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R L Hayes

Publications and source records attributed to R L Hayes.

At least 73 records · Page 4Linked to original sources
PubMed1995

Frequent expression of a mutant epidermal growth factor receptor in multiple human tumors.

The epidermal growth factor receptor has received much interest as a target for various antineoplastic agents, but a complication is that many normal tissues also express this receptor. We have previously identified in human glial tumors an 801-bp in-frame deletion within the epidermal growth factor receptor gene that created a novel epitope at the junction. By using Western blot assays with a mutant-specific antibody as a rapid and sensitive means for detecting this alteration in primary human tumors, it was found that 57% (26 of 46) of high-grade and 86% (6 of 7) of low-grade glial tumors, but not normal brain, express this protein. This altered receptor was also present in 66% (4 of 6) of pediatric gliomas and 86% (6 of 7) of medulloblastomas, 78% (21 of 27) of breast carcinomas, and 73% (24 of 32) of ovarian carcinomas. The fact that this receptor is frequently found in tumors but not in normal tissue makes it an attractive candidate for various antitumor strategies.

Antibodies, Monoclonal↗
PubMed1995

Regional and temporal profiles of c-fos and nerve growth factor mRNA expression in rat brain after lateral cortical impact injury.

Lesion-induced increases in NGF mRNA are thought to be mediated by c-fos gene expression. Conversely, NGF induction of c-fos expression has been reported following administration of exogenous NGF. However, the relationship between c-fos and NGF gene expression after traumatic injury to the intact brain is not known. Thus, we applied in situ hybridization and semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) methods to determine temporal profiles of c-fos and NGF mRNA expression in rat brains after controlled impact to the exposed cortex. Using alternate sections from the same rat brains, in situ hybridization studies showed that in neocortex, c-fos mRNA transiently increased at 30 min, 1 hr, and 3 hr after injury, while there were no increases of NGF mRNA at these postinjury time points. In the hippocampus, in situ hybridization showed that c-fos mRNA increased at 30 min, 1 hr and 3 hr postinjury, while NGF mRNA increased at 1 hr, 3 hr but not at 30 min after injury. RT-PCR studies in hippocampus confirmed that c-fos mRNA increased as early as 5 min after injury, peaked at 30 min postinjury, and remained elevated 5 hr postinjury. Levels of hippocampal NGF mRNA expression increased by 1 hr after injury and plateaued until 3 and 5 hr postinjury. These data are consistent with the possible regulatory role of endogenous c-fos on NGF expression following traumatic brain injury.

Animals↗
PubMed1995

Brief potassium depolarization decreases levels of neurofilament proteins in CNS culture.

Little is known about the effects of brief potassium depolarization that occurs concurrently with transient ischemia, epilepsy and head trauma. To investigate the effect of short-term depolarization on light (NF-L), middle (NF-M), and heavy (NF-H) neurofilament proteins and determine the role played by calcium in that effect, mixed septo-hippocampal cultures were exposed to 60 mM K+ for 6 min, in the presence of 0 to 11.8 mM Ca2+. Twenty-four hours later, neurofilament immunoreactivity in Western blots of depolarized cultures was decreased to 60% or less of control levels. Decreases were Ca2+-dependent, not due to cell loss, and affected both phosphorylated and nonphosphorylated proteins. The phosphorylation state of NF-M and NF-H influenced the degree of loss observed. Changes in the pattern of immunolabelling of neuritic processes were also associated with depolarization. Thus, brief potassium depolarization may contribute to cytoskeletal disruption following brain injury.

Animals↗
PubMed1995

Basal and scopolamine-evoked release of hippocampal acetylcholine following traumatic brain injury in rats.

This study employed in vivo microdialysis in awake, freely-moving Sprague-Dawley rats to examine acetylcholine (ACh) release in the dorsal hippocampus at 14 days following lateral controlled cortical impact. Extracellular levels of ACh were measured prior to and after an intraperitoneal administration of scopolamine (1 mg/kg), which evokes ACh release by blocking autoreceptors. At 14 days post injury there were no differences in basal ACh levels. However, injury produced a significant reduction in scopolamine-evoked ACh release. The data suggest that cholinergic neurotransmission remains chronically compromised, and thus may contribute to previously documented post traumatic spatial memory deficits.

Acetylcholine↗
PubMed1995

Increased cortical nuclear factor-kappa B (NF-kappa B) DNA binding activity after traumatic brain injury in rats.

Nuclear factor-kappa B (NF-kappa B) DNA binding factor is an inducible transcription factor that responds to various cellular signals. Levels of cortical NF-kappa B DNA binding activity were measured in a controlled lateral cortical impact model of traumatic brain injury (TBI) in rats. Using electrophoretic mobility shift assays (EMSAs), we found that NF-kappa B DNA binding activity in cerebral cortex ipsilateral to the injury site increased at 1, 3, 5 and 7 days after injury. Binding activity peaked at 3 days after injury and subsided by 10 days after injury. These data indicate that TBI produces transient increases in NF-kappa B DNA binding activity. Further insights into the role of NF-kappa B in TBI may provide new therapeutic opportunities for head trauma.

Animals↗
PubMed1995

Time course of neurofilament protein loss following depolarization-induced injury in CNS culture.

In septo-hippocampal cell cultures, brief potassium depolarization produces calcium-dependent decreases in neurofilament proteins and loss of fine neuritic processes within 24 h. It is not known whether neurons later exhibit delayed degeneration and die, live with enduring neurofilament loss, or restore neurofilament protein levels. Therefore, we exposed septohippocampal cultures to 6 min potassium depolarization (60 mM) with 2.8-11.8 mM extracellular Ca2+ and evaluated immunoreactivity for low, medium and heavy neurofilament proteins, neuronal number, and neuronal morphology for 10 days. Neuronal number remained unchanged; neurofilament protein levels recovered to between 31% and 99% of control levels, and fine neuritic processes reappeared.

Animals↗
PubMed1995

Improved long term survival after intracavitary interleukin-2 and lymphokine-activated killer cells for adults with recurrent malignant glioma.

BACKGROUND: The median survival for adults with glioblastoma multiforme (GBM) is 12 months, despite surgery, radiation, and chemotherapy. Regimens using interleukin-2 (IL-2) plus lymphokine-activated killer (LAK) cells have been beneficial against systemic cancers, albeit with significant toxicity. METHODS: Nineteen adults with recurrent malignant glioma (5 GBMs, and 4 anaplastic astrocytomas (AA)), Karnofsky performance status 60 or greater, were treated with intracavitary autologous LAK cells plus IL-2 after reoperation. Lymphokine-activated killer cells and IL-2 were given on day 1, and IL-2 alone was given 5 times during a 2-week cycle. This cycle was repeated at 2 weeks to constitute one 6-week course of therapy. Each two-cycle course of treatment was repeated at 3-month intervals for patients with stable disease or response to therapy. At the conclusion of immunotherapy, all patients were offered chemotherapy, generally carmustine or procarbazine, including responders. Corticosteroids were strictly limited during immunotherapy. Sequential reservoir aspirates were obtained for microbiologic and cytologic analyses. RESULTS: The maximal tolerated dose for a 12-dose course of therapy was 1.2 million international units (MIU) per dose. Dose-limiting, cumulative IL-2-related central nervous system (CNS) toxicity was observed at 2.4 MIU per dose. Three responses were confirmed by computed tomography scan during therapy: one complete response (CR) (1 AA), and two partial responses (PR) (2 GBM); as well as a significant increase in GBM survival. One additional CR (GBM) was observed at 17 months. The median survival for immunotherapy patients with GBM was 53 weeks after reoperation (N = 15) (mean, 87.9 +/- 21.4 weeks, standard error for the mean), with 8 of 15 surviving more than 1 year (53%). The median survival for 18 contemporary patients with GBM reoperated and treated with chemotherapy was 25.5 weeks (mean, 27.4 +/- 3.7 weeks), with 1/18 alive at 1 year (> 6%). Six of the 15 patients with GBM had additional surgery or biopsy, and chemotherapy after immunotherapy. The contribution of subsequent chemotherapy to survival cannot be discounted. CONCLUSIONS: Lymphokine-activated killer cells and IL-2 can be administered safely within the CNS resulting in improved long term survival in patients with recurrent glioblastoma. Increased survival was associated with significant biologic changes characterized by a regional eosinophilia, and extensive lymphocytic infiltration. A prospective randomized clinical trial is warranted.

Adult↗
PubMed1995

Calpain inhibitors reduce depolarization induced loss of tau protein in primary septo-hippocampal cultures.

We studied the effects of a 6-min potassium depolarization injury produced by 60 mM KCl and 1.8 mM or 5.8 mM extracellular CaCl2 on tau protein levels in primary rat septo-hippocampal cultures. One day after injury, Western blot analyses revealed a calcium dependent loss of tau protein of approximately 50% of control values. Loss of tau protein was associated with calpain 1 mediated breakdown products to alpha-spectrin. Calpain inhibitors 1 and 2, applied immediately after depolarization injury and available to cultures for 24 h reduced depolarization induced degradation of tau protein to approximately 35% or 25% of control values, respectively. We propose that brief potassium depolarization causes degradation of tau protein, possibly due to calpain activation. Thus, calpain inhibitors could represent a viable strategy for preserving the cytoskeletal structure of injured neurons.

Animals↗
PubMed1995

Rescue of injury-induced neurofilament loss by BDNF gene transfection in primary septo-hippocampal cell cultures.

We employed primary septo-hippocampal cell cultures to determine the ability of liposome-mediated BDNF gene transfection to facilitate recovery of neurofilament loss caused by depolarization injury. After BDNF gene transfection in uninjured cultures, RT-PCR and immunohistochemical staining confirmed increases in BDNF mRNA and protein in transfected cells. Three days after depolarization injury, Western blot and immunohistochemical analyses detected significant loss of neurofilament proteins in non-transfected cultures, while BDNF transfection produced marked increases in neurofilament proteins following either pre-injury transfection or transfection 24 h following injury. Immunohistochemical studies also detected enhanced immunolabeling of BDNF and total neurofilament protein (phosphorylated and non-phosphorylated) in injured neurons following BDNF transfection or administration of exogenous BDNF protein, compared to untransfected, injured controls.

Animals↗
PubMed1995

Social skills in schizophrenia: assessing the relationship between social skills, psychopathology and community functioning.

Social skills training (SST) has been widely used in attempts to rehabilitate chronic schizophrenic patients. The key assumption underlying SST is that social skills deficits are important determinants of the social isolation, poor social role functioning, and low quality of life characteristic of schizophrenic patients. To test this assumption, 89 patients meeting DSM-III-R criteria for schizophrenia were assessed on behavioural and self-report measures of social skills. A structured clinical interview, a self-report inventory of distress during social interaction, and self-monitoring of time in social interaction were used to assess social functioning. Positive psychotic symptoms were assessed in a standardised clinical interview. A structural equation modelling analysis showed that observed social skills predicted social functioning, and that this association was statistically independent of severity of psychotic symptoms. This finding is consistent with the hypothesis that social skills are important in the social functioning of patients with schizophrenia.

Activities of Daily Living↗
PubMed1995

Time course of increased vulnerability of cholinergic neurotransmission following traumatic brain injury in the rat.

We have previously shown that spatial memory changes following experimental traumatic brain injury (TBI) include long-term changes that are (1) 'overt': detected by routine behavioral assessments, or (2) 'covert': undetected in the absence of a secondary pharmacological challenge, such as by the cholinergic antagonist, scopolamine. Our objective in this study was to extend this finding by characterizing the time course of recovery of overt and covert spatial memory performance following two magnitudes of experimental TBI. The Morris water maze was used to assess cognitive performance. Rats received either moderate magnitude (6 m/s, 1.77 mm deformation) or low magnitude (6 m/s, 1 mm deformation) impacts through a lateral craniectomy under isoflurane anesthesia. Sham rats underwent identical surgical procedures but were not injured. To avoid motor deficits, water maze testing started two weeks post-injury. Rats were given four trials per day for seven consecutive days. For each trial, latency to find a hidden platform was timed. On the sixth, rats were injected (i.p.) with scopolamine (1 mg/kg) 15 min prior to maze testing. The next day, rats were retested. This testing regimen was repeated, beginning 4, 6, and 10 weeks post-TBI. Results showed that, while the low-magnitude injury produced no overt spatial memory deficits, the moderate-magnitude group exhibited overt deficits during the first test regimen. Also, while both injury magnitudes produced an enhanced sensitivity to spatial memory impairment by scopolamine at two weeks post-TBI, this covert deficit persisted only in the severe group at 4, 6, and 10 weeks post-TBI. Qualitative light microscopy showed that both injury groups had graded cortical necrosis. However, underlying subcortical structures such as the hippocampus appeared intact, with no overt cellular or parenchymal damage to the neuropil. These data suggest three distinct stages of functional recovery: (1) the initial period when overt deficits are present, (2) a period following recovery from overt deficits within which covert deficits can be reinstated by a pharmacological challenge, and (3) a period following recovery from both overt and covert deficits. Covert deficits can persist long after the recovery of overt deficits and, like other neurological deficits, the rate of recovery is dependent on the magnitude of TBI. Finally, spatial memory deficits can occur in the absence of light microscopic evidence of cell death in the hippocampus.

Acetylcholine↗
PubMed1995

Changes in gene expression following traumatic brain injury in the rat.

This paper reviews changes in gene expression produced by two rodent models of traumatic brain injury: cortical impact injury and fluid-percussion injury. Cortical impact injury produces transient increases in c-fos mRNA expression, which begin as early as 5 min after injury and subsides by 1 day after injury in the cerebral cortex ipsilateral to injury. In addition, AP-1 transcription factor binding is greatly increased in the injured cerebral cortex at 1, 3, and 5 h post-injury. AP-1 binding remains increased for at least 1 day after injury, while SP-1 transcription factor binding activity does not increase. Additional studies have confirmed increases in c-fos mRNA expression in the hippocampus at 30 min, 1 h, and 3 h after injury. These increases in c-fos mRNA in the hippocampus preceded increased levels of NGF mRNA that were detected at 1 and 3 h but not at 30 min following injury. Following fluid-percussion injury, increases in c-fos mRNA can be detected as early as 2 h following injury in the cortex ipsilateral to the site of injury as well as in the hippocampus. Heat-shock protein (hsp72) mRNA is also increased in the ipsilateral cortex and hippocampus following fluid percussion injury. By 24 h post-injury, both c-fos and hsp72 gene expression return to control levels. Severe but not moderate fluid percussion injury produces increased gene expression for glucose-regulated proteins (grp78, grp94) 12 h following injury. Fluid-percussion injury also produces significant increases in expression of both interleukin-1 beta and tumor necrosis factor-alpha in the injured cortex and ipsilateral hippocampus as early as 1 h post-injury, that remains elevated up to 6 h in the injured cortex and hippocampus.

Animals↗
PubMed1995

Cytoskeletal derangements following central nervous system injury: modulation by neurotrophic gene transfection.

This paper reviews important new evidence indicating that traumatic brain injury can produce more widespread derangements to the neuronal cytoskeleton than previously recognized. Although cytoskeletal derangements in axons have long been documented, recent data suggest that traumatic brain injury can produce structural derangements to dendrites and cell bodies as well. Many of these investigations have employed in vivo models to provide important insights into mechanisms possibly mediating the acute loss of cytoskeletal proteins, including disturbances in calcium homeostasis and activation of calcium-dependent proteolytic enzymes. However, we have little understanding of processes mediating the recovery of cytoskeletal proteins following injury. This paper provides recent evidence from in vitro models of central nervous system injury that neurotrophic proteins can enhance the recovery of the neuronal cytoskeleton. Neurotrophin-based therapy could employ either administration of exogenous neurotrophic proteins and/or transfection of cDNA for appropriate neurotrophins.

Animals↗
PubMed1995

Induction of 70-kD heat shock protein in scleractinian corals by elevated temperature: significance for coral bleaching.

In this study, the induction of the 70-kD family of heat shock proteins (hsp70) has been examined in stony coral tissues. In these experiments, the only difference from control conditions has been exposure to a temperature approximating that at which field bleaching in the Caribbean is known to occur, approximately 30 degrees C or 1 degree-2 degrees C above long-term average seasonal maximum temperatures. A constitutive hsp70 has been identified both in the zooxanthellate (hermatypic) coral, Montastrea annularis, and in two corals lacking symbiotic algae, Tubastrea cocchinea and Astrangia danae (Cnidaria, Anthozoa, Scleractinia). Western blots of experimental tissues fractionated by polyacrylamide gel electrophoresis indicate that the initial induction of hsp70 occurs rapidly, within one hour of transfer to water of elevated temperature. Thereafter, the level of hsp70 decreases within 12-24 hours to approximately the constitutive level. In field-bleached specimens of M. annularis, hsp70 is not detected. Since this coral tissue, once bleached to whiteness, contains no 70-kD heat shock protein, we conclude that the process of coral bleaching might include, among other metabolic alterations, a failed heat shock response. In addition to being compromised in other normal functions, the bleached coral would lose the capacity to protect itself against environmental stress. The eventual loss of algae by bleached coral is likely to be consequent to several metabolic changes in the coral tissue. However, the uncoupling of that symbiotic relation is not concomitant with the initial stress response of heat shock protein synthesis.

Animals↗
PubMed1994

Increased expression of c-fos mRNA and AP-1 transcription factors after cortical impact injury in rats.

Levels of c-fos mRNA and AP-1 transcription factors co-expression were measured in a controlled lateral cortical impact model of traumatic brain injury (TBI) in rats. Ipsilateral cerebral cortex and bilateral hippocampal c-fos mRNA increases were revealed by in situ hybridization after lateral cortical impact injury. Based on regional in situ hybridization data, we employed semi-quantitative RT-PCR methods to study the temporal profile of changes in the ipsilateral cortex at the site of injury. We found that TBI produces transient increases of c-fos mRNA expression in the ipsilateral cerebral cortex at 5 min postinjury, which peaks at 1 h postinjury and subsides by 1 day postinjury. Gel shift nuclear protein binding assays showed that AP-1 transcription factor binding was robustly increased in injured cerebral cortex at 1 h, 3 h, 5 h and 1 day after injury. These data indicate that TBI can produce significant increases in c-fos expression and subsequent upregulation of the AP-1 transcription factors. Thus, AP-1 transcription factors modulation of downstream gene expression may be an important component of pathophysiological responses to TBI.

Animals↗
PubMed1994

Traumatic brain injury causes a decrease in M2 muscarinic cholinergic receptor binding in the rat brain.

Numerous studies indicate that an acute, excessive activation of muscarinic acetylcholine receptors (mAChR) contributes to the pathophysiological sequela of TBI. The present study examined the effect of moderate fluid percussion traumatic brain injury (TBI) on binding to M1 and M2 mAChR subtypes in the hippocampal formation and adjacent cortex using quantitative autoradiography. Injured animals along with concurrent controls were sacrificed by in situ freezing at 3 h or 24 h following TBI. Slide-mounted tissue sections were incubated in either [3H]pirenzepine (23 nM) for M1 or [3H]AFDX384 (9 nM) for M2 mAChR subtype labeling. Binding of [3H]pirenzepine to the M1 mAChR subtype was not significantly altered by TBI when compared to sham-injured animals. [3H]AFDX384 binding to the M2 mAChR subtype was significantly decreased at 24 h in hippocampal CA2-3 region and dorsal blade of the dentate gyrus (P < 0.05). The differences observed between M1 and M2 subtypes suggests that these muscarinic subtypes may differentially contribute to the pathophysiology of TBI.

Animals↗
PubMed1994

Increased anticholinergic sensitivity following closed skull impact and controlled cortical impact traumatic brain injury in the rat.

Evidence suggests that prolonged memory deficits in several neurodegenerative diseases are attributable to deficits in central cholinergic neurotransmission. In traumatic brain injury (TBI), such cholinergic deficits also may contribute to prolonged memory disturbances. This study determined whether moderate magnitudes of TBI produced by controlled cortical impact and mild magnitudes of experimental TBI produced by a new closed head impact technique in rats would produce an enhanced vulnerability to the memory disruptive effects of scopolamine, a muscarinic cholinergic receptor antagonist. Water maze performance was used to determine changes in cholinergic hippocampal function following TBI. In the first experiment, rats received a moderate level of TBI by means of a controlled cortical impact. A Morris water maze task assessed spatial memory function on days 30-34 postinjury. During the 5 day assessment period, statistical analyses showed a group main effect for swim latency. Subsequent post hoc analyses indicated that injured rats had significantly longer latencies on days 30 and 31 (p < 0.05, injury vs sham controls). By days 32-35, injured rats showed no statistically significant deficits in spatial memory performance. On day 35, scopolamine (1 mg/kg, IP) was injected into injured rats and sham-injured rats 15 min prior to being retested in the maze. Results showed that although the scopolamine had no effects on the performance of the sham-injured rats, the same dose significantly (p < 0.05) increased the latency to find the hidden platform in the injured group. In the second experiment, rats received a mild concussive closed head impact. Water maze performance was assessed on days 8-12 postinjury. No significant water maze performance deficits were observed. On day 13, injured and uninjured rats were pharmacologically challenged with scopolamine (1 mg/kg) and retested. Similar to the first experiment, injured rats manifested a significantly greater (p < 0.05) sensitivity to scopolamine than sham controls. The results from both experiments suggest that concussive and more severe levels of TBI can produce an enhanced vulnerability to disruption of cholinergically mediated memory function, even when memory function appears normal in the absence of secondary challenges. These data demonstrate that covert deficits can persist after the recovery of normal function. These deficits may be attributable to a decrease in the ability of cholinergic neurons to function properly. These data also provide important insights into features of receptor-coupled disturbances that could contribute to the maintenance of enduring cognitive deficits following TBI.

Acetylcholine↗