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Biomedical subjects

A Armstrong

Publications and source records attributed to A Armstrong.

At least 55 records · Page 3Linked to original sources

The quantity of rhodopsin in human eyes.

The content of rhodopsin in the eyes of 15 donors (30 eyes) was determined. Both retinal and pigment epithelial fractions were collected from each globe, extracted using 1% CTAB, and the rhodopsin difference spectrum of each fraction was obtained separately. The total amount of rhodopsin, obtained by summing the amounts recovered from the retinal and PE fractions, ranged from 2.00 to 11.94 (median: 6.40) nmoles/eye. Previously reported mean values of about 3.5 to 4.0 nmoles per retina have been obtained using a variety of methods. The present higher values, perhaps largely dependent on procedural details described herein, appear plausible given the known concentrations of rhodopsin in rod outer segments, rod outer segment volumes, and number of rods in the human retina.

Adult↗

Mercaptan and dicarboxylate inhibitors of hamster dihydroorotase.

In mammals, dihydroorotase is part of a trifunctional protein, dihydroorotate synthetase, which catalyzes the first three reactions of de novo pyrimidine biosynthesis. Dihydroorotase catalyzes the formation of a peptide-like bond between the terminal ureido nitrogen and the beta-carboxyl group of N-carbamyl-L-aspartate to yield heterocyclic L-dihydroorotate. A variety of evidence suggests that dihydroorotase may have a catalytic mechanism similar to that of a zinc protease [Christopherson, R. I., & Jones, M. E. (1980) J. Biol. Chem. 255, 3358-3370]. Tight-binding inhibitors of the zinc proteases, carboxypeptidase A, thermolysin, and angiotensin-converting enzyme have been synthesized that combine structural features of the substrates with a thiol or carboxyl group in an appropriate position to coordinate a zinc atom bound at the catalytic site. We have synthesized (4R)-2-oxo-6-thioxohexahydropyrimidine-4-carboxylate (L-6-thiodihydroorotate) and have found that this analogue is a potent competitive inhibitor of dihydroorotase with a dissociation constant (Ki) in the presence of excess Zn2+ ion of 0.17 +/- 0.02 microM at pH 7.4. The potency of inhibition by L-6-thiodihydroorotate in the presence of divalent metal ions decreases in the order Zn2+ greater than Ca2+ greater than Co2+ greater than Mn2+ greater than Ni2+; L-6-thiodihydroorotate alone is less inhibitory and has a Ki of 0.85 +/- 0.14 microM. 6-Thioorotate has a Ki of 82 +/- 8 microM which decreases to 3.8 +/- 1.4 microM in the presence of Zn2+. Zn2+ alone is a moderate inhibitor of dihydroorotase and does not enhance the potency of other inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS)

Amidohydrolases↗

Kinetic and biochemical heterogeneity in vertebrate chondroepiphyseal regions during development.

The purpose of this study was to see if kinetic and biochemical heterogeneity could be documented in vertebrate chondroepiphyseal regions as they develop from mesenchymal condensations to cartilage. The kinetics of developing proximal and distal femoral chondroepiphyseal regions were studied from early limb bud stage to newborn animals in chicks, mice, and rabbits with thymidine autoradiography. Proteoglycan synthesis in the proximal femoral chondroepiphyseal region of the rabbit was studied with radioactive sulfate incorporation at 28 days of gestation and at 1 and 4 days after birth. The results indicated that these kinetic and biochemical characteristics of the developing chondroepiphyseal regions became heterogeneous very early in development. This early programming of populations of cells for division and for different biochemical functions existed during the fetal period when heterogeneity has been described histologically but has not been well documented.

Animals↗

Hepatitis B markers in Lancashire police officers.

A total of 284 Lancashire police officers each with a minimum of 5 years experience was tested for evidence of hepatitis B infection. None was hepatitis B surface antigen positive (HBsAg). Three were positive for both antibody to hepatitis B core antigen (anti-HBc) and HBsAg (anti-HBs). Five were positive for anti-HBc alone. Thus the overall prevalence was 2.8% which is within the range reported for blood donors in the UK. There was no association with working in the drug squad or custody office but there was a higher prevalence in those who had worked in the scene-of-crime's squad. However, the numbers were small, and of this group of 28 officers, 2 of the 3 with detectable hepatitis B markers were positive for anti-HBc alone. Therefore for police officers in mixed rural/urban areas of the UK, routine administration of hepatitis B vaccine is not justified although special consideration should be given to those working in selected groups. Further studies are required to ascertain whether there may be an increased risk for police officers working in conurbations.

Adult↗

Elaboration of neutral proteoglycanase by growth-plate tissue cultures.

This report describes the properties of a neutral protease that was synthesized and secreted into medium by intact cartilaginous growth plate in tissue culture. Bovine cartilaginous growth plate was grown for seven days in tissue culture, during which time the chondrocytes remained viable and metabolically active as determined by quantitation of trypan-blue exclusion and incorporation of 3H-cytidine. Protease activity, assayed by viscometry using proteoglycan monomer from cartilage as a substrate, was absent on day 1 but was present at high levels on days 2 through 5. The protease activity did not require activation and was highest at neutral and alkaline pH. Protease activity was abolished by twenty-millimolar EDTA but was unaffected by pepstatin, iodoacetate, and soybean trypsin inhibitor. In contrast to the high levels of activity of neutral protease that were present in tissue cultures of the intact growth plate, no protease activity could be detected when chondrocytes from the cartilaginous growth plate were grown in cell culture, even after sonication of the cells or activation with aminophenyl mercuric acetate or trypsin. Since hypertrophic chondrocytes probably do not survive the disruption of tissue that is involved in establishing cell cultures, these observations suggest that neutral protease is probably released into the medium by the hypertrophic chondrocytes that are present in the cultures of cartilaginous growth-plate tissue. It appears that the organization of the growth plate in tissue culture, as well as the maturation of proliferating chondrocytes into hypertrophic chondrocytes in tissue culture, may be required for synthesis of the neutral protease and its extracellular secretion by hypertrophic chondrocytes.

Alkaline Phosphatase↗

cDNA sequence of human beta-preprotachykinin, the common precursor to substance P and neurokinin A.

The nucleotide sequence of cDNA encoding the human substance P precursor, beta-preprotachykinin (beta-PPT), has been determined. The source of mRNA was a human laryngeal carcinoid tumour that contained a high concentration of immunoreactive substance P. The human beta-PPT polypeptide is 129 amino acids long and contains regions encoding substance P and neurokinin A, each flanked by basic amino acid residues. Residues 72-107 of the human beta-PPT polypeptide encode the sequence of neuropeptide K, an N-terminally extended form of neurokinin A recently isolated from porcine brain.

Animals↗

Effects of gastrin, glutamine, and somatostatin on the in vitro growth of normal and malignant human gastric mucosal cells.

This study evaluated the dose-related trophic effects of glutamine, gastrin, and somatostatin on the in vitro growth of human gastric cancer cells and normal human gastric mucosal cells. Quadruplicate cell cultures were seeded into growth medium with or without glutamine, gastrin, or somatostatin. After 72 hours' incubation, cells were counted and their numbers compared with those of controls. Glutamine and gastrin stimulated the growth of both normal and malignant gastric mucosal cells. Compared with normal cells, the malignant cells responded to these growth factors at lower concentrations. Somatostatin enhanced growth of gastric cancer cells at all concentrations and inhibited growth of normal cells at high concentrations. Further studies on the responsiveness of gastric adenocarcinoma to gastrointestinal tract hormones may elucidate mechanisms of oncogenesis and suggest new therapeutic avenues for patients with gastric cancer.

Adenocarcinoma↗

Characterization and partial purification of a 5,700 dalton form of substance P-like immunoreactivity from the rat hypothalamus.

Analysis of rat hypothalamic extracts by gel filtration and HPLC revealed the presence of a form of substance P-like immunoreactivity with a molecular weight of approximately 5,700 (6k-SP). 6k-SP is probably an N-terminally extended form of substance P; the peptide may be an intermediate in SP biosynthesis, or may function as a biologically active peptide in its own right.

Animals↗

Brucellosis.

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Brucellosis↗

What can experimental colorectal cancer tell us about colorectal cancer in man?

From a survey of the etiology of colorectal cancer, consideration of the basic pathologic characteristics of ulcerative colitis and familial polyposis of the colon suggests that they might be sister diseases. Similarities in the primary and secondary target organs and in fecal steroid metabolism might be more than coincidental. Thus, it would seem reasonable to pay particular attention to several groups of patients who are receiving therapy or have had operations that alter bile acid and/or cholesterol metabolism: 1) patients who have had jejunoileal bypass procedures for morbid obesity or hypercholesterolemia, 2) patients taking cholestyramine and/or other anti-hyperlipidemic agents, 3) patients receiving chenodeoxycholic therapy to dissolve gallstones, 4) patients taking the birth control pill, and 5) young patients who have had a cholecystectomy. Proponents of such therapy and physicians treating patients who develop colonic cancer or polypoid disease after having had such therapy are encouraged to provide accurate reports.

Adult↗

Effect of short-term therapy with propylthiouracil in patients with alcoholic liver disease.

The effect of propylthiouracil (PTU; 300 mg/day) on alcoholic liver disease was evaluated in 133 patients in a short-term randomized double-blind trial. Severity of the disease was assessed by a composite clinical and laboratory index (CCLI). A normalization rate (NR) representing the rate of improvement in CCLI was calculated. Patients with alcoholic hepatitis, with and without cirrhosis, showed a significantly higher NR on PTU (43.6 +/- 4.6) than on placebo (19.8 +/- 3.3; P less than 0.001). A similar effect was observed in patients with abnormal prothrombin (no biopsy): NR was 32.9 +/- 6.9 on PTU and 2.6 +/- 3.7 on placebo (P less than 0.005). The effect of PTU on each clinical and laboratory component of the CCLI was also compared in these two groups. In 38 patients with alcoholic hepatitis and in 25 with abnormal prothrombin, those on PTU showed a greater improvement in 15 of 15 items (P less than 0.001) and 14 of 15 (P less than 0.01), respectively. When patients were divided according to the severity of the disease into those in the lower and upper halves of the CCLI range (81 and 52 patients, respectively), PTU was shown to have a significant effect only in the latter: The NR was 41.4 +/- 3.8 on PTU and 22.5 +/- 4.2 on placebo (P less than 0.005). PTU was ineffective in patients with inactive cirrhosis.

Double-Blind Method↗

Interaction of thyroid peroxidase with concanavalin A covalently coupled to agarose.

We have investigated the interaction between concanavalin A-agarose (Con A-agarose) and thyroid peroxidase, an integral membrane protein found in the 105,000 X g, 1-h particulate fraction of thyroid tissue. An intact form of porcine thyroid peroxidase was obtained by solubilization with the nonionic detergent Triton X-100 and two fragmented, hydrophilic forms of the enzyme were prepared by trypsin treatment of the membrane. The three types of thyroid peroxidase bind to Con A-agarose and can be eluted with alpha-methyl-D-mannoside. The alpha-methyl-D-mannoside eluate of the most purified thyroid peroxidase preparation has been analyzed by polyacrylamide gel electrophoresis. Peroxidase activity corresponds with a glycoprotein band. The binding of thyroid peroxidase to Con A-agarose can be inhibited by sugars in the following order: alpha-methyl-D-mannoside greater than D-mannose greater than alpha-methyl-D-glucoside greater than D-glucose greater than D-galactose. This order of specificity is typical of Con A-sugar interactions. Furthermore, inactivation of the carbohydrate binding site of Con A by demetallization greatly reduces the extent of thyroid peroxidase binding. Reactivation of the carbohydrate binding site by the addition of Ca2+ and Mn2+ to demetallized Con A-agarose restores thyroid peroxidase binding. These and other experiments suggest that htyroid peroxidase is, like several other peroxidases, a glycoprotein. In addition, the interaction between thyroid peroxidase and Con A-agarose may provide a new purification tool for thyroid peroxidase.

Animals↗

Solubilization of thyroid peroxidase by nonionic detergents.

We have examined the ability of nonionic detergents to solubilize thyroid peroxidase from a porcine thyroid particulate fraction, as measured by the release of peroxidase activity into the supernatant fraction after centrifugation at 105,000 X g for 1 hour and the retardation of the supernatant peroxidase of Sepharose 6B. The parameters of peroxidase solubilization by Triton X-100 have been investigated in detail. Under optimum conditions, 60 to 95% of the thryoid peroxidase and about 50% of the total protein is released into the 105,000 X g, 1-hour supernatant. Under the optimum conditions established with Triton X-100, a series of Brij detergents of different chemical structure were equally effective in releasing peroxidase and protein. The protein patterns of the supernatants obtained with these detergents were similar on sodium dodecyl sulfate-polyacrylamide electrophoresis gels, suggesting that the detergents studied release similar membrane proteins. The Triton X-100 and Brij 58 supernatants were chromatographed separately on Sepharose 6B equilibrated with 0.1% Triton X-100 or Brij 58, respectively. In both cases, 75 to 80% of the peroxidase activity was retarded, thereby indicating that the nonionic detergents effect solubilization of the peroxidase rather than dispersal of nonsedimentable membrane fragments. These studies report the first successful solubilization of thyroid peroxidase by nonionic detergents. Together with previous evidence from our laboratory, these experiments indicate that thyroid peroxidase is an integral membrane protein.

Animals↗

Thyroid microsomal membrane proteins. Effects of solubilization on molecular size.

The molecular size of microsomal membrane proteins from frozen porcine thyroids before and after solubilization by proteolytic and non-proteolytic techniques has been investigated by means of polyacrylamide-gel electrophoresis in the presence of 1% sodium dodecylsulfate. When thyroid microsomal membrane proteins are solubilized by non-proteolytic methods such as high pH, n-butanol, or deoxycholate, no major change in the electrophoretic pattern compared to untreated microsomes has been observed, thereby suggesting that these non-proteolytic methods are capable of extracting membrane proteins from thyroid microsomes without altering their molecular size. However, treatment of microsomes with protein-solubilizing levels of trypsin (1-5 mug trypsin per mg thyroid protein) results in degradation of all major proteins with a molecular weight greater than 30 000. The high-molecular-weight proteins are particularly susceptible to attack by trypsin. Thus, these experiments indicate that the use of trypsin to solubilize thyroid microsomal membrane proteins, particularly thyroid peroxidase, will result in fragmented proteins and should be avoided if intact membrane proteins are desired.

Animals↗