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R C Roberts

Publications and source records attributed to R C Roberts.

At least 109 records · Page 6Linked to original sources
PubMed1987

Three high molecular weight protease inhibitors of rat plasma. Reactions with trypsin.

We have compared the reactions of trypsin with human alpha 2-macroglobulin (alpha 2M), and three rat plasma protease inhibitors, alpha 1-macroglobulin (alpha 1M), alpha 1-inhibitor III (alpha 1I3), and alpha 2M. All four of these proteins appear to contain reactive thiol esters. The electrophoretic mobility in agarose gels of human and rat alpha 2M is increased by 1 mol of trypsin, while the mobility of alpha 1M and alpha 1I3 is decreased. Treatment with methylamine causes similar mobility changes, except in the case of rat alpha 2M. Titration of human and rat macroglobulins by repeated small additions of trypsin and by assay of liberated SH groups or enhanced ligand fluorescence revealed a stoichiometry of about 1 mol of trypsin/mol of inhibitor. In contrast, addition of macroglobulin to a fixed amount of trypsin and detection of residual amidase or protease activity revealed a stoichiometry of about 2 mol of trypsin for 1 mol of human alpha 2M, about 1.4 mol for rat alpha 1M, and about 1 mol for rat alpha 2M. One mol of trypsin reacted with 2 or more mol of alpha 1I3 by the criteria of SH groups liberated or protease inhibition. Methylamine-treated rat alpha 2M binds a significant amount of trypsin releasing about 2 mol of SH. Radioactive beta-trypsin was covalently bound to subunits of the purified plasma inhibitors. The Mr of the labeled products with rat and human alpha 2M had molecular weights which suggested trypsin was bound to intact as well as cleaved subunit chains and also to multiple chains via cross-linking. Rat alpha 1M also produced a product which may be an intact subunit alpha chain plus trypsin. Greater than 80% of the trypsin was bound covalently to these inhibitors at low molar ratios.

Acute-Phase Proteins↗
PubMed1987

Three high molecular weight protease inhibitors of rat plasma. Isolation, characterization, and acute phase changes.

Rat blood plasma contains three high molecular weight thiol ester-containing proteinase inhibitors, alpha 1-macroglobulin (alpha 1M), alpha 1-inhibitor III (alpha 1I3), and alpha 2-macroglobulin (alpha 2M). Rat serums have been analyzed using a two-dimensional gel electrophoretic technique which optimizes recovery of high molecular weight proteins. alpha 1M, and (alpha beta)4-tetramer in native solution, separated in the second sodium dodecyl sulfate-containing electrophoretic dimension as a disulfide-linked (alpha beta)2-dimer with an approximate Mr of 360 kDa. alpha 1I3 separated in the gels as a single 190-kDa polypeptide. It is also a monomer in native solution by ultracentrifugation criteria. Native rat alpha 2M is a tetramer, but it separates in the gels as a disulfide-linked dimer with an Mr of approximately 360 kDa. The kinetics of changes in concentration of these proteins during the induction of polyarthritis was also measured by quantitative immunoelectrophoresis. In rats with adjuvant-induced polyarthritis, the concentration of alpha 1I3 dramatically decreases and alpha 2M appears and continues to increase in a biphasic manner for 2 weeks. The alpha 1M concentration remains relatively constant. All three macroglobulins were purified utilizing modern rapid chromatographic techniques, and parallel comparisons of their native physicochemical properties were carried out. The N-terminal sequence of the alpha-chain of rat alpha 1M was also shown to share sequence homology with that of alpha 2M. In agreement, Esnard et al. (Esnard, F., Gutman, N., El Moujahed, A., and Gauthier, F. (1985) FEBS Lett. 182, 125-129) recently reported that alpha 1I3 also contains a thiol ester bond, as do alpha 1M and alpha 2M, since it reacts covalently with [14C]methylamine and is cleaved autolytically at 80 degrees C. We have examined negatively stained preparations of native, trypsin-treated, and methylamine-treated human alpha 2M, rat alpha 2M, and rat alpha 1M in the electron microscope. Trypsin appears to convert globular ring-shaped native molecules to rectangular box-like structures, in agreement with the conclusions of a recent report on human alpha 2M (Tapon-Bretaudiere, J., Bros, A., Couture-Tosi, E., and Delain, E. (1985) EMBO J. 4, 85-89).

Acute-Phase Proteins↗
PubMed1987

Postpartum lymphocytic thyroiditis. Prevalence, clinical course, and long-term follow-up.

A group of 238 women were surveyed for thyroid disease at six and 12 weeks post partum. Twenty-seven (11.3%) of 238 entered into the study were found to have thyroid disease. Fifteen (56%) of 27 had positive microsomal hemagglutinin antibody titers. A spectrum of thyroid disease was found: persistent hypothyroidism (two patients), transient thyrotoxicosis followed by persistent hypothyroidism (one) or transient hypothyroidism (three), euthyroid goiter (five), transient thyrotoxicosis (seven), transient hypothyroidism (three), high-normal thyroxine levels (five), and low-normal thyroxine levels (one). All nine patients who underwent biopsy had active lymphocytic thyroiditis. Three-year follow-up of 25 of the 27 affected individuals revealed that 12 (48%) still had thyroid disease. This study demonstrates that there is a high incidence of postpartum thyroid disease, usually of a transient nature, and that only about one fourth of the cases are detected or clinically obvious.

Cross-Sectional Studies↗
PubMed1987

An electron microscopic study of GABAergic neurons and terminals in the central nucleus of the inferior colliculus of the rat.

Neurons and terminals in the ventral lateral portion of the central nucleus of the inferior colliculus (ICCN) of the rat were labelled immunocytochemically with antisera to GABA or to its synthesizing enzyme, glutamic acid decarboxylase. Four types of GABAergic neuron are described: small, medium-sized and large multipolar neurons, as well as medium-sized bipolar neurons. All sizes of GABAergic multipolar neurons are characterized by highly infolded nuclei, many mitochondria and both asymmetric and symmetric axosomatic synapses. A dense plexus of terminals occurs on the proximal dendrites of GABAergic neurons, and most of these terminals form asymmetric axodendritic contacts. Small GABAergic neurons (diameter less than 15 microns) are multipolar, and have a large nucleus to cytoplasm ratio, prominent nucleoli and usually two to five axosomatic synapses per thin section, with the majority of these contacts being symmetric. Medium-sized GABAergic neurons (15-25 microns in diameter) display both multipolar and fusiform shaped somata, have a more abundant cytoplasm than the small neurons and show about ten axosomatic contacts per thin section. Large GABAergic neurons (diameters greater than 25 microns) have eccentrically located, highly infolded nuclei, abundant cytoplasm and a denser plexus of terminals that form axosomatic synapses than the other cell types. These results indicate that four of the six major cell types in the ICCN are probably GABAergic inhibitory neurons. The axon initial segments of GABAergic neurons in the ICCN all have similar features in that they are contacted by only one or two terminals that form symmetric synapses on their proximal portions and are invested by a glial sheath from 3 to 20 microns from the cell body. Many immunoreactive myelinated axons (approximately 0.5 micron in diameter) are observed and some terminals that arise from these axons form synapses with small neuronal somata. Both these and other labelled terminals are shown to form symmetric synapses. These data suggest a complex circuitry for the GABAergic neurons within the ICCN.

Animals↗
PubMed1987

Characterization of Bdellovibrio bacteriovorus bacteriophage MAC-1.

The bacteriophage MAC-1, which specifically infects Bdellovibrio bacteriovorus, was plaque purified and raised to high titre. The phage was purified by NaCl/polyethylene glycol precipitation, followed by two cycles of isopycnic density gradient centrifugation in CsCl. The purified phage exhibited a density of 1.363 g cm-3 and a sedimentation coefficient of 94S. Nucleic acid isolated from purified phage was resistant to hydrolysis under alkaline conditions and to digestion with RNAase, but it was hydrolysed by DNAase, providing evidence that the phage genome is made up of DNA. The lack of hyperchromic effect upon denaturation, hydrolysis of phage DNA by S1 nuclease, characteristic fluorescent staining with acridine orange, and resistance to digestion with a variety of restriction endonucleases are consistent with the DNA being single-stranded. A buoyant density of 1.722 g cm-3 and a sedimentation coefficient of 17.9S were obtained for the phage DNA. The molecular mass of phage DNA was determined as 1.58 MDa by agarose gel electrophoresis with single-stranded DNA as standards. Electron microscopy of the DNA showed that the genome is circular in nature. In addition, using Southern blots, the two replicative forms, RF1 (supercoiled) and RF2 (circular) have been identified and isolated from infected cell extracts.

Bacteriophages↗
PubMed1987

Inhibition of monosynaptic reflexes in the human lower limb.

1. Presynaptic inhibition of muscle spindle Ia afferents by afferents from the same and other muscles has been studied in the human lower limb. The experiments have utilized conditioning of test monosynaptic reflexes by vibration of both the test and other muscles. 2. The pattern of inhibition invariably includes autogenetic actions. 3. There are powerful effects from flexor to extensor Ia afferents. Actions from flexor to flexor, and from extensor to extensor, are weaker. Actions from extensors to flexors are very weak. 4. The strength of presynaptic inhibition from one muscle type to another weakens as the muscles considered become more anatomically distant. 5. The inhibition studied both by vibration and by electrical conditioning stimulation of nerves becomes weaker during voluntary isometric contraction of the test muscle. It is strongest at rest and during antagonist contraction. 6. Evidence is provided suggesting that descending control is the primary cause of this modulation of inhibition during contraction. 7. Stimulation of afferents in cutaneous nerves reduces group I presynaptic inhibition of Ia afferents.

Adolescent↗
PubMed1986

Regulation of protein synthesis in rabbit reticulocyte lysates. Requirement of initiation factor eIF-2 holoprotein for substrate specificity of heme-regulated protein kinase.

The specificity of the heme-regulated protein kinase (HRI) was investigated further by utilizing the isolated 38,000 Da subunit (alpha subunit) polypeptide of eIF-2 as the substrate. For this purpose, the three subunit polypeptides of eIF-2 (38,000 Da, alpha; 50,000 Da, beta; and 52,000 Da, gamma) were resolved by reversed-phase high performance liquid chromatography (HPLC). Results show that HRI is incapable of phosphorylating the 38,000 Da subunit separated from the other two eIF-2 polypeptides. Data suggest that the substrate specificity of HRI is determined by the quaternary structure assumed by the alpha subunit in association with the other two subunits in the eIF-2 holoprotein.

Animals↗
PubMed1986

Functional activities and nonenzymatic glycosylation of plasma proteinase inhibitors in diabetes.

The functional activity of three of the major plasma protease inhibitors, alpha 1-proteinase inhibitor, antithrombin III and alpha 2-antiplasmin, has been measured in a series of persons with diabetes mellitus and compared with healthy controls. The mean specific functional activity of both diabetic plasma antithrombin III (50.8 +/- 7.0 U/mg) and alpha 1-proteinase inhibitor (35.3 +/- 5.6 mU/mg) was found to be significantly lower than in healthy controls (57.9 +/- 6.0 U/mg) and (42.2 +/- 10.7 mU/mg). No difference was detected in alpha 2-antiplasmin activity or levels. The glycosylated protein fraction of 83% of diabetic plasmas showed the presence of less than 1% of the total alpha 1-proteinase inhibitor when isolated by phenylboronate affinity chromatography. Incubation of normal plasma with 250 nmol/l glucose (a level approximately 6 X higher than encountered in uncontrolled diabetes) resulted in a 17% and 6% decrease in antithrombin III and alpha 1-proteinase inhibitor activities. We conclude that the decrease in specific activity of alpha 1-proteinase inhibitor is not related to nonenzymatic glycosylation, but the decrease in antithrombin III specific activity may be related.

Adult↗
PubMed1986

Anatomical changes of the GABAergic system in the inferior colliculus of the genetically epilepsy-prone rat.

The number of GABAergic neurons as determined by GAD immunocytochemistry and total neurons as determined from Nissl preparations were counted and classified at the light microscopic level in the inferior colliculus (IC) of the genetically epilepsy prone rat (GEPR) and the non-epileptic Sprague-Dawley (SD) strain of rat. GAD-positive neurons are abundant in the IC and a significant increase in the number of GAD-positive neurons occurs in the GEPR as compared to the SD in all three subdivisions. However, the most pronounced difference occurs in the ventral lateral portion of the central nucleus, where there is a selective increase in the small (200%) and medium-sized (90%) GABAergic somata (10-15 microns in diameter and 15-25 microns in diameter, respectively). As determined from Nissl preparations an increase in total numbers of neurons also occurs. Thus, a 100% increase in the number of small neurons and a 30% increase in the number of medium-sized neurons occur in the adult GEPR as compared to the SD rat. A statistically significant increase in the numbers of small neurons also occurred in the IC of the young GEPR. At 4 days of age, a 55% increase in the number of small neurons was found, and at 10 days of age this increase was 105%. The numbers of the medium and large neurons were similar in the older group of rats. These data suggest that the increase in cell number observed in the adult GEPR is not compensatory to the seizure activity, but may either be genetically programmed or be a failure of cell death. Based on other studies of genetic models of epilepsy, we propose that the additional GABAergic neurons may disinhibit excitatory projection neurons in the IC.

Acoustic Stimulation↗
PubMed1986

Reduced trypsin binding capacity of alpha 2-macroglobulin in diabetes.

The plasma proteinase inhibitor, alpha 2-macroglobulin, is usually elevated in diabetes. The trypsin binding capacity and the concentration of alpha 2-macroglobulin in 90 diabetics sera were compared with 30 age- and sex-matched normal sera. The mean alpha 2-macroglobulin concentration determined by radial immunodiffusion was 313 mg/dl for the diabetics as compared to 240 mg/dl for the healthy subjects (significantly higher, p less than 0.01). The mean of the ratio, mol trypsin bound/mol alpha 2-macroglobulin (molar binding ratio) for the Type I diabetics (n = 54), 0.82, was significantly lower than the mean of the healthy subjects, 0.87, or the mean of the Type II diabetics, 0.87. No relationship between the molar binding ratios and the levels of glycosylated hemoglobin was found. The alpha 2-macroglobulin was isolated from the plasma of 11 Type I diabetics and 7 normals. The maximum molar trypsin binding capacities of the diabetic alpha 2-macroglobulin were significantly lower. The mean for the diabetic alpha 2-macroglobulin was 1.72 vs. 1.97 for the normal alpha 2-macroglobulin. These results indicate that the trypsin binding function of alpha 2-macroglobulin is moderately impaired in diabetes. No differences were found in the extent of proteolytic cleavage of the 'bait region' of diabetic alpha 2-macroglobulin, autolytic cleavage or the methylamine reaction at the thiolester site between diabetic and normal alpha 2-macroglobulin. Nonenzymatic glucosylation of normal alpha 2-macroglobulin did not lower the trypsin binding capacity. The nature of the modification of alpha 2-macroglobulin leading to reduced trypsin binding capacity or the physiological significance is not yet known.

Adult↗
PubMed1986

The ultrastructure of the central nucleus of the inferior colliculus of the Sprague-Dawley rat.

Previous studies from this laboratory have indicated that increased numbers of GABAergic neurons, as well as total neurons, occur in the central nucleus of the inferior colliculus (IC) of genetically epilepsy-prone rats (GEPRs) as compared to non-seizuring Sprague-Dawley rats. Since electron microscopic studies of the IC have not been reported for rats, we wanted to determine the ultrastructure of neurons and their processes in this brain region to serve as a basis for future studies on neuronal circuitry in the GEPRs. Both disc-shaped and stellate types were found for each of three size categories: large, medium and small. Thus, six types of neuron were distinguished by differences in somatic size, shape, organelles and dendritic orientation. Large neurons (longest diameter greater than 25 micron), which are the least frequent cell type, contained vast perikaryal cytoplasm, eccentrically located nuclei and abundant granular endoplasmic reticulum (GER) adjacent to the nucleus as well as clustered in the cytoplasm; many axosomatic, symmetric synapses were present. Medium-sized neuronal somata (15-25 micron in diameter) had smooth as well as infolded nuclear membranes and clusters of GER in their cytoplasm but no GER adjacent to the nucleus; synapses were sparse along the surface of their somata. Small neurons (10-15 micron in diameter), which are the most frequent cell type, had scant perikaryal cytoplasm, usually infolded nuclei, frequently two nucleoli, and few or no stacked cisternae of GER in the perikaryal cytoplasm; only infrequent axosomatic synapses were found. Based on previous retrograde and immunocytochemical studies, most large disc-shaped and stellate cells project to the medial geniculate body and are probably excitatory, but some large stellate neurons have been shown to be GABAergic and it is doubtful that such neurons participate in this projection. A dense plexus of terminals that form symmetric synapses covers the soma and proximal dendrites of large neurons, and may provide a strong GABAergic inhibition of this type of projection neuron. Small and medium-sized disc-shaped cells also project to the thalamus but they lack this dense axosomatic plexus. The stellate cells from these same two size categories probably do not project to the thalamus and may be GABAergic local circuit neurons. Other ultrastructural features of IC neurons that were analysed include dendrites, dendritic spines, axon hillocks, initial segments and terminals, as well as the laminae of myelinated axons. Dendrites were either beaded or smooth and few spines were observed.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Characterization of dusts collected from swine confinement buildings.

As part of a project to evaluate health hazards for workers in swine confinement buildings, the air in 21 different buildings was sampled with 37 mm cassette filters with and without cyclone preselectors and with cascade impactors. Filter results yielded a mean total aerosol of 6.3 mg/m3, a mean respirable aerosol of 0.5 mg/m3; the geometric mean diameter was 2.9 microns. Cascade impactor measurements revealed a mean total aerosol of 7.6 mg/m3, a respirable aerosol of 2.5 mg/m3 and a mass median diameter of 9.6 microns. The two major constituents in these aerosols were grain particles and dried fecal matter. The grain particles were larger than fecal particles and proportionately more abundant in finishing buildings where 50 kg X 100 kg animals are housed. Therefore the respirable fraction was less in finishing buildings than in farrowing and nursery buildings. Culturing of settled dusts yielded six different mold species, with the highest counts for Verticillium sp. (5 X 10(2) cfu/mg dry dust) grown at 37 degrees C. Thermophilic Actinomycetes and both gram negative and gram positive bacteria were isolated. Azocasein proteinase activity was found in most dust samples analyzed. This dust had a protein content of about 23% and a mean adsorbed ammonia content of 0.4%.

Agricultural Workers' Diseases↗
PubMed1986

Presynaptic inhibition of monosynaptic reflexes in the lower limbs of subjects with upper motoneuron disease.

Presynaptic inhibition of muscle spindle Ia afferents by group I afferents from the same and other muscles has been studied in the lower limbs of subjects with upper motoneuron lesions. The experiments utilised conditioning of soleus test monosynaptic reflexes during controlled voluntary contraction. The protocol was designed to isolate presynaptic inhibition from postsynaptic components. The relation between estimate of inhibition and test reflex amplitude was examined. The subjects showed less inhibition than controls at all levels of voluntary torque investigated (less than or equal 15 Nm). Two thirds had weak inhibition which did not show the decrease during muscle contraction characteristic of controls. The degree of difference from the normal situation correlated with severity of the clinical sign (weakness of voluntary ankle flexion).

Adult↗
PubMed1985

Increased numbers of GABAergic neurons occur in the inferior colliculus of an audiogenic model of genetic epilepsy.

The numbers of GABAergic neurons as determined by GAD immunocytochemistry and total neurons as determined from Nissl preparations were counted and classified at the light microscopic level in the inferior colliculus (IC) of the genetically epilepsy-prone rat (GEPR) and the non-epileptic Sprague-Dawley (SD) strain of rat. GAD-positive neurons are abundant in the IC in all 3 subdivisions. Several sizes of multipolar neurons as well as medium-sized bipolar or fusiform neurons are GAD-positive. GAD-positive punctate structures that were interpreted to be axon terminals and transversely-sectioned dendrites and preterminal axons are abundant in the IC of both the GEPR and SD. A dramatic increase in the number of GAD-positive neurons occurs in the GEPR as compared to the SD. This increase is most evident in the middle of the rostrocaudal extent of the IC. Although the increase is statistically significant in all subdivisions of the IC, it is most pronounced in the central nucleus, especially the ventral lateral portion. Within the central nucleus, the increase in the number of GAD-positive neurons is due to a selective increase in the small (200%) and medium (90%) cell body size populations (10-15 micron and 15-25 micron in diameter, respectively). Concomitant with this increase in the number of GAD-positive neurons, an increase in total numbers of neurons occurs as determined from Nissl preparations. A 100% increase in the number of small neurons and a 30% increase in the number of medium-sized neurons occur in the GEPR as compared to the SD rat. The proportion of GAD-positive neurons to total neurons is also increased in the GEPR. Approximately 25% of the neurons in the IC in SD rat are GAD-positive, while about 35% of the neurons in the GEPR are GAD-positive. These data demonstrate an anatomical difference in the IC of the GEPR as compared to the SD which appears to be preferential for the GABAergic system.

Acoustic Stimulation↗
PubMed1985

Increased numbers of neurons occur in the inferior colliculus of the young genetically epilepsy-prone rat.

To determine if the increase in the number of neurons observed in the inferior colliculus (IC) of the adult genetically epilepsy-prone rat (GEPR) as compared to the Sprague-Dawley rat was present in the young GEPRs prior to the time at which seizure activity commences, brains from both types of rats 4-10 days of age were studied. A statistically significant increase in the numbers of small neurons occurred in the IC of the young GEPR. At 4 days of age, a 55% increase in the number of small neurons was found in the GEPR as compared to the Sprague-Dawley rat and at 10 days of age this increase was 105%. The numbers of the medium and large neurons were similar in the older group of rats. These data suggest that the increase in cell number observed in the adult GEPR is not compensatory to the seizure activity, but is genetically programmed.

Acoustic Stimulation↗