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Biomedical subjects

M J Wells

Publications and source records attributed to M J Wells.

At least 19 recordsLinked to original sources
PubMed1998

In vivo clearance of ternary complexes of vitronectin-thrombin-antithrombin is mediated by hepatic heparan sulfate proteoglycans.

Thrombin is inhibited by its cognate plasma inhibitor antithrombin, through the formation of covalent thrombin-antithrombin (TAT) complexes that are found as ternary complexes with vitronectin (VN-TAT). To determine whether the metabolism of VN-TAT ternary complexes is different from that previously reported for binary TAT complexes, plasma clearance studies were done in rabbits using human VN-TAT. 125I-VN-TAT was shown to be cleared rapidly from the circulation (t1/2alpha = 3.8 min) in a biphasic manner mainly by the liver. 125I-TAT had a similar initial clearance (t1/2alpha = 5.3 min) but had a significantly faster beta-phase clearance (t1/2beta = 42.8 min versus 85.4 min for VN-TAT; p = 0.005). Protamine sulfate and heparin abolished the rapid initial alpha-phase of 125I-VN-TAT clearance and reduced its liver-specific association and in vivo degradation. Heparin also reduced the alpha-phase clearance of 125I-TAT and was associated with the appearance of high molecular weight complexes, suggesting enhanced complex formation between VN and TAT. 125I-VN-TAT binding to HepG2 cells was reduced by competition with VN-TAT or heparin but to a much lesser extent in the presence of TAT. The binding of VN-TAT to HepG2 cells was not inhibited by competition with the low density lipoprotein receptor-related protein ligand, methylamine-alpha2-macroglobulin. 125I-VN-TAT binding was also inhibited by treating HepG2 cells with heparinase or by growing the cells in the presence of beta-D-xyloside. Finally, both heparin and chloroquine, but not methylamine-alpha2-macroglobulin, reduced the internalization and degradation of VN-TAT by HepG2 cells. Taken together, these data indicate the importance of VN in TAT metabolism and demonstrate that VN-TAT binds to liver-associated heparan sulfate proteoglycans, which mediate its internalization and subsequent intracellular degradation.

Animals↗
PubMed1997

Cytokeratin 18 is expressed on the hepatocyte plasma membrane surface and interacts with thrombin-antithrombin complexes.

During experiments to identify putative hepatic receptors for thrombin-antithrombin (TAT) complexes, a 45-kDa protein was identified by ligand blotting. Following gel purification, amino acid sequencing revealed the 45-kDa TAT-binding polypeptide to be cytokeratin 18 (CK18). The presence of CK18 on the surface of intact rat hepatoma cells was demonstrated by binding of 125I-anti-CK18 antibodies. Anti-CK18 antibodies reduced the binding and internalization of 125I-TAT by rat hepatoma cells. Immunocytochemical analysis, to determine the location of CK18 in vivo, revealed a periportal gradient of CK18 staining; with hepatocytes around the portal triads demonstrating striking pericellular staining. In addition, anti-CK18 IgG associated with perfused livers to a significantly greater extent than preimmune IgG. Taken together, these data provide evidence that CK18 is found on the extracellular surface of hepatocytes and could play a role in TAT removal. Finally, these data, in conjunction with recent reports of CK8 (Hembrough, T. A., Li, L., and Gonias, S. L. (1996) J. Biol. Chem. 271, 25684-25691) and CK1 cell membrane surface expression (Schmaier, A. H. (1997) Thromb. Hemostasis 78, 101-107), indicate a novel role for these proteins as putative cellular receptors or cofactors to cellular receptors.

Animals↗
PubMed1994

Prevalence of antithrombin deficiency in healthy blood donors: a cross-sectional study.

The prevalence of antithrombin (AT) deficiency in the general population has been variously estimated to be between 0.05 and 5 per 1,000 in the population; 2,491 blood donors were screened in an attempt to clarify this issue using plasma samples taken from the blood donor units. From this initial population, 122 individuals were identified as having plasma AT levels lower than 2 standard deviations below the normal mean. Twenty-two samples had evidence that thrombin had been generated during blood collection and the remaining cohort of 100 blood donors were asked to return but only 59 complied. The data obtained from these 59 were compared with that from 51 age- and sex-matched control blood donors. Both groups of subjects were assessed for previous evidence, or family history, of thrombotic events, as well as exposure to risk factors associated with the development of deep vein thrombosis (DVT). All had venous blood samples taken from which the supernatant plasma was immediately removed and quick frozen for later assaying. Only 6 of the 59 subjects with initial low AT levels had repeat AT-Xa levels below 0.80 units/ml (normal range 0.94 +/- 0.14). Upon repeating the AT-Xa determinations on new samples from these six individuals, only three were found again to be low. One was found to have a type 3 AT deficiency (an Arg47Cys substitution). The other two with a low AT level had mean functional AT-Xa levels of 0.61 and 0.71 units/ml, respectively, with correspondingly low AT:Ag levels consistent with a type 1 AT deficiency. Two of these three subjects has been in high risk situations without evidence of having developed DVT and none had evidence of venous reflux on Doppler venography. In addition, none had personal or family histories of previous thrombotic events. These present data indicate that the prevalence of AT deficiency in our blood donor population is 2 per 1,000 (95% confidence intervals: 0.7-6/1,000).

Antithrombins↗
PubMed1993

An antithrombin III assay based on factor Xa inhibition provides a more reliable test to identify congenital antithrombin III deficiency than an assay based on thrombin inhibition.

OBJECTIVES: To determine whether functional antithrombin III (AT-III) levels measured by a factor Xa inhibition (AT-III-Xa) assay identifies AT-III deficient individuals more reliably than functional AT-III levels measured by a thrombin inhibition (AT-III-IIa) assay. STUDY DESIGN: Cross-sectional study. PATIENT POPULATION: Sixty-seven members of a large family with type 2 AT-III deficiency. INTERVENTION: DNA analysis was used as the reference diagnostic standard for AT-III status and subjects were classified as AT-III deficient or non deficient according to these results. Functional AT-III levels were measured in all subjects using: 1) a chromogenic substrate for thrombin and added human thrombin (AT-III-IIa), and 2) a chromogenic substrate for factor Xa and added bovine factor Xa (AT-III-Xa). Functional heparin cofactor II (HC-II) levels were measured using a commercially available kit. The proportions of 125I-alpha-thrombin complexed to AT-III and HC-II were measured by polyacrylamide gel electrophoresis and autoradiography. RESULTS: Thirty-one (46%) individuals were classified as AT-III deficient and 36 (54%) as AT-III non deficient. AT-III-Xa assay measured a significantly lower mean AT-III value and a narrower range for individuals classified as AT-III deficient than the AT-III-IIa assay. Using the AT-III-IIa assay, six subjects had borderline AT-III levels compared to none with the AT-III-Xa assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1992

Molecular cloning and expression of rabbit antithrombin III.

A cDNA containing the complete open-reading frame encoding rabbit antithrombin III (AT-III) was isolated from a rabbit liver cDNA expression library, using a specific antibody as a probe. Sequence analysis showed 84% identity between the deduced amino acid sequences of the rabbit and human proteins. A previously described cell-free expression system was used to verify the identity of the clone. The full-length cDNA was inserted into an expression vector, and messenger RNA (mRNA) transcripts generated. In vitro translation of these transcripts, in the presence of [35S]methionine, in an mRNA-dependent rabbit reticulocyte lysate system resulted in the synthesis of a 51-Kd polypeptide, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This nonglycosylated protein was capable of forming SDS-stable complexes with human alpha-thrombin. Complex formation was significantly enhanced following the deletion of nucleotides encoding the signal peptide, and the resultant generation of a 47-Kd nonglycosylated mature protein product. When the template DNA giving rise to this product was internally truncated, two rabbit AT-III deletion mutants were generated that lacked the ability to interact with thrombin, but retained the ability to bind heparin. Cell-free expression plasmids encoding the human and rabbit AT-III mature molecules were manipulated to produce two interspecies fusion proteins. For the first, human codons were used to replace rabbit codons from residue 369-433, while in the second human codons replaced rabbit codons from residue 217-433. Both fusion proteins exhibited less efficient thrombin-complexing ability than the original cell-free-derived mature rabbit AT-III. Thus, portions of AT-III molecules from the two species, despite their high degree of homology, are not interchangeable. Knowledge of the structure of rabbit AT-III, combined with the availability of the rabbit cDNA, will permit defined experimentation aimed at understanding antithrombin III structure relative to its function in vivo.

Amino Acid Sequence↗
PubMed1987

Smodingium dermatitis.

Smodingium argutum is the plant most commonly responsible for causing acute allergic contact dermatitis in South Africa. When an outbreak of Smodingium dermatitis occurred in a local school the allergenic principle present in this plant was chemically isolated and identified, and its allergenic property proved in the clinic by patch testing. The value of using an extract of fresh Smodingium leaves in lieu of fresh leaves themselves was confirmed, but freeze-dried material was found to be unsuitable for patch-test purposes.

Acute Disease↗
PubMed1987

Flight trial of a helmet-mounted display image stabilisation system.

An image stabilisation system for improving reading performance with a helmet-mounted display (HMD) during whole-body vibration was tested at night in a helicopter. Six subjects read arrays of 50 numerals as quickly and as accurately as possible while flying in three different flight conditions. The mean reading time for each array while stationary on the ground was approximately 21 s, and the mean reading error was 0.4% without stabilisation. In-flight mean reading time increased to approximately 40 s, and reading error was 18% without the stabilisation system. Stabilising the image significantly reduced the mean in-flight reading time to approximately 25 s with a 4% reading error. Data from the flight trial support the results of previous experiments, which suggest that HMD reading performance with vibration and night viewing conditions may be inferior to performance with daylight conditions.

Adult↗
PubMed1984

Anticonvulsant activity of some 4-aminobenzamides.

A series of 4- aminobenzamides of some simple primary and secondary amines were prepared and evaluated for anticonvulsant effects. The compounds were tested in mice against seizures induced by electroshock and pentylenetetrazole ( metrazole ) and in the rotorod assay for neurologic deficit. For those N-alkyl amides tested, 4-amino-N- amylbenzamide (6) was the most potent against maximal electroshock seizures (MES): ED50 = 42.98 mg/kg; however, the N- cyclohexylbenzamide (8) showed the greatest protective index (PI = TD50/ED50), 2.8. The introduction of a second aromatic ring produced more potent compounds, with d,l-4-amino-N-(alpha-methylbenzyl)-benzamide (12) showing the highest level of activity. This compound has an anti-MES ED50 of 18.02 mg/kg in mice when administered intraperitoneally (ip) and a TD50 of 170.78 mg/kg (PI = 9.5) in the same species. These data compare quite favorably with those for phenobarbital and phenytoin in the same assays.

Animals↗
PubMed1984

Benefits of helmet-mounted display image stabilisation under whole-body vibration.

The effects of whole-body vertical vibration in the range 2.5-25 Hz on visual performance with two types of raster scan helmet-mounted display have been determined. The benefit of an image stabilisation system on numeral reading performance during vibration was also assessed with both display systems. Increases in mean reading time of over 130%/m . s-2 R.M.S. and increases in percentage reading error of more than 30%/m . s-2 R.M.S. were recorded with unstabilised displays. With vertical and horizontal image stabilisation, these decrements in performance were reduced to less than 40%/m . s-2 R.M.S. increase in reading time and less than 10%/m . s-2 R.M.S. increase in reading error. Data on the transmission of vibration from the seat to the head and from the head to the helmet were also obtained. These indicate a relation between biodynamic behaviour and visual performance during vibration.

Biomedical Engineering↗
PubMed1980

The effects of extracts from neurosecretory cells in the anterior vena cava and pharyngo-ophthalmic vein upon the hearts of intact free-moving octopuses.

Recordings of pressure and frequency were made from the hearts of free-moving Octopus vulgaris. The effects of extracts from neurosecretory endings in the anterior vena cava (AVC) and the pharyngo-ophthalmic vein (POV), injected through fine cannulae into a branchial heart, efferent branchial vessel or the dorsal aorta, were studied and compared with the effects of acetylcholine, 5-hydroxytryptamine, adrenaline, histamine and tyramine. AVC and POV extracts each produce a different spectrum of effects, unlike those of any of the drugs tested. AVC extract is effective at doses of less than 2% of the material extractable from a single vein per kg, increasing the force and amplitude of the heartbeats. With a natural release point just upstream of the branchial hearts the AVC material must be relevant to the normal performance of the hearts. POV extract is effective only at doses equivalent to several veins per kg, and is unlikely to have a role in cardiac regulation. Section of the visceral nerves did not affect the action of drugs or extracts, indicating that effects were not indirectly mediated via the CNA. Further experiments were made with hearts and the aorta in vitro with effects that did not always parallel those found in vivo. Reasons for these differences are discussed.

Acetylcholine↗
PubMed1980

Nervous control of the heartbeat in octopus.

The circulatory system of cephalopods is based on a trio of hearts, with two pairs of associated ganglia linked to the CNS by a pair of visceral nerves. The beat of the hearts was recorded from free-moving octopuses before and after surgical removal or disconnexion of elements of the nervous system. Severing the visceral nerves does not stop the hearts, which continue to beat in a powerful well co-ordinated manner in isolation from the CNS. The nerves seem to be concerned in raising the cardiac output in exercise, and with stopping the hearts when mantle movements cease, but they are not necessary for the initiation of maintenance of the normal rhythm. Removal of the fusiform ganglia severs all nervous connexions between the ywo gill hearts, and deprives the systemic heart of its nerve supply. The trio of hearts continues to beat as strongly as before. Removal or disconnexion of a cardiac ganglion disrupts the beat of the corresponding gill heart which now tends to contract in an ill-coordinated and rather feeble manner, though at much the same frequency as before; with both cardiacs gone the systemic heart, which contracts only when it is filled, tends to drop in frequency and the mean aortic pressure falls. The system remains rhythmic, however, and the beat may recover, to the point where aortic pressures and frequencies approach those found in intact animals at rest; even octopuses with both fusiform and both cardiac ganglia removed can survive for many hours. From the performance of the isolated branchial heart, the existence of a pulsating vesicle in each cardiac ganglion, the effects of cardiac ganglion removal and the remarkable steadiness of heartbeat frequency shown by intact animals under a variety of conditions, it is argued that the heartbeat rhythm is normally controlled by pacemakers in the branchial heart/ cardiac ganglion complexes, and perhaps, in intact animals, from within the cardiac ganglia themselves. The picture of the control of the heartbeat that emerges from the study of free moving essentially intact animals is quite different from that arising from in vitro and acute preparation studies. It suggests that the conventional wisdom about the control of the heartbeat in cephalopods (and perhaps by implication, in other molluscs) may need to be considerably revised.

Animals↗
PubMed1980

Distortion and adaptation in underwater sound localization.

Sounds should be localized at more medial positions under water than in air, because the values of the interaural temporal and intensity differences are reduced in water. Thirteen blindfolded divers were required to move a pointer towards the apparent location of a sound source under water, and they showed significant errors towards the median plane. In a second experiment, five divers were tested in air before and after underwater training at swimming towards a sound source. The search patterns of these divers showed systematic errors towards the median plane. A comparison of the first and second air tests showed some evidence of an aftereffect away from the median plane, discounting the most lateral angles (80 degrees left and right) where little effect would be expected. It is concluded that both a distortion of localization and some adaptation to the distortion occur under water.

Auditory Perception↗
PubMed1978

Reproduction versus somatic growth: hormonal control in Octopus vulgaris.

1. Octopus vulgaris can be forced into precocious maturity by removal of the subpedunculate lobe from the brain, an operation that releases the optic glands from inhibition, and allows them to secrete a gonadotropin. 2. 14C-leucine was injected into the bloodstream of immature animals and its subsequent incorporation into muscle protein followed by taking successive samples from the arms. The optic glands were then activated, and a further injection of 3H-leucine given and followed by means of further arm samples. 3. Optic gland secretion suppresses protein synthesis in the muscles. This is associated with an increase in the total amino acid pool in the muscles and with a considerable increase in the concentration of free amino acids circulating in the blood. 4. If an ovary is present these events are associated with a rapid growth of the ovary and its ducts, and a loss of weight elsewhere. In ovariectomized animals the ducts grow, but there is no yolk to absorb the large pool of free amino acids, and the animals gain weight by osmotic uptake of water into the muscles. 5. The developing ovary may produce a hormone that increases the release of amino acids from muscle, since the concentration circulating in the blood of intact animals remains at least as high as in ovariectomized octopuses, despite the demands of the developing ovary. 6. These matters are discussed in relation to other evidence for a gonadial hormone and in relation to the 'self-destruct' effect of the optic gland secretion in determining the post-reproductive death of octopuses.

Age Factors↗
PubMed1975

The subfrontal lobe and touch learning in the octopus.

Octopuses with the supraoesophageal lobes of the brain divided longitudinally can be taught to discriminate using the arms on either side. If there is no further lesion the two sides behave alike. Lesions limited to one side did not affect the performance of the contralateral, "control" side. Lesions made in the vertical (n=7) lobes led to a slight drop in the quality of performance in training to take a smooth sphere, in discrimination training (rough vs. smooth spheres) and in subsequent extinction and transfer tests. After removal of the median inferior frontal lobe (n = 10) there were somewhat greater effects in the same direction. Much larger effects followed interference with the subfrontal lobe (n = 20). Removal of parts from this always led to a marked loss of capacity for touch learning, broadly dependent on the amount of tissue removed. Removal of the whole of the subfrontal lobe (n = 6) produced animals that showed, at best, only very slight signs of learning. Such animals can adjust their overall level of response as a result of training but they seem incapable of adjusting response levels to two objects independently. These results are discussed in relation to the function of the subfrontal lobe as a memory store.

Animals↗