Aims: Bordetella bronchiseptica is an etiologic agent of bronchopneumonia and progressive atrophic rhinitis (PAR) in swine. Both toxigenic and nontoxigenic B. bronchiseptica strains have been associated with bronchopneumonia. Monitoring and investigation of outbreaks involving these bacteria require sensitive and accurate identification and reliable determination of the isolates. In the present study, we report the development, optimization and performance characteristics of polymerase chain reaction (PCR) for B. bronchiseptica strains. Methodology and Results: A total of 47 isolates of B. bronchiseptica were biochemically identified from 90 pigs suffering from bronchopneumonia maintained in a semi intensive rearing system of organized piggery in Meghalaya. PCR was employed with filamentous hemagglutinin toxin genes (fhaB and fhaC) and fimbrial toxin genes (fim2 and fim3) primers to identify the specific toxin types of B. bronchiseptica. All the 47 isolates were positive for all the toxin genes. The specifity of designed primer pairs was tested by screening some common bacterial species related to the respiratory tract namely, Pasteurella multocida, Staphylococcus aureus and Streptococcus spp. No DNA amplifications of the organisms tested could be seen in the specificity test. Amplicon mobility in agarose gels indicate the amplicons are highly stable. Conclusion, significance and impact of study: The data presented, establish this PCR as a reliable method for identification and study of adhesins of B. bronchiseptica that may greatly simplify investigations of swine bronchopneumonia and PAR for Indian isolates.

BACKGROUND AND OBJECTIVELeukemic microenvironment has a major role in the progression of leukemia. Leukemic cells can induce reversible changes in microenvironmental components, especially the stromal function which results in improved growth conditions for maintaining the malignant leukemic cells. This study aimed to investigate the survival advantage of leukemic cells over normal hematopoietic cells in stromal microenvironment in long term.

METHODSThe mice were injected intraperitoneally with N-N' ethylnitrosourea (ENU) to induce leukemia; the mice received injection of normal saline were used as control. At 180 days after ENU induction, the mice were killed and the bone marrows were cultured for 19 days. Colony-forming assays were used to analyze the formation of various cell colonies. The expression of Sca-1, CD146, VEGFR2, CD95, pStat3, pStat5, and Bcl-xL in marrow cells were detected by flow cytometry.

RESULTSLong-term leukemic bone marrow culture showed abnormal elongated stromal fibroblasts with almost absence of normal hematopoietic cells. Adherent cell colonies were increased, but CFU-F and other hematopoietic cell colonies were significantly decreased in leukemia group (P<0.001). Primitive progenitor-specific Sca-1 receptor expression was decreased with subsequent increased expression of CD146 and VEGFR-2 in leukemic bone marrow cells. Decreased Fas antigen expression with increased intracellular pStat3, pStat5 and Bcl-xL proteins were observed in leukemic bone marrow cells.

CONCLUSIONSStromal microenvironment shows altered morphology and decreased maturation in leukemia. Effective progenitor cells are decreased in leukemia with increased leukemia-specific cell population. Leukemic microenvironment plays a role in promoting and maintaining the leukemic cell proliferation and survivability in long term.

Animals ; Antigens, Ly ; metabolism ; Bone Marrow Cells ; metabolism ; pathology ; CD146 Antigen ; metabolism ; Cell Count ; Cells, Cultured ; Colony-Forming Units Assay ; Erythroid Precursor Cells ; metabolism ; pathology ; Ethylnitrosourea ; Female ; Fibroblasts ; metabolism ; pathology ; Granulocyte-Macrophage Progenitor Cells ; metabolism ; pathology ; Granulocytes ; metabolism ; pathology ; Hematopoiesis ; Hematopoietic Stem Cells ; metabolism ; pathology ; Leukemia ; chemically induced ; metabolism ; pathology ; Male ; Membrane Proteins ; metabolism ; Mice ; Myeloid Progenitor Cells ; metabolism ; pathology ; Phenotype ; STAT3 Transcription Factor ; metabolism ; STAT5 Transcription Factor ; metabolism ; Tumor Microenvironment ; physiology ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism ; bcl-X Protein ; metabolism ; fas Receptor ; metabolism

Inflammatory bowel disease (IBD), once considered a disease of the Western hemisphere, has emerged as a global disease. As the disease prevalence is on a steady rise, management of IBD has come under the spotlight. 5-Aminosalicylates, corticosteroids, immunosuppressive agents and biologics are the backbone of treatment of IBD. With the advent of biologics and small molecules, the need for surgery and hospitalization has decreased. However, economic viability and acceptability is an important determinant of local prescription patterns. Nearly one-third of the patients in West receive biologics as the first/initial therapy. The scenario is different in developing countries where biologics are used only in a small proportion of patients with IBD. Increased risk of reactivation of tuberculosis and high cost of the therapy are limitations to their use. Thiopurines hence become critical for optimal management of patients with IBD in these regions. However, approximately one-third of patients are intolerant or develop adverse effects with their use. This has led to suboptimal use of thiopurines in clinical practice. This review article discusses the clinical aspects of thiopurine use in patients with IBD with the aim of optimizing their use to full therapeutic potential.