Orientation-Based Control of Microfluidics

Most microfluidic chips utilize off-chip hardware (syringe pumps, computer-controlled solenoid valves, pressure regulators, etc.) to control fluid flow on-chip. This expensive, bulky, and power-consuming hardware severely limits the utility of microfluidic instruments in resource-limited or point-of-care contexts, where the cost, size, and power consumption of the instrument must be limited. In this work, we present a technique for on-chip fluid control that requires no off-chip hardware. We accomplish this by using inert compounds to change the density of one fluid in the chip. If one fluid is made 2% more dense than a second fluid, when the fluids flow together under laminar flow the interface between the fluids quickly reorients to be orthogonal to Earth's gravitational force. If the channel containing the fluids then splits into two channels, the amount of each fluid flowing into each channel is precisely determined by the angle of the channels relative to gravity. Thus, any fluid can be routed in any direction and mixed in any desired ratio on-chip simply by holding the chip at a certain angle. This approach allows for sophisticated control of on-chip fluids with no off-chip control hardware, significantly reducing the cost of microfluidic instruments in point-of-care or resource-limited settings

Effect of cytokine and antigen stimulation on peripheral blood lymphocyte syndecan-1 expression

IntroductionCytokines are not only produced by activated lymphocytes but also interact with a number of cell-surface molecules on the same cells. Syndecan-1 is one such cell-surface molecule, which has the capacity to bind a variety of growth factors as well as cytokines. The aim of this study was to examine the effects of transforming growth factor beta (TGF-beta), interleukin-1 (IL-1), IL-2, IL-4, lipopolysaccharide (LPS) from Porphyromonas gingivalis and tetanus toxoid on syndecan-1 expression by B and T lymphocytes.MethodsB and T lymphocytes were obtained from the peripheral blood of healthy donors. Following exposure to the above growth factors, cytokines and antigens, syndecan-1 expression was determined by flow cytometry.ResultsSubjects could be categorized as high or low expressors of syndecan-1. In the high-responder group TGF-beta1 alone resulted in a significant increase in syndecan-1 expression by both B and T cells. None of the other cytokines and antigens produced a significant response. When analysed in combination, TGF-beta1 in combination with IL-2, IL-4, P. gingivalis LPS and tetanus toxoid all produced significant increases in syndecan-1 expression by B cells. For T cells, combinations of TGF-beta1 with IL-2 and tetanus toxoid resulted in increased syndecan-1 expression.ConclusionsBoth B and T lymphocytes synthesize the cell-surface proteoglycan syndecan-1 and its expression can be modulated by TGF-beta1, either alone or in combination with IL-2, IL-4 and LPS from P. gingivalis and tetanus toxoid. While these may reflect general responses under inflammatory conditions their biological significance requires further investigation.J. F. Manakil, G. J. Seymour, P. M. Bartol

Azurophil granule proteins constitute the major mycobactericidal proteins in human neutrophils and enhance the killing of mycobacteria in macrophages.

Pathogenic mycobacteria reside in, and are in turn controlled by, macrophages. However, emerging data suggest that neutrophils also play a critical role in innate immunity to tuberculosis, presumably by their different antibacterial granule proteins. In this study, we purified neutrophil azurophil and specific granules and systematically analyzed the antimycobacterial activity of some purified azurophil and specific granule proteins against M. smegmatis, M. bovis-BCG and M. tuberculosis H37Rv. Using gel overlay and colony forming unit assays we showed that the defensin-depleted azurophil granule proteins (AZP) were more active against mycobacteria compared to other granule proteins and cytosolic proteins. The proteins showing antimycobacterial activity were identified by MALDI-TOF mass spectrometry. Electron microscopic studies demonstrate that the AZP disintegrate bacterial cell membrane resulting in killing of mycobacteria. Exogenous addition of AZP to murine macrophage RAW 264.7, THP-1 and peripheral blood monocyte-derived macrophages significantly reduced the intracellular survival of mycobacteria without exhibiting cytotoxic activity on macrophages. Immunofluorescence studies showed that macrophages actively endocytose neutrophil granular proteins. Treatment with AZP resulted in increase in co-localization of BCG containing phagosomes with lysosomes but not in increase of autophagy. These data demonstrate that neutrophil azurophil proteins may play an important role in controlling intracellular survival of mycobacteria in macrophages