Streptococcus dysgalactiae is a bacterium that accounts for a notable proportion of both clinical and subclinical intramammary infections (IMIs). Thus, the present study explores the function of milk neutrophils and the lymphocyte profile in mammary glands naturally infected with Streptococcus dysgalactiae. Here, we used 32 culture-negative control quarters from eight clinically healthy dairy cows with low somatic cell counts and 13 S. dysgalactiae-infected quarters from six dairy cows. Using flow cytometry, we evaluated the percentage of milk monocytes/macrophages and neutrophils, expression of CD62L, CD11b and CD44 by milk neutrophils, the levels of intracellular reactive oxygen species (ROS) production and phagocytosis of Staphylococcus aureus by milk neutrophils, and neutrophil viability. Furthermore, the percentages of B cell (CD21+) and T lymphocyte subsets (CD3+/CD4+/CD8−; CD3+/CD8+/CD4−; and CD3+/CD8−/CD4−), and the expression of CD25 by T milk lymphocytes (CD3+) and T CD4+ milk cells were also assessed by flow cytometry using monoclonal antibodies. The present study showed a higher SCC and percentage of milk neutrophils, and a decrease in the percentage of milk monocytes/macrophages from S. dysgalactiae-infected quarters when compared to uninfected ones. We also observed a higher expression of CD11b by milk neutrophils and a tendency toward a decrease in neutrophil apoptosis rate in S. dysgalactiae-infected quarters. In addition, the S. dysgalactiae-infected quarters had higher percentages of milk T cells (CD3+) and their subset CD3+CD8+CD4− cells. Overall, the present study provided new insights into S. dysgalactiae IMIs, including distinct lymphocyte profiles, and a tendency toward an inhibition of apoptosis in milk neutrophils.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008

Extended abstract of a paper presented at Microscopy and Microanalysis 2007 in Ft. Lauderdale, Florida, USA, August 5 – August 9, 2007

Extended abstract of a paper presented at Microscopy and Microanalysis 2007 in Ft. Lauderdale, Florida, USA, August 5 – August 9, 2007

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2006

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2005

Extended abstract of a paper presented at Microscopy and Microanalysis 2006 in Chicago, Illinois, USA, July 30 – August 3, 2006

We studied the protein adsorption and subsequent macrophage adhesion on poly(ethylene glycol) [PEG] hydrogels crosslinked using a focused electron beam. Thin-film gels were patterned on silicon substrates and could be formed with swell ratios (hydrated height/ dry height) anywhere between fifteen and unity. We have shown that laminin does not adsorb onto highly swelling gels but that it does adsorb on heavily-crosslinked low-swelling gels. As part of ongoing research on patterning surfaces to control neurite growth in the context of the inflammatory environment of a spinal cord injury, we are interested in how these gel surfaces interact with macrophages. We show that the high-swelling PEG gels resist macrophage adhesion, but the macrophages adhere to low-swelling gels pre-exposed to laminin. By spatially patterning combinations of low and high swelling gels, we show that macrophage adhesion can be confined to specific locations on a surface.

Central to the successful advance of tissue engineering (e.g. 1, 2) is the development of highly engineered materials which trigger particular body responses to regenerate specific tissue or sets of tissue. Such properties as hydrophilicity, hydrolytic susceptibility, and protein-recognition behavior are critical to the success of tissue scaffolds. Poly(ethylene glycol) [PEG] is often introduced into scaffold polymers to tune hydrolytically controlled properties. Relatively little is known about the local morphology and nature of phase separation in these systems, however. This research studies the development of phase-separated morphology in blends and randommultiblock copolymers of a tyrosine-based poly(psuedo amino acid) [poly(DTE carbonate)] and PEG 1000 (fig. 1). This system exhibits attractive biocompatability, strength and modulus, and resorption behavior which can be controlled by main-chain and pendant-chain chemistries (3, 4).

The nature of phase separation was studied by bright-field TEM (Philips CM30 ST; CM20 FEG TEM/STEM) using solvent-cast thin films.

The common experimental method to determine the total inelastic mean free path i by electron energy-loss spectroscopy (EELS) is by the relation : t/λi= ln(It/IO) [1] where t is the specimen thickness, It, is the total integrated intensity, and Io is the intensity of the zero-loss peak. The accuracy of this measurement depends on the thickness determination. Model geometries like cubes, wedges, and spheres enable accurate thickness determination from transmission images.

Spherical polymers with diameters of order 10-200nm can be made from a number of high-Tg polymers by solvent atomization. This research studied atomized spheres of poly(2-vinyl pyridine) [PVP]. A solution of 0.1% PVP in THF was nebulized. After solvent evaporation during free fall within the chamber atmosphere, solid spherical polymer particles with a range of diameters were collected on holey-carbon TEM grids at the bottom of the atomization chamber.

Arborescent graft polymers are highly branched materials synthesized by grafting multiple generations of macromolecular precursors on a central core [1,2]. They can be synthesized with controllable size and molecular weight depending on the number of generations and the precursor molecular weight. They possess many of the unique structural [3], thermodynamic, and rheological [4] properties of dendrimers but do so on a length scale roughly one order of magnitude higher. This research describes the effect of confinement on the shape of arborescent graft polystyrene (PS).

This work used 3rd generation arborescent graft PS synthesized with 5kg/mole linear polystyrene precursor (G3- 5K). The G3-5K PS sample was mixed to a dilution of approximately 0.005 wt% in toluene. A single drop of solution was placed on either a holey-carbon film or an ultrathin continuous carbon film. The solvent was evaporated under ambient conditions. A Philips CM20 FEG TEM/STEM equipped with a Gatan 666 PEELS and a 679 multiscan CCD camera was used to study the structure of these specimens.

Chemical imaging of polymeric materials is attractive, because of the inherently poor contrast typically afforded by multiphase amorphous polymers in bright-field imaging techniques. Spectrum imaging and energy-filtering approaches are being increasing applied to polymeric materials (1,2, 3).

One recent example from the Stevens group using a Philips CM20 FEG TEM/STEM interfaced to an EmiSpec Vision data acquisition/control system and a Gatan 666 PEELS spectrometer is presented in figure 1. The specimen is a PVP sphere on a holey-carbon TEM grid. The specimen was cooled to approximately -125 °C in a single-tilt cryo stage. Drift correction was implemented by collecting HAADF STEM images from the sub-image region indicated by the dashed white box at periodic intervals. Based on an autocorrelation with the initial image, electronic shifts where imposed to correct for drift. A 3-D spectrum dataset was generated from the 70 x 70 pixel box around the polymer particle at an interpixel spacing of 5 nm and a pixel dwell time of 1.5 sec.