Summary. Investigations of the haemostatic functions in three patients with the Wiskott‐Aldrich syndrome are presented. All patients had severe thrombocytopenia and prolonged bleeding times. The platelets had abnormal morphology with reduced size and variations of shape. Electron microscopy revealed ultrastructural abnormalities with a reduced number of organelles, and many of the platelets contained large numbers of tubules. Platelet electrophoretic mobility in citrated plasma was not reduced by collagen, and platelet aggregation by collagen and ADP was deficient. Biochemical studies revealed a lack of the storage pool of adenine nucleotides. Platelet adhesiveness in vitro in whole blood was reduced. Platelet factor‐3 release by kaolin, ADP and freezing and thawing was normal in one and reduced in another of the patients. Platelet survival studies showed a normal survival time of normal donor platelets in one patient, while autologous platelets had a markedly reduced survival time in two of the patients. The bone marrow contained normal numbers of megakaryocytes. By electron microscopy of the bone marrow, blood platelets were found to be phagocytosed by macrophages and reticulum cells. The main cause of the thrombocytopenia is most probably incrcased peripheral destruction of platelets. It is suggested that the qualitatively defective platelets are recognized in the reticulo‐endothelial system as foreign particles and phagocytosed.

Summary1. Blood platelet aggregation was induced by addition of saline “extract” of tendons to citrated PRP (aggregation time 50—60 seconds). The aggregates were sedimented by centrifugation, and aggregating activity was demonstrated in the supernatant (termed supernat. A) with an aggregation time of about 15 seconds.2. It was demonstrated that the activity of the supernatant was due to ADP released from the platelets.3. The conclusion is drawn that the aggregation reaction initiated by the tendon “extract” can be divided into two steps: 1. release of ADP from the platelets, 2. aggregation caused by released ADP.4. The observations are discussed in relation to the formation of platelet plugs in vivo.

The dura-arachnoid junction is examined in normal animals and in animals subjected to subdural infusion of blood immediately prior to death, simulating acute subdural hemorrhages. The Norwegian landrace pig is used as the experimental animal. Horseradish peroxidase (HRP) has been added to the injected blood to serve as a macromolecular tracer. The material is studied by light and electron microscopy. Special attention is given to the level of the induced subdural cleavage plane, the total distribution of the infused blood, and the natural sites of drainage. The dura-arachnoid junction, identified here as the subdural compartment (the dural border layer of others), consists of an avascular tissue with flake-like, relatively electron-lucent cells stacked upon each other in several layers with narrow intercellular clefts. Under normal conditions there is no evidence of a so-called "subdural space." When under the present experimental conditions bleeding takes place into this cellular tissue, it splits without any particular, predestined cleavage plane, although most often close to the fibrous matter of the dura. The bleeding extends throughout the cerebral and spinal parts of the compartment and also along the spinal nerve roots. Contamination of the subarachnoid space occurs only in some cases subjected to large infusions and apparently only at spinal levels. The HRP diffuses into the dura, but does not traverse the arachnoid barrier layer.

Compstatin, a newly described C3-binding peptide, inhibits complement activation by blocking C3 convertase-mediated cleavage of C3. As the complement activation is an essential part of the rejection reaction, we evaluated the ability of Compstatin to delay or prevent hyperacute rejection in an ex vivo xenograft model. Pig kidneys were perfused with fresh human blood containing either Compstatin (n=6) or a control agent (n=6). Graft survival and activation of complement, leukocytes and platelets both in the fluid-phase and in the tissue were examined. The survival of the Compstatin-perfused kidneys (median, 380 min) was significantly (P=0.0036) longer than that of the controls (median, 90 min). The classical complement pathway (C1rs-C1inhibitor and C4bc) was significantly and equally activated in both groups during the first 60 min. C3 activation products increased fivefold and terminal complement complex eightfold in the control group, but no increase occurred in the Compstatin group during this period. Immunohistochemistry showed less C3 and fibrin deposition and immune electron microscopy showed less terminal SC5b-9 complement complex deposition in the Compstatin group. A significant change in total white cells, neutrophils, myeloperoxidase, and expression of the surface activation markers CD11b (CR3) and CD35 (CR1) and CD62L (L-selectin) was observed in both groups. Leukocyte activation was lower in the Compstatin group but the difference was not statistically significant. There were no differences in platelet counts, thrombospondin, soluble P-selectin or beta-thromboglobulin between the groups. We conclude that Compstatin prolongs graft survival and suggest that it may be a useful agent for attenuating hyperacute rejection by inhibiting C3 and thus terminal complement pathway activation.

H. pylori was shown to give rise to two colony forms: at normal pH the population was dominated by L colonies. One strain was chosen for further studies. Bacteria from L colonies retained VacA toxin and urease, did not invade or adhere to epithelial cells, and contained normal quantities of phosphatidylethanolamine. In a small frequency, spontaneous S colonies were formed. Bacteria from these colonies released VacA and urease, adhered to and invaded epithelial cells and contained increased amounts of lysophosphatidyl ethanolamine and phosphatidyl serine. After addition of HCl to the culture medium (pH6), almost only S colonies were formed. The results demonstrate that environmental factors, such as HCl, can change the bacterial cell wall, and thereby enhance expression of virulence factors of H. pylori in vitro. A similar in vivo variation would have implications for our understanding of the interaction between HCl secretion in the gastric mucosa and H. pylori in the development of peptic ulcer disease.

Generalized argyria was precipitated in a patient by treating gingival erosions with a solution of silver nitrate for several months. High silver concentrations were measured in skin biopsies. treatment with penicillamine did not increase the urinary silver excretion, indicating that silver is deposited in tissues in a chemically stable and apparently inert form. Electron microscopy showed that in the kidney, silver was deposited mainly in the basal membranes as electron-dense particles. These particles were studied by using X-ray emission spectrometry and electron diffraction. the particles consisted of Ag2Se in the low temperature orthorhombic alfaform. The lattice parameters are: a = 0.433 nm, b = 0.693 nm and c = 0.784 nm. This selenide complex seems to be remarkably non-toxic, since the renal function of the patient was unaffected and only negligible reactive changes were observed in kidney biopsies.

Image analysis quantitation of interstitial collagen with Sirius red corresponded well to light microscopic semiquantitative assessment of interstitial fibrosis. In prediction of long-term graft function, the semiquantitative method was superior, indicating that accumulation of matrix molecules other than fibrillary collagens, oedema and inflammation are also important in graft prognosis.

SummaryRabbit blood platelet aggregates were produced in vitro by addition of rabbit tendon “extract” to citrated platelet-rich plasma. The aggregating activity was not due to presence of adenosine diphosphate in the “extracts” and was unrelated to blood coagulation. The aggregating principle was destroyed by heating of the “extract” to 60° C for 15 minutes or by incubation with collagenase for 1 hour at 37° C. The aggregating effect was associated with particles which were sedi- mented by centrifugation for 30 minutes at 10,000 G. By means of electron microscopy the particles were identified as cross-striated collagen fibrils.