The scope of mycoplasma contamination within the biopharmaceutical industry.

Mycoplasma is well recognized as one of the most prevalent and serious microbial contaminants encountered within the manufacturing of biopharmaceuticals from the research phase to clinical development and production. The potential for mycoplasma contamination within cell culture systems was first identified by Robinson et al. in 1956.

Calcium/calmodulin-dependent protein kinase II-delta isoform regulation of vascular smooth muscle cell proliferation.

There is accumulating evidence that Ca(2+)-dependent signaling pathways regulate proliferation and migration of vascular smooth muscle (VSM) cells, contributing to the intimal accumulation of VSM that is a hallmark of many vascular diseases. In this study we investigated the role of the multifunctional serine/threonine kinase, calmodulin (CaM)-dependent protein kinase II (CaMKII), as a mediator of Ca(2+) signals regulating VSM cell proliferation. Differentiated VSM cells acutely isolated from rat aortic media express primarily CaMKIIgamma gene products, whereas passaged primary cultures of de-differentiated VSM cells express primarily CaMKIIdelta(2), a splice variant of the delta gene.

Adhesion-dependent activation of CaMKII and regulation of ERK activation in vascular smooth muscle.

Cell adhesion-dependent activation of ERK1/2 has been linked functionally to focal adhesion dynamics. We previously reported that in adherent vascular smooth muscle (VSM) cells, CaMKII mediates ERK1/2 activation in response to Ca(2+)-mobilizing stimuli. In the present study, we tested whether CaMKII regulates ERK1/2 signaling in response to VSM cell adhesion.

Measurement of high-molecular-weight hyaluronan in solid tissue using agarose gel electrophoresis.

The size of hyaluronan in solid tissue was measured using a combination of agarose gel electrophoresis and a radiometric assay. Radiolabeled hyaluronan binding proteins, used in the radiometric assay, were also used to detect hyaluronan after transfer to a nylon membrane following gel electrophoresis. Lane intensity on the autoradiograph was linearly related to the amount applied to the gel between 10 and 100ng.

Relationship between lymph and tissue hyaluronan in skin and skeletal muscle.

The size of hyaluronan was compared between tissue and lymph using a combination of agarose gel electrophoresis and radiometric assay. Prenodal lymph was collected from heel skin and the gastrocnemius muscle in anesthetized rabbits. The major fraction of hyaluronan in both tissues had a molecular weight >4 million.

Ischemia-reperfusion does not cause significant hyaluronan depolymerization in skeletal muscle.

The size of hyaluronan in lymph and gastrocnemius muscle was measured to test the hypothesis that reperfusion of ischemic skeletal muscle leads to hyaluronan depolymerization. Prenodal lymph was collected from the gastrocnemius muscle in anesthetized rabbits. Hyaluronan size was measured using a combination of agarose gel electrophoresis and a radiometric assay.