THE BLOCKAGE OF AXOPLASMIC FLOW OF PROTEINS BY COLCHICINE AND CYTOCHALASINS A AND B

Abstract— Colchichine blocks axoplasmic flow of proteins in chicken sciatic nerve. The slow component is more effectively blocked than the fast. The colchicine effect on slow flow is independent of the time delay between colchicine injection and that of the [¹⁴C]-leucine used to measure flow, over a period extending from 2 h after to 9 days before the leucine. It is still effective, but to a lesser degree, after a period of 27 days. There is little effect on the fast component when the colchicine is administered after leucine. When given before leucine the effect is not pronounced up to a time interval of 1 day. Maximum blockage was obtained with longer intervals of up to 27 days. These results are discussed in relation to the involvement of the microtubules in both slow flow and rapid flow.
The effect of both cytochalasins A and B on slow and rapid protein flow has also been studied. Neither drug had any significant effect on slow flow. Cytochalasin A reduced the amount of protein flowing at the rapid rate to a small extent when administered 2 h before [¹⁴C]leucine. Cytochalasin B also caused a similar reduction and this effect was independent of dose over the ranges of doses used. The cytochalasin B diminished the incorporation of amino acid into protein in the spinal cord and it has been concluded that these results are due to a membrane effect which reduces uptake of amino acid rather than a direct effect on neurofilaments in the axons.     

Activated lymphocytes trigger lymphoblast extravasation.

Intraperitoneal stimulation of adoptively sensitized rats with bacterial antigen promotes the localization of lymphoblasts at the site of antigen deposition. Lymphoblast extravasation activity (LEA) is generated only when specifically immune donor lymphocytes and the recipients of these cells share at least on Ag-B haplotype. However, if the specificity criteria for its formation are satisfied, LEA promotes the local development of lymphoblasts of all available specificities and irrespective of their Ag-B genotype. Allogeneic lymphoblasts do not participate actively in the delayed inflammatory reaction even when they are passively recruited into exudates. The results suggest that LEA is a T cell-derived mediator that amplifies the delayed type hypersensitivity reaction by directing recently activated lymphocytes into lesions.     

Methimazole and generation of oxygen radicals by monocytes: potential role in immunosuppression.

A study was conducted investigating the possibility that the immunosuppressive action of methimazole (the active metabolite of the antithyroid drug carbimazole) might be due to an effect on the production of oxygen radicals by monocytes. Techniques comprised measurement of luminol dependent chemoluminescence in monocytes and a spectrophotometric assay for production of hydrogen peroxide. The results showed definite inhibition of formation of oxygen radicals by resting and stimulated monocytes, which may explain the immunosuppressive action of the drug in Graves'' disease. The findings also suggest that the formation of oxygen radicals and the initiation of the immune response may be closely related.     

Identification of ARAP3, a novel PI3K effector regulating both Arf and Rho GTPases, by selective capture on phosphoinositide affinity matrices.

We show that matrices carrying the tethered homologs of natural phosphoinositides can be used to capture and display multiple phosphoinositide binding proteins in cell and tissue extracts. We present the mass spectrometric identification of over 20 proteins isolated by this method, mostly from leukocyte extracts: they include known and novel proteins with established phosphoinositide binding domains and also known proteins with surprising and unusual phosphoinositide binding properties. One of the novel PtdIns(3,4,5)P3 binding proteins, ARAP3, has an unusual domain structure, including five predicted PH domains. We show that it is a specific PtdIns(3,4,5)P3/PtdIns(3,4)P2-stimulated Arf6 GAP both in vitro and in vivo, and both its Arf GAP and Rho GAP domains cooperate in mediating PI3K-dependent rearrangements in the cell cytoskeleton and cell shape.     

Long-term anaesthesia with alfentanil and midazolam for lung transplantation in the dog

An anaesthetic regime was developed for lung transplantation in the dog using a continuous infusion of alfentanil and midazolam. This combination of agents provided excellent analgesia and also produced loss of consciousness. Cardiovascular stability was well maintained over a 24-h period of anaesthesia following lung transplantation. Although no animals were allowed to recover from anaesthesia in the present series, the regime described is likely to be suitable for recovery anaesthesia, particularly since both of the agents used can be reversed with specific antagonists.