Neuroprotective effect of nimesulide, a preferential COX-2 inhibitor, against pentylenetetrazol (PTZ)-induced chemical kindling and associated biochemical parameters in mice

Brain cyclooxygenases (COX), the rate-limiting enzyme in prostaglandin synthesis, is rapidly and transiently induced by convulsions in hippocampal and cortical neurons. Previous studies have explored the protective effect of naproxen (non-selective COX-inhibitor) or rofecoxib (selective COX-2 inhibitor) against chemical kindling in mice. With this background, the present study was designed to explore the possible effect of nimesulide (a preferential COX-2 inhibitor) against pentylenetetrazol (PTZ)-induced kindling epilepsy in mice. To induce kindling, PTZ was injected in a subconvulsive dose (40 mg/kg, i.p.) every other day for 15 days. Nimesulide (2.5 or 5 mg/kg, p.o.) was administered each day 45 min before either PTZ or vehicle challenge. The intensity of kindling was assessed immediately after PTZ administration according to a prevalidated scoring scale. On 16th day i.e. 24 h after the last dose of PTZ, animals were sacrificed and various biochemical parameters were assessed in the whole brain. Compared with normal control group, PTZ-kindled mice had significantly higher levels of malondialdehyde, nitrite, myeloperoxidase but had lower levels of reduced glutathione in the whole brain homogenate. Chronic treatment with nimesulide (2.5 or 5 mg/kg, p.o.) for 15 days showed significant decrease in kindling score and could play a role in controlling the accompanying biochemical alterations due to PTZ. These results suggested that nimesulide, a preferential COX-2 inhibitor offered neuroprotection against PTZ-induced kindling in mice.     

Lactoferrin Interaction with Actinobacillus actinomycetemcomitans

The interaction of lactoferrin with Actinobacillus actinomycetemcomitans was examined in a ¹²⁵I-labeled protein binding assay. The binding of human and bovine lactoferrins reached maximum within 1 h. Lactoferrin binding to the bacterium was pH-dependent and reversible. Scatchard analysis indicated the existence of two different types of binding sites on the bacterium, one with a high affinity constant k_α=8.8×10⁻⁷ M) and the other with a low one (k_α=1.8×10⁻⁶ M). Bacteria in the exponential phase of growth showed higher binding than cells in the stationary phase. Bacteria grown in medium containing serum and/or lysed erythrocytes bound lactoferrin to a lesser extent. Heat-inactivated serum, lysed erythrocytes and other proteins such as mucin and laminin inhibited lactoferrin binding to A. actinomycetemcomitans in a competitive binding assay. Sodium dodecyl sulfate polyacrylamidegel electrophoresis and Western blot analysis of the cell envelope as well as the outer membrane of A. actinomycetemcomitans revealed lactoferrin-reactive protein bands at 29 kDa and 16.5 kDa. The 29-kDa band displayed a heat-modifiable lactoferrin-reactive form with a molecular weight of 34 kDa. Neither proteinase K-treated cell envelope nor lipopolysaccharide of this bacterium showed reactivity with lactoferrin. These data suggests a specific interaction of lactoferrin with outer membrane proteins of A. actinomycetemcomitans .     

Thromboelastography in abruptio placentae.

The purpose of this study was to document thromboelastographic (TEG) changes in abruptio placentae and to compare these results with that of conventional tests used to monitor coagulation. This was a prospective study of 30 patients with abruptio placentae. All coagulation investigations including the TEG were performed on admission, immediately following delivery, and after periods of 4 hours and 24 hours. Results showed that standard coagulation tests detected coagulation abnormalities except the platelet count returned to normal limits within 24 hours of delivery. There was strong correlation between fibrinogen levels and the TEG parameters, ma and k time (r=0.8). There was moderate correlation between platelet count and ma (r=0.6). In conclusion, minor abnormalities in the clotting profile are clinically unimportant. The TEG does not detect such minor abnormalities because of its inherent ability to test the coagulation cascade as a whole. Major abnormalities are clinically relevant and the TEG detect 75% of them. Further, the TEG establishes the diagnosis of hypercoagulability an early sign of disseminated intravascular coagulation. Although standard laboratory tests are still necessary to detect coagulation abnormalities on admission, the TEG is a useful test in large obstetric units where laboratory results are not immediately available for the purposes of monitoring and treating ongoing coagulation defects.     

Particle agglutination assays for rapid detection of fibronectin, fibrinogen, and collagen receptors on Staphylococcus aureus.

Latex beads (0.8-micron diameter; Difco Laboratories) were coated with fibronectin, fibrinogen, collagen type I, or denatured collagen (gelatin) and evaluated in a particle agglutination assay (PAA) for the rapid detection of fibronectin, fibrinogen, or collagen binding to Staphylococcus aureus. These assays were compared with a commercial test for detecting the binding of fibrinogen and immunoglobulin G (Staphaurex). Bacterial cells (approximately 10(10) cells per ml) suspended in 0.02 M potassium phosphate buffer (pH 6.8) caused the clumping of standard fibronectin, collagen, gelatin, and fibrinogen latex suspensions within 2 min on glass slides. The test results were scored semiquantitatively from strongly positive ( ) to weakly positive (+) and negative (-) reactions. The negative PAA reactions corresponded to a median value of 11.5% relative to the binding of 125I-labeled protein to strain Cowan 1, indicating the high sensitivity of the test. The reactions with fibronectin and fibrinogen latex suspensions and with Staphaurex were optimal for cells grown on tryptic soy and brain heart infusion broth media. Blood agar was optimal for reactions with collagen and gelatin latex suspensions. Media containing high salts (mannitol salt agar and staphylococcus medium 110) enhanced the tendency of cells to autoaggregate. These assays were also clinically evaluated on 187 S. aureus isolates. The PAA reagents were stable, and the assays were highly specific, sensitive, and reproducible, thus making PAA suitable for the rapid screening of the binding of various bacterial pathogens to serum and connective-tissue proteins.