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Biomedical subjects

M L Thomas

Publications and source records attributed to M L Thomas.

At least 145 records · Page 8Linked to original sources

Purification and characterization of the human interferon-gamma receptor from placenta.

Purification of the human interferon-gamma (IFN-gamma) receptor was facilitated by identification of human placenta as a large-scale receptor source. When analyzed in radioligand binding experiments, intact placental membranes and detergent-solubilized membrane proteins expressed 1.3 and 5.9 X 10(12) receptors per mg of protein, respectively, values that were 13-163 times greater than that observed for U937 membranes. Two protocols were followed to purify the IFN-gamma receptor from octyl glucoside-solubilized membranes: (i) sequential affinity chromatography over wheat germ agglutinin- and IFN-gamma-Sepharose and (ii) affinity chromatography over columns containing receptor-specific monoclonal antibody and wheat germ agglutinin. Both procedures resulted in fully active preparations that were 70-90% pure. Purified receptor migrated as a single molecular species of 90 kDa either when analyzed on silver-stained NaDodSO4/polyacrylamide gels or when subjected to electrophoretic transfer blot analysis using a labeled IFN-gamma receptor-specific monoclonal antibody. The identity of the 90-kDa component as the receptor was confirmed by demonstrating its ability to specifically bind 125I-labeled IFN-gamma following NaDodSO4/PAGE and transfer to nitrocellulose. Certain receptor preparations converted into a 55-kDa fragment either during purification or upon storage at 4 degrees C. On the basis of N-Glycanase digestion studies, the IFN-gamma receptor appeared to contain 17 kDa of N-linked carbohydrate. The ligand binding site, the epitope for the receptor-specific monoclonal antibody, and all of the N-linked carbohydrate could be localized to the 55-kDa domain of the molecule.

Epitopes↗

Lateral thigh varicose veins: a phlebographic study.

The phlebographic findings in nine patients with primary varicose veins on the lateral aspect of the thigh are described. These were shown by varicography to connect primarily with the deep femoral vein via communicating veins: the symptoms were either cosmetic or mild aching in the thigh. Surgical treatment, if indicated, consists of subfascial ligation or sclerosis of the varices, or flush ligation of the incompetent communicating veins at their connections with the deep femoral vein.

Adult↗

The effect of low dietary calcium and calcium supplementation on calcium metabolism and bone in the immature, growing rat.

The data presented here were obtained from a series of experiments designed to determine 1) whether normal growth and bone development could be maintained in young, growing rats (3-9 weeks of age) on a diet containing 0.1% Ca, and 2) whether Ca presented in a bolus would be utilized as effectively as the same amount of Ca distributed throughout the diet. Weanling female rats were raised to 9 weeks of age on diets containing 0.4% P and either 0.5% or 0.1% Ca. One group of animals on the 0.1% Ca diet was given oral supplements of CaCO3 twice each day to supply the same amount of Ca consumed by age-matched animals on the 0.5% Ca diet. We found that animals consuming diet containing 0.1% Ca grew at the same rate as animals receiving 0.5% Ca, or 0.1% Ca + supplement for up to 9 weeks of age when the experiment was terminated. Measurement of femur length indicated that long bone length was the same for all animals. However, the 0.1% Ca group exhibited mild hypocalcemia (9.1 mg/dl vs 10.4 for controls), a 2.6-fold elevation in immunoreactive parathyroid hormone, and an increase of similar magnitude in circulating levels of 1,25-dihydroxyvitamin D. Bones from the 0.1% Ca group contained less than half as much Ca as bones from the 0.5% Ca group, and exhibited significant decreases in mid-shaft diaphyseal thickness, % trabecular volume of the distal metaphysis and breaking strength (torsion testing). These results suggest that while a diet containing 0.1% Ca is able to maintain normal growth, bone mineralization is compromised.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Subcutaneous calcium heparin versus intravenous sodium heparin in treatment of established acute deep vein thrombosis of the legs: a multicentre prospective randomised trial.

One hundred patients with phlebographically proved acute deep vein thrombosis of the legs were prospectively randomised into two treatment groups to compare the safety and efficacy of subcutaneous calcium heparin versus intravenous sodium heparin administered by constant infusion pump. The dose of heparin was determined by daily measurement of the kaolin cephalin clotting time. Treatment was maintained for up to 14 days, after which phlebography was repeated. Of 49 patients who received subcutaneous calcium heparin, two showed an increase in thrombus size, while eight showed complete lysis. In the 47 patients who received intravenous sodium heparin thrombus increased in size in 13 while only one showed evidence of complete lysis. These differences were significant. There were no significant differences between the two groups in the incidence of serious complications, although almost half of those receiving intravenous heparin had some minor problem with the constant infusion pump and just over half of those receiving subcutaneous heparin had some bruising at the injection site. This study showed that subcutaneous calcium heparin was more effective in helping lyse existing thrombus and preventing its propagation than intravenous sodium heparin.

Clinical Trials as Topic↗

Early diagnosis of femorodistal graft stenoses.

Detection and early correction of stenoses following femorodistal bypass grafting may prevent graft occlusion. We studied 43 grafts (36 saphenous vein and 7 human umbilical vein) prospectively by peroperative angiography and then by clinical assessment, post-exercise Doppler ankle pressures and digital subtraction angiography at 1 week, 3 months, 6 months and 1 year. There were 11 grafts inserted for critical ischaemia, 16 for rest pain and 16 for disabling claudication. Four patients died within the year with five patient grafts and these were excluded from the final results. Eight grafts failed during the first year; four within 1 month and a further four subsequently, with no warning alteration of symptoms, ankle Doppler pressure or digital subtraction angiography. Four additional grafts were jeopardized by symptomatic stenoses. Six grafts developed an asymptomatic stenosis of greater than 50 per cent without a change in exercising ankle Doppler pressure. Three symptomatic stenoses were successfully revised but the fourth occluded on conservative management. Three asymptomatic stenoses have also been successfully revised and three have been treated conservatively. Close surveillance of femorodistal grafts is justified by this study but a deterioration in ankle brachial Doppler index was only useful in the patients with recurrent symptoms and was unable to detect half the stenoses of greater than 50 per cent. The detection of these by intravenous digital subtraction angiography might help to avoid graft failure.

Adult↗

Changes in the apparently normal limb in unilateral venous ulceration.

Forty-seven patients with unilateral venous ulceration have been investigated to determine if any abnormalities were present in the contralateral limbs which had not had lipodermatosclerosis or ulceration. Ascending phlebography in the non-ulcerated limbs showed post-thrombotic changes in 28 per cent and incompetent lower leg communicating veins in 19 per cent. This incidence was not significantly different to the limbs with healed ulceration (45 and 23 per cent respectively, chi 2 test, P = 0.10). Half volume refilling time measured by foot volumetry suggested that 79 per cent of the non-ulcerated limbs had evidence of deep vein incompetence or incompetent lower leg communicating veins, which was again similar to the incidence in the previously ulcerated limbs (85 per cent). Transcutaneous oxygen readings, expressed as a ratio of a reading at a standard site in the gaiter region of the leg over a reading from the upper arm, were significantly lower in non-ulcerated limbs (mean 0.84 +/- 0.26 s.d.) than in a cohort of age and sex matched controls (mean 1.02 +/- 0.14, Student's t test, P less than 0.001), and significantly higher than in previously ulcerated limbs (mean 0.68 +/- 0.31, P less than 0.01). Abnormalities in venous anatomy and function have been shown, in conjunction with evidence of reduced oxygen diffusion, through the gaiter skin before overt skin changes develop.

Adult↗

Structural variants of human T200 glycoprotein (leukocyte-common antigen).

Structural variation in the primary structure of human T200 glycoprotein has been detected. Three cDNA variants have been characterized each of which encode T200 molecules that differ in size as a result of sequence differences in their amino-terminal regions. The largest form of the molecule is distinguished from the smallest by an insert of 161 amino acids, after the first eight amino-terminal residues. The other variant has an insert at the same location of 47 amino acids identical to residues 75-121 in the larger insert. Both extra domains are rich in serine and threonine residues and are likely to display multiple O-linked oligosaccharides. These structural variants which probably arise by cell-type-specific alternative splicing provide a molecular basis for the previously observed structural and antigenic heterogeneity of T200 glycoprotein. In addition to the variable amino-terminal region, the external domain of human T200 glycoprotein consists of a second cysteine-rich region of about 400 amino acids, a single transmembrane-spanning region and a large cytoplasmic domain of 707 amino acids shared by all of the structural variants and highly conserved between species. The gene encoding human T200 is located on the long arm of chromosome 1.

Amino Acid Sequence↗

B-cell variant of mouse T200 (Ly-5): evidence for alternative mRNA splicing.

The complete cDNA sequence of mouse T200 glycoprotein from the pre-B-cell line 70Z/3 has been determined. The deduced protein sequence differs from that previously reported for a T-cell form of the molecule [Saga, Y., Tung, J.-S., Shen, F.-W. & Boyse, E. A. (1986) Proc. Natl. Acad. Sci. USA 83, 6940-6944] by the insertion of 139 amino acid residues in the amino-terminal region of the molecule. RNA transfer blotting using a cDNA probe encoding this sequence established that the predominant T200 mRNA species from B cells and cytotoxic T-cell clones but not T-helper cell clones or thymocytes contain all or part of the insert. Overlapping genomic clones of murine T200 were isolated and analysis of the intron-exon structure at the 5' end of the gene provides evidence that variants of T200 glycoprotein are generated by alternative mRNA splicing.

Amino Acid Sequence↗

Limb muscle compression: an alternative technique for intravenous digital subtraction angiography.

A simple, muscle-compression technique to introduce a bolus of contrast medium from hand or foot injections for intravenous digital subtraction angiography (DSA) is described. Fifty millilitres of the non-ionic contrast medium iopamidol 300 was used for each projection. In 18 out of the 20 examinations an acceptably diagnostic arteriogram was obtained. It is suggested that the compression technique is a useful alternative for intravenous DSA when conventional venous access is impossible.

Adult↗

Effects of ovariectomy on duodenal calcium transport in the rat: altered ability to adapt to low-calcium diet.

Studies were undertaken to determine whether ovariectomy (ovx) would alter the ability of female rats to adapt to low dietary Ca intake by exhibiting an in duodenal active Ca transport. Intact and ovx female rats were fed diets containing 1.5, 0.50, or 0.02% Ca prior to measuring active Ca transport using everted duodenal sacs in vitro. In some experiments, ovx animals were pair-fed to intact animals of the same age consuming the same diet. When ovx animals were allowed to eat ad lib, we found that both growth rate and duodenal active Ca transport increased relative to age-matched, intact controls. However, when growth of ovx animals was maintained at the control rate by pair-feeding, ovx per se did not affect intestinal active Ca transport. Ovx did not alter circulating levels of 1,25-dihydroxyvitamin D (1,25(OH)2D). We found that intact females responded to the low-Ca (0.02%) diet with increased circulating 1,25(OH)2D levels and increased intestinal active Ca transport. Ovx animals exhibited the same increase in circulating concentrations of 1,25(OH)2D in response to low-Ca diet, but did not demonstrate increased duodenal active Ca transport. When ovx animals consumed the diet ad lib, they became larger and exhibited higher Ca transport rates than intact animals fed the high-Ca diet, but there was no difference in Ca transport between ovx animals fed diets containing different Ca contents. The results of these experiments demonstrate that in female rats, the ability to adapt to altered dietary Ca intake is dependent on intact ovarian function and is not necessarily directly related to circulating concentrations of 1,25(OH)2D.

Acclimatization↗