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Biomedical subjects

J C Brown

Publications and source records attributed to J C Brown.

At least 163 records · Page 9Linked to original sources

Calculation of a "narrowed" autocorrelation function.

A method of calculating an autocorrelation function with extremely narrow peaks is described. This is done by including terms in the autocorrelation expression corresponding to delays at 2 tau, 3 tau, etc., in addition to the usual term with delay tau. Implications in the frequency domain are explored. Graphs of this autocorrelation function for a number of violin sound samples, including a two-octave scale, vibrato, and glissando, are presented. Graphs of the autocorrelation function for some synthetic sound samples are also included.

Acoustics↗

Use of Ar+ plasma etching to localize structural proteins in the capsid of herpes simplex virus type 1.

Partially cored herpes simplex virus type 1 (HSV-1) capsids (B capsids) were eroded in a low-energy (0.5-keV) Ar+ ion plasma under conditions in which the outermost structural proteins were expected to be degraded before more internal ones. After various periods of etching, the proteins remaining intact were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and determined quantitatively by densitometric scanning of the stained gels. The results showed that the major capsid polypeptide (VP5) and two other capsid proteins, VP19 and VP23, were degraded rapidly beginning as soon as capsids were exposed to the ion plasma. In contrast, significant lags were observed for erosion of VP21, VP22a, and VP24, suggesting that these proteins were available to accelerated ions only after other, more external structures had been damaged or eroded away. The results suggest that VP5, VP19, and VP23 are exposed on the surface of the capsid, while VP21, VP22a, and VP24 are found inside the capsid cavity. The experiments are consistent with the view that VP5 constitutes the major structural component of the hexavalent capsomers. It is proposed that VP19 and VP23 may form other surface structures such as the pentavalent capsomers, the capsid floor, or the intercapsomeric fibers.

Animals↗

Nucleocapsid mass and capsomer protein stoichiometry in equine herpesvirus 1: scanning transmission electron microscopic study.

The Brookhaven scanning transmission electron microscope was employed to measure the masses of two nucleocapsid species (of light and intermediate densities) of equine herpesvirus 1. These were found to be 196.7 +/- 9.2 and 229.0 +/- 9.5 megadaltons (MDa), respectively. Biochemical assays showed that neither nucleocapsid contained any significant amount of DNA (less than 0.2% [wt/wt]). Taking into account data on protein composition, we conclude that the difference between their masses is essentially contributed by viral protein 22 (46 kDa), which is an integral component of the maturable intermediate nucleocapsid but not of the abortive light nucleocapsid. In view of earlier ultrastructural information on capsomer symmetry, our mass determinations are consistent only with the 150 hexavalent capsomers being hexamers of the 148-kDa major capsid protein.

Animals↗

Role of organic anions in renal response to dietary acid and base loads.

Bicarbonate is formed when organic anions are oxidized systemically. Therefore, changes in organic anion excretion can affect systemic acid-base balance. To assess the role of organic anions in urinary acid-base excretion, we measured urinary excretion in control rats, NaHCO3-loaded rats, and NH4Cl-loaded rats. Total organic anions were measured by the titration method of Van Slyke. As expected, NaHCO3 loading increased urine pH and decreased net acid excretion (NH4+ + titratable acid - HCO3-), whereas NH4Cl loading had the opposite effect. Organic anion excretion was increased in response to NaHCO3 loading and decreased in response to NH4Cl loading. We quantified the overall effect of organic ion plus inorganic buffer ion excretion on acid-base balance. The amounts of organic anions excreted by all animals in this study were greater than the amounts of NH4+, HCO3-, or titratable acidity excreted. In addition, in response to acid and alkali loading, changes in urinary organic anion excretion were 40-50% as large as changes in net acid excretion. We conclude that, in rats, regulation of organic anion excretion can contribute importantly to the overall renal response to acid-base disturbances.

Acid-Base Equilibrium↗

Computer-assisted audits of blood component transfusion.

Comprehensive review of clinical blood transfusion practice at a tertiary-care medical center is complicated by the extraordinary number of patients that receive such therapy. Computer-assisted review of the key objective data used in making the decisions about transfusion is necessary to evaluate the process. Use of 15,873 units of red blood cells, 3,641 units of plasma, 2,619 pools of platelets or pheresis units, and 259 pools of cryoprecipitate was screened by comparing pre-transfusion and post-transfusion blood counts with the medical staff's evaluation criteria. On this basis, 81.4% of transfusion episodes (TEs) were considered fully justified. Medical records were selected for audit from the cases in which the transfusion decisions could not be justified by on-line information. Abstracted data subsequently justified 82 of 139 audited cases; 68.4% of the comments pertaining to the remaining 57 cases adequately explained the transfusion decision. Thus, nearly 96% of the TEs were justifiable as determined by peer review.

Blood Preservation↗

Hyperimmunization alters Fc gamma antigenicity. Linkage to glycosylation.

Rabbits were hyperimmunized with streptococcal vaccine to produce hyperimmune (HI) IgG, and both HI IgG and autologous preinoculation (PI) IgG were used to prepare Fc gamma. mAb raised against these, and deglycosylated PI Fc gamma preparations were subsequently used to determine if antigenic differences existed between autologous HI and PI Fc gamma. Although the majority of mAb examined did not discriminate between the two sources of Fc gamma, several mAb exhibited remarkable specificity for autologous HI Fc gamma. We did not detect any mAb which exhibited the converse specificity. Of the mAb chosen for study, all except one appeared specific for determinants located in C gamma 2; the exception specifically reacted with pFc', and did not discriminate between the two Fc preparations. Importantly, partial deglycosylation of autologous PI Fc gamma led to equivalent reactivity with previously HI-specific mAb. The results of this study provide immunologic evidence that abnormal C gamma 2 structures exist prior to and reach serologically detectable levels during the hyperimmune response in these animals, apparently as a result of glycosylation pattern alterations within the Fc region. Such autoantigenic differences could account for the induction of RF under these conditions, and perhaps in certain human arthritic diseases as well.

Alkylation↗

NMR-invisible lactate in blood plasma.

Resonances for lactate are broadened in 500 MHz 1H NMR spectra of human blood plasma and only about one-third is visible in Hahn spin-echo spectra. Similar effects are observed for some other carboxylate anions. Lactate added to the high-Mr fraction of plasma can give rise to peaks which are too broad to observe in either single-pulse or spin-echo spectra. Addition of agents such as NH4Cl of SDS dramatically increases the intensities of lactate peaks. Some glycoproteins appear to broaden lactate resonances.

Ammonium Chloride↗

Existence and coexistence of calcitonin gene-related peptide, vasoactive intestinal polypeptide- and somatostatin-like immunoreactivities in spinal cord motoneurons of developing embryos and post-hatch chicks.

By use of immunocytochemical methods, it is shown that immunoreactive calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and somatostatin (SOM) are present in motoneurons in the chicken spinal cord. While CGRP-like immunoreactivity (LI) is present in numerous motoneurons both before and after hatching, SOM- and VIP-LI markedly decline at the end of the embryonic period. Evidence is also provided for coexistence of some of these peptides in certain spinal motoneurons.

Animals↗

Effect of galanin on endocrine secretions from the isolated perfused rat stomach and pancreas.

Nerves containing galanin immunoreactivity have been shown to be present in the stomach and pancreas. The present experiments were designed to test the effect of galanin on the release of gastric somatostatin-like immunoreactivity (SLI) and immunoreactive gastrin (IR-G), pancreatic SLI and immunoreactive insulin (IR-I) from the isolated perfused rat stomach and pancreas respectively. Galanin (2 X 10(-10)-5 X 10(-8) M) inhibited gastric SLI and IR-G release dose dependently. At a concentration of 10(-8) M, galanin also suppressed IR-I release stimulated by gastric inhibitory polypeptide (2 X 10(-10) M) in the presence of 8.9 X 10(-3) M glucose. Pancreatic SLI release under this condition was not altered. Thus the present study suggests that galanin might be involved in the neural regulation of gastric and pancreatic endocrine secretions.

Animals↗

Factors affecting 1H NMR spectra of blood plasma: cancer, diet and freezing.

Single pulse and Hahn spin-echo 400 and 500 MHz 1H NMR spectra from normal subjects and cancer patients are reported. No significant difference was found between the average -CH3 and -(CH2)n- lipoprotein linewidths of the two groups in contrast to a previous report. Such measurements are shown to be complicated by freezing and thawing of samples which broadens lipoprotein resonances, and by eating which sharpens them. Deconvolution of the composite lipoprotein signals enables these effects to be interpreted. Other notable features of plasma spectra are discussed including resonances for 'acute-phase' glycoproteins, ketone bodies and other small molecules.

Acute-Phase Proteins↗

TRH-like immunoreactivity in endocrine cells and neurons in the gastro-intestinal tract of the rat and guinea pig.

By use of the indirect immunofluorescence technique, the cellular localization of thyrotropin-releasing hormone (TRH) was studied in the gastrointestinal tract of rats and guinea pigs of different ages. TRH-like immunoreactivity (LI) was observed in many pancreatic islet cells of young rats and guinea pigs but only in single cells of 6-month-old rats. In aged guinea pigs, a reduction in the number of TRH-positive cells was evident; however, numerous strongly fluorescent cells were still present. In the guinea pig, TRH-LI was in addition observed in gastrin cells in the stomach. TRH-positive nerve fibers occurred in the myenteric plexus of the oesophagus, stomach and intestine of the rat, and in the muscle layers of the guinea pig. These results suggest a functional role of TRH both as hormone and neuroactive compound in various portions and sites of the gastro-intestinal tract of the rat and guinea pig.

Animals↗

Autoradiographic studies of the gamma-aminobutyric acid (GABA) system in the rat pancreas.

The beta-cells of the pancreatic islets have been shown to contain gamma-aminobutyric acid (GABA) together with insulin. Autoradiographic analysis indicated that high affinity GABA binding sites (GABA receptors) are not present in the pancreas. High affinity GABA uptake sites are present, not in beta-cells, but in a few cells on the periphery of the islets. These observations cast doubt on the suggestion that GABA has a paracrine role in the pancreas.

Animals↗

Use of Ar+ plasma etching to localize structural proteins in viruses: studies with adenovirus 2.

The experiments described here were undertaken to test the idea that low energy Ar+ plasma etching could be employed as the basis of a method to order viral structural polypeptides according to their physical proximity to the virus surface. Since low energy (500 eV) Ar+ ions do not penetrate deeply into virus surfaces, one expects that the outermost proteins will be damaged before internal ones when intact virions are irradiated. To test this expectation, we exposed adenovirus 2 to a 500-eV Ar+ plasma and then employed sodium dodecyl sulfate-polyacryl-amide gel electrophoresis to assess the extent of damage to the major structural polypeptides. Gel analyses showed that the proteins exposed on the virus surface (proteins II, III, and IV) were degraded rapidly during the first 10 s of irradiation while protein VII, the major core polypeptide, was almost completely protected. Proteins located between the capsid and the core, such as proteins IIIa and VI, were degraded at intermediate rates. Quantitative measurements demonstrated that the observed decay rate differences were not due simply to differences in protein target size; distance to the virion surface made an important contribution. The plasma etching technique, therefore, appears to have considerable potential for the structural analysis of viruses and other macromolecular assemblies where the proximity of individual proteins to the particle surface is unknown.

Adenoviridae↗

Cysteamine-induced reduction in gastrointestinal somatostatin: evidence for a region-specific loss in immunoreactivity.

Administration of cysteamine (beta-mercaptoethylamine; 2-aminoethanethiol) to rats has been shown to decrease the levels of somatostatin-like immunoreactivity (SLI) in the gastrointestinal tract and pancreas but its mode of action is unclear. In the current study the effect of cysteamine on gastrointestinal and pancreatic SLI has been studied using two antisera with different regional specificities. In addition, the in vitro effect of cysteamine on SS-14 and SS-28 has been studied by high-performance liquid chromatography (HPLC). Characterization of the two antisera (AS 26.3.2 and AS 1001) with a range of analogs of SS-14 revealed that both were directed against the midportion of the molecule but that AS 1001 was also sensitive to changes at the N- and C-termini. Tissue extracts from cysteamine-treated rats measured with AS 26.3.2 showed no significant change for the stomach, jejunum or pancreas but duodenal levels were reduced. With AS 1001 SLI levels were reduced in all tissues. Gel permeation chromatography of stomach extracts measured with AS 1001 showed a reduction in both SS-14 and SS-28. With AS 26.3.2 an increase in SLI eluting prior to the SS-14 peak occurred explaining why no significant reduction in total SLI was detected. With duodenal extracts the elution profiles with AS 1001 reflected the large reduction in total SLI whereas with AS 26.3.2 a smaller reduction occurred. Both SS-14 and SS-28 were reduced. HPLC analysis of SS-14 and SS-28 following incubation with cysteamine in vitro showed a time-dependent decrease in both somatostatin species with absorbance at 280 nm was measured. New peptide peaks which developed were not all detectable by radioimmunoassay with either antibody. The results suggest that cysteamine causes a change in the structure of somatostatin which probably first involves a reduction of the disulphide bridge and then the N- and C-terminal regions of the molecule thus making it unmeasurable by antisera sensitive to changes in these regions.

Animals↗

Enteroinsular axis.

A physiological role for GIP as an insulinotropic hormone involved in the enteroinsular axis has been established and ingestion of glucose, fatty acids and certain amino acids will produce an increase in circulating IR-GIP levels. The insulinotropic action of GIP is glucose concentration dependent in normal animals. A role for GIP in NIDDM is equivocal although several studies have demonstrated elevated serum levels. Animal models have indicated a disturbance of GIP receptor function associated with hyperinsulinaemia, i.e. lowering of the minimum glucose concentration at which GIP is insulinotropic.

Diet↗

Plasma enteroglucagon and small bowel cytokinetics in rats fed soluble nonstarch polysaccharides.

The influence of dietary fiber on plasma glucagon and enteroglucagon (EG) and on ileal crypt cell production rate (CCPR) was assessed in the rat. A fiber-free semisynthetic diet and diets containing insoluble cellulose, Na-carboxymethylcellulose (CMC), guar gum, pectin, gum arabic, methylcellulose (MC), and hydroxypropylmethylcellulose (HPMC) at 10 g/100 g were fed for 14 d. Animals given soluble polysaccharides had plasma EG levels significantly higher than animals given insoluble cellulose, but only those fed the most viscous gums showed evidence of increased mucosal cell proliferation in the distal ileum. It was concluded that viscous nonfermentable polysaccharide gums stimulate the release of EG in the rat by slowing the absorption of nutrient, whereas the nonviscous polysaccharide gum arabic, or its breakdown products, probably directly stimulate mucosal endocrine cells. The failure of gum arabic to stimulate CCPR despite increased plasma EG indicated that EG alone is not a sufficient stimulus for increased mucosal cell proliferation.

Animals↗

Isolation from porcine intestinal extracts of a cholecystokinin-like peptide and a peptide with homology to cytochrome oxidase polypeptide VII and chymodenin.

A number of different forms of cholecystokinin (CCK) exist in the brain and intestine. Gel permeation and ion exchange chromatography and high performance liquid chromatography have been used to isolate a peptide from partially purified porcine intestinal extracts with N-terminal homology to porcine brain CCK-58. This peptide contracted both the guinea pig ileum longitudinal muscle and gallbladder muscle and these responses were inhibited by dibutyryl cyclic GMP or proglumide. The potency was approximately 1/100 of that of CCK-8. The reason for this low potency is unclear, but it is possible that a critical part of the biologically active region is modified or that it is a truncated form of CCK-58. A further peptide was isolated with a sequence homologous to cytochrome oxidase polypeptide VII and chymodenin.

Amino Acid Sequence↗