peptones and Peritonitis

Fucosylated chondroitin sulfate

Topics: Animals; Anti-Inflammatory Agents; Cartilage; Chondroitin Sulfates; Circular Dichroism; Cucumaria; Disease Models, Animal; Female; Humans; Magnetic Resonance Spectroscopy; Molecular Structure; Peptones; Peritonitis; Rats; Rats, Wistar; Salmo salar; Structure-Activity Relationship; Treatment Outcome

Nuclear factor-kappa B (NF-κB) is a multipotent transcription factor that plays a pivotal role in immune reactions, inflammation, and possibly hematopoiesis as well. Mobilization of neutrophilic granulocytes during inflammation is a highly regulated process, but one that is incompletely understood. We studied the in vivo activity of NF-κB in mouse organs and cells, with a focus on bone marrow, during acute inflammation. NF-κB activity was studied in transgenic mice expressing a luciferase reporter expressed in a NF-κB activation-dependent fashion. Acute peritoneal inflammation was induced by lipopolysaccharide (LPS), the casein digest bacto-tryptone, or the insoluble polysaccharide zymosan. Organs were removed and blood, bone marrow, and peritoneal cells were separated using density gradient centrifugation. NF-κB activity in organ homogenates and cell lysates was quantified. These three inflammatory agents increased NF-κB activity to a variable extent within the inflamed peritoneal cavity, liver, and spleen, with LPS being the strongest stimulus. LPS, but not bacto-tryptone or zymosan, activated NF-κB in lung and bone marrow, the latter activity mainly observed in density fractions rich in immature bone marrow cells. NF-κB activation was prominent at 6 h after induction of peritonitis, fading at 24 h, as expected for an acute phase phenomenon. From this proof-of-principle study with luciferase reporter mice dependent on NF-κB activation, we suggest that, in steady-state mice, mobilization of bone marrow granulocytes to an inflammatory site can occur without discernible activation of NF-κB in bone marrow.

Topics: Animals; Bone Marrow; Female; Gene Expression; Genes, Reporter; Lipopolysaccharides; Liver; Luciferases; Lung; Macrophages, Peritoneal; Mice; Mice, Transgenic; Neutrophil Infiltration; Neutrophils; NF-kappa B; Peptones; Peritonitis; Spleen; Zymosan

To assess the effects of inflammation on the generation of circulating DNA from dead and dying cells, plasma DNA levels were determined in BALB/c mice, administered apoptotic or necrotic Jurkat cells following induction of peritonitis by treatment with thioglycollate (TG), peptone (PT), or sodium periodate (NaIO(4)). In mice receiving TG or NaIO(4), plasma DNA levels following intraperitoneal administration of Jurkat cells were significantly reduced compared with controls, whereas they were not affected in mice receiving PT. To determine the basis of these differences, the cellular composition of peritoneal fluids prior to the administration of the dead cells was analyzed. Among agents tested, TG administration led to the largest increase in cells, both neutrophils and monocytes. As shown by flow cytometry, the exudates contained apoptotic neutrophils and macrophages, with the highest levels in the TG-induced exudates. Analysis of DNA and caspase 3 in the fluids also showed differences. TG exudates showed increases in DNA and caspase 3, while NaIO(4)-induced exudates had an increase only in DNA. Fluid from PT-treated mice did not have increases in DNA or caspase 3. Together, these results indicate that prior inflammation can affect the generation of blood DNA from apoptotic or necrotic cells, although this effect may vary depending on the composition of the exudates with respect to cells as well as DNA.

Topics: Animals; Apoptosis; Cells; DNA; Female; Humans; Inflammation; Jurkat Cells; Mice; Mice, Inbred BALB C; Peptones; Periodic Acid; Peritoneum; Peritonitis; Thioglycolates; Time Factors

The potency of the oligosaccharides SiaLe(x), SiaLe(a), HSO(3)Le(x), and HSO(3)Le(a), their conjugates with polyacrylamide (PAA, 40 kD), and other monomeric and polymeric selectin inhibitors has been compared with that of the polysaccharide fucoidan. The following assay systems were used: 1) a 96-well assay based either on the use of recombinant E-, P-, and L-selectins or an analogous assay with natural P-selectin isolated from human platelets; 2) a platelet-based P-selectin cell assay; and 3) a rat model of peritoneal inflammation. IC(50) values for the neoglycoconjugate SiaLe(a)-PAA were 6, 40, and 85 microM for recombinant E-, P-, and L-selectins, respectively; all monomeric inhibitors were about two orders of magnitude weaker. PAA-conjugates, containing as a ligand tyrosine-O-sulfate (sTyr) in addition to one of the sialylated oligosaccharides, were the most potent synthetic blockers in vitro. Compared with fucoidan, the most potent known P- and L-selectin blocker, the bi-ligand glycoconjugate HSO(3)Le(a)-PAA-sTyr displayed similar inhibitory activity in vitro towards L-selectin and about ten times lower activity towards P-selectin. All of the tested synthetic polymers displayed a similar ability to inhibit neutrophil extravasation in the peritonitis model (in vivo) at 10 mg/kg. The data provide evidence that monomeric SiaLe(x) is considerably more effective as a selectin blocker in vivo than in vitro, whereas the opposite is true for fucoidan and the bi-ligand neoglycoconjugate HSO(3)Le(a)-PAA-sTyr.

Topics: Acrylic Resins; Acute Disease; Animals; E-Selectin; Female; Glycoconjugates; Humans; L-Selectin; Neutrophils; Oligosaccharides; P-Selectin; Peptones; Peritonitis; Polymers; Polysaccharides; Rats; Recombinant Proteins; Selectins

Neutrophil recruitment into systemic inflammatory sites in vivo is thought to be initiated by selectin-mediated endothelial adherence. The effect of fucoidan (natural sulfated polymer of L-fucose) on the selectin dependent PMN migration into rat peritoneum following the induction of inflammation by peptone injection was studied. Peritonitis was characterized by an increase in the total cell number (from 45.3 x 10(6) to 91.6 x 10(6)/rat), and by highly elevated PMN content (from 0.2% to 58%) in the rat peritoneal cavity 3 h after peptone injection. Intravenous administration of fucoidan was found to reduce, in a dose-dependent manner, neutrophil migration into peritoneum. Fucoidan in a dose as low as 0.8 mg per rat caused 96.8% reduction of neutrophil extravasation. The inhibitory effect of fucoidan was also dependent on the time intervals between the peptone and fucoidan injections. The maximal inhibitory effect of fucoidan was observed within the first 15 min after the induction of peritonitis and it was maintained at a level of 80% during 1.5 h. Administration of fucoidan 2.5 h after peptone injection had practically no effect on PMN extravasation. Since P-selectin is known to play a key role at the earlier stages of PMN extravasation, it was suggested that the inhibitory effect of fucoidan was mostly due to its interaction with P-selectin. The in vitro experiments demonstrated the high affinity of fucoidan for both isolated P-selectin and P-selectin in plasma membranes of activated platelets.

Topics: Animals; Biotinylation; Blood Platelets; CA-19-9 Antigen; Carbohydrate Metabolism; Cell Movement; Female; Gangliosides; Humans; Kinetics; Neutrophils; Oligosaccharides; P-Selectin; Peptones; Peritonitis; Polysaccharides; Rats; Rats, Wistar; Sialyl Lewis X Antigen

Topics: Animals; Body Temperature; Exudates and Transudates; Hot Temperature; Irritants; Peptones; Peritonitis; Phagocytosis; Rabbits