGE > ribotyping > RAPD. Newly developed molecular methods were used to trace ingested probiotic strains L. rhamnosus GG (LGG) and B. lactis Bb12 in the GI-tract. The identity of LGG colonies was verified using a species-specific PCR and Bb12 was detected using the PCR-DGGE method. Both probiotic strains colonised the gut transiently and they were no longer detected in the faeces one week after the end of the administration in most subjects. The synbiotic approach with galactooligosaccharide (GOS) did not prolong the persistence of Bb12. Furthermore, LGG was found to attach in vivo to colonic mucosae and, although the attchment was temporary, to remain for more than a week after discontinuation of LGG administration. PCR-DGGE method was used to monitor qualitative changes in adult faecal Bifidobacterium populations in response to B. lactis Bb12 and/or GOS administration. In most subjects two weeks administration of Bb12 and/or GOS did not affect the qualitative composition of indigenous bifidobacterial populations, while Bb12 transiently colonised the gut. Qualitative molecular analysis was used to study the bacterial, bifidobacterial and lactobacilli populations in faeces of breast-fed and formula-fed infants before and after weaning. Genus and group-specific PCRs combined with DGGE and subsequent sequencing of the amplified 16S rDNA fragments revealed no difference in the prevalence or species distribution of Bifidobacterium and Lactobacillus between the two groups of infants. In general, DGGE patterns of 16S rDNA showed equal complexity of bacterial communities in breast-fed and formula-fed infants. Equally intensive changes occurred in the faecal microbiota in infants of both groups due to weaning.</p
Going Beyond Counting First Authors in Author Co-citation Analysis
Full text linkThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
High Intake of Sugar and the Balance between Pro- and Anti-Inflammatory Gut Bacteria
Full text linkThe so-called Western diet is rich in saturated fat and sugars and poor in plant-derived fibers, and it is associated with an increased risk of metabolic and cardiovascular diseases, as well as chronic (low grade) inflammation. The detrimental effects of poor diet are in part mediated by gut microbiota, whose composition, functionality and metabolic end products respond to dietary changes. Recent studies have shown that high intake of sugars increase the relative abundance of Proteobacteria in the gut, while simultaneously decreasing the abundance of Bacteroidetes, which can mitigate the effects of endotoxin, as well as reinforce gut barrier function. Thus, a high sugar intake may stagger the balance of microbiota to have increased pro-inflammatory properties and decreased the capacity to regulate epithelial integrity and mucosal immunity. Consequently, high dietary sugar can, through the modulation of microbiota, promote metabolic endotoxemia, systemic (low grade) inflammation and the development of metabolic dysregulation and thereby, high dietary sugar may have many-fold deleterious health effects, in addition to providing excess energy
Variations on the Author
Full text link“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Appropriate Similarity Measures for Author Cocitation Analysis
Full text linkWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
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