Biomedical subjects
Publications and source records attributed to M D White.
We measured the effect of arthroscopic lavage and debridement of the osteoarthritic knee by comparing objective measurements of thigh muscle function before and after operation. There was some improvement in quadriceps isokinetic torque at six and 12 weeks after joint lavage but not after debridement. Neither method significantly relieved the patients' symptoms.
Despite many attempts to relate components of physique to core temperature cooling rate, no consistent relationship has emerged. The inconsistencies among the reported findings may arise from unaccounted thermoregulatory responses or incomplete physique assessment or both. A study was designed to examine this relationship in the range of rectal temperatures (Tre) within which shivering and sweating are absent, defined as the null zone, thus minimizing the contribution of these effector responses. Twenty healthy subjects (10 male and 10 female), representing a variety of physiques, participated in the study. The anthropometric protocol included 5 heights, body mass, 8 skinfolds, 10 girths, and 4 breadths. This permitted derivation of a body surface-area-to-mass ratio and estimates of adipose and muscle tissue masses using a cadaver-validated mass fractionation model. Subjects were heated in a 40 degrees C bath followed immediately by cooling in a 30.6 degrees C bath. During the cooling, forehead sweating rate and oxygen uptake were monitored to establish the boundaries of the Tre null zone. In addition, on-line recordings were made of Tre, skin temperature, and surface heat flux at six sites. The rate of cooling of Tre (Tre), in the range of temperatures between thresholds for sweating and shivering, was correlated to the components of physique. Estimates of adipose and skeletal muscle tissue masses did not correlate to Tre in the range of core temperatures investigated. However, total mass exhibited a significant correlation (r = 0.5, P less than or equal to 0.05) with Tre, but gender seemed to distort this relationship, possibly due to differences in adipose tissue distribution.
In many transplant centers, the T lymphocytes of transplant patients are routinely monitored, both to predict and diagnose the cellular events that result from transplantation and to evaluate the effectiveness of immunosuppressive therapy. During the course of this monitoring, we have observed anomalous results: the number of T cells said to be present varies greatly depending on the surface marker used for the assay. It is shown, in this study, that these "anomalous" results can be predicted from a knowledge of the spectrum of specificity of the anti-lymphocyte antibody therapy being administered. Because polyclonal antibodies have varied and unstated specificities, it is difficult to interpret results obtained from monitoring T-cell numbers or subset ratios without some knowledge of the specificities of the drugs being used.
The effects of blood transfusion, blood components, and surgical trauma on the growth of an experimental sarcoma have been examined. Recipient animals were inbred adult female WAB rats, which received allogeneic transfusions from inbred adult female PVG rats, syngeneic blood from inbred colony-mates, or saline infusions. Small volume transfusions (1-4 ml) of whole blood had no effect on tumour growth, but growth of the MC7 sarcoma was significantly enhanced following allogeneic transfusions of 5 ml whole blood, or when 4 ml was combined with sham laparotomy. Maximal enhancement of tumour growth occurred when 4 ml transfusions of allogeneic washed cells were given, but allogeneic plasma was also able, to a lesser degree, to enhance tumour growth. These data confirm that blood transfusion may enhance growth of the MC7 sarcoma, that the effect may be dose dependent, and synergistic with the immunosuppression of surgery. Many components of an allogeneic transfusion may be responsible for this effect.
The 'hollow cylinder' protein (Harris, J.R. (1968) Biochim. Biophys. Acta 150, 534-537) has been purified from human erythrocyte membranes. The molecular weight of the native protein, as determined by analytical ultracentrifugation, was found to be 747,000. By means of sodium dodecyl sulphate gel electrophoresis, the purified protein was shown to be composed of three different low molecular weight polypeptides of average molecular weight 25,000. This study provides convincing evidence that the spectrin tetramer is not responsible for the characteristic electron microscopic appearance of the hollow cylinder protein.
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The interaction of ApU, Aps4U, Aps4Up, ApAps4Up and Gps4U with RNase A was studied by CD difference spectroscopy. The use of 4-thiouridine (s4U) containing oligonucleotides enables to distinguish between the interaction of the different components of the ligand with the enzyme. The mode of binding of the oligonucleotides to the enzyme is described. From this mode of binding it is explained why Aps4U, for example, inhibits RNase A, while s4UpA serves as a substrate.
Forty different oligonucleotides were investigated as possible inhibitors of the depolymerizing activity of RNase A. The strongest inhibitors among the diribonucleoside 2'-5' mono- phosphates were: G2'-5'G, C2'-5'G and U2'-5'G, and among the diribonucleoside 3'-5' monophosphates: ApU, ApC and GpU. Of the eight trinucleotides investigated, ApApUp, ApApCp and ApGpUp were the strongest inhibitors. All four dinucleotides studied (ApUp, ApCp, GpUp and GpCp) were very strong inhibitors, ApUp being the strongest one. The results show that the nature of the various bases in the oligonucleotide has an effect on the degree of inhibition, and that the 3' phosphomonoester group increases the binding of the oligonucleotide to RNase A. These inhibitors can be used in physicochemical and biochemical studies of ribonuclease.
A ouabain-insensitive ATPase activity associated with the water-soluble proteins of the human and bovine erythrocyte membrane is demonstrated by means of activity-staining in polyacrylamide gels. The ATPase activity from both sources had an absolute requirement for Mg2+, activity becoming easily detectable at 0.2 mM Mg2+. At low Mg2+ concentrations added Ca2+ appeared to decrease the intensity of the ATPase stain. The activity is unaffected by monovalent cations, does not hydrolyse p-nitrophenyl phosphate and is not inhibited by 2 : 4 dinitrophenol. The ATPase has an apparent molecular weight of approximately 100 000 as determined by electrophoresis in acrylamide gels containing dodecyl sulphate.
An efficient method to grow Escherichia coli W to high cell concentrations on the pilot scale is described and discussed. The method involves growth linked introduction of glucose and ammonia to the culture, sparging with oxygen, and maintenance of aerobic conditions by gradually decreasing the temperature in the culture in order to keep the oxygen demand within the limits of the capacity of supply. Under these conditions the linear rate of cell mass production is actually the result of exponential growth with a gradually decreasing growth-rate constant. About 10 kg packed cells were produced in a 50 liter working-volume fermentor in one run of 13 hr. The concentration of the cells at the end of the growth was about 47 g dry cells/liter. The expenditure for nutrients was minimal and the controls were of simple automatic nature. From the determined yield constants for glucose, nitrogen, phosphorus, and oxygen it may be inferred that the cells grown by this method are similar to those grown exponentially at constant temperature.
A novel procedure for the separation of oligonucleotides from pancreatic RNase-digest of RNA is described. The method involves a group-separation of uracil-containing and of cytosine-containing nucleotides on Dowex 50W. The obtained groups are further separated on DEAE-Sephadex A-25 by a linear gradient of NH4HCO3.