Biomedical subjects

J C Brown

Publications and source records attributed to J C Brown.

At least 127 records · Page 7Linked to original sources

PubMed1991

The use of beta-galactosidase fusion proteins encoding the early region 1 transforming proteins of adenovirus type 12 to examine the humoral response in tumour-bearing animals.

Sera from 26 rats bearing tumours induced by wild-type (wt) and mutant human adenovirus type 12 (Ad12), or by cells transformed with these viruses, were analysed for antibodies against the early region 1 (E1) transforming proteins. Six Ad12-beta-galactosidase fusion proteins encoding different regions of the Ad12 E1 proteins were constructed. The sera from the tumour-bearing animals reacted most strongly with the fusion protein encoding the N terminus of the E1A protein. Tumour-bearing rats exposed to the E1B 54K and 19K proteins showed strong reactions with the N terminus of the 54K protein and the C terminus of the 19K protein. Monospecific polyclonal antisera were raised against five of the fusion proteins by immunization of rats and rabbits; these sera cross-reacted with the purified native protein. No antibodies could be obtained which recognized a fusion protein containing amino acids 136 to 268 of the 54K protein. The fusion proteins were also used to purify monospecific antisera from tumour-bearer sera using affinity chromatography.

Adenoviridae Infections↗

PubMed1991

Investigations in the amplitude of sounded piano tones.

The relationship between final hammer velocity and maximum amplitude of radiated piano sound was investigated. Piano tones with varying hammer velocities were produced by a computer-monitored acoustic piano containing optical sensors and solenoids, and the sounded tones were recorded and digitized for analysis. Maximum amplitudes over the duration of the sounded tones were linearly proportional to piano hammer velocities for a range of frequencies and hammer velocities. Changes in room acoustics did not alter the linear relationship. Measurements of maximum amplitudes of individual tones and combined tones (dyads) also indicated a linear relationship between the sum of the maximum amplitudes of the individual tones and the maximum amplitude of the dyads. These findings indicate that the principle of superposition holds for peak amplitudes of sounded piano tones. Findings are discussed with regard to production and perception of musical dynamics.

Computer Graphics↗

PubMed1991

Structure of the herpes simplex virus capsid: effects of extraction with guanidine hydrochloride and partial reconstitution of extracted capsids.

Viral B capsids were purified from cells infected with herpes simplex virus type 1 and extracted in vitro with 2.0 M guanidine hydrochloride (GuHCl). Sodium dodecyl sulfate-polyacrylamide gel analyses demonstrated that extraction resulted in the removal of greater than 95% of capsid proteins VP22a and VP26 while there was only minimal (less than 10%) loss of VP5 (the major capsid protein), VP19, and VP23. Electron microscopic analysis of extracted capsids revealed that the pentons and the material found inside the cavity of B capsids (primarily VP22a) were removed nearly quantitatively, but extracted capsids remained otherwise structurally intact. Few, if any, hexons were lost; the capsid diameter was not greatly affected; and its icosahedral symmetry was still clearly evident. The results demonstrate that neither VP19 nor VP23 could constitute the capsid pentons. Like the hexons, the pentons are most likely composed of VP5. When B capsids were treated with 2.0 M GuHCl and then dialyzed to remove GuHCl, two bands of viral material were separated by sucrose density gradient ultracentrifugation. The more rapidly migrating of the two consisted of capsids which lacked pentons and VP22a but had a full complement of VP26. Thus, VP26 must have reassociated with extracted capsids during dialysis. The more slowly migrating band consisted of torus-shaped structures approximately 60 nm in diameter which were composed entirely of VP22a. These latter structures closely resembled torus-shaped condensates often seen in the cavity of native B capsids. The results suggest a similarity between herpes simplex virus type 1 B capsids and procapsids of Salmonella bacteriophage P22. Both contain an internal protein (VP22a in the case of HSV-1 B capsids and gp8 or "scaffolding" protein in phage P22) that can be extracted in vitro with GuHCl and that is absent from mature virions.

Animals↗

PubMed1991

Insulin secretion and islet endocrine cell content at onset and during the early stages of diabetes in the BB rat: effect of the level of glycemic control.

Although it is agreed that autoimmune destruction of pancreatic islets in diabetic BB rats is rapid, reports of endocrine cell content of islets from BB diabetic rats at the time of onset of diabetes vary considerably. Because of the rapid onset of the disease (hours) and the attendant changes in islet morphology and insulin secretion, it was the aim of this study to compare islet beta-cell numbers to other islet endocrine cells as close to the time of onset of hyperglycemia as possible (within 12 h). As it has been reported that hyperglycemia renders the beta cell insensitive to glucose, the early effects of different levels of insulin therapy (well-controlled vs. poorly controlled glycemia) on islet morphology and insulin secretion were examined. When measured within 12 h of onset, insulin content of BB diabetic islets, measured by morphometric analysis or pancreatic extraction, was 60% of insulin content of control islets. Despite significant amounts of insulin remaining in the pancreas, 1-day diabetic rats exhibited fasting hyperglycemia and were glucose intolerant. The insulin response from the isolated perfused pancreas to glucose and the glucose-dependent insulinotropic hormone, gastric inhibitory polypeptide (GIP), was reduced by 95%. Islet content of other endocrine peptides, glucagon, somatostatin, and pancreatic polypeptide, was normal at onset and at 2 weeks post onset. A group of diabetic animals, maintained in a hyperglycemic state for 7 days with low doses of insulin, were compared with a group kept normoglycemic by appropriate insulin therapy. No insulin could be detected in islets of poorly controlled diabetics, while well-controlled animals had 30% of the normal islet insulin content.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1991

Effect of a purified somatostatin monoclonal antibody and its Fab fragments on gastrin release.

The effect of a purified somatostatin monoclonal antibody, SOMA 10, and its Fab fragments on gastrin release have been examined in the isolated perfused rat stomach. SOMA 10 was purified from ascites fluid by ammonium sulfate precipitation followed by hydroxylapatite (HAP) chromatography. This gave a preparation of greater than 90% purity, as assessed by fast affinity and size exclusion high-performance liquid chromatography (HPLC). HAP-purified SOMA 10 had a binding capacity of 760 mmol somatostatin/mol antibody, a dissociation constant of 2.2 nM, and recognized amino acid residues 5-12 of the somatostatin molecule. Fab fragments of SOMA 10 were prepared and purified by protein A affinity chromatography, and the purity assessed by HPLC and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Single passage perfusion of SOMA 10 (100 micrograms/ml) into the gastric vasculature caused a paradoxical decrease in basal gastrin release. However, recirculation of the antibody (20 micrograms/ml) caused an increase in cumulated gastrin release. Perfusion of Fab fragments of SOMA 10 (66 micrograms/ml) also increased gastrin secretion. These results support the suggestion that somatostatin exerts a tonic restraint on gastrin release and that penetration of the antibody to the site of somatostatin release is necessary for it to have an effect.

Animals↗

PubMed1991

Modulation of acetylcholine-stimulated insulin release by glucose and gastric inhibitory polypeptide.

The interactions of glucose, acetylcholine and gastric inhibitory polypeptide in the regulation of insulin secretion were examined using the in situ perfused rat pancreas. Acetylcholine (1 x 10(-6) M) had no effect on the release of immunoreactive insulin in the presence of 2.2 x 10(-3) M glucose. However, in the presence of 4.4, 6.6 or 8.9 x 10(-3) M glucose, the same concentration of acetylcholine stimulated insulin secretion approximately fourfold. At the highest glucose concentration tested (17.8 x 10(-3) M), the stimulatory effect of acetylcholine on insulin release was less pronounced. The insulin response to acetylcholine was potentiated by the presence of gastric inhibitory polypeptide. This potentiation was most marked when the peptide was present at a concentration of 1 x 10(-9) M, whereas the effect of concomitant infusion of acetylcholine and 2 x 10(-10) M gastric inhibitory polypeptide was only slightly greater than additive. Both atropine (1 x 10(-6) M) and hexamethonium (1 x 10(-4) M) inhibited the insulin response to acetylcholine. However, neither of these cholinergic antagonists had a significant effect on glucose- or gastric inhibitory polypeptide-stimulated insulin secretion. These results demonstrate that the insulinotropic action of acetylcholine is glucose-dependent. A synergistic interaction may exist between acetylcholine and gastric inhibitory polypeptide at the beta-cell which does not involve glucose or gastric inhibitory polypeptide acting on cholinergic receptors in the pancreas. Cholinergic stimulation of insulin secretion in the perfused rat pancreas appears to be mediated by both muscarinic receptors on the beta-cell and nicotinic receptors, presumably on intrapancreatic ganglia.

Acetylcholine↗

PubMed1991

Comparison of the enteroinsular axis in two strains of obese rat, the fatty Zucker and the JCR:LA-corpulent.

The 'fatty' Zucker and more recently the JCR:LA-cp 'corpulent' have been studied extensively as genetic models of the hyperinsulinemia, insulin resistance and abnormal fat metabolism of obesity. It has been hypothesized that an abnormal enteroinsular axis leading to hypersecretion of the insulin releasing hormone gastric inhibitory polypeptide (GIP) could contribute to the hyperinsulinemia of obesity, although this has been controversial. The present study was undertaken to compare the enteroinsular axis in fatty Zucker and JCR:LA-cp rats. The in vivo GIP and insulin responses to an oral glucose challenge, as well as glucose tolerance, were compared in lean and obese phenotypes of both strains as well as the sensitivity of the perfused pancreas to the secretagogues glucose, arginine and GIP. In addition, the effect of perfusate glucose concentration on the beta cell response to GIP was assessed in both strains. Tissue samples from the pancreas were taken for immunocytochemical analysis of comparative size and composition of pancreatic islets. Our results indicate that corpulent rats are hyperGIPemic when compared to fatty Zuckers and that hyperinsulinemia (both in vivo and in vitro) is more severe in the JCR:LA-cp than in the fatty Zucker, as is the degree of insulin resistance (as evidenced by glucose intolerance). Islets of corpulent rats were found to be larger than those of fa/fa rats as well as having a larger area occupied by beta cells. It was concluded that GIP may contribute to fasting hyperinsulinemia in the Zucker rat (as a result of a defective glucose threshold for the insulinotropic action of GIP), whereas the hyperGIPemia of the JCR:LA-cp rat may contribute to the massive nutrient-stimulated hyperinsulinemia observed in the male phenotype of this strain.

Administration, Oral↗

PubMed1990

Angiocentric immunoproliferative lesion/T-cell non-Hodgkin's lymphoma and the acquired immune deficiency syndrome: a case report and review of the literature.

The lesions known as lymphocytic vasculitis, polymorphic reticulosis (midline malignant reticulosis, lethal midline granuloma), lymphomatoid granulomatosis, and angiocentric lymphoma form what have been collectively termed the angiocentric immunoproliferative lesions (AIL). Because of recent reports demonstrating clonal rearrangements of the T-cell receptor in these lesions, the AIL are now thought to represent a continuous spectrum of post-thymic T-cell non-Hodgkin's lymphoma (NHL). NHL associated with the acquired immune deficiency syndrome (AIDS) represents intermediate or high-grade B-cell malignancies in HIV-infected patients that may be etiologically related to the Epstein-Barr virus (EBV). There have been reports of EBV-associated T-cell NHL, AIL, and large granular lymphocyte (LGL) proliferations, as well as HIV-associated T-cell neoplasia, LGL/T-cell proliferations, and AIL. We describe a case of polymorphic reticulosis (lethal midline granuloma) arising in an HIV-infected individual, who later progressed to AIDS, and review the literature on HIV-associated and EBV-associated T-cell neoplasia, LGL/T-cell proliferations, and AIL. The etiology of this AIL/T-cell NHL, especially in relation to EBV and HIV, is discussed.

Acquired Immunodeficiency Syndrome↗

PubMed1990

Identification of the B1 and B2 subunits of human placental laminin and rat parietal-yolk-sac laminin using antisera specific for murine laminin-beta-galactosidase fusion proteins.

Antisera raised against fusion proteins consisting of murine laminin B1 and B2 subunit sequences fused to the C-terminus of Escherichia coli beta-galactosidase were tested for their subunit specificity on Western blots of deglycosylated murine Engelbreth-Holm-Swarm (EHS) laminin. The antisera raised against B2 subunit sequences (anti-XLB2.1 and anti-XLB2.2) bound only to the EHS laminin B2 subunit. One of the antisera raised against B1 subunit sequences (anti-XLB1.2) was specific for the B1 subunit, whereas two others (anti-XLB1.1 and anti-XLB1.3) cross-reacted with the EHS laminin B2 subunit. Gold-labelled heparin-albumin was shown to bind specifically to the A subunit of deglycosylated EHS laminin on Western blots. These reagents were used to identify the homologous subunits in rat parietal-yolk-sac laminin and human placental laminin. The anti-(fusion protein) antisera identified the B1 and B2 subunits of the rat laminin, and these were similar in size to the murine EHS B subunits. Human placental laminin gave bands of 400, 340, 230, 190 and 180 kDa on reducing SDS/PAGE. The anti-(fusion protein) antisera identified the 230 and 190 kDa bands as the B1 and B2 subunits respectively. Gold-labelled heparin-albumin bound to the 400, 340 and 190 kDa bands of human placental laminin and so did not unambiguously identify a single A subunit. The human placental laminin may contain a mixture of isoforms, with alternative subunits substituting for the A subunit.

Amino Acid Sequence↗

PubMed1990

Measurement of association constants in ELISA. Reactions between solid-phase antibody and fluid-phase biotinylated antigen.

An ELISA procedure which utilized an avidin-peroxidase amplification reaction to detect and quantify the amount of fluid-phase biotinylated antigen bound to solid-phase antibody was developed to determine antibody association constants. The methodology required is uncomplicated, avoids the use of radioisotopes, and is theoretically amenable for use with any protein which can be biotinylated, and any receptor protein which can be immobilized on plastic wells. A plot of known immobilized biotinylated antigen concentrations, which ranged from approximately 100 ng/ml to 1000 ng/ml, versus avidin-peroxidase conjugate product formation, was used to establish a standard curve from which the amount of antibody-associated antigen in binding assays could be determined. Antibody association constants obtained with this method ranged from 10(5) to 10(9) M-1.

Animals↗

PubMed1990

The biological effects and digestible energy value of a sugar-beet fibre preparation in the rat.

A sugar-beet fibre preparation (SBF) was incorporated into a semi-synthetic rat diet at a level of 100 g/kg. The material caused no feed aversion, and gain of live weight was unimpaired. SBF appeared to be slightly more fermentable than wheat bran and only marginally less effective as a faecal bulking agent when compared with equalized intakes of non-starch polysaccharide. SBF did not stimulate mucosal cell turnover in the small intestine. Some enlargement of the caecum was observed in animals given SBF, but it was no greater than that of animals given wheat bran. Animals given SBF had a lower serum cholesterol concentration than both the fibre-free controls and those given wheat bran. This hypocholesterolaemic effect was less than that of guar gum however. The partial digestibility of energy for SBF was 0.64, and its partial digestible energy value was 11.3 kJ (2.7 kcal)/g. The partial digestibility of energy for non-starch polysaccharide in SBF was estimated to be 0.53 and its partial digestible energy value was 9.1 kJ (2.2 kcal)/g. This value was not significantly different from that expected for unavailable carbohydrate in mixed human diets.

Animals↗

PubMed1990

Energy balance and energy values of alpha-amylase (EC 3.2.1.1)-resistant maize and pea (Pisum sativum) starches in the rat.

Apparent and partial digestible energy values for alpha-amylase (EC 3.2.1.1)-resistant, retrograde starches, isolated from cooked maize and pea starches (RMS and RPS respectively), were determined in male Wistar rats (about 180 g) during a 28-29 d balance period with ten animals per treatment. The starches were provided as supplements (100 g/kg diet) to a semi-synthetic basal diet (B), and their effects on the apparent digestibilities of nitrogen and fat, and on gains of live weight, fat and lean tissue were examined. Diet B alone was the control; sucrose (Su) and Solka-floc cellulose (SFC) were also examined for reference. Apparent digestibilities for Su, SFC, RMS and RPS were 1.0, 0.16, 0.98 and 0.89 respectively. Whereas the apparent digestibilities of gross energy, N and fat in the diet were unaffected by supplementation with Su, each was decreased by supplementation with SFC, RMS and RPS. Partial digestible energy values calculated from the intakes and faecal losses of energy in the basal and supplemented diets were 15, 12.4 and 0.8 kJ/g for RMS, RPS and SFC respectively. These values were smaller than corresponding apparent digestible energy values calculated from the apparent digestibility of the supplement and its gross energy value. Only the Su and starch supplements increased the intake of apparent digestible energy and the gain of live weight. Both starches and Su increased total energy (and fat) deposition to almost similar extents. It is concluded that the resistant starches contribute significant dietary energy, enhance growth and elevate fat deposition to extents almost similar to Su.

Animal Feed↗

PubMed1990

Effect of dietary homocysteine on copper status in rats.

The effect on copper status of diets containing homocysteine, an intermediate in the transsulfuration pathway of methionine metabolism, was investigated in rats. Two groups of six male weanling Sprague-Dawley rats were provided with deionized water and pair-fed diets that were adequate (14.0 mg/kg) or deficient (1.3 mg/kg) in Cu to groups fed diets similarly adequate or deficient in Cu but containing DL-homocysteine (10 g/kg). Hemoglobin concentration, tissue Cu levels and the activities of the Cu-dependent enzymes--ceruloplasmin, superoxide dismutase and cytochrome c oxidase--were markedly lowered by Cu-deficient diets and by homocysteine. These dietary treatments also lowered the activity of glutathione peroxidase and produced concomitant increases in the activity of manganese-dependent superoxide dismutase and iron levels in the liver. However, dietary homocysteine decreased hepatic Mn and low Cu diets decreased cardiac iron content. Moreover, both dietary treatments significantly lowered kidney Fe levels. Homocysteine increased heart, liver and kidney weights (g/100 g body tissue) and greatly elevated the level of thiobarbituric acid reactive substances (TBARS) in heart tissue. These results indicate that dietary homocysteine can markedly lower Cu status in rats and result in tissue redistribution of Fe and increased cardiac levels of TBARS, a measure of lipid peroxidation.

Animals↗

PubMed1990

The lumbar lordosis below Harrington instrumentation for scoliosis.

This retrospective study evaluates lumbar lordosis in 43 patients before and after Harrington instrumentation into the lumbar spine. The authors measured overall lumbar lordosis, lordosis of unfused lumbar levels, and sagittal vertical axis. Lordosis decreased progressively in lower levels of fusion. The increase in lordosis below the fusion did not compensate for the overall loss of lordosis. The sagittal vertical axis moved forward, producing a subtle, asymptomatic form of flat back syndrome.

Humans↗

PubMed1990

Evidence that the Fc region of autologous rabbit IgG isolated before and after hyperimmunization is structurally different: recognition by rheumatoid factor and monoclonal antibodies.

Autologous hyperimmune (HI) and pre-inoculation (PI) rabbit Fc gamma populations were found to be conformationally different by spectroscopic measurements, and antigenically different by measurements which examined rheumatoid factor (RF) and monoclonal antibody (MoAb) binding specificity for both populations in ELISA. Circular dichroism spectra of HI rabbit Fc gamma (prepared from animals after hyperimmunization with streptococcal vaccine) were both qualitatively and quantitatively different, particularly in the 225-228 nm range, in comparison to both homologous normal and autologous PI Fc gamma. Binding studies in ELISA showed that affinity constants obtained for reactions of both rabbit RF and various murine MoAb with HI IgG and Fc were approximately 10-fold higher relative to those observed for PI IgG and Fc. Enzymatic deglycosylation of HI and PI Fc gamma led to elimination of CD spectral differences. Further, association constants obtained for RF and MoAb reactions with deglycosylated (sialic acid and galactose removed) PI Fc gamma were equivalent to those obtained in the presence of untreated HI Fc gamma. Together, these results suggest the complex oligosaccharide structure of rabbit IgG may play a significant role in the expression of Fc gamma determinants, and alteration of this structure under hyperimmune or other conditions may be related to induction of an RF response.

Animals↗

PubMed1990

Three-dimensional structures of maturable and abortive capsids of equine herpesvirus 1 from cryoelectron microscopy.

Cryoelectron microscopy and three-dimensional computer reconstruction techniques have been used to compare the structures of two types of DNA-free capsids of equine herpesvirus 1 at a resolution of 4.5 nm. "Light" capsids are abortive, whereas "intermediate" capsids are related to maturable intracellular precursors. Their T = 16 icosahedral outer shells, approximately 125 nm in diameter, are indistinguishable and may be described in terms of three layers of density, totalling 15 nm in thickness. The outermost layer consists of protruding portions of both the hexon and the penton capsomers, rising approximately 5 nm above a midlayer of density. The innermost layer, or "floor," is a 4-nm-thick sheet of virtually continuous density except for the orifices of the channels that traverse each capsomer. Hexon protrusions are distinctly hexagonal in shape, and penton protrusions are pentagonal. The structures of the three kinds of hexons (distinguished according to their positions on the surface lattice) are closely similar but differ somewhat in their respective orientations and in the shapes of their channels. The most prominent features of the midlayer are threefold nodules ("triplexes") at the trigonal lattice points. By analogy with other viral capsids, the triplexes may represent trimers of another capsid protein, possibly VP23 (36 kilodaltons [kDa]) or VP26 (12 kDa). Intermediate capsids differ from light capsids, which are empty, in having one or more internal components. In individual images from which the shell structure has been filtered away, these components are seen to have dimensions of 20 to 30 nm but to lack a visible substructure. This material--which is smeared out in the reconstruction, implying that its distribution is not icosahedrally symmetric or necessarily consistent from particle to particle--consists of aggregates of VP22 (46 kDa). From several lines of evidence, we conclude that this protein is located entirely within the capsid shell. These aggregates may be the remnants of morphogenetic cores retained in capsids interrupted in the process of DNA packaging.

Animals↗

PubMed1990

Characterization of immunoreactive motilin from the rat small intestine.

Immunocytochemistry, radioimmunoassay, chromatography, and biological assay using a rabbit isolated duodenal muscle strip preparation were used in attempting to characterize motilin from the rat small intestine. Several different antisera and monoclonal antibodies directed against natural porcine motilin were used. A variety of fixation techniques using Bouin's, paraformaldehyde, and benzoquinone with different staining methods including, fluorescein-conjugated second antibody, peroxidase-antiperoxidase or peroxidase-conjugated second antibody techniques were used. All methods failed to detect immunoreactive motilin cells in the rat small intestine. The same antisera were used in radioimmunoassays for motilin to evaluate extracts of rat intestinal tissue. Two of these detected immunoreactive motilin in gut extracts, and these antisera showed a different distribution for the peptide. Samples containing immunoreactive motilin obtained from cation exchange chromatography on SP-Sephadex-G25 were concentrated and assayed for biological activity in a rabbit duodenal muscle strip preparation. Desensitization of duodenal tissue to porcine motilin could be demonstrated by pretreatment with this peptide. The biological activity of partially purified rat intestinal immunoreactive motilin was not prevented by pretreatment of the tissue with motilin. Further purification of this preparation on Bio-Gel P-10 yielded an immunoreactive motilin peak that co-eluted with natural porcine motilin. Rat intestinal immunoreactive motilin did not co-elute with natural porcine motilin following high pressure liquid chromatography on a Waters microBondapak C18 reversed-phase column using a linear gradient of water-acetonitrile (10-45%) over 30 min. Although of similar molecular size, rat motilin is probably structurally dissimilar to other mammalian motilins.

Animals↗

PubMed1990

Clinical features of patients attending a gender-identity clinic.

Of 106 patients attending a gender-identity clinic, 73% satisfied DSM-III criteria for transsexualism. These DSM-III positives had a significantly younger age of onset and were less likely to experience sexual arousal with cross-dressing than those who did not satisfy DSM-III criteria. They were also significantly more likely to fulfil a definition of 'core transsexualism'. 'Core transsexualism' may represent a subgroup within DSM-III criteria for transsexualism and its defining features in this study were an early age of onset, low sexual activity, lack of sexual arousal with cross-dressing and homosexual orientation. Of the whole sample, 23% were referred for gender reassignment surgery, of whom 100% were DSM-III positive and 60% were 'core transsexuals'.

Adult↗