Ristocetin- and thrombin-induced platelet aggregation at physiological shear rates: differential roles for GPIb and GPIIb-IIIa receptor

We recently reported that washed platelets (WP) activated with ADP and expressing surface-bound vWF aggregated in flow through small tubes or in a cylindrical couette device at physiological shear rates of G = 300 s(-1)-1000 s(-1) in the absence of exogenous ligands, with GPIb-vWF partially, and activated GPIIb-IIIa totally required for the aggregation. We have now extended these studies to aggregation of platelets "activated" with ristocetin or thrombin. Washed platelet suspensions with added soluble vWF and ristocetin (0.3-0.75 mg/ml), or activated with thrombin (0.01-0.5 U/ml) but no added ligand, were sheared in a coaxial cylinder device at uniform shear rate, G = 1000 s(-1). The collision capture efficiency (alphaG) with which small aggregates form (= experimental/calculated initial rates of aggregation) was correlated with vWF platelet binding assessed by flow cytometry. The vWF-GPIb interaction was exclusively able to support ristocetin-mediated shear aggregation of metabolically active platelets, with very few vWF monomer equivalents bound per platelet (representing < or = 10 molecules of 10 million Da) required to yield high capture efficiencies (alphaG = 0.38+/-.02; n = 11), suggesting rapid and stable bond formations between vWF and GPIb. However, platelet surface-expressed vWF, generated by addition of thrombin to washed platelets, was found to mediate platelet aggregation with alphaG = 0.08+/-.01 (n = 6), surprisingly comparable to that previously reported for WP and ADP activation. Blocking the GPIIb-Illa receptor decreased alphaG by 95+/-3% (n =3), while a monoclonal antibody to the vWF site on GPIb caused a 49+/-7% (n = 8) decrease in alphaG. The partial role for GPIb thus appears to reflect a facilitative function for increasing contact time between flowing platelets, and allowing engagement of the GPIIb-IIa receptor to yield stable attachment.     

Application of polymerase chain reaction for the correlation of Salmonella serovars recovered from greyhound feces with their diet

The polymerase chain reaction was employed to correlate Salmonella serovars isolated from fecal material of greyhounds suffering from gastroenteritis with those isolated from the diet fed to the greyhounds prior to onset of diarrhea. Kennels around the Abilene, Kansas, area were contacted and supplied with materials needed to collect a portion of the diet each day. With the onset of diarrhea, the kennels were instructed to ship the fecal material and diet from the previous 10 days to the laboratory for testing. Forty-one fecal samples and corresponding diets were screened for Salmonella, Clostridium perfringens, Campylobacter jejuni, Staphylococcus aureus, Staphylococcus intermedius, and pathogenic (piliated) Escherichia coli by direct culture using standard procedures. The fecal material was also screened for coronavirus and parvovirus using electron microscopy. Thirty-five "normal" fecal samples were screened for all of the above mentioned microorganisms as a control. In addition, the fecal material was screened for E. coli verotoxins I and II and clostridial enterotoxins. A total of 61 Salmonella isolates were recovered from the 41 samples of feces and diet submitted for testing; 31 were recovered from the feces and 30 from the diet. Four Salmonella isolates were recovered from the normal fecal samples. Results obtained by PCR, plasmid profiles, antigenic analysis, and antibiogram profiles indicated that 16 of the 31 isolates recovered from the fecal material were the same strain as that recovered from the diet.     

Diagnostic value of determining hemoglobin in cerebrospinal fluid in acute cerebral circulatory disorders

The results of hemoglobin determination in the CSF by the method of fluorescence microscopy in 195 patients speaks in favor of relative diagnostical importance of hemoglobin in the supra fall-out CSF for establishing the character of the stroke. The following circumstances testify to this fact: 1) in an artificial admixture of blood in the CSF in some cases there may be hemolysis of "passing" erythrocytes; 2) in a genuine admixture of blood, tests of hemoglobin in the centrifugate of the CSF may be negative since in some cases the transition of hemoglobin into bilirubin in the subarachmoid space occurs more rapidly than the hemolysis of erythrocytes. Thus, the hemoglobin does not accumulate in the liquid in quantities, sufficient enough to be measured by the existing methods; 3) in a large amount of artificial blood admixture, even without a hemolysis of "passing" erythrocytes the hemoglobin tests in the centrifugate of the CSF may be positive at the expense of hemoglobin of the blood plasma.     

Production of anti-erythrocyte antibodies by leukemic and nonleukemic B cells in chronic lymphocytic leukemia patients

We have assessed the specificity of antibodies from the leukemic B cells of five patients with both chronic lymphocytic leukemia and autoimmune hemolytic anemia (CLL-AHA). Leukemic cells from one patient displayed surface immunoglobulin with heavy and light chain isotypes identical to that of the patient's anti-red blood cell (RBC) antibodies, and the leukemic cells secreted antibodies in vitro with anti-RBC activity. However, in the remaining patients, the leukemic cells displayed surface immunoglobulin with light chain isotypes different from that of the patient's anti-RBC antibodies and secreted antibodies in vitro with no detectable anti-RBC activity. Thus, there are two distinct classes of CLL-AHA patients, differentiated by the presence or absence of an anti-RBC antibody-producing leukemic B cell clone. The apparent heterogeneity in the source of pathogenic anti-RBC antibodies may impact the treatment response of the two classes of CLL-AHA patients.     

Intracerebral haemorrhage in a young adult: consider amphetamine abuse

We present the case of a young female who suffered a massive intracerebral bleed following the ingestion of a small quantity of amphetamine (speed). Physicians should be aware that amphetamine abuse can lead to cerebrovascular events in young adults.     

Reduced speed of sound in tibial bone of haemodialysed patients: association with serum PTH level

BACKGROUND: In end-stage renal disease, average bone mineral density has been reported to be normal or only modestly reduced, more so in the cortical bone. The purpose of the present study was to explore the potential use of quantitative ultrasound, a method reflecting both quantitative and qualitative properties of bone, in assessing bone status in patients on maintenance haemodialysis. METHODS: We studied 71 patients (age 17-81 years, time on dialysis 0-18 years). The speed of sound waves (tSOS; m/s) propagating along the cortical bone has been determined at the tibial shaft. tSOS results were expressed as Z scores, i.e. units of standard deviations from age- and sex-matched normal mean values, and correlated with relevant clinical and biochemical variables. RESULTS: SOS Z score averaged -2. 0 (range -6.8 to 0.6; P<0.001) and was negative in 93% of the patients. Significant inverse correlations were found between SOS Z score and both time on dialysis (r=-0.52; P<0.0001) and serum PTH (r=-0.39; P=0.0002). Markedly reduced SOS Z score, below -2, was found in 80% of the patients whose PTH levels exceeded 34 pmol/l (five times the upper normal limit), compared with 43% of the patients whose PTH levels were below 34 pmol/l(P=0.04). Compared to patients without bone pain (n=51), subjects with bone pain (n=20) had somewhat lower SOS Z scores -2.5+/-2.0 versus -1.8+/-1.4; P=0. 08), but this could be accounted for by longer time on dialysis. CONCLUSIONS: tSOS is substantially reduced in the majority of haemodialysed patients and is related to time on dialysis and serum PTH level. The clinical value of this novel method needs further exploration.     

Actinoplanic acids A and B as novel inhibitors of farnesyl-protein transferase

Actinoplanic acids A and B are macrocyclic polycarboxylic acids that are potent reversible inhibitors of farnesyl-protein transferase. Actinoplanic acids A and B were isolated from Actinoplanes sp. MA 7066 while actinoplanic acid B was isolated from both MA 7066 and Streptomyces sp. MA 7099. Actinoplanic acids A and B are competitive with respect to farnesyl diphosphate and are selective inhibitors of farnesyl-protein transferase because they do not inhibit geranylgeranyl-protein transferase type 1 or squalene synthase. MA 7066 is believed to be a novel species of actinomycetes while MA 7099 is believed to be a novel strain of Streptomyces violaceusniger on the basis of morphological, biochemical and chemotaxonomic characteristics as well as its production of actinoplanic acids.     

Improvement of some pharmaceutical properties of drugs by cyclodextrin complexation. 4. Chlorpromazine hydrochloride

This study is a retrospective review of admissions, discharge records and blood culture results of neonates admitted to the Neonatal Intensive Care Unit of Korle Bu Teaching Hospital in Accra, from the first of January 1991 to the 31st of December 1992. During this two year period there were 443 positive blood cultures. Ninety percent of the blood cultures were from babies born in Korle Bu Teaching Hospital, thus making the incidence of neonatal bacteraemia 22.2 per 1000 live births. The overall mortality rate was 37.2%. Gram negative bacteria accounted for 70.9% and Gram positive bacteria for 29.1% of all neonatal bacteraemia. The most common isolates were Enterobacter species 29.6%; Streptococcus faecalis 14.4%; Staphylococcus aureus 10.8%; Acinetobacter species 9.5%; Klebsiella species 9% and Escherichia coli 8.8%. It is concluded that the incidence of neonatal bacterial sepsis is high in our hospital and is associated with a very high mortality rate. There is thus an urgent need to institute appropriate preventive and therapeutic measures.     

Accidental hypothermia: incidence, risk factors and clinical course of patients admitted to hospital

This study was initiated to identify the incidence, risk factors and outcome predictors of patients admitted to hospital in the Netherlands because of accidental hypothermia. Information about these patients was available for study through the National Health Care Data Bank. Between 1987 and 1990, 612 accidental hypothermic patients were admitted: 185 hypothermic patients also suffered from submersion (HYPSUBS), but this was not the case in the remaining 427 patients (HYPNOTSUBS). Patients in the HYPNOTSUBS group were older (average age 55.2 years versus 38.9 years; p < 0.001), remained longer in hospital (average 20.8 days versus 9.2 days; p < 0.001) and had a higher death rate than those in the HYPSUBS group (16.9% versus 5.9%; p < 0.001). In HYPNOTSUBS, increasing age correlated with increases in the length of hospital stay and death rate. This relationship was not found in HYPSUBS. Trauma was the major associated problem in both groups; these patients had the highest death rate (22.8% versus 16.7%; not significant). Death occurred within 2 days in 54% of HYPNOTSUBS non-survivors and 73% of HYPSUB non-survivors. HYPNOTSUBS admitted to university hospitals showed a lower death rate (5.9%) compared with HYPNOTSUBS admitted to non-university hospitals with less than 400 beds (13.4%) or more than 400 beds (21.7%). In contrast, the death rate in HYPSUB was higher in university hospitals (14.3%) than in non-university hospitals with less than 400 beds (5.2%) or more than 400 beds (3.6%). We observed that the incidence of accidental hypothermia is low at 1.1 per 100,000 inhabitants per year. We concluded that HYPNOTSUBS and HYPSUB are different groups of patients with respect to demographic data, risk factors and prognostic factors. Old age is an important unfavourable prognostic factor in HYPNOTSUB but not in HYPSUB. Hypothermia with trauma is an unfavourable combination in both groups. Almost half of the HYPNOTSUBS non-survivors died after more than 2 days. Because body temperature will have returned to normal by then, this must be the result of late complications. Most HYPSUB non-survivors died during the first 2 days, probably as a direct result of the submersion injury.