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Biomedical subjects

D W Walker

Publications and source records attributed to D W Walker.

At least 19 recordsLinked to original sources
PubMed2005

Effects of age and pregnancy on the circulatory activin response of sheep to acute inflammatory challenge by lipopolysaccharide.

The release of activin A in response to intravenous injection of the bacterial cell-wall component lipopolysaccharide (LPS) was investigated in an ovine model of acute inflammatory challenge in newborn and adult sheep, and in non-pregnant and pregnant ewes. Neonatal lambs (<20 days of age) showed a quantitatively similar response in terms of circulating concentrations of activin A, its binding protein follistatin and the cytokine interleukin-6 compared with adult ewes challenged with an equivalent dose (300 ng/kg bodyweight) of LPS. The fever response and plasma tumour necrosis factor-alpha release in response to LPS, however, were significantly (P < 0.01) less in lambs than in the adult group. Pregnant ewes in the last trimester of gestation had similar responses to LPS, in all aspects measured, compared with their non-pregnant counterparts, apart from an ablated fever response. Although the adult and neonatal sheep responded to LPS, a similar response was not apparent in the fetal circulation, possibly due to a protective effect of the placenta. A 10-fold increase in the dose of LPS (from 300 ng to 3 microg/kg bodyweight) given to neonatal lambs elicited an increase in several cytokine responses measured, with a significant (P< 0.05) increase in follistatin release. In contrast, the amount of activin released by the increased dose of LPS was similar to that invoked by the lower dose. The effect of tolerance to LPS was investigated by giving a second challenge of LPS 5 days after the initial injection. In all animals studied, there was an ablated (P < 0.05) response to the subsequent LPS injection, apart from a similar temperature-response profile. These data provide further evidence that activin A concentrations in the bloodstream are acutely responsive to inflammatory challenge in post-natal life and suggest that the response forms a significant component of the innate immune system.

Activins↗
PubMed2003

Chronic ethanol treatment reduces the magnitude of hippocampal LTD in the adult rat.

Human alcoholics and animals that have received chronic ethanol treatment (CET) display memory deficits. Previous work in our laboratory has shown that CET produces damage to the hippocampus and a reduction in the magnitude of hippocampal long-term synaptic potentiation. In the present report we examined the effects of CET on hippocampal long-term depression (LTD). We used in vitro hippocampal slices to examine LTD after rats received 38-41 weeks of paired feeding on liquid diets containing ethanol or isocaloric sucrose. Stimulation delivered through electrodes in the CA3-CA1 Schaffer collateral pathway activated synaptic population responses in CA1. LTD of CA1 stratum radiatum evoked field potential slope was not induced by 900 single pulses at 1 Hz, but was elicited by 900 pulse pairs separated by 50 or 200 msec delivered at 1 Hz (pLFS50, pLFS200). LTD evoked by pLFS200, but not by pLFS50, was significantly reduced in slices from ethanol-treated rats. The N-methyl D-aspartate (NMDA) receptor antagonist 2-amino-5-phosphonopentanoic acid (AP5) (50 micro M) blocked LTD induced by pLFS50 and pLFS200 equally, but the L-type calcium channel blocker nimodipine (10 micro M) had no effect on either type of LTD. Thus, direct effects on these channels cannot explain how CET selectively reduces the magnitude of pLFS200 LTD. Finally, we describe a novel and robust LTD of the presynaptic afferent volley that is resistant to CET, NMDAR antagonists, GABA-A receptor blockade, and blockade of L-type calcium channels.

Animals↗
PubMed2003

A comparison of desiccant materials used for monitoring atmospheric tritium concentrations in a semi-arid climate.

Atmospheric tritium monitoring involves the collection of tritiated water vapor by collecting atmospheric moisture from air that is drawn through a bed of desiccant material. This study is a comparison between molecular sieve and silica gel adsorbent media used for atmospheric moisture sampling conducted in the semi-arid climate of the Idaho National Engineering and Environmental Laboratory. Water vapor was collected simultaneously using two columns containing different desiccant materials (one column containing molecular sieve and the other containing silica gel). Data collected during air sampling periods were compared with meteorological data collected, and atmospheric moisture collection efficiencies were determined. Breakthrough of atmospheric moisture past the desiccant material was suspected with both media at elevated temperatures indicating that smaller sample volumes, lower volumetric flow rates, or longer adsorbent columns should be used during summer when ambient temperatures are elevated.

Air Pollutants, Radioactive↗
PubMed2003

A field test of electret ion chambers for environmental remediation verification.

In 1999, the State of Idaho INEEL Oversight Program performed a field test to determine if electret ion chambers could be used as a replacement to soil sampling in field survey releases at a remediated radioactively contaminated site. This study compared exposure rates measured using electret ion chambers with gamma spectroscopic analysis results of soil samples collected at the remediated site. Exposure rate measurements were made in 1999 using electret ion chambers at the same locations that INEEL Oversight Program sampled soil in 1998 following MARSSIM protocol. The waste site was divided into nine survey units based upon site history including previous site surveys and physical boundaries. Exposure rate measurements at the remediation site compared well with exposure rate measurements made at reference background locations used for routine environmental monitoring by the State of Idaho indicating that the remedial action met cleanup criteria. A poor correlation between exposure rate measurements and 137Cs concentrations and other discrepancies were observed during this study with respect to measurements made during the 1998 final site survey.

Biodegradation, Environmental↗
PubMed2002

The effects of chronic ethanol consumption on neurotrophins and their receptors in the rat hippocampus and basal forebrain.

Damage to the basal forebrain frequently results in deficits in learning and memory. Mnenonic dysfunction also occurs following prolonged ethanol consumption in humans and in animal models of chronic ethanol intake, accompanied by specific abnormalities in synaptic transmission between the basal forebrain and hippocampus. The integrity of at least some of the reciprocal neuronal connections between these brain regions is influenced by target-derived neurotrophic factors. We used a semiquantitative reverse transcription polymerase chain reaction technique to measure the messenger RNA for neurotrophins BDNF and NGF, and for their receptors trkB, trkA, and the low affinity receptor, p75(NTR) in the hippocampus and basal forebrain of rats after 28 weeks of alcohol consumption without malnutrition. This chronic ethanol treatment (CET) resulted in a marked and selective reduction in basal forebrain trkA mRNA. Western blotting revealed a similar reduction of basal forebrain trkA protein. CET effects on basal forebrain trkA may reflect impaired NGF signaling that could compromise septohippocampal synaptic connections, cholinergic differentiation, and emergent functional abilities dependent on these properties.

Alcohol Drinking↗
PubMed2002

Concurrent quantification of quinolinic, picolinic, and nicotinic acids using electron-capture negative-ion gas chromatography-mass spectrometry.

Quinolinic, picolinic, and nicotinic acids and nicotinamide are end products of the kynurenine pathway from l-tryptophan and are intermediates in the biosynthesis of nicotinamide adenine dinucleotide. These compounds are involved in complex interrelationships with inflammatory and apoptotic responses associated with neuronal cell damage and death in the central nervous system. To facilitate the study of these compounds, we have utilized gas chromatography-mass spectrometry in electron capture negative ionization mode for their concurrent trace quantification in a single sample. Deuterium-labeled quinolinic, picolinic, and nicotinic acids were used as internal standards and the compounds were converted to their hexafluoroisopropyl esters prior to chromatography. Nicotinamide was readily quantified after conversion to nicotinic acid using gas-phase hydrolysis-a process which did not affect the deuterated internal standards. The on-column limit of quantification was less than 1 fmol for each of the analytes and calibration curves were linear. A packed column liner was used in the gas chromatograph inlet to effectively eliminate sample interference effects in the analysis of trace (femtomolar) levels of quinolinic acid. The method enables rapid and specific concurrent quantification of quinolinic, picolinic, and nicotinic acids in tissue extracts and physiological and culture media.

Animals↗
PubMed2001

Effect of finasteride on behavioural arousal and somatosensory evoked potentials in fetal sheep.

This study examines the effect of inhibiting the synthesis of gamma-aminobutyric acid(A) (GABA(A)) agonist steroids on behavioural activity and somatosensory evoked potentials (SEP) in late gestation fetuses. Pregnane steroid production was suppressed by infusion of the 5alpha-reductase inhibitor, finasteride in chronically catheterised fetal sheep, 130-135 days gestation. Finasteride treatment (160 mg in 10 ml of vehicle over 2 h) significantly increased the incidence of fetal arousal during the period 4-10 h after commencing the infusion (P<0.05, n=6), whereas other behavioural parameters were not effected. In three of four animals, finasteride produced an increase in the amplitude of the N22 peak of the SEP during high voltage electrocortical activity. We conclude that suppression of pregnane steroid synthesis, by inhibition of the 5alpha-reductase enzyme, increases arousal activity in the fetus which is consistent with a reduction in GABA(A) receptor mediated inhibition.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed2001

Peroxide mediates ethanol-induced cytotoxicity in PC12 cells.

Pheochromocytoma (PC12) cell cultures exhibited a loss of cells and increase in intracellular oxidative stress when exposed to ethanol (EtOH) for 24 h. Catalase, an enzyme that hydrolyzes hydrogen peroxide (H(2)O(2)) to O(2) and H(2)O can attenuate EtOH-induced cell loss and oxidative stress in PC12 cells. This study provides the first clear evidence that oxidative stress in the form of elevated intracellular H(2)O(2) is a primary mechanism of EtOH neurotoxicity in PC12 cells.

Animals↗
PubMed2001

Regional changes in kynurenic acid, quinolinic acid, and glial fibrillary acidic protein concentrations in the fetal sheep brain after experimentally induced placental insufficiency.

OBJECTIVE: This study was undertaken to examine the effects of chronic embolization of the umbilical circulation during late gestation on regional concentrations of quinolinic acid and kynurenic acid (neuroactive products of tryptophan catabolism) and of the astrocyte-associated glial fibrillary acidic protein in the fetal brain. STUDY DESIGN: Pregnant ewes bearing fetuses with long-term catheter placement were treated daily with injections of either saline solution (n = 4; control group) or mucopolysaccharide microspheres (n = 5; embolized group) into the umbilical circulation through a femoral artery catheter between 120 and 140 days' gestation. The fetuses in the embolized group received sufficient microspheres each day to reduce and maintain the femoral arterial PO2 at < or =12 mm Hg. Autopsies were performed at 140 days' gestation to obtain the fetal brain for chemical analysis. RESULTS: Umbilical embolization resulted in nonacidemic hypoxia and hypoglycemia at 140 days' gestation. Quinolinic acid concentrations in the embolized group were significantly increased in the medulla, pons, midbrain, hypothalamus, and hippocampus, whereas kynurenic acid concentrations in the embolized group were reduced in the hippocampus and hypothalamus. There were significant reductions in glial fibrillary acidic protein contents in the occipitoparietal cortex, hippocampus, and pons in the embolized group. CONCLUSION: Placental compromise during late pregnancy had effects on kynurenine metabolism and astrocyte function in some regions of the fetal sheep brain. We suggest that these changes increase the vulnerability of the brain to asphyxial injury during late gestation and the perinatal period.

Animals↗
PubMed2001

Kynurenine production and catabolism in fetal sheep with embolized or nonembolized placentas.

OBJECTIVE: The effect of maternal tryptophan loading on fetal plasma and brain, kynurenic acid, and quinolinic acid concentrations was compared in late gestation fetal sheep with either chronically embolized or nonembolized placentas. STUDY DESIGN: The placentas of 4 ewes were embolized by daily injection of mucopolysaccharide microspheres into the umbilical artery from 120 days gestation in amounts sufficient to reduce the fetal arterial PO2 to < or = 12 mm Hg. Four fetuses with nonembolized placentas were the control group. At 135 to 138 days gestation, the ewe received an infusion of tryptophan (100 mg/kg, intravenously) or an equivalent volume of saline solution (100 mL) over 2 hours. Maternal and fetal arterial blood samples were obtained between 2 and 48 hours from the start of the infusion for the measurement of plasma tryptophan and kynurenine metabolites. Brains were then obtained from embolized and nonembolized fetuses 24 hours after a further maternal tryptophan loading experiment and from nonembolized non-tryptophan-treated fetuses for analysis of regional kynurenic acid and quinolinic acid content. RESULTS: Maternal tryptophan infusion resulted in a significant increase of kynurenine in fetal plasma, but this increase was significantly smaller in fetuses with an embolized placenta compared with a nonembolized placenta. Both kynurenic acid and quinolinic acid levels increased significantly in fetal plasma, with no differences between the groups. Kynurenic acid and quinolinic acid levels were increased in all regions of the fetal brain after maternal tryptophan loading, but these increases were greater in the fetuses with an embolized placenta, compared with a nonembolized placenta. CONCLUSION: Fetal tryptophan and kynurenine metabolism is significantly altered when placental function is chronically compromised in late gestation. The decreased production of kynurenine from tryptophan may result from the compromise of hepatic function in the fetus, whereas the increased production of kynurenic acid and quinolinic acid in the brain is likely to reflect alterations of metabolism of tryptophan and kynurenine to these neuroactive products by glial cells in the fetal brain.

Animals↗
PubMed2000

Localization of p450scc and 5alpha-reductase type-2 in the cerebellum of fetal and newborn sheep.

Prenatally, neuroactive steroids that modulate GABAergic activity may be synthesized de novo within the fetal brain. We have examined changes in immunoreactivity staining for the steroidogenic enzymes cholesterol P450 side-chain cleavage (P450scc), and 5alpha-reductase type-2 in the cerebellum of late gestation (130-145 days gestation) fetal sheep and newborn lambs (1-4 weeks of age). Both enzymes were predominantly localized in the Purkinje cell body and dendrites of the fetal and newborn cerebellum, with weaker immunoreactivity in a few cells of the inner granular layer. P450scc immunoreactivity was present in Purkinje neurons expressing either of the neuronal microtubule associated proteins MAP1b/5 or MAP2a/b, but was absent from GFAP and HNK-1 positive cells. Soma of Purkinje neurons were also immunopositive for 5alpha-reductase type-2 in the fetuses, but expression decreased to just detectable levels in the 1-2 and 2-4 week old lambs. Both MAP1b/5- and MAP2a/b-positive Purkinje neurons showed 5alpha-reductase type-2 expression in the fetus, whereas the residual 5alpha-reductase staining in the newborn lamb was present only in MAP2a/b-positive Purkinje neurons. Allopregnanolone in the cerebellum decreased from 21.8+/-1.9 ng/g wet weight in fetuses at 140-145 days gestation to 6.7+/-0.5 ng/g in 2-4 week old lambs (P<0. 05). Thus, synthesis of neuroactive steroids from cholesterol is possible in cerebellar neurons in late gestation and persists into neonatal life, 5alpha-reductase type-2 expression is greater in the fetus compared to the neonate, and allopregnanolone concentrations in the cerebellum decrease significantly after birth.

Animals↗
PubMed2000

Extension of life-span with superoxide dismutase/catalase mimetics.

We tested the theory that reactive oxygen species cause aging. We augmented the natural antioxidant systems of Caenorhabditis elegans with small synthetic superoxide dismutase/catalase mimetics. Treatment of wild-type worms increased their mean life-span by a mean of 44 percent, and treatment of prematurely aging worms resulted in normalization of their life-span (a 67 percent increase). It appears that oxidative stress is a major determinant of life-span and that it can be counteracted by pharmacological intervention.

Aging↗
PubMed2000

Characterisation of GABA(A) receptors in fetal, neonatal and adult ovine brain: region and age related changes and the effects of allopregnanolone.

Progesterone metabolites acting via GABA(A) receptors suppress central nervous system (CNS) activity. The aim of the present study was to examine binding characteristics of GABA(A) receptors in fetal, newborn and adult sheep brains using [(35)S]TBPS, and to determine the effects of allopregnanolone on this binding. Receptor affinity (K(D)) and density (B(MAX)) in the brainstem were not different in fetal, newborn (1-2 days old) and adult brains. In the hypothalamus K(D) and B(MAX) increased significantly in the fetus between 85 and 128 days gestation, and were then similar to postnatal and adult values. In the frontal cortex K(D) and B(MAX) increased progressively between 85 days and term ( approximately 147 days gestation), and were then not different from postnatal and adult values. The K(i) values for the GABA(A) receptor antagonist picrotoxin was similar at all ages. Allopregnanolone inhibited [(35)S]TBPS binding in the presence of 5 microM GABA, but enhanced binding in the absence of GABA. These results show that (i), functional GABA(A) receptors are present in the fetal brain from at least 85 days gestation; (ii), 3alpha-pregnane steroids modify receptor affinity in the late gestation fetal brain; and (iii) there are region-specific changes in GABA(A) receptor binding parameters. Steroid modulation of the GABA(A) receptor in the fetal brain is likely to influence fetal CNS activity in late gestation.

Aging↗
PubMed2000

Ethanol-induced alterations in the expression of neurotrophic factors in the developing rat central nervous system.

Neonatal rats were exposed to ethanol throughout gestation, or during the early postnatal period (postnatal days 4-10 (P4-10)), and enzyme-linked immunoabsorbent assays were subsequently conducted in order to assess nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) protein content in hippocampus, septum, cortex/striatum and cerebellum. These determinations revealed that following prenatal ethanol treatment, there were significant ethanol-induced increases in NGF in P1 cortex/striatum, but no changes in any of the three neurotrophic factors (NTFs) in the other brain regions. Cortex/striatal NGF protein returned to control levels by P10. Following early postnatal exposure, BDNF was elevated in hippocampus and cortex/striatum (assessed on P10), and NGF was also enhanced in cortex/striatum at this age. Hippocampal and cortex/striatal BDNF returned to control levels by P21, but cortex/striatal NGF levels remained enhanced at this age. This NTF did not differ in ethanol and control animals by P60, however. The possible significance of elevated levels of NTFs as a function of ethanol exposure is discussed, and it is speculated that while such alterations could play a protective role, increases in these substances during critical developmental periods could also prove to be deleterious, and could even contribute to certain of the neuropathologies which have been observed following developmental ethanol exposure.

Animals↗
PubMed2000

Effect of a neuroactive steroid infused into the cerebral ventricles of fetal sheep in utero using small infusion volumes.

Placental progesterone metabolites may influence fetal behaviour during late gestation. We have investigated the effect of the neuroactive metabolite, pregnanolone, on the sleep/awake behaviour of fetal sheep. These studies can only be effectively performed in unanesthetized animals with chronically implanted leads and catheters. The relatively large internal volumes of these catheters raises problems for cerebroventriclear infusions. Therefore, we developed a device with negligible deadspace ( approximately 10 microl) consisting of a ventricular cannula attached to a small cap which allowed the instillation of microl volumes of neuroactive steroids into a cerebral ventricle of fetal sheep in utero. Two catheters attached to the cap allow it to be filled in a push-pull configuration. The smaller internal diameter of the ventricular cannular prevents the solution entering the ventricle until the outflow catheter is occluded and solution is forced through the probe. Using this device, the infusion of 5beta-pregnane-3alpha-ol-20-one (pregnanolone, 1 mg), in 35% hydroxypropyl-beta-cyclodextrin, resulted in a marked suppression of fetal behaviour parameters indicative of arousal, while vehicle alone did not affect these parameters. We conclude that neuroactive steroids can be infused into the cerebroventricular system of chronically catheterised fetal sheep in small volumes and that pregnanolone may act directly to influence fetal behaviour.

Animals↗
PubMed2000

Effect of tibial bone resection on the development of fast- and slow-twitch skeletal muscles in foetal sheep.

To determine if longitudinal bone growth affects the differentiation of fast- and slow-twitch muscles, the tibial bone was sectioned at 90 days gestation in foetal sheep so that the lower leg was permanently without structural support. At 140 days (term is approximately 147 days) the contractile properties of whole muscles, activation profiles of single fibres and ultrastructure of fast- and slow-twitch muscles from the hindlimbs were studied. The contractile and activation profiles of the slow-twitch soleus muscles were significantly affected by tibial bone resection (TIBX). The soleus muscles from the TIBX hindlimbs showed: (1) a decrease in the time to peak of the twitch responses from 106.2 +/- 10.7 ms (control, n = 4) to 65.1 +/- 2.48 ms (TIBX, n = 5); (2) fatigue profiles more characteristic of those observed in the fast-twitch muscles: and (3) Ca2+ - and Sr2+ -activation profiles of skinned fibres similar to those from intact hindlimbs at earlier stages of gestation. In the FDL, TIBX did not significantly change whole muscle twitch contraction time, the fatigue profile or the Ca2+ - and Sr2+ -activation profiles of skinned fibres. Electron microscopy showed an increased deposition of glycogen in both soleus and FDL muscles. This study shows that the development of the slow-twitch phenotype is impeded in the absence of the physical support normally provided by the tibial bone. We suggest that longitudinal stretch is an important factor in allowing full expression of the slow-twitch phenotype.

Animals↗