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Biomedical subjects

J H Rand

Publications and source records attributed to J H Rand.

At least 55 records · Page 3Linked to original sources
PubMed1994

von Willebrand factor binding to collagen in patients with end-stage renal disease.

Bleeding abnormalities are common in patients with end-stage renal disease (ESRD). Although von Willebrand factor (vWF) abnormalities have been suspected in patients with ESRD, none have been clearly defined. Because vWF function is related to its collagen-binding capacity, we investigated whether this parameter might be altered in patients with ESRD. We measured vWF binding to type III collagen and levels of vWF antigen and ristocetin cofactor in 20 patients undergoing hemodialysis before and after routine hemodialysis sessions, in 10 patients with ESRD who had not previously undergone dialysis, and in 22 healthy, nonsmoking persons who served as controls. We found significant increases of vWF antigen levels in all patients with ESRD (undialyzed: mean, 6.4 +/- 3.0 U/ml, p < 0.001; before dialysis: mean, 5.1 +/- 4.0 U/ml, p < 0.001; after dialysis: mean, 4.8 +/- 3.4 U/ml, versus 0.81 +/- 0.26 U/ml in controls, p < 0.001). The ristocetin cofactor levels were increased in the patients who had not undergone dialysis (mean, 1.30 +/- 1.26 U/ml, p = 0.04), whereas both before and after hemodialysis groups (means, 0.71 U/ml and 0.75 U/ml, respectively) were not significantly different from controls (mean, 0.72 +/- 0.30 U/ml, p = 0.96 and p = 0.8, respectively). Patients with ESRD who had not undergone dialysis showed no difference in vWF binding to collagen (mean, 1.21 +/- 0.63 U/ml) compared with the control group (mean, 1.16 +/- 0.13).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Co-localization of von Willebrand factor and type VI collagen in human vascular subendothelium.

The binding of von Willebrand factor (vWF) to subendothelium constitutes an important initial step in the process of platelet adhesion to exposed subendothelium following blood vessel injury. We previously demonstrated that vWF is present in human vascular subendothelium and recently found that a 150 kd vWF-binding protein, which we extracted from subendothelium, is type VI collagen. Although we have established that vWF and type VI collagen bind in vitro, it is not known whether these two proteins are associated in the vascular subendothelium in situ. We, therefore, 1) investigated the morphological effects of our biochemical extraction procedure on human umbilical veins by scanning and transmission electron microscopy, 2) analyzed the subendothelial extract by immunofluorescence for the presence of vWF and collagens and by electron microscopy for morphological characteristics, and 3) localized vWF and type VI collagen in subendothelium by immunofluorescence and by single- and double-label immunoelectron microscopic studies with protein A-conjugated gold particles. We found that the surface exposed following de-endothelialization is composed of microfibrils and contains very little fibrillar collagen. The subendothelium is stripped after sodium dodecyl sulfate-urea extraction, and the extract itself contains immunoreactive vWF and type VI collagen but no immunoreactive type I or III fibrillar collagens. Immunofluorescence and immunoelectron microscopic studies showed that vWF and type VI collagen are both present in subendothelium, where both co-localized to microfibrils. In conclusion, vWF that binds to type VI collagen in vitro, also co-localizes with type VI collagen in subendothelium, where both are associated with microfibrils. Type VI collagen, therefore appears to serve as a biologically significant binding site for vWF in vivo and may thereby play a role in mediating platelet adhesion to exposed subendothelium following vascular injury.

Collagen↗
PubMed1992

Modest doses of nitroglycerin do not interfere with beef lung heparin anticoagulation in patients taking nitrates.

The results of a prior clinical report suggested that nitroglycerin may interfere with the anticoagulant effect of heparin. Therefore, 30 adult patients undergoing cardiac surgery were studied in a controlled, prospective fashion. Thirteen patients on chronic nitrate therapy received an intraoperative nitroglycerin infusion at 1 micrograms/kg/min intravenously. Seventeen patients received no preoperative or intraoperative nitrates (control group). Heparin, 300 units/kg, was administered to all patients in three consecutive doses: 40 units/kg, 80 units/kg, and 180 units/kg. The activated coagulation time and activated partial thromboplastin time were measured prior to heparin, and 5 minutes after each heparin dose. There were no differences in automated activated coagulation times or in activated partial thromboplastin times between the groups at any measurement period. The study is limited in that only patients on chronic nitrates were included in the treatment group and that only a modest dose of nitroglycerin was used. However, it is concluded that a modest dose of intravenous nitroglycerin does not interfere with the anticoagulant effect of boluses of beef lung heparin in patients undergoing cardiac surgery.

Adult↗
PubMed1992

Reoperation, emergency and urgent open cardiac surgery in Jehovah's Witnesses.

Progressive advances in perfusion technology and perioperative supportive management have made it possible for members of the Jehovah's Witnesses religious group to undergo open cardiac operations with remarkable safety. However, hospital mortality remains high in (1) patients requiring reoperation (in whom both technical and bleeding problems tend to be more frequent) and (2) patients with significantly compromised cardiac performance requiring urgent or emergency operation. Employing a number of perioperative measures designed to minimize blood loss and maintain hematocrit levels (including use of the recently available recombinant human erythropoietin in two patients whose cases are reported herein), 13 reoperations and five urgent or emergency operations were performed. The one death in the entire series occurred in a patient (reoperation group) who died of a cerebrovascular accident of presumed embolic etiology, having undergone combined debridement of a stenotic heavily calcified aortic valve and a second coronary artery revascularization procedure. None of the patients required surgical exploration for bleeding. We suggest that currently available methodology permits Jehovah's Witnesses to undergo reoperation, emergency surgery, or urgent open cardiac operation at a level of risk not dissimilar to that seen in patients who permit use of homologous blood and products in their treatment.

Adult↗
PubMed1991

Desmopressin acetate is a mild vasodilator that does not reduce blood loss in uncomplicated cardiac surgical procedures.

Desmopressin acetate (DA) is a synthetic analog of vasopressin that may improve perioperative coagulation in cardiac surgical patients. Twenty-seven adult patients with good left ventricular function and normal preoperative coagulation profiles scheduled to undergo elective cardiac surgery participated in the double-blinded, placebo-controlled study. The 14 patients in the DA group received the drug over 10 minutes (starting 15 minutes after protamine administration). The 13 patients in the placebo group received an equal volume of saline. Preoperative template bleeding time was longer in the placebo group (P = 0.04). Otherwise, there were no statistically significant differences between the groups in demographics, coagulation variables, renal concentrating function, blood loss, or transfusion requirements at any study interval. The only significant hemodynamic differences detected were an increase in cardiac output in the DA group and a corresponding decrease in systemic vascular resistance. Five of 13 patients who received DA required treatment for hypotension, whereas none of 12 patients who received placebo required treatment during the infusion (P = 0.008). The authors conclude that DA causes mild vasodilation, but does not reduce blood loss or transfusion requirements in patients undergoing primary uncomplicated cardiac surgical procedures.

Anesthesia, Intravenous↗
PubMed1991

Alterations of platelet function induced by interleukin-2.

We recently reported that thrombocytopenia and bleeding are often limiting effects of immunotherapy with interleukin-2 (IL-2). In order to understand the mechanisms that lead to this unexpected clinical toxicity, we studied the effects of IL-2 on in vitro platelet function. When platelet aggregation was studied using whole blood (impedance, electrical) aggregometry, inhibition of aggregation was detected within 1 min of the addition of exogenous IL-2 to whole blood. IL-2-induced platelet secretion was quantified by radioimmunoassay (RIA) of PF4, BTG, and TXB2 independent of the addition of an aggregating agonist (ADP). Platelet secretion and inhibition of aggregation were an indirect consequence of a cellular effect of IL-2 on mononuclear cells, since aggregation was normal when whole blood was depleted of mononuclear cells and its reconstitution with autologous mononuclear cells led to a cell concentration-dependent inhibitory effect of aggregation and release of alpha-granule components in the presence of IL-2. In order to understand the mechanism of platelet secretion mediated by IL-2-activated mononuclear cells, we quantified the release of eicosanoid products from cultures of mononuclear cells exposed to IL-2 and found a significant increase in TXB2. Our results indicate that platelet secretion, indirectly initiated by IL-2-activated cells, is followed by inhibition of aggregation. These findings may not only have important implications for the planning of clinical immunotherapy trials with IL-2, but may also provide a novel link for a better understanding of the relationships between the hemostatic and the immune systems.

Blood Platelets↗
PubMed1991

150-kD von Willebrand factor binding protein extracted from human vascular subendothelium is type VI collagen.

We have previously shown that von Willebrand factor (vWF), a glycoprotein which plays a critical role in the adhesion of platelets to injured blood vessels, is present within vascular subendothelium. We investigated the identity of the subendothelial binding site(s) for vWF by examining vWF binding to subendothelial constituents and solubilized a 150-kD protein with SDS-urea that bound vWF. This protein had an amino-acid composition similar to that of the type VI collagen alpha-1/alpha-2 chains, was recognized by specific polyclonal antibodies against type VI collagen, and had a similar acidic isoelectric point. Furthermore, we found that purified type VI collagen also bound vWF. Thus, we have identified the extracted 150-kD protein as type VI collagen. This protein may play a significant role in the binding of vWF to vascular subendothelium in vivo.

Amino Acids↗
PubMed1990

von Willebrand factor antigen is not an accurate marker of rat and guinea pig liver endothelial cells.

To determine whether von Willebrand Factor (vWF) is a valid marker of liver endothelial cells, we determined vWF immunoreactivity in rat and guinea pig liver sections and in smears of elutriated nonparenchymal cells isolated from these two species. In frozen sections, positive staining for vWF was seen only in the endothelium lining large hepatic vessels in both species, and no immunoactivity was detected in the sinusoids. On the other hand, immunohistochemical staining for vimentin (a marker of mesenchymal cells) showed positive reaction throughout the vascular and sinusoidal endothelial cells in both the rat and guinea pig liver. In fractions of elutriated rat and guinea pig nonparenchymal liver cells, which included almost exclusively liver endothelial cells, only 25-40% of the cells displayed a positive reaction for vWF. However, when these same fractions were stained for vimentin, 70-95% of the cells exhibited immunoreactivity. Most of the vWF-negative cells were not red and white blood cells, biliary epithelial and Kupffer cells, and hepatocytes, and had ultrastructural features of sinusoidal endothelial cells. We conclude that in both the rat and guinea pig, liver sinusoidal endothelial cells do not exhibit vWF immunoreactivity. Thus, in routine immunohistochemical assays, vWF is not an accurate marker of rat and guinea pig liver endothelial cells. Vimentin is more appropriate for this purpose, provided that other mesenchymal cells are separated or independently identified.

Animals↗
PubMed1989

Effects of endotoxin on iron uptake by the hepatocyte.

Administration of endotoxins is often followed within 12-24 h by marked hypoferremia. Because the hepatocyte is the major site of both iron storage and transferrin synthesis, we have investigated the effects of an Escherichia coli endotoxin (lipopolysaccharide, LPS) on these parameters on isolated hepatocytes from normal Wistar rats (ND), rats previously treated intraperitoneally with 2.5 mg/kg (LD) or 25 mg/kg (HD) LPS, and control rats injected intraperitoneally with sterile saline (CD). No effects were observed on iron uptake from transferrin by ND cells incubated in vitro with up to 350 micrograms LPS/10(7) hepatocytes. There was also no significant difference in iron uptake between CD, HD, and LD hepatocytes 1 h after LPS injection. However, hepatocytes isolated 24 h after LPS administration took up iron significantly faster than controls. The uptake of non-transferrin-bound iron was also increased in HD and LD hepatocytes at 24 h but only in HD cells at 1 h. Transferrin binding was not altered in LPS-treated cells from ND rats but was depressed in cells from LPS-treated rats both at 1 h and at 24 h after injection. Transferrin receptor recycling was significantly increased at 24 h in cells from both LD and HD rats. Transferrin and total protein synthesis were also depressed at 1 h in LPS-treated rats, returning to normal values at 24 h. Direct preincubation of ND cells, however, failed to increase synthesis except at the highest concentrations of LPS. We conclude that LPS has an immediate (although indirect) effect on protein synthesis by the hepatocyte but not on iron uptake.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Unusual venous thrombosis associated with protein C deficiency.

A patient in whom massive puerperal thrombophlebitis developed had protein C deficiency. In patients who initially have thrombosis that is unusual either in its nature or extent, investigation may, as in this case, reveal the presence of a hypercoagulable state.

Adult↗
PubMed1987

Isolation and partial characterization of plasma membrane fatty acid binding proteins from myocardium and adipose tissue and their relationship to analogous proteins in liver and gut.

We describe a general method for isolating a class of 40 kDa plasma membrane fatty acid binding proteins which have been identified previously only in rat liver and jejunum. Proteins extracted with 2 M salt from rat adipocyte and cardiac myocyte plasma membranes were subjected to preparative isoelectric focusing. Fractions with pI's greater than or equal to 9.0 were further purified by oleate-agarose affinity chromatography and HPLC. Each tissue yielded a single 40 kDa protein which co-chromatographed with [3H]-oleate on gel permeation HPLC, and reacted on Western blots with an antibody to the corresponding hepatic membrane protein. Related plasma membrane fatty acid binding proteins have now been isolated from each of the major sites of fatty acid transport.

Adipose Tissue↗
PubMed1987

In vitro multimerization of human von Willebrand factor from its subunits.

von Willebrand factor (vWF) is a multimeric glycoprotein composed of subunits having an estimated molecular mass of 220-240 kDa which are linked by disulphide bonds. Intact, unreduced vWF, has ristocetin co-factor activity, which is lacking in the reduced subunits. To determine whether reduced vWF subunits are capable of being re-multimerized vWF was purified, reduced and then subsequently dialysed in the absence of reducing agents. (1) Purified vWF, (2) reduced vWF, and (3) reduced and dialysed vWF were compared for size composition and for ristocetin co-factor activities. SDS-PAGE showed that the purified vWF failed to enter the gels, while the reduced vWF migrated to a position consistent with a mass of approximately 240 kDa. Reduced and subsequently dialysed vWF subunits multimerized into larger forms, most of which did not enter the gels. No residual 240 kDa material was detectable following remultimerization. vWF multimer analysis confirmed the loss of monomer in the reduced and dialysed material and the appearance of new multimers with a size range smaller than the native material. Reduced vWF showed no detectable ristocetin co-factor activity, while the re-multimerized material regained some activity. Thus, we deduce that vWF can spontaneously re-multimerize from its reduced subunits and regain a small but measurable quantity of ristocetin co-factor activity.

Humans↗
PubMed1987

Localization of surface vWF on resting and stimulated platelets.

We used immunoelectron microscopic localization techniques to investigate whether platelets stimulated by ADP or ristocetin in the plasma milieu bind von Willebrand factor (vWF) to their surfaces. We found by both peroxidase- and ferritin-based methods that unstimulated platelets lack vWF on their surfaces, whereas platelets that are stimulated with ADP or ristocetin have vWF associated with their surfaces. The specificity of the findings was confirmed by absorption studies using severe von Willebrand disease (vWD) and hemophilic plasmas. The anti-vWF antibodies were blocked by incubation with hemophilic plasma but not by incubation with severe vWD plasma. Thus, in the plasma environment, in the presence of fibrinogen, vWF becomes associated with the platelet surface subsequent to stimulation with ADP or ristocetin.

Adenosine Diphosphate↗
PubMed1986

Thrombocytopenia following peritonitis in surgical patients. A prospective study.

Thrombocytopenia is commonly found in patients with serious infection. To investigate this phenomenon, 14 consecutive patients (68 +/- 10 years) who underwent laparotomy for bowel perforation and culture-proven peritonitis were prospectively studied. Ten noninfected laparotomy patients served as a control group. None of the 10 control patients developed thrombocytopenia. Of the infected group, 12 of 14 patients (85%) developed thrombocytopenia (less than 100,000/mm3). One patient (9%) developed disseminated intravascular coagulation (DIC). Of the remaining 11 patients with thrombocytopenia, platelet counts fell from preoperative level of 350,000 +/- 166,000 to 54,000 +/- 30,000 (p less than 0.001) and reached this nadir 4.3 +/- 2 days after surgery. There was no statistically significant difference in prothrombin time, partial thromboplastin time, or fibrinogen levels before versus after operation in this group. Bleeding times in seven patients were 5.5 +/- 2 minutes, and bone marrow examination in five patients with platelet counts of less than 50,000/mm3 revealed normal or increased megakaryocytes. No patient in this group bled, had medications held, or received platelet transfusions. Platelet counts increased greater than 100,000/mm3 at a mean of 8.9 +/- 4.1 days after operation. It is concluded that thrombocytopenia is common following surgery for intra-abdominal infection, is not usually associated with DIC, clinical bleeding, or coagulation abnormalities, does not commonly result from bone marrow suppression, and is transient and does not require routine platelet transfusions.

Disseminated Intravascular Coagulation↗
PubMed1982

Subendothelial deposition of von Willebrand's factor requires the presence of endothelial cells.

The presence of FVIII/VWF in human aortic subendothelium has been previously established. The present study was undertaken to determine the origin of this FVIII/VWF. Indirect immunofluorescence microscopy was employed to detect FVIII/VWF in rabbit aortas, after passage of a balloon catheter. Immediately after this de-endothelialization, FVIII/VWF was found to be present directly luminal to the internal elastic lamina. However, in the subsequent 5 days to 2 months, no FVIII/VWF was found on the luminal surface of exposed, nonre-endothelialized neointima. After endothelial cell regrowth, however, FVIII/VWF was again seen on the luminal surface. Thus these studies reveal that the deposition of subendothelial FVIII/VWF in neointima requires the presence of endothelial cells and that relatively trivial amounts of plasmatic FVIII/VWF are deposited on neointimal connective tissue after de-endothelialization.

Animals↗