Preparation of α‐Oxo Semicarbazone Oligonucleotide Microarrays

This unit describes the preparation of α‐oxo aldehyde functionalized oligodeoxynucleotides, the preparation and characterization of semicarbazide glass slides, and the fabrication of α‐oxo semicarbazone microarrays by site‐specific ligation of α‐oxo‐aldehyde oligodeoxynucleotides to the semicarbazide glass slides. The α‐oxo aldehyde group COCHO is extensively used in ligation chemistry for the preparation of large molecular constructs. It is stable toward air oxidation and mainly present in aqueous solution in the hydrated form COC(OH)2. It reacts efficiently with hydrazine derivatives, in particular, with the semicarbazide group. The reaction occurs spontaneously in water at pH 5.5. Site‐specific immobilization of glyoxylyl oligodeoxynucleotides on semicarbazide glass slides allows the preparation of high‐quality microarrays that can be used directly in hybridization experiments.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/143637/1/cpnc1206.pd

Philosophy in Science: Can philosophers of science permeate through science and produce scientific knowledge?

Most philosophers of science do philosophy ‘on’ science. By contrast, others do philosophy ‘in’ science (‘PinS’), i.e., they use philosophical tools to address scientific problems and to provide scientifically useful proposals. Here, we consider the evidence in favour of a trend of this nature. We proceed in two stages. First, we identify relevant authors and articles empirically with bibliometric tools, given that PinS would be likely to infiltrate science and thus to be published in scientific journals (‘intervention’), cited in scientific journals (‘visibility’) and sometimes recognized as a scientific result by scientists (‘contribution’). We show that many central figures in philosophy of science have been involved in PinS, and that some philosophers have even ‘specialized’ in this practice. Second, we propose a conceptual definition of PinS as a process involving three conditions (raising a scientific problem, using philosophical tools to address it, and making a scientific proposal), and we ask whether the articles identified at the first stage fulfil all these conditions. We show that PinS is a distinctive, quantitatively substantial trend within philosophy of science, demonstrating the existence of a methodological continuity from science to philosophy of science

The visibility of philosophy of science in the sciences, 1980–2018

In this paper, we provide a macro level analysis of the visibility of philosophy of science in the sciences over the last four decades. Our quantitative analysis of publications and citations of philosophy of science papers, published in 17 main journals representing the discipline, contributes to the longstanding debate on the influence of philosophy of science on the sciences. It reveals the global structure of relationships that philosophy of science maintains with science, technology, engineering and mathematics (STEM) and social sciences and humanities (SSH) fields. Explored at the level of disciplines, journals and authors, this analysis of the relations between philosophy of science and a large and diversified array of disciplines allows us to answer several questions: what is the degree of openness of various disciplines to the specialized knowledge produced in philosophy of science? Which STEM and SSH fields and journals have privileged ties with philosophy of science? What are the characteristics, in terms of citation and publication patterns, of authors who get their philosophy of science papers cited outside their field? Complementing existing qualitative inquiries on the influence of specific authors, concepts or topics of philosophy of science, the bibliometric approach proposed in this paper offers a comprehensive portrait of the multiple relationships that links philosophy of science to the sciences

Genomic island excisions in Bordetella petrii

<p>Abstract</p> <p>Background</p> <p>Among the members of the genus <it>Bordetella B. petrii </it>is unique, since it is the only species isolated from the environment, while the pathogenic Bordetellae are obligately associated with host organisms. Another feature distinguishing <it>B. petrii </it>from the other sequenced Bordetellae is the presence of a large number of mobile genetic elements including several large genomic regions with typical characteristics of genomic islands collectively known as integrative and conjugative elements (ICEs). These elements mainly encode accessory metabolic factors enabling this bacterium to grow on a large repertoire of aromatic compounds.</p> <p>Results</p> <p>During <it>in vitro </it>culture of <it>Bordetella petrii </it>colony variants appear frequently. We show that this variability can be attributed to the presence of a large number of metastable mobile genetic elements on its chromosome. In fact, the genome sequence of <it>B. petrii </it>revealed the presence of at least seven large genomic islands mostly encoding accessory metabolic functions involved in the degradation of aromatic compounds and detoxification of heavy metals. Four of these islands (termed GI1 to GI3 and GI6) are highly related to ICE<it>clc </it>of <it>Pseudomonas knackmussii </it>sp. strain B13. Here we present first data about the molecular characterization of these islands. We defined the exact borders of each island and we show that during standard culture of the bacteria these islands get excised from the chromosome. For all but one of these islands (GI5) we could detect circular intermediates. For the <it>clc</it>-like elements GI1 to GI3 of <it>B. petrii </it>we provide evidence that tandem insertion of these islands which all encode highly related integrases and attachment sites may also lead to incorporation of genomic DNA which originally was not part of the island and to the formation of huge composite islands. By integration of a tetracycline resistance cassette into GI3 we found this island to be rather unstable and to be lost from the bacterial population within about 100 consecutive generations. Furthermore, we show that GI3 is self transmissible and by conjugation can be transferred to <it>B. bronchiseptica </it>thus proving it to be an active integrative and conjugative element</p> <p>Conclusion</p> <p>The results show that phenotypic variation of <it>B. petrii </it>is correlated with the presence of genomic islands. Tandem integration of related islands may contribute to island evolution by the acquisition of genes originally belonging to the bacterial core genome. In conclusion, <it>B. petrii </it>appears to be the first member of the genus in which horizontal gene transfer events have massively shaped its genome structure.</p

Transcriptomic and proteomic profiles of vascular cells involved in human abdominal aortic aneurysm

Diseases & disorder

Identification of novel genes potentially involved in somatic embryogenesis in chicory (Cichorium intybus L.)

<p>Abstract</p> <p>Background</p> <p>In our laboratory we use cultured chicory (<it>Cichorium intybus</it>) explants as a model to investigate cell reactivation and somatic embryogenesis and have produced 2 chicory genotypes (K59, C15) sharing a similar genetic background. K59 is a responsive genotype (embryogenic) capable of undergoing complete cell reactivation <it>i.e</it>. cell de- and re-differentiation leading to somatic embryogenesis (SE), whereas C15 is a non-responsive genotype (non-embryogenic) and is unable to undergo SE. Previous studies <abbrgrp><abbr bid="B1">1</abbr></abbrgrp> showed that the use of the β-D-glucosyl Yariv reagent (β-GlcY) that specifically binds arabinogalactan-proteins (AGPs) blocked somatic embryo production in chicory root explants. This observation indicates that β-GlcY is a useful tool for investigating somatic embryogenesis (SE) in chicory. In addition, a putative AGP (DT212818) encoding gene was previously found to be significantly up-regulated in the embryogenic K59 chicory genotype as compared to the non-embryogenic C15 genotype suggesting that this AGP could be involved in chicory re-differentiation <abbrgrp><abbr bid="B2">2</abbr></abbrgrp>. In order to improve our understanding of the molecular and cellular regulation underlying SE in chicory, we undertook a detailed cytological study of cell reactivation events in K59 and C15 genotypes, and used microarray profiling to compare gene expression in these 2 genotypes. In addition we also used β-GlcY to block SE in order to identify genes potentially involved in this process.</p> <p>Results</p> <p>Microscopy confirmed that only the K59, but not the C15 genotype underwent complete cell reactivation leading to SE formation. β-GlcY-treatment of explants blocked <it>in vitro </it>SE induction, but not cell reactivation, and induced cell wall modifications. Microarray analyses revealed that 78 genes were differentially expressed between induced K59 and C15 genotypes. The expression profiles of 19 genes were modified by β-GlcY-treatment. Eight genes were both differentially expressed between K59 and C15 genotypes during SE induction and transcriptionally affected by β-GlcY-treatment: <it>AGP </it>(DT212818), <it>26 S proteasome AAA ATPase subunit 6 </it>(<it>RPT6</it>), <it>remorin </it>(<it>REM</it>), <it>metallothionein-1 </it>(<it>MT1</it>), two non-specific lipid transfer proteins genes (<it>SDI-9 and DEA1</it>), <it>3-hydroxy-3-methylglutaryl-CoA reductase </it>(<it>HMG-CoA reductase</it>), and <it>snakin 2 </it>(<it>SN2</it>). These results suggest that the 8 genes, including the previously-identified <it>AGP </it>gene (DT212818), could be involved in cell fate determination events leading to SE commitment in chicory.</p> <p>Conclusion</p> <p>The use of two different chicory genotypes differing in their responsiveness to SE induction, together with β-GlcY-treatment represented an efficient tool to discriminate cell reactivation from the SE morphogenetic pathway. Such an approach, together with microarray analyses, permitted us to identify several putative key genes related to the SE morphogenetic pathway in chicory.</p

The RPGRIP1-deficient dog, a promising canine model for gene therapy

PURPOSE: To evaluate the RPGRIP1-deficient miniature longhaired dachshund (MLHD) dog as a potential candidate for gene therapy. METHODS: Six RPGRIP1-deficient MLHD dogs from our dog colony have been observed for two years using a variety of noninvasive procedures. These included bilateral full-field electroretinograms (ERG) to evaluate retinal function, fundus photographs to evaluate retinal vascularization, and optical coherence tomographs (OCT) to evaluate retinal thickness. We also performed histological examination of hematoxylin- and eosin-stained retinal sections as well as sections labeled in situ by the terminal dUTP nick end labeling (TUNEL) method. RESULTS: ERG findings showed that as early as 2 months of age, cone function was lost while rod function was preserved. However, by 9 months of age, both cone and rod functions could not be detected. Functional visual assessment based on the ability to avoid obstacles showed that vision was retained up to the age of 11 months. Both OCT and histopathology studies revealed a progressive thinning of the outer nuclear layer (ONL) over the first 2 years of age. TUNEL labeling identified apoptotic photoreceptor cell death as the cause of this thinning of the ONL. CONCLUSIONS: A treatment strategy should consist in initiating gene therapy as early as possible after birth to prevent or delay the loss of rod function. In the MLHD, successful subretinal delivery of a therapeutic vector is feasible at 2 months of age and may prevent or delay the loss of rod function

Exact two-time correlation and response functions in the one-dimensional coagulation-diffusion process by the empty-interval-particle method

The one-dimensional coagulation-diffusion process describes the strongly fluctuating dynamics of particles, freely hopping between the nearest-neighbour sites of a chain such that one of them disappears with probability 1 if two particles meet. The exact two-time correlation and response function in the one-dimensional coagulation-diffusion process are derived from the empty-interval-particle method. The main quantity is the conditional probability of finding an empty interval of n consecutive sites, if at distance d a site is occupied by a particle. Closed equations of motion are derived such that the probabilities needed for the calculation of correlators and responses, respectively, are distinguished by different initial and boundary conditions. In this way, the dynamical scaling of these two-time observables is analysed in the longtime ageing regime. A new generalised fluctuation-dissipation ratio with an universal and finite limit is proposed.Comment: 31 pages, submitted to J.Stat.Mec

Detection of small RNAs in Bordetella pertussis and identification of a novel repeated genetic element

Background: Small bacterial RNAs (sRNAs) have been shown to participate in the regulation of gene expression and have been identified in numerous prokaryotic species. Some of them are involved in the regulation of virulence in pathogenic bacteria. So far, little is known about sRNAs in Bordetella, and only very few sRNAs have been identified in the genome of Bordetella pertussis, the causative agent of whooping cough. Results: An in silico approach was used to predict sRNAs genes in intergenic regions of the B. pertussis genome. The genome sequences of B. pertussis, Bordetella parapertussis, Bordetella bronchiseptica and Bordetella avium were compared using a Blast, and significant hits were analyzed using RNAz. Twenty-three candidate regions were obtained, including regions encoding the already documented 6S RNA, and the GCVT and FMN riboswitches. The existence of sRNAs was verified by Northern blot analyses, and transcripts were detected for 13 out of the 20 additional candidates. These new sRNAs were named Bordetella pertussis RNAs, bpr. The expression of 4 of them differed between the early, exponential and late growth phases, and one of them, bprJ2, was found to be under the control of BvgA/BvgS two-component regulatory system of Bordetella virulence. A phylogenetic study of the bprJ sequence revealed a novel, so far undocumented repeat of ~90 bp, found in numerous copies in the Bordetella genomes and in that of other Betaproteobacteria. This repeat exhibits certain features of mobil