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Background Several pathogens have been debated to play a role in inflammatory bowel disease (IBD) including Crohn's disease (CD). None of these pathogens have been investigated together in same clinical samples. We developed a multiplex PCR and multi-color fluorescent in situ hybridization (m-FISH) protocols for simultaneous detection of CD-associated pathogens including Mycobacterium avium subspecies paratuberculosis (MAP), Klebsiella pneumoniae, and adherent-invasive Escherichia coli strain LF82. Methods The multiplex PCR is based on 1-h DNAzol.sup.[R] extraction protocol modified for rapid extraction of bacterial DNA from culture, blood, and intestinal biopsies. Oligonucleotide primers sequences unique to these pathogens were evaluated individually and in combinations using bioinformatics and experimental approaches. m-FISH was based on fluorescent-tagged oligonucleotides and confocal scanning laser microscopy (CSLM). Results Following several attempts, the concentration of the oligonucleotide primers and DNA templates and the PCR annealing temperatures were optimized. Multiplex PCR analyses revealed excellent amplification signal in trials where a single primer set and combinations of two and three primers sets were tested against a mixture of DNA from three different bacteria or a mixture of three bacterial cultures mixed in one tube before DNA extraction. Slides with individual and mixtures of bacterial cultures and intestinal tissue sections from IBD patients were tested by m-FISH and the CSLM images verified multiplex PCR results detected on 3% agarose gel. Conclusion We developed a 4-h multiplex PCR protocol, which was validated by m-FISH images, capable of detecting up to four genes from major pathogens associated with CD. The new protocol should serve as an excellent tool to support efforts to study multi-pathogens involved in CD and other autoimmune disease. Keywords: Crohn's disease, Ulcerative colitis, MAP, m-FISH, Multiplex PCR, Author(s): Robert C. Sharp[sup.1] , Ebraheem S. Naser[sup.1] , Karel P. Alcedo[sup.1] , Ahmad Qasem[sup.1] , Latifa S. Abdelli[sup.1] and Saleh A. Naser[sup.1] Background Inflammatory bowel disease (IBD), which consists [...]

Cell therapy is emerging as a new strategy to circumvent the adverse effects of heart disease. Many experimental and clinical studies investigating the transplantation of cells into the injured myocardium have yielded promising results. Moreover, data from these reports show that transplanted stem cells can engraft within the myocardium, differentiate into major cardiac cell types, and improve cardiac function. However, results from clinical trials show conflicting results. These trials demonstrate significant improvements in cardiac function for up to 6 months. However, these improved functions were diminished when examined at 18 months. In this review, we will discuss the current literature available on cell transplantation, covering studies ranging from animal models to clinical trials. Key words: cardiomyopathy, cardiac myocytes, adult and embryonic stem cells, cell transplantation therapy, clinical trials. La therapie cellulaire apparait comme une nouvelle strategie pour contrer les effets indesirables des maladies du coeur. De nombreuses etudes experimentales et cliniques sur la greffe de cellules dans le myocarde endommage ont donne des resultats prometteurs. De plus, les donnees provenant de ces etudes montrent que les cellules souches greffees peuvent s'implanter dans le myocarde, se differencier en types de cellules cardiaques majeurs et ameliorer la fonction cardiaque. Toutefois, les essais cliniques ont des resultats contradictoires. Ces essais demontrent des ameliorations considerables de la fonction cardiaque en moins de 6 mois, mais une diminution de ces ameliorations apres 18 mois. Dans la presente synthese, nous discuterons de la documentation actuelle sur la greffe de cellules, des etudes sur les modeles animaux aux etudes sur les essais cliniques. Mots-cles : cardiomyopathie, myocytes cardiaques, cellules souches adultes et embryonnaires, therapie cellulaire, essais cliniques. [Traduit par la Redaction], Introduction Development and progression of heart disease (HD) involves a variety of pathological modifications that contribute to altered heart structure and function. Myocardial infarction (MI) and related coronary artery diseases [...]