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Biomedical subjects

J R Baker

Publications and source records attributed to J R Baker.

At least 271 records · Page 15Linked to original sources

Fructosamine in diabetic pregnancy.

Fructosamine, an indicator of glycosylated serum protein, was measured in 79 non-diabetic pregnant women and 20 women with gestational diabetes. The test provided a clear discrimination between groups; it detected 17 (85%) of the women with gestational diabetes and gave only 4 (5%) false-positive results. 19 women with established diabetes before pregnancy had very high levels. Maternal fructosamine at 29 weeks' gestation correlated significantly with both fasting blood glucose levels at the time and birthweight ratio. Levels of fructosamine in cord blood were significantly higher in gestational diabetic than in normal pregnancies, suggesting a possible additional role for fructosamine in retrospectively detecting the hyperglycaemic fetus. Fructosamine estimation is fully automated and may provide a simple, inexpensive means to screen for diabetes in pregnancy.

Adult↗

Clinical usefulness of estimation of serum fructosamine concentration as a screening test for diabetes mellitus.

Fructosamine, a putative measure of serum glycosylated proteins, was measured in 74 subjects referred for oral glucose tolerance tests. A normal range (mean (2 SD] of 1.6 (0.4) mmol/l (40(10) mg/100 ml) derive from results obtained in 83 healthy non-diabetic volunteers permitted the detection of 15 out of 17 (88%) subjects with proved diabetes and yielded only five (9%) false positive diagnoses. Fructosamine concentrations correlated significantly (p less than 0.001) with fasting plasma glucose concentrations (r = 0.76) and glycosylated haemoglobin concentrations (r = 0.70). A longitudinal study suggested that fructosamine concentration was an index of intermediate term (one to three weeks) blood glucose control. Fructosamine concentration was not related to uraemia and did not depend on albumin or total protein concentrations, provided that serum albumin concentrations remained above 30 g/l. Estimation of fructosamine concentrations is a fully automated procedure and may provide a simple means of screening for diabetes mellitus.

Adolescent↗

Identification of a monoclonal antibody that specifically recognizes corneal and skeletal keratan sulfate. Monoclonal antibodies to cartilage proteoglycan.

Monoclonal antibodies were raised against proteoglycan core protein isolated after chondroitinase ABC digestion of human articular cartilage proteoglycan monomer. Characterization of one of the monoclonal antibodies (1/20/5-D-4) indicated that it specifically recognized an antigenic determinant in the polysaccharide structure of both corneal and skeletal keratan sulfate. Enzyme immunoassay analyses indicated that the mouse monoclonal IgG1 recognized keratan sulfate in native proteoglycan aggregate and proteoglycan monomer preparations isolated from hyaline cartilages of a wide variety of animal species (human, monkey, cow, sheep, chicken, and shark cartilage). The 1/20/5-D-4 monoclonal antibody did not recognize antigenic determinants on proteoglycan isolated from Swarm rat chondrosarcoma. This finding is consistent with several biochemical analyses showing the absence of keratan sulfate in proteoglycan synthesised by this tissue. A variety of substructures isolated after selective cleavage of bovine nasal cartilage proteoglycan (Heinegård, D., and Axelsson, J. (1977) J. Biol. Chem. 252, 1971-1979) were used as competing antigens in radioimmunoassays to characterize the specificity of the 1/20/5-D-4 immunoglobulin. Substructures derived from the keratan sulfate attachment region of the proteoglycan (keratan sulfate peptides) showed the strongest inhibition. Both corneal and skeletal keratan sulfate peptides as competing antigens in radioimmunoassays showed similar inhibition when compared on the basis of their glucosamine content. Therefore, the 1/20/5-D-4 monoclonal antibody appears to recognize a common determinant in their polysaccharide moieties. Chemical desulfation of the keratan sulfate reduced the antigenicity of the glycosaminoglycan. The antibody did not recognize determinants present in dermatan sulfate, heparin, heparin sulfate, or hyaluronic acid.

Amino Acids↗

Case of equine goitre.

A brief review of the literature on equine goitre is presented, together with a case of congenital goitre in a foal. This animal showed localised swelling of the neck, hyperextension of the lower limbs and initial difficulty in sucking. The goitre was removed surgically and the hyperextension successfully treated with surgical shoes and bandaging. The cause was probably iodine deficiency coupled with a goitrogen in the diet.

Animal Nutritional Physiological Phenomena↗

On the fidelity of the lactoperoxidase method of cell membrane radioiodination: an electron microscopic autoradiographic study.

The radiolabelling of human peripheral blood lymphocytes by lactoperoxidase-catalysed iodination using 2 different sources of hydrogen peroxide has been compared using electron microscopic autoradiography. A method of statistical analysis of the autoradiographs has permitted precise identification of radioactive sources, in particular cellular compartments, taking into account cross-fire of Auger electrons producing silver grains in compartments other than those from which they are emitted. Our data confirm the postulates of previous investigators that the majority of radioiodine is located at the plasma membrane of cells labelled by enzymic iodination. The results further suggest that the glucose-glucose oxidase system for generation of hydrogen peroxide permits a greater degree of specific radiolabelling of plasma membrane proteins with less damage than equivalent lactoperoxidase iodination reactions promoted by exogenously added hydrogen peroxide.

Autoradiography↗

Fructosamine: a new approach to the estimation of serum glycosylprotein. An index of diabetic control.

The development of a novel manual method designed to measure serum glycosylprotein as an index of diabetic control is described. The method relies on the ability of ketoamines (fructosamines) to act as reducing agents in alkaline solution. Conditions are described for a simple colorimetric procedure which permits assay of both a synthetic fructosamine and purified albumin while severely limiting the contribution of interfering substances. Applied to whole sera, the measurement is linear with volume of serum assayed. It allows clear discrimination of normal and diabetic populations (p less than 0.001), and is significantly correlated with fasting blood glucose concentration (r = 0.72) and with a thiobarbituric acid procedure for measuring glycosylprotein-derived hydroxymethylfurfural (r = 0.58). The method is rapid (at least 12 samples per hour) and demands only simple equipment.

Blood Glucose↗

Clinical importance of the reversible fraction of haemoglobin A1c in Type 2 (non-insulin-dependent) diabetes.

Two studies demonstrating the clinical relevance of the labile fraction of haemoglobin A1c are presented. Haemolysates, dialysed at 37 degrees C against 0.9% NaCl, showed an average decrease of haemoglobin A1c value of 7.0% of total glycosylated haemoglobin, post-dialysis. There was a close correlation between the pre- and post-dialysis haemoglobin A1c values (r = 0.9644). In four Type 2 (non-insulin-dependent) diabetic subjects, blood glucose and pre- and post-dialysis haemoglobin A1c values were determined at weekly intervals over an 18-week period, including 6 weeks when oral hypoglycaemic therapy was withheld. During this time, pre- and post-dialysis haemoglobin A1c values responded synchronously to changes in blood glucose concentration. The mean coefficient of variation of the pre- and post-dialysis haemoglobin A1c values did not differ significantly over the 18-week period. It is concluded that the routine elimination of the labile fraction of haemoglobin A1c is not necessary to assess control in Type 2 diabetes.

Blood Glucose↗

Partial characterization and clinical correlation of circulating human immunoglobulins directed against thyrotrophin binding sites in guinea pig fat cell membranes. Development of a direct enzyme immunoassay.

To obviate several problems inherent in indirect thyroid-stimulating hormone (TSH) receptor antibody assays, we developed an enzyme-linked immunosorbent assay (ELISA) that measures antibodies binding to guinea pig fat cell membrane, which contain high concentrations of TSH receptors. Solubilized guinea pig fat cell membranes were adsorbed to plastic microtiter plates and served as the solid-phase antigen. Test sera and affinity-purified alkaline phosphatase-conjugated anti-human IgG were co-incubated with membranes, after which p-nitrophenyl phosphate was added. Results were read when a positive control reached a standard color change (OD405nm). Specificity of this assay was demonstrated by the inability of albumin, insulin, TSH subunits, propranolol, or dexamethasone to block binding 30. normal subjects had a mean OD value of 0.080 +/- 0.050 (SD). 23 of 25 untreated Graves' patients had OD values at least 2 SD above the normal mean (Grave's mean +/- SD; 0.46 +/- 0.33, P less than 0.001) and in each case 10(-6) M TSH inhibited the binding by at least 60%, suggesting that the immunoglobulins were directed at the TSH receptor. Seven of 25 serum samples from patients with Hashimoto's disease, seven of 23 serum samples from patients with transient hyperthyroidism (subacute thyroiditis or painless thyrotoxic thyroiditis), and two of 10 samples from patients with thyroid carcinoma had significant elevations in the titers of membrane-directed immunoglobulins. Graves' patients who were treated with ablative therapy at least 6 mo earlier and who were euthyroid when restudied continued to have abnormally elevated membrane-directed immunoglobulins in six of eight samples studied. Further studies involved the substitution of affinity-purified alkaline phosphatase anti-IgM antisera for the anti-IgG antisera routinely used. Seven of 12 serum samples from patients with Graves' disease had significant elevations in binding which in every instance was inhibited by greater than 60% by 10(-6) M TSH. In sum, the present results indicate that (a) we have developed a sensitive, specific, reproducible, convenient ELISA for the measurement both of the total amount of circulating membrane-directed antibodies and of TSH-displaceable membrane-directed immunoglobulins. (b) This ELISA detected significant elevations in TSH-displaceable guinea pig membrane binding in 23 of 25 untreated Graves' patients as well as in approximately 30% of patients with Hashimoto's thyroiditis and subacute thyroiditis. (c) Elevated membrane directed antibodies may continue to be present many months or years after restoration of the euthyroid state. (d) Circulating membrane binding IgM immunoglobulins have been detected in patients with Graves' disease. Further studies using this ELISA should prove useful in a variety of investigative and clinical studies.

Adipose Tissue↗

Immunological methods for the detection and determination of connective tissue proteoglycans.

In this paper we report the use of immunological methods for specifically detecting and determining proteoglycan in cartilage and other connective tissues. Antibodies (polyclonal and monoclonal) have been raised against specific components of cartilage proteoglycan aggregates (i.e., proteoglycan monomer and link protein). Radioimmunoassay procedures and immunohistochemical procedures have been developed and used to demonstrate the occurrence of cartilage-like proteoglycan and link protein in bovine aorta. Similarly, immunofluorescent studies have been used to analyze proteoglycan distribution in skin. Using antibodies specific for chondroitin-4-sulfated proteoglycan, their presence was demonstrated in dermal connective tissue and connective tissue surrounding nerve and muscle sheaths. However, chondroitin-4-sulfated proteoglycan was completely absent in the epidermis of skin and areas surrounding invaginating hair follicles. These immunological procedures are currently being used to complement conventional biochemical analyses of proteoglycans found in different connective tissue matrices.

Animals↗

Mechanism of entry and development of Trypanosoma dionisii in non-phagocytic cells.

Cultures of 'buffalo' (bison) lung (BL) cells were infected with epimastigotes or trypomastigotes of Trypanosoma (Schizotrypanum) dionisii derived from cultures in vitro, fixed after various periods of incubation at 37 degrees C and examined by light or electron microscopy. Few if any epimastigotes entered the BL cells, but many trypomastigotes did so; they adhered to the cell surface within 2 h and then appeared to sink into furrows on the cell surface until engulfed in parasitophorous vacuoles. Cytochalasin D (5-10 micrograms ml-1) completely, but reversibly, inhibited entry of trypomastigotes without affecting parasite motility. It was concluded that entry depended on the interaction of stage-specific components on the trypomastigote's surface with receptors on the BL cells, and that this interaction induced active uptake of the protozoa by a phagocytic process not involving pseudopod formation. Soon after entry of the trypomastigotes into BL cells, the membranes of the parasitophorous vacuoles disintegrated and the parasites, which were now lying free in the cytoplasm of the host cell, transformed into amastigotes (micromastigotes) during the next 24-48 h. Replication then occurred, followed by transformation, beginning after 3 days, through a transitional promastigote phase to small intracellular trypomastigotes at 7 days. The promastigotes had a characteristic curved protrusion extending from the lip of the flagellar pocket (or reservoir) into the host cell's cytoplasm. Trypomastigotes, released into the supernatant medium by rupture of the plasma membranes of the BL cells after 8 days, could re-invade other cells.

Animals↗

Characterization of glycosaminoglycans associated with Rauscher murine leukemia virions.

Host-derived sulfated components that copurify and are physically associated with the envelope of Rauscher murine leukemia virions grown in JLS-V9 cells were characterized by digestion with chondroitinase ABC and chondroitinase AC II, as well as nitrous acid degradation. A dermatan-sulfate-chondroitin-sulfate copolymer and heparin or heparan sulfate wee shown to be associated with the virions. Competitive binding studies indicated a specificity of the virions for association with heparan sulfate. The physiological importance of the association is discussed.

Animals↗