Search results for "rt-PCR"

Expression of cytokeratins in human umbilical cord at term

2009

Background. Cytokeratins are the proteins that costitute intermediate filaments. In the cytoplasm, keratin filaments extend from the nucleus to the plasma membrane. This link is important for the structural organization of the cytoplasm and for many cellular processes such as mitosis, cell movement, cell differentiation and adhesion. The pattern of expression of cytokeratins is frequently organ or tissue specific and depends mainly on the type of epithelium, the level of differentiation and the stage of development. The expression of cytokeratins in umbilical cord has been investigated by many groups, but these previous studies gave only fragmentary informations: each single study focused o…

Expression of Remodelling Enzymes (MMP-2, MMP-9) in Normal and Pathological Oral Mucosa Samples

2008

Introduction. The oral cavity contains many structures compressed in a small area. It is exposed to chronic or recurrent, physical and chemical trauma leading to mucosal reactions (e.g. hyperplasia, dysplasia and tumors). For a long time, our group has been studying the expression of some metalloproteinases (MMP-2, MMP-9) in healthy and pathological human oral mucosa through immunohistochemistry and RT-PCR to investigate the entity of the remodelling process in the different cases. Matherials and Methods. In this paper we investigate a sample of normal oral mucosa and four pathological samples (mucocele, fibroma, dysplasia and carcinoma). We identified the localization of MMP-2 and MMP-9 th…

Immunohistochemical and transcriptional expression of the matrix metalloproteinases MMP-2 and MMP-9 in normal and pathological human oral mucosa

2009

The oral cavity is exposed to chronic or recurrent, physical and chemical trauma that could lead to mucosal reactions (e.g. hyperplasia, dysplasia and tumors). The objective of this study is to investigate the expression and the possible changes of the two matrix metalloproteinases MMP-2 and MMP-9 in normal and pathological human oral mucosa samples. Normal oral mucosa samples and three different types of pathological conditions (hyperplasia, dysplasia and carcinoma) were used for this study. Immunohistochemical analysis was used to evaluate protein expression for the two enzymes, while Reverse Transcription ? Polymerase Chain Reaction (RT-PCR) was used to investigate gene expression. Image…

Molecular study of porcine transmissible gastroenteritis virus after serial animal passages revealed point mutations in S protein

2010

Porcine respiratory coronavirus is related genetically to porcine transmissible gastroenteritis virus with a large deletion in S protein. The respiratory virus is a mutated form that may be a consequence of the gastroen- teritis virus's evolution. Intensive passages of the virus in its natural host may enhance the appearance of mutations and therefore may contribute to any attenuated form of the virus. The objective of this study was to characterize the porcine transmissible gastroenteritis virus TMK22 strain after passages in piglets from 1992 until 2007. A typical experimental infection, molecular characterization, and serological analysis were also carried out to further char- acterize a…

Detection and identification Fabavirus species by one-step RT-PCR and multiplex RT-PCR

2014

The genus Fabavirus of the family Secoviridae comprises a group of poorly characterized viruses. To date, only five species have been described: Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV-2), Lamium mild mosaic virus (LMMV), Gentian mosaic virus (GeMV) and Cucurbit mild mosaic virus (CuMMV). The development is described of two RT-PCR procedures for the detection and identification of Fabavirus species: a one-step RT-PCR using a single pair of conserved primers for the detection of all fabaviruses, and a one-step multiplex RT-PCR using species-specific primers for the simultaneous detection and identification of the above-mentioned species of the genus Fabavirus. These m…

Anti-inflammatory Function of High-Density Lipoproteins via Autophagy of IκB Kinase

2015

Background & Aims: Plasma levels of high-density lipoprotein (HDL) cholesterol are frequently found decreased in patients with inflammatory bowel disease (IBD). Therefore, and because HDL exerts anti-inflammatory activities, we investigated whether HDL and its major protein component apolipoprotein A-I (apoA-I) modulate mucosal inflammatory responses in vitro and in vivo. Methods: The human intestinal epithelial cell line T84 was used as the in vitro model for measuring the effects of HDL on the expression and secretion of tumor necrosis factor (TNF), interleukin-8 (IL-8), and intracellular adhesion molecule (ICAM). Nuclear factor-κB (NF-κB)-responsive promoter activity was studied by …

Localisation et visualisation de transcrits fongiques in situ: Une méthode originale qui combine biologie moléculaire et microscopie confocale

2011

National audience; Les Gloméromycètes, formant la symbiose mycorhizienne à arbuscules, sont des champignons biotrophes obligatoires qui sont intimement associés aux tissus végétaux qu’ils colonisent. Les techniques utilisées pour suivre l’expression des gènes fongiques dans les racines mycorhizées ne permettent pas de définir le profil spatio-temporel de leur activité. Afin de tracer l’activité transcriptionnelle fongique dans les racines mycorhizées, une méthode innovante, basée sur la RT-PCR in situ, a donc été développée. Cette technique permet de localiser, en microscopie optique confocale à balayage laser, des transcrits fongiques, à l’aide d’amorces spécifiques marquées par un fluoroc…

Essential oils as multi-target compounds for novel food safety strategies

This research work has been developed in the context of the potencial applications of essential oils as multi-target compounds for novel food safety strategies, such as active packaging technologies. Essential oils are aromatic oils derived from plants and usually extracted by hydrodistillation when intended to be used in food. There is an increasing interest in the antimicrobial properties of essential oils due to the possibility of using them to replace ‘synthetic’ preservatives in food or, in general, to reduce viable numbers of pathogens along the food chain.

Globularia nudicaulis, a new host of Cucumber mosaic virus

2006

) is a perennial, foundnaturally on European mountains at altitudes between 900 and 2000 m.In June 2004, G. nudicaulis plants, with a yellow mosaic and/or variega-tion on malformed leaves, were noted among plant species cultivated in theBotanical Garden at Bologna University, Italy. No elongated virus-likeparticles were observed in affected-leaf extracts by transmission electronmicroscopy using a leaf dip method. By applying a protein A sandwichenzyme-linked immunosorbent assay (PAS-ELISA) technique ( Edwards Csystemic symptoms were observed in Nicotiana tabacum , N. benthamiana ,N. glutinosa, N. clevelandii and Capsicum annuum, and Cucumis sativusand C. melo. Reverse transcription-polymera…

Teschoviruses and sapeloviruses in faecal samples from wild boar in Spain

2013

Teschovirus and Sapelovirus are two genera of the Picornaviridae family, comprising highly variable and heterogeneous enteric viruses, commonly found in faecal samples from domestic pigs. Although both of them are also known to infect wild boar, studies on their presence in these wild suids are scarce. The present study aimed at determining the presence of porcine teschovirus (PTV) and sapelovirus (PSV) in free-living wild boar populations, as well as to study their relationships with similar viruses present in pigs. Fresh faecal samples (n = 63) from wild boar were collected in Doñana Biological Reserve (SW Spain) during 2007 and 2011, and analysed using multiplex RT-PCR for the simultaneo…