Biomedical subjects

J A McNulty

Publications and source records attributed to J A McNulty.

At least 19 recordsLinked to original sources

PubMed2001

Assessment of melatonin's ability to regulate cytokine production by macrophage and microglia cell types.

Evidence in support of melatonin's role as an immunomodulator is incomplete and, in some cases, contradictory. The present studies determined whether melatonin modulates the activity of stimulated macrophages. In vitro lipopolysaccharide (LPS, 10-1000 ng/ml) treatment of alveolar, splenic and peritoneal macrophages isolated from mice and/or rats resulted in a dose-dependent increase in interleukin-1beta (IL-1beta) and tumor necrosis factor (TNF-alpha) secretion. Treatment with melatonin (10(-10)-10(-6) M) prior to the addition of LPS, had no effect on IL-1beta or TNF-alpha release. Additionally, melatonin had no effect on stimulated BV2 microglial cell line cytokine secretion. To determine whether melatonin had an indirect effect on macrophage cytokine release via T cells, melatonin was added to unfractionated mouse spleen cells. Again, melatonin showed no priming effect on LPS-stimulated spleen cells. These results suggest that melatonin has no direct or indirect effect on mouse and rat macrophages. In vivo studies, where melatonin was continuously available in the drinking water, showed that melatonin did not have a priming effect on LPS-stimulated mouse peritoneal macrophages. These findings suggest that melatonin is not an important modulator of macrophage and microglia function.

Adjuvants, Immunologic↗

PubMed2001

Microglia play a role in mediating the effects of cytokines on the structure and function of the rat pineal gland.

The role of the pineal gland in regulating immune function has been extensively investigated. However, there is little information about possible feedback mechanisms of immunological factors on pineal gland neuroendocrine functions. Therefore, experiments were designed to test the effects of cytokines (interferon-gamma, IFN-gamma, interleukin-1 beta, IL-1 beta; tumor necrosis factor-alpha, TNF-alpha; transforming growth factor-beta 1, TGF-beta 1) on pinealocytes and the role of pineal microglia in mediating these cytokine effects in the pineal gland of the rat. Our studies showed that IFN-gamma enhanced 5-hydroxytryptamine (5-HT) content (measured by high-performance liquid chromatography, HPLC) and increased pinealocyte process length in pineal cultures. IL-1 beta treatment decreased 5-HT content in both cell and organ culture, but exhibited no effect on pinealocyte process length. 5-HT content and process length were decreased by TNF-alpha treatment. IFN-gamma and IL-1 beta exhibited no significant effect in the absence of microglia in cell cultures. In contrast, TNF-alpha caused a further decline in 5-HT content even in the absence of microglia in the cultures. The effects of TNF-alpha were probably due to toxic effects, since an increased number of pyknotic nuclei were observed in treated cultured explants. TGF-beta 1 treatment caused aggregation of pinealocytes in cultures and suppressed process length and 5-HT content. In conclusion, cytokine effects on pinealocytes may be mediated by microglia (IFN-gamma and IL-1 beta) or act directly on pinealocytes (TNF-alpha). The presence of IL-1 beta and TGF-beta 1 protein in the pineal gland and the suppressive effect of TGF-beta 1 on pinealocytes in cultures further suggest that endogenous cytokines play regulatory roles in response to peripheral homeostatic changes.

Animals↗

PubMed2001

TGF-beta1 and IL-6 expression in rat pineal gland is regulated by norepinephrine and interleukin-1beta.

The pineal gland is part of the neuroendocrine system that modulates immune functions. Because the gland is outside the blood-brain barrier, it is accessible to direct feedback from circulating cytokines that affect the synthesis and secretion of melatonin. Recent studies have suggested that intrinsic immunoregulatory cytokines mediate these neuro-immune interactions under the control of sympathetic innervation to the pineal. This study focused on the expression of transforming growth factor-beta1 (TGF-beta1) and interleukin-6 (IL-6), two cytokines that have important regulatory functions on both neurons and immune cells. Northern blot RNA analysis showed that TGF-beta1, but not IL-6, was expressed in freshly dissected rat pineal glands from neonatal age (1-day-old) into adults. Immunocytochemistry for TGF-beta1 in adult glands revealed localization of this protein in astrocyte-like cells. The sympathetic neurotransmitter norepinephrine (NE) increased transcript levels for both TGF-beta1 and IL-6 in adult pineal organ cultures. The effect of NE on IL-6 expression was not found in dispersed cell cultures established from neonatal pineal glands. The immunoregulatory molecule interleukin-1beta (IL-1beta) up-regulated the expression of both IL-6 and TGF-beta1 in adult pineal organ cultures, but not in neonate pineal organ cultures. These findings suggest that TGF-beta1 and IL-6 have intrinsic regulatory roles in the pineal gland and that both neural and immune factors are important mechanisms of regulation.

Adrenergic alpha-Agonists↗

PubMed2000

Evaluation of Web-based computer-aided instruction in a basic science course.

PURPOSE: To demonstrate the applicability of server statistics, in combination with user surveys, to evaluate utilization of Web-based computer-aided instruction (CAI) in the undergraduate medical curriculum. METHOD: Individual user surveys with students' names provided information about computer literacy prior to the course and use of CAI during the course. Utilization of specific web-based CAI developed for the course was recorded by server software and the daily logs correlated with course content. Regression analyses were used to measure correlation of server access logs of individual students versus information from user surveys and performances in the course based on in-course examinations. RESULTS: There was no correlation between computer literacy of students at matriculation and their subsequent levels of use of CAI in the curriculum. Utilization of CAI developed for specific course objectives coincided closely with course content, which is an indication of the effectiveness of the applications in achieving their curricular objectives. In contrast, student use of tutorials coincided most closely with in-course examinations. Students' responses to surveys were generally substantiated by server statistics, but discrepancies were sufficiently large (10% to 20%) to call into question the validity of these surveys. Significant differences in CAI utilization correlated with the performances of students in the course. CONCLUSIONS: This study demonstrates an important advantage of web-based applications to collect and evaluate CAI utilization efficiently and objectively at both the level of the class and the level of the individual student.

Computer Literacy↗

PubMed1999

Enhanced expression of neutrophil NADPH oxidase components in intestine of rats after burn injury.

We have evaluated the accumulation of neutrophils in the gut and their infiltration into the intestinal extravascular spaces in rats subjected to a 25% total body surface area scald burn. The accumulation of neutrophils was assessed via measurements of myeloperoxidase (MPO) activity in the intestinal homogenates, and the immunohistochemical localization of neutrophil NADPH oxidase component proteins (p47phox and p67phox) within the intestinal extravascular spaces determined neutrophil tissue infiltration. MPO measurements demonstrated a 12- and 21-fold increase above the control value in the intestinal tissue at day 1 and day 3 post-burn, respectively, suggesting that a substantial total tissue accumulation of neutrophils occurs in the gut after burn injury. The immunohistochemical staining procedures showed both a definitive presence of the neutrophil in the intestinal extravascular spaces and an enhanced immunoreactivity in neutrophils accumulating in intestine after burn injury. There was no evidence of either the presence of neutrophils in the extravascular regions or any significant neutrophil immunoreactivity to NADPH oxidase component proteins in the intestines of sham control rats. These findings indicate that burn injury causes an enhanced migration of circulating neutrophils into the intestinal interstitial spaces and an upregulation of NADPH oxidase activity in the infiltrating neutrophils.

Animals↗

PubMed1999

Interleukin-1beta expression in the pineal gland of the rat.

Increasing evidence of the neuroimmunomodulatory role of the pineal gland prompted the present study of pineal gland expression of the immunoregulatory cytokine, interleukin (IL)-1beta. IL-1beta was constitutively expressed in the adult gland, with mRNA levels higher in glands collected during the photophase than in those collected during the scotophase of the light:dark cycle. IL-1beta was up-regulated in pineal cultures, after treatment with either norepinephrine (NE) or interferon (IFN)/lipopolysaccharide (LPS). Although both astrocytes and microglia expressed IL-1beta, important differences were found in the cellular expression of this cytokine under in vivo and in vitro conditions. Increased IL-1beta expression by NE ex vivo and the decline in IL-1 expression at night, when NE levels are elevated, can be explained by immunocytochemical data showing that astrocytes are the predominant cell type expressing this cytokine in vivo, whereas IL-1beta-positive cells are predominantly microglia in pineal explants and dispersed cell cultures. These results are consistent with the hypothesis that cytokines secreted by pineal glia (astrocytes and microglia) may have an important regulatory role in the pineal gland.

Animals↗

PubMed1997

Microglia in the pineal gland of the neonatal rat: characterization and effects on pinealocyte neurite length and serotonin content.

Microglia in the pineal gland of 1-day-old Sprague-Dawley rats were examined by OX-42 immunocytochemistry and DiI-acetylated-LDL uptake in pineal cell suspension and were found to comprise 3-5% of the total cells in the pineal gland of the neonates. In order to investigate the effects of microglia on pinealocyte structure and function, microglia-depleted and microglia-enriched pineal cell cultures were generated from 1-day-old neonate by fluorescence activated cell sorting (FACS). After 7 days of culture, tissues were processed for either immunocytochemistry for pinealocyte S-antigen and serotonin or high performance liquid chromatography to measure serotonin. Morphometric analysis of immunoreacted cells revealed that pinealocyte neurite length was enhanced in microglia-depleted cultures and was inhibited in a microglia-enriched environment (ANOVA, P < 0.001). Serotonin content of pineal cultures decreased in microglia-depleted cultures and was elevated in microglia-enriched cultures (ANOVA, P < 0.001) without any significant change in pinealocyte numbers. These findings are consistent with a working hypothesis that microglia function to mediate neuroendocrine-immune interactions of the gland.

Animals↗

PubMed1997

Enriched immune-environment of blood-brain barrier deficient areas of normal adult rats.

The circumventricular organs (CVOs) in the brain are without a blood-brain barrier (BBB) and as such directly exposed to blood plasma constituents and blood-borne pathogens. In light of previous studies showing discrepancies regarding the immunocompetence of these organs, we initiated the present study to provide a comprehensive immunohistochemical analysis of the cellular expression of immune-associated antigens within the pineal gland, area postrema and the subfornical organ. In all CVOs, subpopulations of cells morphologically similar to complement receptor type 3 immunoreactive microglial/macrophage cells expressed major histocompatibility complex (MHC) class II antigen, leucocyte common antigen (LCA/CD45), as well as CD4 and ED1 antigen. Based on morphological criteria the MHC class II antigen expressing cells could be grouped into a major population of classical parenchymal and perivascular ramified microglial cells and a minor population presenting itself as scattered or small groups of rounded macrophage-like cells. CD4 and ED1 antigen were expressed by both cell types. CD45 was preferentially expressed by macrophage-like cells. MHC class I antigen was expressed by the vascular endothelium in both BBB-protected and BBB-deficient areas and was additionally present as a lattice-like network throughout the BBB-deficient parenchyma in all CVOs. The results suggest that the BBB-free areas of the brain besides being constantly surveyed by blood-borne macrophages, possess an intrinsic immune surveillance system based on resting and activated microglial cells, which may function as a non-endothelial, cellular barrier against blood-borne pathogens.

Animals↗

PubMed1997

A note on the time dependence of pineal gland research publications.

Counts of the number of publication titles containing the search truncation "pinea*" were compiled via Current Contents over the time period 1978-1994. These counts and their time dependence were examined for autocorrelations and frequency spectral components. Such analyses were carried out irrespective of either author, research laboratory, funding, or other factors. Interestingly, the results show the research publication rate in pineal studies to be linked to community history. In particular, the interactions affecting output among community members operate on a few characteristic time scales ranging from one to several years.

Animals↗

PubMed1997

Pinealocyte ultrastructure in mutant mice that lack sympathetic innervation to the pineal gland.

Pinealocyte synaptic ribbons (SR) and dense-cored vesicles (DCV) were quantitated in mice that have developmental deficiencies in the sympathetic innervation of the pineal gland due to a null mutation for the p75 low-affinity NGF receptor (NGFR). SR exhibited a significant increase whereas there was a significant decline in the frequency of DCV in mutant mice. These findings support the hypothesis that pinealocyte SR and DCV are regulated by neural mechanisms associated with sympathetic system. Ultrastructural characteristics of pinealocytes in mutants included degenerative changes that culminated in the breakdown of cellular components and the accumulation of flocculent-containing vesicles within the cytoplasm. Ribosomal lamellar complexes were also commonly seen in pinealocytes of mutant rats. Although the mutant pineal gland exhibited signs of metabolic imbalances, the cytoarchitecture of the gland (e.g., vascular compartment) and differentiation of the cells were generally unaffected by developmental deficiencies in the gland's innervation.

Animals↗

PubMed1997

Relationship of short-term verbal memory to the need for hyperbaric oxygen treatment after carbon monoxide poisoning.

It has long been known that carbon monoxide (CO) poisoning has a spectrum of effects on cognitive functioning, including memory, perception, and attention. The current study reports the investigation of the effects of CO poisoning on short-term verbal memory, both rote and context aided. Impairment was measured before and after hyperbaric oxygen (HBO) treatment. Twenty-six patients who had been admitted for emergency treatment after exposure to significant CO poisoning were tested using a measure of short-term recall for word lists with no or varying degrees of internal context-aided structure. Impairment of context-aided memory (but not rote memory) has been previously reported to be associated with low relative frontal volume in psychiatric patients. Carbon monoxide poisoning was significantly associated with impairment of context-aided memory, with the degree of pretreatment impairment predicting the number HBO treatments judged to be necessary on the basis of clinical monitoring of the patient. In patients with poisoning of moderate severity, pretreatment performance in context-aided memory improved after the first HBO treatment. The implications of these findings for the effects of CO poisoning on frontal area function are discussed. The memory measure used in this study appears to have considerable potential usefulness in the clinical assessment of the severity of CO poisoning in patients treated in an emergency setting.

Adolescent↗

PubMed1996

Regulation of synaptic ribbons in rat pineal gland explants by norepinephrine and sympathetic neurons in a co-culture model.

The hypothesis that synaptic ribbons in the mammalian pinealocyte are influenced by adrenergic mechanisms was tested in the present study using a co-culture model of pineal glands and superior cervical ganglia from neonatal rats. Pineal gland explants survived and showed a high degree of differentiation when cultured for up to 30 days regardless of the presence or absence of superior cervical ganglia. Pineal glands also had neurotrophic properties promoting the survival and neurite extension from superior cervical ganglia. Synaptic ribbons were a common ultrastructural feature in all pineal cultures. There was a significant decline in synaptic ribbon numbers when co-cultured with superior cervical ganglia for both 7 and 30 days. A similar significant decrease in synaptic ribbon frequency was observed after treatment of pineal explants with norepinephrine (10(-5 )M) for both 7 and 30 days. These findings are consistent with the hypothesis that neural mechanisms play an important role in regulating synaptic ribbon numbers, at least during early development. The study also illustrates the utility of pineal gland-superior cervical ganglia co-cultures as a model for future investigations of neuron-target interactions in the pineal.

Animals↗

PubMed1995

Neurotrophic effects of the pineal gland: role of non-neuronal cells in co-cultures of the pineal gland and superior cervical ganglia.

The pineal gland (PG) is a source of several trophic factors. In this study, PG and superior cervical ganglia (SCG) from Sprague-Dawley neonates (1-day-old) were co-cultured to test the hypothesis that endogenous release of PG NGF (or an NGF-like cytokine) is sufficient to promote survival of SCG neurons. Neuronal density of SCG neurons was significantly enhanced when co-cultured with PG for 7 days compared to SCG cultured alone. SCG survival and neurite formation in PG co-cultures was less than in SCG treated with exogenous NGF (100 ng/ml). The neurotrophic effect of PG co-cultures was abolished when 1% anti-NGF was added to the medium. Co-cultures of SCG neurons with established 7-day PG cultures induced extensive SCG neurite formation within 24 hr compared to SCG co-cultured with 1-day PG cultures. This suggests that PG neurotrophic effects are due to PG non-neuronal cells (nnc) that proliferate to confluency by 7 days in culture. S-antigen-positive pinealocytes did not proliferate in culture. There was decreased SCG survival when neurons were seeded onto PG cultures that had been previously killed by drying, which suggests that the neurotrophic effects of nnc are not substrate-dependent. Immunocytochemical characterization of PG nnc revealed a heterogenous mixture of astrocytes, macrophage/microglia, and fibroblasts. These findings support the hypothesis that NGF is actively secreted by PG and that nnc are the principal source of this neurotophin.

Animals↗

PubMed1994

Glutamate inhibition of the adrenergic-stimulated production of melatonin in rat pineal gland in vitro.

The effect of L-glutamate on the adrenergic-stimulated release of melatonin in the rat pineal gland was examined using an in vitro perfusion system. L-Glutamate by itself had no effect on melatonin secretion whereas L-glutamate administered prior to (-)-isoproterenol (beta-adrenergic agonist) and L-phenylephrine (alpha-adrenergic agonist) inhibited melatonin production by 42%. L-Glutamate did not inhibit melatonin secretion when glands were stimulated with (-)-isoproterenol alone. D-Glutamate, as well as the L-glutamate agonists kainate, N-methyl-D-aspartate, quisqualate, and trans-1-aminocyclopentane-1,3-dicarboxylic acid, had no effect on the (-)-isoproterenol- and L-phenylephrine-stimulated secretion of melatonin, which suggests that the inhibitory effects of glutamate are not mediated via any of the known glutamate receptor subtypes. The possibility that L-glutamate may be converted to another neuroactive compound (GABA) prior to the addition of (-)-isoproterenol and L-phenylephrine is suggested by the observation that simultaneous administration of L-glutamate with (-)-isoproterenol and L-phenylephrine did not inhibit melatonin production.

Amino Acids↗

PubMed1994

Hyperoxia increases airway cell S-phase traversal in immature rats in vivo.

Exposure of 21-day-old Sprague-Dawley rats to hyperoxia (> 95% O2 for 8 days) causes thickening of the airway epithelial and smooth muscle layers. To test the hypothesis that hyperoxic exposure increases airway layer DNA synthesis, we labeled the nuclei of cells undergoing S-phase by administering the thymidine analog bromodeoxyuridine (BrdU). BrdU was administered on days 3 and 4, 5 and 6, or 7 and 8 of air or O2 exposure, and the lungs were harvested immediately thereafter. Histologic sections were stained with an avidin-biotin-immunoperoxidase stain that revealed BrdU incorporation into nuclei, and a hematoxylin counterstain. After 4 days of air or O2 exposure, there was no difference in BrdU fractional labeling between control and hyperoxic animals. Thereafter, fractional BrdU labeling of the small airway (circumference < 1,000 microns) epithelium and smooth muscle layer was significantly increased in O2-exposed animals (P < 0.01, unpaired t test). The fractional labeling of larger, central airway smooth muscle layer cells was also increased after 8 days of O2 exposure (P < 0.01). In another cohort of O2-exposed animals, measurements of airway layer dimensions demonstrated increases in small airway epithelial and smooth muscle layer thickness that paralleled the time course seen for BrdU incorporation. We conclude that O2 exposure of immature rats increases airway epithelial and smooth muscle layer cellular DNA synthesis. These data suggest that hyperplasia of airway epithelial and smooth muscle layer cells may contribute to hyperoxia-induced airway remodeling.

Animals↗

PubMed1993

Immunocytochemical demonstration of nerve growth factor (NGF) receptor in the pineal gland: effect of NGF on pinealocyte neurite formation.

Nerve growth factor receptor immunoreactivity (NGFRI) in the pineal gland was examined both light and electron microscopically using the monoclonal antibody 192IgG. NGFRI was located on sympathetic fibers and on perivascular cells resembling macrophage/microglia. A pineal gland dispersed cell culture model confirmed the presence of NGFRI in cells that exhibited processes of varying lengths and were distributed among pinealocytes and other flat cells. Pinealocytes in dispersed cell culture were identified immunocytochemically by their expression of S-antigen, their round shape and small size and their tendency to extend neurites in the direction of the flat cells in culture. The length of pinealocyte neurites showed a significant increase when cultured in the presence of NGF (25 ng/ml), suggesting that trophic factors, mediated by these macrophage/microglial cells, are important to the morphogenesis of these neuroendocrine cells. Neurotrophic activation of these neuroendocrine macrophage/microglia may have neuro-immunomodulatory implications leading to expression of proteins encoded by the major histocompatibility complex.

Animals↗

PubMed1993

Immunocytochemical and electron-microscopic characterization of macrophage/microglia cells and expression of class II major histocompatibility complex in the pineal gland of the rat.

Interstitial cells in the pineal gland of the rat were characterized immunocytochemically using the monoclonal antibodies MRC OX-42 and ED1 for macrophages/microglia, and MRC OX-6, which recognizes major histocompatibility complex (MHC) class II antigen. A polyclonal antibody against GFAP was used to identify astrocytes. Cells immunopositive for OX-42 and/or ED1 were distributed throughout the gland; they extended processes primarily along the perivascular spaces and occasionally within the parenchyma of the gland. Ultrastructurally, these OX-42-positive cells were characterized by a nucleus with sparse heterochromatin and cytoplasmic vacuoles/lysosomes. Cells expressing MHC class II antigen had a distribution and morphology similar to OX-42-immunopositive cells, suggesting that pineal macrophages/microglia play a role as antigen-presenting cells. GFAP-positive astrocytes were concentrated at the proximal end of the pineal where the pineal stalk enters the gland. The occurrence of antigen-presenting cells in this circumventricular neuroendocrine gland has important functional implications as these cells may be mediators of neuroimmunomodulatory mechanisms, and involved in certain disease states such as autoimmune pinealitis.

Animals↗

PubMed1993

Characterization of a [3H]glutamate binding site in rat pineal gland: enhanced affinity following superior cervical ganglionectomy.

Glutamate, an excitatory neurotransmitter/neuromodulator involved in cell-to-cell communication within the central nervous system, is now believed to play a role in neuroendocrine function. In this study we describe a single, saturable, stereospecific, and temperature-, time-, and pH-dependent binding site for glutamate in the pineal gland of the rat (Kd = 612 +/- 23 nM, Bmax = 3.17 +/- 0.33 pmol/mg protein). After removal of the sympathetic innervation to the pineal gland, [3H]glutamate binding displayed a higher apparent affinity (Kd = 412 +/- 28 nM) (P < 0.05) without a change in binding site number (Bmax = 3.60 +/- 0.24 pmol/mg protein). No difference in [3H]glutamate binding site number was observed in pineal glands obtained from animals sacrificed during the middle of the light and dark periods. These data suggest a possible modulatory role for a glutamate binding site in pineal gland function.

Animals↗