Biomedical subjects

J A Dent

Publications and source records attributed to J A Dent.

45 records · Page 3Linked to original sources

PubMed1987

Interfragmentary screw fixation of fractures of the tibial shaft.

The records of 54 patients treated with interfragmentary screws were reviewed a mean of 2 years after surgery. Fifty-one fractures were of moderate severity (Ellis, 1950), 31 had a ratio of fracture length to tibial diameter of 2 or more and 17 were comminuted. An anatomical reduction, achieved initially in 44 fractures, was lost before union occurred in 14 (32 per cent), despite plaster splintage. Excellent results, with anatomical healing, were achieved in 30 patients (55 per cent) and good results with less than 5 degrees malalignment in 6 (11 per cent). There was a statistically highly significant association of satisfactory results with initial anatomical operative reduction of the fracture, a ratio of fracture length to tibial diameter of 2 or more and the absence of comminution. The orientation and number of screws, delayed operation, the degree of initial displacement of the fracture and the patient's age were not shown to affect the outcome. Delayed union occurred in 4 patients (7.4 per cent) and non-union in 3 (5.5 per cent). Deep infection occurred twice (3.7 per cent). As more satisfactory results could be expected for these fractures using other techniques, we conclude that this method cannot be recommended.

Adolescent↗

PubMed1987

Proteolytic degradation of von Willebrand factor after DDAVP administration in normal individuals.

The infusion of 1-deamino-8-D-arginine vasopressin (DDAVP) in normal individuals is followed by an increase in factor VIII/von Willebrand factor (vWF) in plasma, by an increase in intensity of all sizes of multimers, and by the appearance of larger multimers of vWF than those seen in the resting state. Since the larger multimers are rapidly cleared and proteolysis is known to cause disaggregation of large multimers, we evaluated the degree of vWF proteolysis after DDAVP administration. DDAVP was infused into eight normal adult volunteers, and the relative proportions of the intact 225 kilodalton (kDa) subunit and the 189, 176, and 140 kDa vWF fragments were compared before and at different times after DDAVP infusion. The relative proportion of the 176 kDa fragment was increased, whereas that of the other species was decreased, thereby indicating that proteolytic fragmentation had occurred. However, plasmin did not appear to be responsible because the vWF fragments characteristically produced by this enzyme could not be detected. Concomitant analysis of vWF multimeric structure showed that these changes were accompanied by an increase in the relative proportion of the satellite bands, which suggests that they were proteolytically generated. Proteolysis may explain, at least in part, rapid clearance of larger vWF multimers released by DDAVP.

Adolescent↗

PubMed1986

Isolation and characterization of a collagen binding domain in human von Willebrand factor.

von Willebrand factor binds to fibrillar type I collagen in a rapid, temperature-independent, reversible, specific, and saturable manner. Evaluation of binding isotherms by Scatchard-type analysis demonstrated that 6-18 micrograms of von Willebrand factor bind per mg of collagen, with Ka between 2 and 8 X 10(8) M-1. Five distinct tryptic fragments, purified under denaturing and reducing conditions and representing over 75% of the molecular mass of the von Willebrand factor subunit, were tested for their capacity to inhibit the von Willebrand factor-collagen interaction. Complete inhibition was obtained with a 52/48-kDa fragment at a concentration of approximately 1 microM. The location of this fragment in the subunit was established to be between Val-449 and Lys-728. Fifteen monoclonal antibodies against the 52/48-kDa fragment inhibited von Willebrand factor binding to collagen. Six antibodies against other portions of the von Willebrand factor subunit had no inhibitory effect. The tryptic fragment was a competitive inhibitor of von Willebrand factor binding to collagen and, therefore, recognizes the same interaction site as the intact molecule. These studies precisely define a domain in the von Willebrand factor subunit that interacts with type I collagen.

Antibodies, Monoclonal↗

PubMed1986

Subunit composition of plasma von Willebrand factor. Cleavage is present in normal individuals, increased in IIA and IIB von Willebrand disease, but minimal in variants with aberrant structure of individual oligomers (types IIC, IID, and IIE).

We have evaluated the subunit composition of plasma von Willebrand factor (vWF) and found evidence that cleavage is present in normal individuals, increased in IIA and IIB von Willebrand disease (vWD), but decreased or absent in variants with aberrant structure of individual oligomers. vWF was rapidly purified from plasma on an analytical scale by monoclonal antibody immunoaffinity chromatography in the presence of protease inhibitors. After reduction and electrophoresis in 5% polyacrylamide gels containing sodium dodecyl sulfate, fragments of 189, 176, and 140 kD, as well as the predominant 225-kD subunit, were identified in plasma vWF from 25 normal individuals. The vWF polypeptides were detected by immunoblotting with a mixture of 55 anti-vWF monoclonal antibodies followed by 125I-rabbit anti-mouse antibody and autoradiography. In five individuals with type IIA and five individuals with type IIB vWD, the proportions of 176 and 140-kD fragments were increased relative to the intact 225-kD subunit, as determined by excising each band and quantitating incorporated radioactivity. In contrast, these fragments were either not detectable or were present in only trace amounts in variants with abnormal structure of individual oligomers (types IIC and IID, and a new variant, type IIE vWD). The results reported here provide evidence that absence of large vWF multimers in these two groups of variants results from different mechanisms. In addition, they demonstrate that partial cleavage of the plasma vWF subunit is a normal event.

Antibodies, Monoclonal↗

PubMed1986

A new variant of type II von Willebrand disease with aberrant multimeric structure of plasma but not platelet von Willebrand factor (type IIF).

A patient with a lifelong bleeding disorder was diagnosed as having Type II von Willebrand disease. The larger multimers of von Willebrand factor were absent from her plasma but present in platelets. A high-resolution electrophoretic technique was used to study the complex structure of individual von Willebrand factor multimers. In normal plasma, each multimer could be resolved into five bands: a more intense central one and four less intense, two moving faster and two slower than the central band. In normal platelets, each multimer could also be resolved into five bands. The central one had a mobility similar to that in plasma, whereas the four satellite bands had a mobility that differed from that of the corresponding plasma bands. In the patient, platelet von Willebrand factor antigen content and ristocetin cofactor activity were normal, and von Willebrand factor showed the same structure of individual multimers as seen in normal platelets. On the other hand, plasma von Willebrand factor antigen and ristocetin cofactor activity were decreased, and the structure of individual von Willebrand factor multimers was different from that of normal plasma and similar to that seen in normal and patient's platelets. After infusion of 1-deamino-8-D-arginine vasopressin, the largest von Willebrand factor multimers, as well as new satellite bands with a mobility similar to those in normal plasma, appeared in the patient plasma, and the levels of von Willebrand factor antigen and ristocetin cofactor activity became normal. Yet no relevant change in the prolonged bleeding time was observed. This new variant of von Willebrand disease, therefore, is characterized by the presence of a dysfunctional von Willebrand factor molecule that exhibits unique structural abnormalities in plasma but appears to be normal in platelets. The designation of Type IIF is proposed for this type of von Willebrand disease in accordance with the terminology that has been previously used.

Bleeding Time↗

PubMed1986

The heterogeneity of type IIA von Willebrand's disease: studies with protease inhibitors.

The absence of large von Willebrand factor (vWF) multimers from plasma is a characteristic of Type IIA von Willebrand's disease (vWD) and is thought to contribute to the clinical expression of this disorder. Recently, three IIA patients have been reported in whom intermediate and large multimers could be restored if blood were collected in 5 mm EDTA, 6 mmol/L N-ethylmaleimide, and 1 mmol/L leupeptin. This suggested that absence of large multimers resulted from in vitro proteolysis. We have now collected blood from ten Type IIA vWD patients in these inhibitors but were not able to detect large multimers in the plasma of any of them. In addition, intermediate-sized multimers were reduced or completely absent in all. The inclusion of inhibitors in the citrate anticoagulant, as compared to citrate alone, was found to increase the relative proportion of intermediate multimers in some patients but had no effect in others, and in none did it restore large multimers to plasma. The results with platelet vWF were more varied. Four patients showed an absence or decrease of large multimers, whereas in seven patients large multimers were present. When compared with citrate anticoagulant alone, the inclusion of inhibitors in the anticoagulant had little or no effect on the platelet multimeric pattern. 1-Deamino-8-D-Arginine Vasopressin (DDAVP) was administered to six patients from five families. Two patients from one family showed complete correction and a third patient showed almost complete correction of her bleeding time. Two patients showed minimal correction and one showed no detectable correction. An increase in multimer size after DDAVP tended to be associated with correction of the bleeding time. However, in no case did the largest multimers appear in plasma even in patients with complete bleeding time correction. The presence or absence of inhibitors in the anticoagulant had little or no effect on the multimeric pattern after DDAVP. These results indicate that Type IIA vWD is a heterogeneous disorder in which absence of largest and intermediate multimers is an in vivo phenomenon.

Bleeding Time↗

PubMed1986

Subunit composition of plasma von Willebrand factor in patients with the myeloproliferative syndrome.

In order to evaluate the role of proteolysis in acquired von Willebrand's disease (vWD) associated with the myeloproliferative syndrome, we have determined the relative quantity of von Willebrand factor (vWF) fragments as compared with the intact 225 kDa subunit in four patients. The plasma vWF of each individual lacked large multimers; each had a prolonged bleeding time; and both platelet and leukocyte counts were elevated. Plasma was obtained from blood drawn into 1 mmol/L leupeptin, 6 mmol/L N-ethylmaleimide, and 5 mmol/L EDTA to prevent in vitro proteolysis. vWF was isolated from plasma by immunoadsorbent chromatography, reduced, subjected to SDS-5% polyacrylamide gel electrophoresis, and immunoblotted with a mixture of 55 anti-vWF monoclonal antibodies. In three patients with essential thrombocytosis (ET) the 176 and 140 kDa fragments were increased in proportion to the intact 225 kDa subunit indicating increased proteolysis. Treatment of one ET patient with CCNU (Lomustine) decreased the platelet count and, to a lesser extent, the white blood cell count. This was associated with a correction of the bleeding time, a partial correction of the multimeric abnormality, and a lessening of vWF cleavage. In a patient with polycythemia rubra vera (PRV) the proportion of the 176 kDa fragment was increased to the upper limit of normal but there was no definite evidence of increased proteolysis. These studies provide evidence that proteolysis plays a role in the acquired von Willebrand's disease associated with the myeloproliferative syndrome. However, other mechanisms must also be considered.

Bleeding Time↗

PubMed1981

Development of the mossy fibers of the dentate gyrus: I. A light and electron microscopic study of the mossy fibers and their expansions.

The postnatal development of the axons of the dentate granule cells--the so-called mossy fibers--was studied at the light microscopic level in Timm and Golgi preparations and also by transmission electron microscopy. In the Timm-stained material, there was a distinctive coloration in the hilus and incipient stratum lucidum, indicating the presence of mossy fibers, on the first postnatal day. Over the next two weeks, the stained areas became more extensive, the size and density of the stained particles increased, and the particles became more intensely stained. These signs of progressive development of the mossy fibers appeared to reflect, temporally and topographically, the developmental gradients followed by their parent granule cells. The Golgi material confirmed the presence of mossy fibers in the hilus on the first postnatal day. Fasciculi of mossy fibers were observed in the stratum lucidum of the 3-day-old hippocampus, and although these immature axons were devoid of large synaptic expansions, they did have prominent growth cones at their termini. Small expansions along the lengths of the axons first appeared on day 7 and these grew to approximately an adult size and complexity by about day 14. The postsynaptic component of the mossy fiber synapse, the "thorny excrescence," did not begin to emerge from the proximal portion of the pyramidal cell dendrites until sometime after day 9. At the electron microscopic level we observed, on the first postnatal day, small, immature mossy fiber expansions which made both symmetric and asymmetric contacts directly with dendritic shafts. These profiles, which were only one tenth the size of mature expansions, grew rapidly between postnatal days 1 and 9 and increased their mean area by a factor of five. On or about day 9, as the "thorny excrescences" emerged, the asymmetric synapses came to be associated with these spinous processes. Taken together, the Golgi and electron microscopic analyses support the suggestion that mossy fibers establish synaptic contact with pyramidal cell dendrites early in the postnatal period, several days before there is any indication of spine development. Furthermore, the "thorny excrescences" develop after the more typical, pedicellate spines have appeared on the distal pyramidal cell dendrites. Finally, while it is clear that the mossy fibers in our 21-day-old material are, for the most part, fully matured, a more subtle and protracted development of the system, long into adulthood, is indicated by the increased area and density of stained particles in the Timm preparations from adult animals.

Aging↗

PubMedDate not supplied

Fractures in early childhood: osteogenesis imperfecta or child abuse?

We studied the radiographs of 194 fractures sustained by 39 children less than 5 years of age with osteogenesis imperfecta (OI) and compared them with radiographs of 84 fractures sustained by 69 normal children of the same age. A great variety of fracture patterns was observed in undoubted OI. Spiral and transverse fractures were the most common. Metaphyseal fractures accounted for 15% of the fractures. The results of this study do not support the view that a particular fracture pattern renders the diagnosis of OI unlikely.

Child Abuse↗