Suppression of cold ischemic injury in stored kidneys by the antimicrobial peptide bactenecin

BACKGROUND: Cold ischemic injury plays an important role in short- and long-term function of kidneys after transplant. Antimicrobial peptides have not previously been studied for their impact on cold ischemia in transplanted kidneys. METHODS: Bactenecin (L- and D-forms) was added to University of Wisconsin (UW) preservation solution for 3-day cold storage of dog kidneys. Effects on membrane permeability were studied in synthetic liposomes and in kidney cortex tissue slices. The role of bactenecin as a tissue mitogen and direct cytoskeletal stabilizer were studied with cultured cells and in vitro. RESULTS: Bactenecin (both L- and D- forms) resulted in significant decreases in postoperative serum creatinine and time required for return of creatinine to the normal range showing the effect was independent of chirality. Bactenecin permeabilized synthetic liposomes and altered kidney cortex tissue slice membrane permeability characteristics, irrespective of chirality. Neither did bactenecin act as a mitogen for either primary renal tubule or Madin-Darby canine kidney (MDCK) cells stored in UW solution, nor did it appear to directly affect cytoskeletal dynamics. CONCLUSIONS: These results show that the antimicrobial peptide bactenecin can improve the quality of static cold storage of kidneys. The mechanism of its action is independent of receptor binding and does not appear to involve either an effect on the cytoskeleton or via activity as a mitogen. Current evidence best supports the hypothesis that bactenecin protects against cold ischemic injury by a controlled permeabilization of the membranes of the kidney during cold storage.     

Human stored blood inositol phospholipids.

The total amount of phospholipids of the stored blood erythrocytes does not change during the first week of storage. After the second and the third week of storage the changes are only insignificant. During the fourth week this amount decreases by 25%. Detailed analysis of inositol phospholipids shows that balance of the phosphorylation-dephosphorylation cycle is shifted towards dephosphorylation. The decrease in phosphatidylinositol-4,5-bisphosphate (PtdIns-4,5-P2) is accompanied by the increase in phosphatidylinositol-4-phosphate (PtdIns-4-P) and phosphatidylinositol (PtdIns). The total amount of inositol phospholipids does not change. The increase in PtdIns-4-P is accompanied by the appearance of echinoidal forms of erythrocytes. The results of this study suggest that PtdIns-4-P can be considered as one of the important factors which determine the shape of erythrocytes.     

Potassium and blood vessels.

Intraarterial infusion of potassium causes vasodilation whereas reduction of the potassium concentration in the inflowing blood causes vasoconstriction. These responses have always been puzzling since the predicted effects from the Nernst equation are just the reverse. Recent evidence suggests that they result form effects on the activity of sarcolemmal Na, K ATPase in the vascular smooth muscle cell which influence the electrogenic NaK pump. The possible relevance of these findings is discussed.     

Fructose formation in stored blood

At the high glucose concentrations used in the collection and storage of donor blood the activity of the fructose-forming polyol pathway (Reaction I and II) could act to deplete NADPH and thus GSH, thereby exposing the cells to oxidative stress. Fructose levels were found to be high in red cells and the supernatant plasma of blood collected into CP2D, which contains 258 mM glucose. Elevated fructose was not produced by the polyol pathway, but was formed by the autoclaving process. A high fructose concentration sufficient to account for the fructose in donor red cells was also found in the CP2D anticoagulant and in samples of autoclaved glucose.     

Superficial anastomoses of blood vessels in the human heart.

We investigated superficial interarterial, intra-arterial, arterio-venous and veno-venous anastomoses in 100 hearts from men and women taken at random who died at 20-85 years of age. By means of careful dissection and injection into the blood vessels of a differently stained 12% solution of vinilit in acetone, we studied the number and localization of the anastomoses and their appearance. Interarterial anastomoses, i.e. anastomoses between the ramification of the left and the right coronary artery, were detected in 33 hearts. Intra-arterial anastomoses, i.e. anastomoses between the ramifications of one and the same artery, were seen in 5 hearts. Arterio-venous anastomoses were found in 39 hearts. Veno-venous anostomoses were found in 49 hearts. The veno-venous anastomoses display a great many variations, being commonly the regular finding in human veins.     

Extra-organic lymphatic vessels and regional lymph nodes of the kidneys in dogs

An anatomical investigation of extraorganic lymphatic vessels and regional renal lymph nodes has been performed in 70 dogs. The regional lymph nodes in the right kidney are stated to be quantitatively constant, as well as cranial and caudal lateroaortal lymph nodes in the left kidney in regard to the middle left lateroaortal nodes, that get lymph from the left kidney parenchyma. One middle left lateroaortal lymph node is found in 47 animals examined, two lymph nodes--in 17 animals. In 6 cases a lymphatic vessel, that gets lymph from the renal parenchyma and independently runs into the cistern of the thoracic duct is found for the first time. The variant revealed is an exception from the rule known in lymphology: lymph in its way from periphery to the central collector runs, at least, through one lymph node.     

The composition of the expressed sap from cold stored potatoes

Analysis of the chemical composition of expressed sap from potatoes stored at low temperatures (2°) indicates that an increased amount of inorganic phosphate is present in the juice from the cytoplasm and vacuole.     

Time course of the age-related alterations in stored blood

The extent and time course of the impairments occurring in whole blood and erythrocyte cells stored under blood bank conditions were studied by monitoring the reduction of MAL-6 spin label added to the media containing whole blood or erythrocyte cells using electron spin resonance (ESR) technique. Impairments forming in the erythrocyte cells incubated for various times at 37 degrees C were also studied. Erythrocyte cells were found to undergo changes during the storage or incubation, leading to fast decay of MAL-6 spin labels signal height. The extent of the changes depends on storage or incubation time. However, the reduction in incubated or artificially aged erythrocyte (AAE) cells was faster than the reduction in whole blood (WB) and aged erythrocyte (AE) cells stored under blood bank conditions. Two exponential curves attributed to the liquid and cellular parts of a given samples were found to be described best in the reduction of MAL-6 spin label in WB, AE and AAE.     

The fine structure of cephalopod blood vessels II. The vessels of the nervous system

Summary Blood vessels of the perioesophageal nerve ganglia (brain) of Octopus vulgaris and the stellate ganglia of Sepia officinalis are described. The vessels have an incomplete endothelium, a complete basement membrane and a complete investment of pericytes. The pericytes are joined by specialised membrane junctions but these are not tight junctions. The main type of neuron/vessel arrangement is one where there is a collagen-filled space between the pericytes and the surrounding glial cells. Axons or neurons are sometimes applied directly to the vessel pericytes and in the neuropil, pericytes contact glial cells that ensheath bundles of axons. Blood spaces between neurons are also present.We would like thank Professor J. Z. Young and Dr. E. G. Gray for encouragement and advice, Mrs. Jane Astafiev for drawing Fig. 11, Mr. S. Waterman for photographic assistance and Miss Cheryl Martin for secretarial and other assistance.