Search PubMed⌕ Search

Biomedical subjects

R C Roberts

Publications and source records attributed to R C Roberts.

At least 73 records · Page 4Linked to original sources
PubMed1993

Differential effects of suicide transport lesions of the striatonigral or striatopallidal pathways on subsets of striatal neurons.

In the basal ganglia, centrally active suicide transport agents produce apparently selective lesions of the striatopallidal and striatonigral pathways based on receptor binding and neuropeptide mRNA studies. In the present study we sought to determine the selectivity of suicide transport lesions for specific subsets of striatal neurons. Using immunohistochemical methods, the neostriata of adult rats were examined 10 days after an injection of volkensin into the substantia nigra or an injection of OX7-saporin into the globus pallidus. Ricin, a suicide transport agent active in the peripheral but not the central nervous system, was injected into each target as a control. Adjacent sections were processed for (1) Nissl stain to assess neuronal density, both overall and for large interneurons, (2) NADPH-diaphorase (NADPH-d) histochemistry, to mark medium-sized aspiny interneurons, (3) enkephalin immunocytochemistry, to label striatopallidal neurons, or (4) substance P immunocytochemistry, to label striatonigral neurons. Ricin injections produced no change in the densities of these subsets of striatal cells. In animals receiving volkensin or OX7-saporin injections, analyses of Nissl material revealed that the striata ipsilateral to the toxin injections appeared normal and did not exhibit shrinkage or gliosis; however, a quantitation analysis revealed a moderate decrease in cell density (12-16% loss, P < 0.01). The densities of both large and NADPH-d-containing striatal interneurons were unchanged after lesions in either target. Following nigral injections with volkensin, the density of striatal substance P-labeled cells decreased (26% loss, P < 0.01), while the density of enkephalin-labeled cells did not decrease significantly (11% decrease, P > 0.1). After pallidal injections with OX7-saporin, the density of striatal enkephalin-labeled cells decreased (20% loss, P < 0.01), while that of substance P-labeled cells remained unchanged. These data show that nigral volkensin and pallidal OX7-saporin injections differentially lesion striatonigral and striatopallidal projection neurons and spare striatal interneurons. This study provides further evidence for the selectivity, specificity, and utility of suicide transport agents to study brain structure and function.

Animals↗
PubMed1993

Intrastriatal injections of quinolinic acid or kainic acid: differential patterns of cell survival and the effects of data analysis on outcome.

There is controversy about the extent to which lesions of the rat striatum with excitatory amino acids mimic the cellular pathology seen in Huntington's Disease (HD). We sought to resolve this debate by determining with cell counts in adjacent sections the patterns of survival of medium spiny and aspiny striatal neurons using enkephalin immunohistochemistry and NADPH-diaphorase histochemistry as markers of these cell populations, respectively. Results showed that 2 weeks after quinolinic acid lesions, cell loss was qualitatively similar for the two cell groups. However, by varying the size of the sampling area for quantitative analyses and its distance from the lesion zone, the outcome of the statistical analyses varied enormously. Thus, a relative sparing of NADPH-diaphorase-labeled cells compared to enkephalin-labeled cells could be detected quantitatively in transition areas bordering the lesion under some but not all analytical conditions. Kainic acid lesions depleted both cell populations similarly, except in regions of transition farthest from the lesion, where enkephalin-containing neurons were more resistant than NADPH-diaphorase-containing cells. The size of the transition area around the lesion also differed depending upon excitotoxin and cell population. These results help to reconcile the controversy and suggest that with highly specified quantitative conditions quinolinic acid-induced injury of the striatum can resemble the histopathology of HD.

Analysis of Variance↗
PubMed1993

Line bisection errors in visual neglect: misguided action or size distortion?

The rightward line bisection errors made by patients with visuospatial neglect can be explained as due to a spatially misdirected response, which would be predicted on either of two accounts. An alternative view, however, is that such patients actually misperceive the left half of a horizontal line as being shorter than the right half. We have tested this possibility directly in three neglect patients, by giving them prebisected lines: they were found to judge a central transection mark as lying nearer to the left end of the lines. We were also able to test one of the patients on a series of size comparisons using computer-generated patterns. She was found to judge horizontal lines as shorter in the left half of visual space than in the right. This was also true for comparisons of the areas of nonsense figures. However she did not make such constant errors when comparing the lengths of vertical lines. It is suggested that an attentional deficit in left hemispace may result in the underestimation of horizontal extent. This would act in combination with misdirected reaching to determine the magnitude of line bisection errors.

Aged↗
PubMed1993

Cognitive brain potentials in a three-stimulus auditory "oddball" task after closed head injury.

Event-related potentials (ERPs) were recorded in a three stimulus oddball task from 16 patients who had sustained a severe closed head injury at least 6 months before testing, and from 16 control subjects. The stimuli comprised a random sequence of frequent non-target tones (P = 0.70), rare target tones (P = 0.15), and rare novel sounds (P = 0.15). The task requirement was to respond promptly to each target tone. From a latency of 200 msec onwards, the ERPs evoked by frequent non-targets were substantially more negative-going in the head-injured than in the control group. When this difference in the ERPs to the frequent tones was taken into account, there was no evidence to suggest that either the latency or the amplitude of the target-evoked N2 and P3b components differed between the groups. The novel stimuli evoked a prominent P3a component. The amplitude and scalp distribution of this component differed little between the groups, but its peak latency was reliably longer in the head-injured subjects. The findings in respect of the N2 and P3b components suggest that impairments in early processing of task-relevant stimuli are not an invariant feature of closed head injury. The findings regarding P3a suggest that, in the majority of patients, head injury has only a limited effect on the neural systems underlying involuntary shifts of attention.

Acoustic Stimulation↗
PubMed1993

The effect of age and voluntary contraction on presynaptic inhibition of soleus muscle Ia afferent terminals in man.

Transmission in a pathway mediating presynaptic inhibition of soleus Ia afferent terminals was assessed from the reduction of soleus muscle H reflex amplitude brought about by a short conditioning vibration pulse applied to the anterior tibialis muscle. In a group of young subjects (aged between 21 and 37 years) the mean reduction in reflex size with the limb relaxed was to 29.2% of control, whereas in a group of older subjects (aged between 59 and 74 years) the reduction was significantly less, to 55.0% of control (P < 0.001, Student's t test). In the younger group the amount of presynaptic inhibition elicited was inversely related to the plantar-flexor torque produced by tonic contraction of gastrocnemius-soleus; often at higher torques no reduction in reflex size was seen. In contrast the older group showed only a slight reduction in presynaptic inhibition with increasing torque, so that at torques of 5 and 7 N m the amount of presynaptic inhibition elicited was similar in the younger and older groups. The older subjects were not able to sustain the higher torques produced by the younger subjects. These results suggest that the control of transmission in this spinal pathway changes with increasing age.

Adult↗
PubMed1992

Immunocytochemical localization of kynurenine aminotransferase in the rat striatum: a light and electron microscopic study.

Kynurenine aminotransferase is the biosynthetic enzyme for kynurenic acid, an antagonist of excitatory amino acid receptors. Because of the possible role of kynurenic acid in basal ganglia diseases, the distribution of kynurenine aminotransferase immunoreactivity was examined in the adult rat striatum at the light and electron microscopic levels. Kynurenine aminotransferase immunoreactivity was detected in glial cells and in neurons. The preadsorption control vastly reduced or eliminated specific staining at both the light and electron microscopic levels. Kynurenine aminotransferase positive glial cells were abundant and contained a robust and homogeneous distribution of reaction product in both the nucleus and cytoplasm. The majority of neurons, both medium and large, were immunostained and exhibited granular kynurenine aminotransferase immunoreactivity in the cytoplasm of somata and proximal dendrites. At the ultrastructural level, kynurenine aminotransferase immunoreactive astrocytic processes were apparent throughout the neuropil where they often encircled capillaries and surrounded axospinous synapses. Reaction product was associated with the cytoplasmic matrix, filaments, rough endoplasmic reticulum, and the nucleus. In neurons, the majority of label occurred in round membrane-bound cytoplasmic organelles located adjacent to the Golgi apparatus, rough endoplasmic reticulum, and the cell or nuclear membranes. Cisternae and vesicles were identifiable in some of the labeled profiles. Polyribosomes and rough endoplasmic reticulum were also labeled. These data provide an anatomical basis for biochemical studies that have suggested the presence of striatal kynurenine aminotransferase in both astrocytes and neurons.

Animals↗
PubMed1992

Effect of stereotactic thalamic lesion on essential tremor.

The cause of essential (low-frequency) tremor is unknown and its relation to physiological (high-frequency) tremor is unclear. We assessed essential tremor in one patient before and after a stereotactic thalamic lesion. The procedure changed the size of the tremor in the right hand but not in the left. Persistence of a low-frequency component suggested that essential tremor was an additional feature superimposed on physiological tremor in this patient. The focus of essential tremor seems to be an autonomous central generator that is independent of physiological tremor mechanisms.

Adult↗
PubMed1992

Methionine oxidation and inactivation of alpha 1-proteinase inhibitor by Cu2+ and glucose.

The effect of glucose/Cu2+ incubation on (a) pure methionine oxidation, (b) the oxidation of active-site methionine in alpha 1-proteinase inhibitor (alpha 1PI) and (c) the resulting activity and structural changes of this inhibitor was investigated. While no methionine was oxidized during a 24 day, 37 degrees C incubation with 0.01 M EDTA and 100 mM glucose, 64.2% oxidation occurred in 6 days when 0.01 mM Cu2+ was added to the 100 mM glucose. The first-order rate constant for oxidation in 10 mM glucose, 0.01 mM Cu2+ was 0.0218 day-1. Oxidation was inhibited by catalase, but accelerated by ascorbate ion. The active-site methionyl residue of alpha 1PI was oxidized 71.3% after a 4 day incubation in 100 mM glucose, 0.01 mM Cu2+ (pH 7.45), 0.1 M phosphate buffer. The elastase and trypsin inhibiting activities were lowered to 3.1 and 1.5% of control samples during this incubation. The inclusion of 1 mM DETAPAC, a transition metal chelator, resulted in a 98 + % retention of activity. Intrinsic fluorescence (350 nm excitation, 415 nm emission) of alpha 1PI increased 576% over control for the sample incubated in 100 mM glucose, 0.01 mM Cu2+ and SDS-PAGE revealed protein fragment molecular weights of 44.4 and 39.8 kDa. These studies suggest that both methionine oxidation and free radical induced fragmentation contribute to loss of alpha 1PI activity during glucose/Cu2+ incubations.

Binding Sites↗
PubMed1992

Relationship of thrombohemorrhagic complications to endothelial cell function in patients with chronic myeloproliferative disorders.

Thrombotic and hemorrhagic disorders are common complications of the myeloproliferative disorders. Endothelial cells release both procoagulant and profibrinolytic factors, which may contribute to these hemorrhagic or thrombotic complications. The pre- and postvenous stasis levels of the procoagulant and profibrinolytic factors produced by endothelial cells were correlated with the occurrence of complications in polycythemia rubra vera (PRV) patients (n = 29) and essential thrombocythemia (ET) patients (n = 17) compared with normal patients (n = 17). Tissue plasminogen activator (tPA) activity, plasminogen activator inhibitor (PAI) activity, von Willebrand's factor (vWF) activity and antigen (vWF:Ag), and factor VIII activity were measured. The resting tPA activity was significantly higher in the two disease groups compared with normal controls, but no difference between the levels of tPA and either complication within the disease groups was observed. Significantly elevated tPA following venous stasis was observed in the patients of both disease groups who had bleeding complications. Significant decreases, compared with the normal group, in both resting and postvenous stasis levels of PAI were observed in the disease groups regardless of complication history. The subjects from both disease groups with thrombotic complications had significantly elevated resting vWF and both resting and postvenous stasis vWF:Ag levels compared with normal controls. The endothelial cell is likely to be responding to abnormal hemostasis rather than being primarily involved in the genesis of a hyper- or hypocoaguable state.

Adult↗
PubMed1992

Transcription and autoregulation of the stabilizing functions of broad-host-range plasmid RK2 in Escherichia coli, Agrobacterium tumefaciens and Pseudomonas aeruginosa.

The broad-host-range plasmid RK2 has been shown to encode several proteins important for its maintenance within bacterial populations of a number of Gram-negative bacteria. Their genes are organized into two operons: parCBA and parD. These operons have been proposed to be transcribed from two divergent promoters, p-parCBA and p-parD, located within a sequence of approximately 150 bases. In this report we identify and characterize the sequences required for regulated transcription from these promoters in Escherichia coli, Agrobacterium tumefaciens and Pseudomonas aeruginosa. Both of these promoters are repressed by their own gene products in the same manner in all three bacteria tested, with ParA functioning as the primary repressor of p-parCBA and ParD functioning as the repressor of p-parD. The binding regions of these proteins were determined through deletion analyses, DNA mobility shift assays, and an examination of the effect of mutations in this region. Based on these observations, the ParA protein appears to bind to either two inverted repeat or two direct repeat sequences, one downstream from the transcriptional initiation site and the other upstream of the p-parCBA -35 box. The ParD protein appears to bind to one inverted repeat sequence, located between the -35 and -10 boxes of p-parD.

Agrobacterium tumefaciens↗
PubMed1992

The effects of scopolamine on event-related potentials in a continuous recognition memory task.

Event-related potentials (ERPs) were recorded during a task requiring continuous recognition memory for visually-presented words. Twelve subjects each performed the task twice, once following the administration of scopolamine, and once after receiving a saline placebo. In the placebo condition, correctly detected "old" words (i.e., words that had been presented once before during the task) evoked more positive-going ERPs than did "new" words. Scopolamine caused a substantial impairment in task performance, but did not reduce the size of these old-word/new-word ERP differences. It is concluded that old/new ERP effects are unlikely to reflect cholinergically-mediated neural activity underlying normal recognition memory.

Adult↗
PubMed1992

A region of the broad-host-range plasmid RK2 causes stable in planta inheritance of plasmids in Rhizobium meliloti cells isolated from alfalfa root nodules.

We demonstrate for the first time that the broad-host-range stabilization loci from plasmid RK2 cause total retention of plasmids in cells of Rhizobium meliloti during symbiosis with alfalfa. Two derivatives of plasmid RK2, pRK290 and a 7.3-kb mini-RK2 plasmid, were stabilized in R. meliloti cells isolated from root nodules by the insertion of a 3.2-kb DNA fragment or a smaller 0.8-kb DNA fragment derived from the RK2 stabilization region.

Aerobiosis↗
PubMed1992

Definition of a minimal plasmid stabilization system from the broad-host-range plasmid RK2.

The stable inheritance of the broad-host-range plasmid RK2 is due at least in part to functions within a region located at coordinates 32.8 to 35.9 kb, termed the RK2 par locus. This locus encodes four previously identified genes in two operons (parCBA and parD; M. Gerlitz, O. Hrabak, and H. Schwab, J. Bacteriol. 172:6194-6203, 1990, and R. C. Roberts, R. Burioni, and D. R. Helinski, J. Bacteriol. 172:6204-6216, 1990). The parCBA operon is functional in resolving plasmid multimers to monomers. Analysis of the plasmid stabilization capacity of deletions within this region, however, indicates that this multimer resolution operon is required for stabilization only in certain Escherichia coli strains and under specific growth conditions. The deletion analysis further allowed a redefinition of the minimal functional region as 790 bp in length, consisting of the parD gene (243 bp) and its promoter as well as sequences downstream of parD. This minimal region stabilizes an RK2-derived minireplicon in several different gram-negative bacteria and, at least in E. coli, in a vector-independent manner. By insertional mutagenesis, both the parD gene and downstream (3') regions were found to be required for plasmid stabilization. The downstream DNA sequence contained an open reading frame which was subsequently shown by transcriptional and translational fusions to encode a protein with a predicted size of 11,698 Da, designated ParE. Since the parDE operon requires the presence of the parCBA operon for efficient stabilization under certain growth conditions, the potential role of multimer resolution in plasmid stabilization was tested by substituting the ColE1 cer site for the parCBA operon. While the cer site did function to resolve plasmid multimers, it was not sufficient to restore stabilization activity to the parDE operon under growth conditions that require the parCBA operon for plasmid stability. This suggests that plasmid stabilization by the RK2 par locus relies on a complex mechanism, representing a multifaceted stabilization system of which multimer resolution is a conditionally dispensable component, and that the function(s) encoded by the parDE operon is essential.

Amino Acid Sequence↗
PubMed1992

Phosphorylation of the rodent negative acute-phase protein alpha 1-inhibitor-III by the insulin receptor tyrosine kinase.

alpha 1-Inhibitor-III (alpha 1I3), a broad range proteinase inhibitor, member of the alpha-macroglobulin family, is abundant in normal rat plasma. Insulin-dependent tyrosine phosphorylation of a monomeric 195K glycoprotein (pp195) was observed in wheatgerm agglutinin (WGA)-Sepharose-purified insulin receptor preparations from rat liver and muscle. Phosphorylation of pp195 in vitro required a basic poly-amino acid, i.e. poly-L-lysine. We present evidence identifying pp195 as alpha 1I3. In situ perfusion with saline essentially removed pp195 from rat livers. Addition of normal rat plasma to liver homogenates or to WGA eluates restored insulin-stimulated phosphorylation of pp195; plasma from streptozotocin-diabetic rats was much less effective. Liver-derived pp195 copurified with an abundant plasma protein, with the characteristics of alpha 1I3, on size exclusion and ion-exchange chromatography. An approximately 195K protein, comigrating with alpha 1I3, was markedly diminished in plasma from diabetic rats, and alpha 1I3 concentration was decreased by approximately 70% upon immunoblot analysis. Highly purified alpha 1I3 was phosphorylated by muscle- or liver-derived insulin receptors in the presence of 1 microM poly-L-lysine and comigrated with pp195 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. alpha 1I3 phosphorylation was half-maximal at approximately 70 nM and was stimulated by insulin 7-fold. Hindlimb perfusion removed more than 90% plasma albumin but only approximately 20% pp195 from muscles. alpha 1I3 messenger RNA was identified in liver but not in muscle. A specific antibody against alpha 1I3 immunoprecipitated phosphorylated pp195 in WGA-purified insulin receptor preparations from nonperfused liver and from saline perfused and nonperfused muscle. alpha 1I3 is bound and internalized by alpha-macroglobulin receptors; whether it is phosphorylated in vivo is unknown. Hepatic alpha 1I3 synthesis may diminish in diabetic rats.

Acute-Phase Proteins↗
PubMed1991

Event-related potentials related to recognition memory. Effects of unilateral temporal lobectomy and temporal lobe epilepsy.

Event-related potentials (ERPs) were recorded during a continuous recognition memory task for visually presented words, with a 6 item lag between the first and second presentation of each word. The subjects consisted of: (i) a control group of patients with primary generalized epilepsy; (ii) patients who had undergone either a left or a right anterior temporal lobectomy; (iii) unoperated patients with either left or right unilateral temporal lobe epilepsy. In the controls, ERPs to detected 'old' words were reliably more positive-going in the interval 300 to 600 ms post-stimulus than were ERPs to 'new' items. In both left and right lobectomy patients, 'old/new' ERP differences in the same latency range were significantly smaller than in the controls, and did not differ significantly from zero. At midline electrodes, old/new effects in the temporal lobe epilepsy patients were of similar magnitude to those of the controls. In contrast to the control data, the old/new effects in the temporal lobe epilepsy patients were asymmetric, in that they were smaller over the hemisphere ipsilateral to the seizure focus than over the contralateral hemisphere. No relationship was found across subjects between the magnitude of old/new ERP effects and verbal memory performance. In a second task, occasional non-words had to be discriminated against a background of sequentially presented words, some of which were repetitions of the immediately preceding item. ERPs evoked by repeated words were more positive-going than were those to first presentations; this effect was reliable, and of equivalent size, in all patient groups. It is concluded that in the recognition task, old/new ERP effects are dependent on temporal lobe functioning, but that the anterior temporal lobe is not the principal locus of the generators of these effects. The cognitive processes reflected by these effects do not appear to be strongly lateralized to one hemisphere, and neither do they seem to be necessary for normal verbal memory.

Adolescent↗
PubMed1991

Normal P300 following extensive damage to the left medial temporal lobe.

Event-related potentials (ERPs) were recorded during auditory and visual "oddball" tasks from a patient with a severe verbal memory deficit due to a low grade infiltrating glioma which involved the full extent of the left medial temporal lobe. In both sensory modalities, the patient's oddball-evoked P300s were symmetrical and of normal amplitude. These findings are difficult to reconcile with the hypothesis that the hippocampus, or any other medial temporal structure, makes a substantial contribution to the scalp P300.

Adult↗