The Common Shrew (Sorex araneus): A neglected host of tick-borne infections?

Although the importance of rodents as reservoirs for a number of tick-borne infections is well established, comparatively little is known about the potential role of shrews, despite them occupying similar habitats. To address this, blood and tick samples were collected from common shrews (Sorex araneus) and field voles (Microtus agrestis), a known reservoir of various tick-borne infections, from sites located within a plantation forest in northern England over a 2-year period. Of 647 blood samples collected from shrews, 121 (18.7%) showed evidence of infection with Anaplasma phagocytophilum and 196 (30.3%) with Babesia microti. By comparison, of 1505 blood samples from field voles, 96 (6.4%) were positive for A. phagocytophilum and 458 (30.4%) for Ba. microti. Both species were infested with the ticks Ixodes ricinus and Ixodes trianguliceps, although they had different burdens: on average, shrews carried almost six times as many I. trianguliceps larvae, more than twice as many I. ricinus larvae, and over twice as many nymphs (both tick species combined). The finding that the nymphs collected from shrews were almost exclusively I. trianguliceps highlights that this species is the key vector of these infections in this small mammal community. These findings suggest that common shrews are a reservoir of tick-borne infections and that the role of shrews in the ecology and epidemiology of tick-borne infections elsewhere needs to be comprehensively investigated

Interleukin-2 promoter activation in T-cells expressing activated Ha-ras.

Antigen triggering of the T-cell receptor results in an accumulation of activated GTP-bound p21ras protein. To assess the role of ras protein in T-cell activation we have cotransfected the murine thymoma line EL4 with a construct capable of expressing a constitutively active, oncogenic form of Ha-ras and a reporter construct containing the human interleukin-2 promoter fused upstream of the bacterial gene for chloramphenicol acetyltransferase. We show that the ras oncoprotein contributes to interleukin-2 promoter activation. Its pattern of synergism with a calcium ionophore or the lymphokine interleukin-1 indicates that it replaces a signal mediated by protein kinase C. Interleukin-2 promoter activity in the presence of ras oncoprotein was inhibited by H7, a potent inhibitor of protein kinase C, but not by HA1004, an inhibitor of cyclic nucleotide-dependent kinase, suggesting that protein kinase C mediates the ras effect. In addition, we show that in these cells, expression of activated ras results in activation of a synthetic promoter containing several copies of an NF kappa B binding site

Flow cytometry and cell sorting

While flow cytometry is a critical single cell analytical technique in biomedical science, the technology of flow cytometry associated cell sorting is equally important. Physical separation of cells analyzed by flow cytometry was recognized as an important goal even in the field’s beginning, and many of the earliest cytometers were also cell sorters. Isolation of cells based on flow cytometric analysis has formed the foundation of immune cell differentiation and development and continues to grow importance as techniques for genomic and proteomic analysis expand. This brief review will describe both the historical development and current state of cell sorting. The multiple mechanisms for cell sorters will be covered, and critical aspects of cell sorting will be discussed. Newer technologies for cell sorting including microfluidic technologies will also be considered

Cyclosporin A blocks calcium-dependent pathways of gene activation

We have used an interleukin-2 (IL-2) promoter-CAT fusion gene to study activation of IL-2 gene expression by IL-1, phytohemagglutinin (PHA), phorbol myristate acetate (PMA), and calcium ionophore in the murine thymoma line EL4 and the human lymphoma line Jurkat. The two cell lines respond differently to combinations of these stimuli. IL-1 in combination with suboptimal concentration of PMA induced chloramphenicol acetyltransferase (CAT) activity in EL4. In Jurkat cells, IL-1 failed to synergize with PMA or PHA. Cotransfection with the IL-2/CAT gene and a construct capable of expressing murine T-cell type IL-1 receptors converted Jurkat cells to IL-1 responsiveness. IL-1 in combination with PHA but not with PMA resulted in induction of CAT activity in these cells. Induction of IL-2/CAT activity by all stimuli in both cell lines was blocked by the presence of EGTA in the culture medium. EGTA did not inhibit IL-1/PMA activation of an SV40 early promoter-CAT fusion gene in either EL4 or Jurkat cells; therefore, calcium was not required for IL-1 or PMA signal transduction. Jurkat cells were shown to differ from EL4 in their requirement for calcium mobilization. Two different calcium-dependent pathways of gene activation were distinguished, both of which were blocked by the immunosuppressive drug cyclosporin A

A Simultaneous X-ray Diffraction-Differential Scanning Calorimetry Study into the Phase Transitions of Mefenamic Acid

In this study, the polymorphic transitions of mefenamic acid (MA) were studied by synchrotron X-ray powder diffraction combined with differential scanning calorimetry (XRD-DSC). The initial material was found to be phase-pure form I which, when heated, produces two endotherms that can be observed by DSC at 162.72 and 219.55°C. The former was found to correspond to a solid–solid enantiotropic transition from form I to a mixture of forms II and III. The latter is the melting point of form II. As form I is heated, significantly greater unit-cell expansion is seen in the a direction than in b and c, which can be explained by the stronger intermolecular interactions in the bc plane. Refinements of the reported MA structures against the patterns collected during heating revealed that at 175°C there exists a mixture of forms I, II and III, whereas only forms II and III remain at 205°C. However, reflections are observed at both temperatures which cannot be fitted with the known forms of MA. It is hypothesized that a new form of MA is produced upon heating. The stability of MA after the enantiotropic transition temperature is II > III > I, which differs from the previously reported II > I > III