Immunohistopathological appearance of three different types of injury in human skin.

OBJECTIVE AND DESIGN: To determine whether underlying mechanisms of inflammation, like cellular infiltrates, expression of adhesion molecule and cytokine patterns are similar under different conditions of injury. Skin biopsies were taken of three different groups of patients in which local inflammation of the skin might occur. MATERIAL, PATIENTS: Skin activation was studied as a result of incision during surgery under aseptic conditions, as a result of a local bacterial infection and as a result of blunt trauma, resulting in a femoral fracture, without disruption of the epithelial barrier. METHODS: Skin biopsies were snap frozen for immunohistochemical analysis. RESULTS: Incision of the skin resulted in a granulocyte infiltrate, paralleled by E-selectin expression. As a result of infection granulocytes were observed and monocyte/macrophage and T cell numbers were increased. Furthermore, E-selectin, VCAM-1 and ICAM-1 expression increased and cytokine expression markedly changed compared to normal skin. Skin taken at the site of the femoral fracture showed no signs of inflammation. CONCLUSION: Different stimuli lead to different local inflammatory responses in human skin.

Tumor necrosis factor alpha (TNFalpha) in human skin: a comparison of different antibodies for immunohistochemistry.

Conflicting results have been reported regarding the localization and presence of TNFalpha in normal human skin. To study TNFalpha expression, we tested a panel of antibodies directed against human TNFalpha. First, antibodies were tested for immunoreactivity on cytospots of isolated LPS-stimulated peripheral blood mononuclear cells. Second, antibodies were tested to detect recombinant TNFalpha in Western blots. Some antibodies were found to be unable to detect recombinant TNFalpha in the blots. However, most antibodies were able to bind TNFalpha protein, but they did not bind to other irrelevant proteins that were also present in the blots. Finally, antibodies were tested on cryosections of normal human skin. Antibodies that did not react with TNFalpha in the blots were incubated with TNFalpha before the staining procedure to see whether these antibodies specifically bound TNFalpha. We found that, although all the antibodies bound TNFalpha, there were clear differences in staining patterns. This indicates that these antibodies may recognize distinct epitopes or different forms of TNFalpha. The differences found in this study and those reported previously could be the result of differences in the concentration of antibody used, the staining procedure or specificity of the antibody itself. So, for unambiguous interpretation of data, it is important to know the characteristics of the antibodies used.

Local cellular inflammation as a result of elective standardized vascular surgery.

During surgery, incision of the skin under aseptic conditions is performed. Despite the absence of noxious agents, an inflammatory response may be induced. We studied the local inflammatory response in human skin as a result of surgical intervention, under aseptic conditions. Elective standardized vascular surgery served as a model. A series of skin biopsies was taken from the wound edge at different time points after first incision. Biopsies, directly taken at first incision were considered to represent normal skin. Additional biopsies were taken at 30 min after the start of surgery and just before closure of the wound, maximally 270 min after surgery. Kinetics of recruitment of cells, expression of adhesion molecules and the presence of pro-inflammatory cytokines was studied. Granulocytes were observed at first at 30 min after incision of the skin and their number increased in time. This granulocyte infiltration is paralleled by E-selectin expression on endothelial cells, which also was observed at first at 30 min after surgery with a further increase in number in time. Incision of the skin did not change P-selectin, ICAM-1, VCAM-1, TNFalpha, IL1alpha, IL1beta, IL6 and IL8 expression. These results show that incision of the skin under aseptic conditions during elective standardized vascular surgery induces local nonspecific cellular inflammation.

Peritonitis, moderate ascites and hepatitis due to infection with Chlamydia trachomatis and Epstein-Barr virus in a young woman. Diagnosis by polymerase chain reaction from peritoneal tissue.

Perihepatitis with ascites is a well-known presentation of pelvic inflammatory disease due to Chlamydia trachomatis. Diagnosis is based on the presence of IgM antibodies or positive culture from cervical samples or ascites. We describe a young woman with a concomitant infection of C. trachomatis and Epstein-Barr virus presenting with fever, hepatitis and ascites. Cultures remained negative and other tests were--at first--inconclusive. Diagnosis was ultimately established by polymerase chain reaction (PCR) and RNA in situ hybridisation of peritoneal tissue obtained at laparoscopy.

Comparison of the effect of temperature on kidney cortex mitochondria from rabbit, dog, pig, and human: Arrhenius plots of ADP-stimulated respiration.

The effect of temperature on the rate of ADP-stimulated respiration of mitochondria from dog, rabbit, pig, and human kidney cortex mitochondria was plotted according to the Arrhenius relationship. The temperature at which the plot demonstrated a break was at 15 degrees C for mitochondria from dog, pig, and human kidneys. The discontinuity occurred at 10 degrees C or less for mitochondria from rabbit kidneys. This difference suggests that mitochondria from rabbit kidneys undergo a lipid-phase transition at lower temperatures than for other species commonly used in experimental renal preservation. The implications of this difference suggest caution in using results obtained with rabbit kidneys for comparison to results obtained from hypothermic renal preservation of other species kidneys. Apparent fluidization of dog kidney mitochondrial membranes with adamantine abolished the discontinuity in the Arrhenius plot.