Effect of a hexosylceramide fraction of the hemodialysate Solcoseryl on wound-healing angiogenesis.

Rabbit ear chambers were used to investigate the effects of a locally applied hexosylceramide fraction (Hex-Cer) of the hemodialysate Solcoseryl on wound-healing angiogenesis. The transparent methacrylate ear chambers were inserted under full aseptic precautions. Immediately after implantation the chambers in control rabbits were filled with physiological saline while the chambers in experimental rabbits were injected with a solution containing either 0.3 or 2.5 micrograms/ml of Hex-Cer, respectively. On the second postoperative day the chambers were reinjected with the corresponding solutions. Thereafter the chambers were examined three times weekly. The onset and rate of the neovascular response were measured by using a standard dissecting microscope equipped with a camera. Vessel growth in ear chambers treated with Hex-Cer at the lower concentration showed very little difference from that seen in control chambers; in both groups the first appearance of new capillaries occurred on an average of 18 days after implantation, and the chambers became fully filled with vessels by the 30th day. However, in chambers treated with Hex-Cer at the higher concentration, the first invasion of capillaries was detected 3 days earlier than in controls (P less than 0.05). Correspondingly, in the chambers treated with Hex-Cer complete vascularization was achieved 7 days earlier than in the control chambers (P less than 0.01). It is concluded that locally applied Hex-Cer exerted an accelerating effect on wound-healing angiogenesis.

Effect of a hexosylceramide fraction of the hemodialysate Solcoseryl on experimental granulation tissue.

This work was carried out to study the effects of a hexosylceramide fraction (Hex-Cer) of the hemodialysate Solcoseryl on developing granulation tissue in rats. Subcutaneously implanted cylindrical hollow viscose-cellulose sponges were used as an inductive matrix for the growth of granulation tissue. In the control animals, the implants were treated daily by withdrawing 1 ml of wound fluid from the central dead space of the implant and by injecting back 1 ml of physiological saline. In the experimental animals, the aspirated wound fluid was replaced by a corresponding volume of a solution containing 0.08 microgram of Hex-Cer in physiological saline. Analyses of the wound fluid and granulation tissue were carried out 4, 10 and 21 days after the implantation. Statistically significant increases in the mean amounts of granulation tissue DNA and RNA were observed in the Hex-Cer group as compared with the controls, indicating an augmented cellularity. Concurrently, the mean amounts of collagen hydroxyproline in the Hex-Cer group were significantly higher than the respective control values. Similarly, on days 4 and 10, the amounts of uronic acids were higher in the Hex-Cer group than in the controls, reflecting an enhanced accumulation of glycosaminoglycans. The tissue hemoglobin reflecting the degree of vascularization rose gradually as the healing progressed, the mean amounts being generally higher in the Hex-Cer group than in the controls. Wound fluid pO2, pCO2 and pH as well as wound fluid hemoglobin and lactate concentrations showed no essential differences between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Induction of antimyelin and antioligodendrocyte antibodies by vaccinia virus. An experimental study in the mouse.

The appearance of serum antibodies binding to specific brain antigens was monitored in mice inoculated intracerebrally with a dermotropic or a neurotropic strain of vaccinia virus. Antibodies were measured with a binding assay using [125I]protein. A. Inoculation with the neurotropic strain caused an induction of serum antibodies binding to the myelin membrane, the myelin basic protein and oligodendrocytes while no induction of binding antibodies to neurons was observed. The dermotropic strain failed to elicit the formation of binding antibodies to brain antigens.

Actin filament bundles in vaccinia virus infected fibroblasts.

When monolayer cultures of mouse 3T3 cells were infected with vaccinia virus, a rapid decrease of microfilament bundles was observed. The major disappearance of the cellular microfilaments occurred within the first hour after vaccinia virus infection, which is the period of uncoating of the virus core and early viral protein synthesis. The microfilament bundles of infected cells were significantly preserved in the presence of puromycin which inhibits viral protein synthesis.

Strain and host-cell dependence of vaccinia virus proteins.

The neurotropic strain of vaccinia virus (WR) and the dermotropic strain of vaccinia virus (Elstree) which replicated on mouse L-fibroblasts (a cell line of dermotropic origin) were highly purified, dissociated and chemically labelled by [125I]iodine. The proteins of the two virus strains were compared by two-dimensional gel electrophoresis in the nanogram range, patterned after a modified procedure of O'Farrell. A total number of 113 protein spots were resolved in the radioautograms. Vaccinia virus Elstree contained five polypeptides with same molecular weights but different isoelectric points and three polypeptides with different molecular weights and different isoelectric points as compared with vaccinia virus WR. Vaccinia virus WR which replicated on rat adrenal pheochromocytoma PC12 (a cell line of neurotropic origin) contained four additional proteins as compared with the same strain which replicated on the mouse L-fibroblasts.

Phosphorylation of myelin basic protein by vaccinia virus.

Vaccinia virus phosphorylates myelin basic protein in the myelin membrane in vitro. In the presence of vaccinia virus cores maximally 1.5 mol and in the presence of intact virus 0.7 mol phosphate residues were incorporated into 1 mol of myelin basic protein in the myelin membrane. The peptides of myelin basic protein which were phosphorylated by the vaccinia virus kinase were clearly all different from the peptides which were phosphorylated by the endogenous myelin protein kinase. The viral modification of the encephalitogenic protein and its significance to immunological events is discussed.

Mechanisms in the pathogenesis of post-infectious vaccinia virus encephalomyelitis in the mouse.

After intracranial replication of a neurotropic strain of vaccinia in mouse brain, analysis of the purified virus preparation reveals the presence of at least one host protein on the virus which was identified as the myelin basic protein. Vaccinia virus Elstree, a dermotropic virus may substitute for complete Freund's adjuvant (CFA) in inducing experimental allergic encephalomyelitis (EAE). Guinea pigs challenged with virus-myelin emulsions without CFA developed clinical and histological signs of EAE.