Biomedical subjects

D O Carpenter

Publications and source records attributed to D O Carpenter.

At least 19 recordsLinked to original sources

PubMed1999

Sources of reactive oxygen species production in excitotoxin- stimulated cerebellar granule cells.

Reactive oxygen species (ROS) production in rat cerebellar granule cells in the presence of the excitotoxins N-methyl-d-aspartate (NMDA) and kainic acid (KA) and by the protein kinase C activator phorbol myristate acetate (PMA) was Ca2+-dependent and resulted in decreased cell viability. Exposure of stimulated cells to rotenone (a respiratory chain inhibitor) did not decrease ROS levels and did not affect short-term cell viability. In cells stimulated by NMDA and KA, exposure to indomethacin (a cyclooxygenase inhibitor) and nialamide (a monoamine oxidase inhibitor) caused a decrease in ROS levels and increased cell viability occurred in NMDA-treated cells. In contrast, PMA-stimulated neurons did not show decreased ROS levels when exposed to indomethacin and nialamide. These studies suggest that there is a multiplicity of routes for Ca2+-dependent ROS production in neurons but that ROS generation by cyclooxygenase and monoamine oxidase is not controlled by protein kinase C.

Animals↗

PubMed1999

Hazardous wastes in eastern and central Europe: technology and health effects.

Issues of hazardous waste management are major concerns in the countries of eastern and central Europe. A National Institute of Environmental Health Sciences-supported conference was held in Prague, Czech Republic, as a part of a continuing effort to provide information and promote discussion among the countries of eastern and central Europe on issues related to hazardous wastes. The focus was on incineration as a means of disposal of hazardous wastes, with discussions on both engineering methods for safe incineration, and possible human health effects from incineration by-products. Representatives from government agencies, academic institutions, and local industries from 14 countries in the region participated along with a few U.S. and western European experts in this field. A series of 12 country reports documented national issues relating to the environment, with a focus on use of incineration for hazardous waste disposal. A particularly valuable contribution was made by junior scientists from the region, who described results of environmental issues in their countries.

Environmental Health↗

PubMed1998

Vitamin E enhances Ca(2+)-mediated vulnerability of immature cerebellar granule cells to ischemia.

The effects of vitamin E on lipid peroxidation, intracellular free Ca2+ concentration ([Ca2+]i), and cell death were investigated in the postischemic immature cerebellum. Deprivation of oxygen and glucose for 10-min in a suspension of freshly dissociated granule cells from the cerebellum of 9-day-old male rat pups resulted in a recovery-induced consumption of cell nonenzymatic antioxidants (ascorbic acid, glutathione, and alpha-tocopherol) and development of membrane lipid peroxidation as measured by the thiobarbituric acid method. The rate of lipid peroxidation of the postischemic cells was stimulated, not reduced, by treatment of the cells with vitamin E (5-30 microM alpha-tocopherol phosphate). In flow-cytometric studies a 10-min period of ischemia resulted in a small increase in intracellular calcium concentration, lipid peroxidation products and cell death, but in the presence of alpha-tocopherol the same treatment caused a dramatic increase in cell death, accompanied by a large increase in [Ca2+]i and lipid peroxidation products. Pretreatment of the cells with a mixture of three antioxidants (vitamin C/rutin/ubiquinol-10, 10/5/1) or nickel (Ni2+) reduced the alpha-tocopherol-induced increases in [Ca2+]i, and cell death. Hydrogen peroxide (1 mM) and the water-soluble analogue of vitamin E, trolox (50 microM), mimicked the effect of vitamin E on lipid peroxidation in the postischemic cells. Pretreatment of the cells with the intracellular Ca2+ chelator BAPTA-AM, reduced both the alpha-tocopherol-induced increase in [Ca2+]i and cell death. The effect of vitamin E on [Ca2+]i was age dependent and decreased abruptly during maturation of the cerebellum between the first and second weeks of life. Results of in vitro treatment of the immature cerebellar cells with the water-soluble form of vitamin E (alpha-tocopherol phosphate) suggest that, after consumption of cellular co-antioxidants, vitamin E may be converted to an alpha-tocopheroxyl radical, which act as a toxic prooxidant as cellular bioenergetics deteriorate.

Animals↗

PubMed1998

PCBs reduce long-term potentiation in the CA1 region of rat hippocampus.

Prenatal exposure to polychlorinated biphenyls (PCBs) has been associated with a lower IQ in childhood. We have examined the effects of acute exposure to PCB mixtures and two single congeners on synaptic transmission between Schaffer collaterals and CA1 neurons of the rat hippocampus as well as posttetanic potentiation (PTP), paired pulse facilitation (PPF), and long-term potentiation (LTP). PTP and PPF represent transient increases in transmitter release immediately after stimulation, while LTP is a measure of long-term changes in synaptic plasticity that has been related to learning and memory. LTP, but neither PTP nor PPF, was reduced by Aroclor 1016 in a dose-dependent fashion at concentrations that had little effect on general synaptic transmission. The more highly chlorinated Aroclor 1254 at low concentrations specifically blocked LTP, but at higher concentrations also reduced synaptic transmission. The mono-ortho PCB congener 2,4,4'-trichlorobiphenyl and the coplanar congener 3,3',4,4'-tetrachlorobiphenyl also blocked LTP without effect on PTP or PPF. We conclude that PCBs selectively impair the process of LTP in CA1 neurons of the hippocampus.

Animals↗

PubMed1998

Environmental pollution and child health in central and Eastern Europe.

For the last 50 years, the economic and industrial development of the nations of Central and Eastern Europe has been achieved at the cost of environmental degradation. The health risks posed by this pollution to children and the steps necessary to ameliorate such risks are only beginning to be investigated. At a recent conference in Poland, sponsored in part by the National Institute of Environmental Health Sciences, participants from 11 countries in the region, together with scientists from Western Europe and the United States, met to share information regarding pediatric environmental health in Central and Eastern Europe, to consider methodologic issues in the design and conduct of such studies, and to discuss preventive strategies. This report summarizes the deliberations, outlines problem areas such as heavy metals and air pollution, delineates research and training needs to help Central and Eastern Europeans deal more effectively with such problems, and recommends specific future actions and collaborative efforts.

Air Pollution↗

PubMed1998

Human health and chemical mixtures: an overview.

Unlike laboratory animals, people are rarely exposed to a single hazardous chemical. However, most of the information documenting adverse human health effects from environmental and occupational contaminants has come from studies focused on exposure to single chemicals, and there is little information available on how two or more contaminants affect humans. Most information on the effects of mixtures comes from animal systems and limited investigations of isolated human cells in culture, even though the study of mixtures in such systems has also been neglected. Two or more compounds may show additive, antagonistic, or synergistic interactions or may act on totally different systems and thus not interact. Furthermore, even a single chemical may have multiple effects and affect more than one organ system. Effects may vary with age, and metabolites may have totally different actions from the parent compound. This paper will review the variety of health effects in humans that may result from environmental contaminants and discuss how such contaminants may interact with each other. We will also present examples on how different contaminants interact from toxicologic studies of polychlorinated biphenyls performed as part of our Albany, New York, Superfund Basic Research Program project.

Animals↗

PubMed1998

Polychlorinated biphenyls and human health.

Polychlorinated biphenyls (PCBs) were manufactured and used widely for many years. Because they are very persistent in both the environment and biological systems, there has been significant global contamination. This review presents a summary of known or suspected health effects of various PCB congeners, documented on the basis of both human and animal studies. As our knowledge increases several important points become apparent. PCBs interfere with many biological functions, including the immune system, the nervous system, and several endocrine system, and the fetus appears to be particularly vulnerable to these actions. PCBs cause certain cancers in animals. PCBs are mixtures of multiple congeners, differing on the basis of the numbers and positions of chlorines around the biphenyl ring, and it is becoming increasingly apparent that different congeners may have very different actions. These observations suggest that the potential human health hazards from PCB exposure have been underappreciated.

Animals↗

PubMed1998

Lead potentiates cytokine- and glutamate-mediated increases in permeability of the blood-brain barrier.

We have measured the transendothelial electrical resistance across the blood-brain barrier (BBB) with a microelectrode technique and determined the effects of subcutaneous injections (five injections over ten days) of lipopolysaccharide (LPS, 100 ng/g), recombinant mouse interleukin-6 (IL-6, 5 ng/g), and/or inorganic lead (lead, 2.5 5 micrograms/g) on the ion permeability of arterioles in the temporoparietal cortex of anaesthetized mice between 10 and 40 days of age. In controls the electrical resistance increased with age. It was decreased in animals treated with IL-6, but unaffected by lead at the different ages studied. In IL-6 treated mice, repeated neonatal exposure to lead (five injections between 2 and 10 days after birth) caused a delay in the increase in arteriole resistance with age. LPS injections caused a 36% increase in ion permeability of the BBB in twenty-day-old mice, and lead potentiated this effect of LPS. Intra-arterial injections of glutamate did not alter vascular resistance, but topical applications of glutamate on the cerebrum caused a reversible decrease in the resistance in mice not treated with lead, and an irreversible decrease in mice treated with lead. Injections of glutamate in the lumen of arterial vessels in the parietal and temporoparietal brain areas of mice pretreated with lead and LPS, plus a topical application of glutamate, caused depolarization of neurons in the temporoparietal cortex. These results suggest that disruption of the BBB can allow serum glutamate to penetrate the brain, causing further disruption of the BBB, and that lead irreversibly potentiates this cascade of harmful events.

Animals↗

PubMed1998

Lipopolysaccharide and interleukin-6 enhance lead entry into cerebellar neurons: application of a new and sensitive flow cytometric technique to measure intracellular lead and calcium concentrations.

The distribution of intracellular ionized lead (Pb) and calcium in dissociated cerebellar cells of ten-day-old mice was measured by flow cytometry. There are no fluorescent probes specific for lead, whereas commonly used fluorescent calcium indicators bind heavy metals with greater affinity than they do calcium, which impedes discrimination of lead- and calcium-induced fluorescence changes. Therefore, we developed a method to determine [Pb2+]i and [Ca2+]i by employing a combination of the calcium indicator fluo-3 and the heavy-metal chelator TPEN. Using these methods, we studied the effects of multiple in vivo exposure (five subcutaneous injections over 10 days) to lipopolysaccharide (LPS, 100 ng/g), recombinant mouse interleukin-6 (IL-6, 5 ng/g) and/or inorganic lead (lead, 2.5 micrograms/g) on lead and calcium concentrations. Control cells had [Cai] of 112 nM. Lead exposure alone had little effect on [Ca2+]i and resulted in a mean [Pb2+]i of about 7 pM, and did not alter cell volume. A significant fraction of cells (about 44% of living cells) from animals treated with lead plus LPS were swollen, as determined by analysis of the light scattering pattern, and there was a small increase in the number of dead cells, identified with the nucleic acid stain, 7-aminoactinomycin. While [Ca2+]i was not significantly increased in animals treated with either only LPS or IL-6, lead and calcium concentrations were increased in animals exposed to lead and LPS or IL-6 in both the non-swollen and swollen cells, with a mean value of (Pb2+)i of 32 pM and (Ca2+)i of 155 nM in cells not swollen. Electrophysiological analysis showed that LPS injections caused decreases in the membrane potential of endothelial cells of the blood-brain barrier (BBB) and lead potentiated the effect of LPS. IL-6 mimicked the effects of LPS, but was less potent. Thus these experiments indicate a synergistic interaction between lead and cytokines on biophysical properties of both neurons and endothelial cells of the BBB.

Animals↗

PubMed1998

Time series analysis of Aplysia total motion activity.

We rely on various nonparametric time series techniques to quantify several autocorrelated Aplysia total motion activity series. The effect of light on total motion activity has been removed. A general model will be proposed for total motion activity, which is found to display a circadian rhythm, and the toxic effect of PCBs on total motion activity will be assessed. No underlying assumptions are made about the form of the circadian rhythm. Several types of circadian total motion activity patterns are discovered. We form the hypothesis that different data series may be separated into three distinct groups of common patterns of circadian total motion activity. We also investigate if in fact only a single innate oscillator regulates total motion activity.

Animals↗

PubMed1997

Metabotropic GABA receptors regulate acetylcholine responses on snail neurons.

1. The A type of acetylcholine response of Helix neurons is downmodulated by low concentrations of GABA that do not elicit any measurable change in membrane potential or conductance. 2. We find that these physiological actions are associated with an increase in both intracellular cyclic AMP levels and 45Ca2+ influx. 3. The modulation of the acetylcholine response by GABA is blocked when the neurons are injected with EGTA to prevent a rise in intracellular Ca2+ concentration or when tolbutamide, an inhibitor of protein kinase A, is applied. 4. These results are consistent with the effects of GABA being mediated by a metabotropic GABA receptor that is activated at very low GABA concentrations and mediates modulation of the acetylcholine response via regulation of intracellular Ca2+ and cyclic AMP levels.

Acetylcholine↗

PubMed1997

Effects of inorganic mercury and methylmercury on the ionic currents of cultured rat hippocampal neurons.

1. The effects of inorganic Hg2+ and methylmercuric chloride in the ionic currents of cultured hippocampal neurons were studied and compared. We examined the effects of acute exposure to the two forms of mercury on the properties of voltage-activated Ca2+ and Na+ currents and N-methyl-D-aspartate (NMDA)-induced currents. 2. High-voltage activated Ca2+ currents (L type) were inhibited by both compounds at low micromolar concentrations in an irreversible manner. Mercuric chloride was five times as potent as methylmercury in blocking L-channels. 3. Both compounds caused a transient increase in the low-voltage activated (T-type) currents at low concentrations (1 microM) but blocked at higher concentrations and with longer periods of time. 4. Inorganic mercury blockade was partially use dependent, but that by methylmercury was not. There was no effect of exposure of either form of mercury on the I-V characteristics of Ca2+ currents. 5. Na(+)- and NMDA-induced currents were essentially unaffected by either mercury compound, showing only a delayed nonspecific effect at a time of overall damage of the membrane. 6. We conclude that both mercury compounds show a relatively selective blockade of Ca2+ currents, but inorganic mercury is more potent than methylmercury.

Animals↗

PubMed1997

Time course and involvement of protein kinase C-mediated phosphorylation of F1/GAP-43 in area CA3 after mossy fiber stimulation.

1. Protein kinase C (PKC) activity and phosphorylation of F1/growth associated protein (GAP)-43, a PKC substrate, have been proposed to play key roles in the maintenance of long-term potentiation (LTP) at the synapses of Schaffer collateral/commissural on pyramidal neurons in CA1 (Akers et al., 1986). We have studied in the involvement of PKC and PKC-dependent protein phosphorylation of F1/GAP-3 in in vitro LTP observed at the synapses of mossy fiber (MF) on CA3 pyramidal neurons of rat hippocampus by post hoc in vitro phosphorylation. 2. After LTP was induced in CA3 in either the presence or absence of D-2-amino-5-phosphonovaleric acid (AP5), an NMDA receptor antagonist, the CA3 region was dissected for in vitro phosphorylation assay. In vivo phosphorylation of F1/GAP-43 was increased in membranes at 1 and 5 min after tetanic stimulation (TS) but not at 60 min after TS. 3. The degree of phosphorylation of F1/GAP-43 in the cytosol was inversely related to that in membranes at each time point after LTP. 4. The similar biochemical changes obtained from either control slices or AP5-treated slices indicate that LTP and the underlying biochemical changes are independent of the NMDA receptor. Immunoreactivity of the phosphorylated F1/GAP-43 in LTP slices was not significantly different from control, indicating that results from western blotting and post hoc in vitro phosphorylation are consistent. 5. Post hoc in vitro phosphorylation of F1/GAP-43 was PKC-mediated since phosphorylation of F1/GAP-43 was altered by the PKC activation cofactors, Ca2+, phosphatidylserine and phorbol ester. 6. Calmodulin (CaM) at > 5 microM inhibited phosphorylation, consistent with the presence of CaM-binding activity at the site on F1/GAP-43 acted upon by PKC. 7. We conclude that phosphorylation of F1/GAP-43 is associated with the induction but not the maintenance phase of MF-CA3 LTP.

2-Amino-5-phosphonovalerate↗

PubMed1997

Biochemical and physiological evidence that carnosine is an endogenous neuroprotector against free radicals.

1. Carnosine, anserine, and homocarnosine are endogenous dipeptides concentrated in brain and muscle whose biological functions remain in doubt. 2. We have tested the hypothesis that these compounds function as endogenous protective substances against molecular and cellular damage from free radicals, using two isolated enzyme systems and two models of ischemic brain injury. Carnosine and homocarnosine are both effective in activating brain Na, K-ATPase measured under optimal conditions and in reducing the loss of its activity caused by incubation with hydrogen peroxide. 3. In contrast, all three endogenous dipeptides cause a reduction in the activity of brain tyrosine hydroxylase, an enzyme activated by free radicals. In hippocampal brain slices subjected to ischemia, carnosine increased the time to loss of excitability. 4. In in vivo experiments on rats under experimental hypobaric hypoxia, carnosine increased the time to loss of ability to stand and breath and decreased the time to recovery. 5. These actions are explicable by effects of carnosine and related compounds which neutralize free radicals, particularly hydroxyl radicals. In all experiments the effective concentration of carnosine was comparable to or lower than those found in brain. These observations provide further support for the conclusion that protection against free radical damage is a major role of carnosine, anserine, and homocarnosine.

Animals↗

PubMed1997

Flow cytometric measurements of neuronal death triggered by PCBs.

We have examined the effects of several individual polychlorinated biphenyl (PCB) congeners and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on viability and intracellular calcium concentration in acutely dissociated cerebellar granule cell neurons obtained from 7-14 day old rat pups, using a flow cytometer. Cell death was determined using the fluorescent dye, 7-aminoactinomycin-D (7-AAD), while intracellular calcium concentration was determined using Fluo-3-AM. When acutely isolated neurons were exposed to 2,4,4' trichlorobiphenyl (2,4,4', there was a dose- and time-dependent loss of viability beginning within 5 min. At a concentration of 100 microM, 80% of neurons were dead within 45 min. Loss of viability was preceded by an apparent increase in cell granularity. No significant cell death was observed upon exposure to the highly dioxin-like coplanar PCB, 3,4,5,3',4', at a concentration of 100 microM or by TCDD at a concentration of 0.1 microM. A slight loss of viability was seen with the ortho 2,6,2',6' and the coplanar 3,4,3',4', but for both it was small. All of the PCB congeners caused an early, transient increase in intracellular free calcium concentration, although TCDD did not. A small, late increase in intracellular free calcium was seen with 2,4,4', probably reflecting events leading to loss of viability. We conclude that at least one PCB congener, 2,4,4', is very neurotoxic and can cause rapid death of cerebellar granule cells.

Animals↗

PubMed1996

The translocation and involvement of protein kinase C in mossy fiber-CA3 long-term potentiation in hippocampus of the rat brain.

The detailed mechanisms underlying long-term potentiation (LTP) are not known. In hippocampal CA1, translocation of protein kinase C (PKC) activity from cytosol to membrane and subsequent phosphorylation of growth associated protein (GAP)-43 have been demonstrated to be critical events for the maintenance phase of LTP. LTP in mossy fiber (MF)-CA3 pathway and the Schaffer collateral/commissural (SC)-CA1 pathway differ in a number of ways: SC-CA1 LTP depends on NMDA receptors while MF-CA3 LTP does not, and SC-CA1 LTP is primarily postsynaptic while MF-CA3 LTP is primarily presynaptic. The role of PKC in MF-CA3 LTP has not been studied. We investigated the role of PKC in CA3 and show that PKC inhibitors prevent LTP, but that PKC activators produce a reversible synaptic potentiation, indicating that PKC activation is an essential but not sufficient component of LTP in CA3. Then using antibodies against specific PKC isozymes we have determined the membrane vs. cytosolic distribution of various PKC isozymes in slices subjected to low or tetanic stimulation, or perfused with phorbol esters (PDAc). Compared with control, LTP and PDAc slices show greater PKC-alpha and -epsilon immunoreactivity in the membrane fraction, indicating that both LTP and phorbol ester treatment induce translocation of PKC-alpha and -epsilon from cytosol to membrane. However, with PKC-beta and PKC-gamma the only detectable translocation from cytosol to membrane was in the phorbol ester-treated slices. Thus, while phorbol ester treatment causes translocation of PKC-alpha, -beta, -gamma and -epsilon, the only detectable translocation associated with CA3 LTP is that of PKC-alpha and -epsilon.

Analysis of Variance↗

PubMed1996

Flow-cytometric estimation on glutamate- and kainate-induced increases in intracellular Ca2+ of brain neurons: a technical aspect.

Effects of glutamate and kainate on the intracellular Ca2+ concentration ([Ca2+]i) in a large population (several thousand) of dissociated cerebellar granule cell neurons were evaluated using a flow-cytometer and a combination of two fluorescent dyes, fluo-3-AM for estimating [Ca2+]i and ethidium bromide for removing neurons that had compromised membranes from the cell population examined. The number of neurons responding to glutamate or kainate in augmenting the fluo-3 fluorescence increased in a dose-dependent manner. The number of neurons responding to kainate was much greater than that to glutamate. CNQX, a blocker of non-NMDA receptors, completely blocked the response elicited by kainate while the complete blockade of this glutamate-induced response was made by a combination of MK-801, a NMDA receptor blocker, and CNQX. Nicardipine, a calcium antagonist, decreased the number of neurons responding to glutamate and kainate, suggesting involvement of voltage-dependent calcium channels. These results indicate that the flow-cytometric measurement of glutamate and kainate responses has the potential to provide answers to such questions as what percentage of the population of neurons respond to these amino acids and what is the resulting distribution of [Ca2+]i.

Aniline Compounds↗

PubMed1996

Propylthiouracil treatment reduces long-term potentiation in area CA1 of neonatal rat hippocampus.

Rat pups were made hypothyroid by exposure to propylthiouracil in drinking water beginning at 1 week of age, and the degree of long-term potentiation (LTP) in hippocampal area CA1 determined from brain slices of animals ranging in age from 2 to 6 weeks. Serum T3 levels were less than 20% of that of age matched controls after 3 weeks of treatment, and remained at that level. Relative to the age-matched controls, LTP was reduced significantly after 2 weeks of treatment. These observations are consistent with the conclusion that LTP magnitude is a reflection of cognitive function, which is known to be depressed in hypothyroid conditions in both animals and man.

Administration, Oral↗