Structure and Anti-Inflammatory Activity of a New Unusual Fucosylated Chondroitin Sulfate from Cucumaria djakonovi.

Fucosylated chondroitin sulfate CD was isolated from the sea cucumber Cucumaria djakonovi collected from the Avachinsky Gulf of the eastern coast of Kamchatka. Structural characterization of CD was performed using a series of non-destructive NMR spectroscopic procedures. The polysaccharide was shown to contain a chondroitin core [→3)-ß-d-GalNAc-(1→4)-ß-d-GlcA-(1→]n where about 60% of GlcA residues were 3-O-fucosylated, while another part of GlcA units did not contain any substituents. The presence of unsubstituted both at O-2 and O-3 glucuronic acid residues in a structure of holothurian chondroitin sulfate is unusual and has not been reported previously. Three different fucosyl branches Fucp2S4S, Fucp3S4S and Fucp4S were found in the ratio of 2:1:1. The GalNAc units were mono- or disulfated at positions 4 and 6. Anti-inflammatory activity of CD was assessed on a model of acute peritoneal inflammation in rats. About 45% inhibition was found for CD, while a structurally related linear chondroitin sulfate SS from cartilage of the fish Salmo salar demonstrated only 31% inhibition, indicating that the presence of sulfated fucosyl branches is essential for anti-inflammatory effect of chondroitin sulfates of marine origin.

Lethal doses of oxbile, peptones and thiosulfate-citrate-bile-sucrose agar (TCBS) for Acanthaster planci; exploring alternative population control options.

Effective control of outbreaks of Acanthaster planci represents the most immediate and practical intervention to reverse sustained declines in coral cover on reefs in the Indo-Pacific. This study explored the minimum doses of oxbile, oxgall, and thiosulfate-citrate-bile-sucrose agar (TCBS) that result in reliable and comprehensive mortality when injected into adult A. planci. The minimum doses required to induce 100% mortality among starfish (n=10) were 4 g l(-1) of oxbile, 8 g l(-1) of oxgall and 22 g l(-1) of TCBS. Moreover, there was no evidence of unintended side effects for other coral reef organisms (e.g., scleractinian corals, echinoderms and fishes) when using oxbile, oxgall, or TCBS at minimum doses. The effectiveness of peptones in killing crown-of-thorns starfish was also tested, but inconsistency in the results revealed that these proteins are unreliable.

Mononuclear phagocytic cells in peritoneal exudates of guinea pigs: a comparison of inducing agents.

Peritoneal macrophages are used for immunologic assessment of the lymphokine, migration inhibitory factor (MIF). For this purpose, these cells are induced intraperitoneally in animals with inflammatory agents such as mineral oil (MO) and peptone water (PW). Purpose of the present study was two-fold: To assess the changes in biologic properties of guinea pig macrophages induced peritoneally with MO or PW as compared to control cells after administration of normal saline (NS); and to examine the suitability of induced macrophages to respond to MIF in vitro. Peritoneal exudate cells were harvested from guinea pigs 7 days following intraperitoneal injection of MO, PW or NS. They were enumerated in hemocytometers, their differential counts determined by Wright's stain and morphologic characteristics assessed by scanning electron microscopy. Functional activation of peritoneal macrophages was determined by lysosomal enzyme functions, as well as by yeast phagocytosis in vitro. Random cell migration and responses to migration inhibitory factor (MIF) were determined in Mackaness chambers. Mineral oil injection resulted in significantly higher yield of peritoneal macrophages. Greater than 70% of peritoneal exudate cells were macrophages in all three groups. Spread out structures and ruffled borders were seen in electron micrographs of MO induced cells. Such structures were less evident in PW induced cells and were absent in controls. Acid phosphatase (ACP) and beta-N-acetylglucosaminidase (B-NAG) activities as well as yeast phagocytosis significantly increased in MO and PW induced cells. Random migration and responses to MIF in Mackaness chambers remained comparable in the three experimental groups.

Avian peritoneal exudate cells: a comparison of stimulation protocols.

The influence of different stimulation protocols on the induction of peritoneal exudate cells (PECs) and adherent cells, and on the percentage of different adherent cell types was examined in the chicken and Japanese quail. The results suggest that different protocols may be selected to maximize isolation of specific PEC populations for immunological studies. In the chicken, starch, peptone, glycogen, and Sephadex G-40 were all equally effective and superior to saline in generating PECs. While a single injection of Sephadex produced the highest yield of adherent cells with a maximum percentage of macrophages, repeated injections of Sephadex led to dramatic increases in non-adherent PECs (lymphocytes). In contrast, a single injection of starch was optimum for generating non-macrophage adherent cells (primarily heterophils). Since responses of the Japanese quail to stimulation with starch and saline were similar to those observed for the chicken, it is suggested that these protocols may be generally applicable for use with avian species.