Differential temporal expression of matrix metalloproteinases after spinal cord injury: relationship to revascularization and wound healing.

OBJECT: Matrix metalloproteinases (MMPs), particularly MMP-9/gelatinase B, promote early inflammation and barrier disruption after spinal cord injury (SCI). Early blockade of MMPs after injury provides neuroprotection and improves motor outcome. There is recent evidence, however, that MMP-9 and MMP-2/gelatinase A participate in later wound healing in the injured cord. The authors therefore examined the activity of these gelatinases during revascularization and glial scar formation in the contused murine spinal cord. METHODS: Gelatinase activity was evaluated using gelatin zymography 24 hours after a mild, moderate, or severe contusion injury. The active form of MMP-2 was not detected, whereas MMP-9 activity was evident in all SCI groups and rose with increasing injury severity. The temporal expression of gelatinases was then examined using gelatin zymography after a moderate SCI. The active form of MMP-9 was most prominent at 1 day, extended through the early period of revascularization, and returned to control by 14 days. The active form of MMP-2 appeared at 7 days postinjury and remained elevated compared with that documented in sham-treated mice for at least 21 days. Increased MMP-2 activity coincided with both revascularization and glial scar formation. Using in situ zymography, gelatinolytic activity was detected in the meninges, vascular elements, glia, and macrophage-like cells in the injured cord. Results of immunolabeling confirmed the presence of gelatinase in vessels during revascularization and in reactive astrocytes associated with glial scar formation. CONCLUSIONS: These findings suggest that although MMP-9 and -2 exhibit overlapping expression during revascularization, the former is associated with acute injury responses and the latter with formation of a glial scar.

The use of flow cytometry to assess neutrophil infiltration in the injured murine spinal cord.

Inflammatory cells, including neutrophils, are likely candidates in promoting early cell death after spinal cord injury. We describe a simple and reliable method for obtaining neutrophils from the injured murine spinal cord for flow cytometric quantification. Mice were subjected to either a moderate or severe spinal cord contusion injury and euthanized 24 h later. The area of maximal damage, designated the epicenter, was prepared for assessment of myeloperoxidase (MPO) activity, quantitative immunocytochemistry, or quantification of immunolabeled neutrophils by flow cytometry. For flow cytometry, a cell suspension was prepared from the epicenter by gentle mechanical disruption. After centrifugation, the pellet was resuspended, immunolabeled for neutrophils, and analyzed. There was no detectable MPO activity in the injured spinal cord. In contrast, neutrophil infiltration was confirmed by immunocytochemistry and found to be significantly greater in the more severely injured group. Flow cytometry, using a standard neutrophil marker, revealed a similar significant increase in immunolabeled cells in the more severely injured group. However, when cell viability was determined in the neutrophil labeled population, no significant difference in the numbers of live neutrophils were noted between the two injured groups. Together, these findings demonstrate an effective method for the detection and quantification of viable neutrophils in the injured murine spinal cord.

The lactic acid bacteria: a literature survey.

The purpose of this review article on the lactic acid bacteria grew from an early curiosity and a desire to convey and impart the broad scope of literary information on their functions as starter cultures, in the manufacture of fermentation products such as dairy products and alcoholic beverages, as well as their contribution to better health. This review article is an attempt to empower the reader and to circumvent the difficult task in acquiring and elucidating a large body of information. The intent is to familiarize the reader with the various lactic species, their habitat or source, associated food, physiological characteristics, colonial morphology, biochemical characteristics, culture media (enrichment, nonselective, and selective), classic description, and taxonomy. This review provides information on Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Carnobacterium, and Enterococcus. Trends are presented, such as the use of nisin to extend food shelf-life and the current research premise that Probiotic strains may alter the intestinal flora and thus prevent intestinal wall penetration by pathogens.