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Biomedical subjects

D R Gardner

Publications and source records attributed to D R Gardner.

At least 55 records · Page 3Linked to original sources

Pyrrole detection and the pathologic progression of Cynoglossum officinale (houndstongue) poisoning in horses.

Houndstongue (Cynoglossum officinale), a noxious weed that contains pyrrolizidine alkaloids (PAs), infests pastures and fields in the western United States and Europe. The purpose of this study was to develop techniques to better diagnose PA poisoning and describe the progression of gross and microscopic lesions caused by houndstongue intoxication. Six horses were gavaged daily with a suspension of houndstongue containing 5 or 15 mg/kg total PA for 14 days. Two horses were treated similarly with ground alfalfa as controls. Liver biopsy samples and serum biochemical and hematologic values were evaluated biweekly. Within 7 days after dosing, horses treated with 15 mg/kg PA developed severe liver disease characterized by altered bile acid metabolism, elevated serum enzymes, and extensive hepatocellular necrosis with minimal periportal fibrosis and biliary hyperplasia. The condition of these animals continued to deteriorate, and they were euthanized. For several weeks after dosing, horses treated with 5 mg/kg PA were depressed, had transient elevations of serum enzymes and bile acids, and developed minimal periportal hepatocellular necrosis with fibrosis. The biochemical changes resolved by 6-8 weeks; however, the histologic disease persisted with extensive megalocytosis by week 14. Throughout the study, the rate of hepatocellular proliferation remained constant. Biliary cells had an increase in mitotic rate that correlated with the histologic changes. Hepatic tissue-bound pyrroles (PA metabolites) were identified in necropsy samples of treated animals using gas chromatography/mass spectrometry and photometrically with Ehrlich's reagent. These findings suggest that pyrrole extraction and identification are useful in documenting PA exposure and that houndstongue is extremely toxic to horses.

Alkaloids↗

Serum alpha-mannosidase activity and the clinicopathologic alterations of locoweed (Astragalus mollissimus) intoxication in range cattle.

Subclinical intoxication of livestock with Astragalus and Oxytropis species (locoweeds) results in decreased animal feed conversion, reduced weight gains, and reproductive failure. Sensitive diagnostic methods to definitively diagnose and monitor intoxication are needed to minimize these losses and better manage locoweed-infested pastures and rangelands. Sera from cattle grazing locoweed were evaluated for alpha-mannosidase activity, serum biochemical values, electrolytes, and thyroid hormone concentrations. As the cows began to ingest locoweed, the mean serum alpha-mannosidase activities dropped significantly (400.0 microM to 72.5 microM). Changes in other serum chemistry values were less specific; however, individual animals (generally those ingesting more locoweed) had elevated levels of alkaline phosphatase (ALP), aspartate aminotransferase, and lactate dehydrogenase, with decreased serum total protein (5.8 +/- 0.8 g/dl) and albumin (2.3 +/- 0.3 g/dl). Mean serum thyroid concentrations (both T4 and T3) were lower in animals that were ingesting locoweed. The calculated swainsonine dose correlated statistically with serum alpha-mannosidase activity, ALP, albumin, Cl, CO2, and thyroid hormone T3. This correlation suggests that serum alpha-mannosidase activity along with potential changes in ALP, albumin, and thyroid hormone concentrations is a sensitive indicator of locoweed exposure and intoxication. These parameters may also be useful for monitoring intoxication and allowing subclinically affected cattle to be removed from infested areas before irreversible damage occurs.

Animals↗

Effect of feeding ponderosa pine needle extracts and their residues to pregnant cattle.

Solvents including ethanol, methylene chloride, acetone, hexane, and hot and cold water were used in separate procedures to extract needles and bark from ponderosa pine (Pinus ponderosa L.) to determine which system was optimal for the extraction of abortifacient compound(s). The extracts and residues therefrom were fed by gavage to pregnant cows beginning at day 250 of gestation. After methylene chloride extraction, the pine needle residue fed by gavage failed to abort 4 cows. Subsequently, 4 cows fed the methylene chloride extractable material, adsorbed on ground hay, but free of methylene chloride, aborted (days 253-260 of gestation) and all 4 cows retained fetal membranes. Only 2 of 6 cows fed the pine needle residue after ethanol extraction aborted (days 260 and 261 of gestation) and both retained their fetal membranes, while the other 4 cows calved normally. Hot and cold water extracts did not cause abortion, whereas the extracted plant residue did. One cow fed the plant residue after acetone extraction calved normally as did 1 of 2 cows fed residue after hexane extraction. One cow fed hexane extract absorbed on ground hay also calved normally. These results demonstrate that abortifacient compound(s) present in pine needles are most efficiently extracted by methylene chloride.

Abortifacient Agents↗

A review of the electrophysiological, pharmacological and single channel properties of heart ventricle muscle cells in the snail Lymnaea stagnalis.

Although a considerable body of information has accumulated describing the pharmacological properties of a wide range of molluscan muscle types, the physiological bases underlying these properties have not been thoroughly investigated. At present, little is known about the types of ion channels and their regulation in molluscan muscle cell membranes. Voltage-clamp, and more recently, patch-clamp techniques have revealed molluscan muscles possess a complex array of channel types with various pharmacological and electrophysiological properties. The gating properties of these channels and their modulation by chemical agents, however, are still poorly understood. This review summarizes some aspects of molluscan muscle function with particular reference to the heart ventricle muscle of the pond snail, Lymnaea stagnalis.

Animals↗

Activation of a divalent cation-conducting channel in heart ventricle muscle cells of the snail Lymnaea stagnalis by the molluscan cardioactive peptide FMRFamide.

Ion channels with characteristics of Ca2+ channels have been found in isolated heart ventricle cells of the snail Lymnaea stagnalis. Although spontaneous Ca2+ or Ba2+ currents were seen only occasionally, spontaneous inward Na+ currents were readily observed in the absence of patch pipette Ca2+ between membrane potentials of -100 mV and +20 mV. These currents were blocked by 2 mM Ni2+, 2 mM Co2+ and 10 microM Ca2+. The channels usually ceased conducting within a few minutes after seal formation with the patch pipette and could not be re-activated with depolarizing voltage steps. However, at the cell's resting potential, 10(-8) to 10(-6) M of the molluscan cardioactive peptide FMRFamide or its analogue FLRFamide2+ applied to the cell membrane away from the patch pipette, induced unitary Ba2+ currents or, in the absence of Ca2+ in the patch pipette, Na+ currents. This suggests that a secondary messenger is involved in the FMRFamide-induced activation of these channels rather than a direct activation of a channel-receptor complex by the peptide.

Amino Acid Sequence↗

The peptide FMRFamide activates a divalent cation-conducting channel in heart muscle cells of the snail Lymnaea stagnalis.

1. Isolated Lymnaea stagnalis heart ventricle cells contain cation-conducting channels with properties characteristic of Ca2+ channels. These channels, which carry inward Na+ currents in the absence of Ca2+, are activated by the molluscan cardioactive peptides FMRFamide and FLRFamide, and are blocked by Co2+ ions. 2. FMRFamide also activated inward Ba2+ currents at the cell's resting potential. These currents, which were also blocked by Co2+ ions, reversed at a membrane potential of +70 mV. 3. Both sodium and barium currents were initiated when the peptides were applied to the cell outside of the patch pipette indicating that a secondary messenger is likely to be involved in the FMRFamide response.

Animals↗

Non-voltage-gated calcium channels in snail heart ventricle cells.

1. Two recently identified channel types in Lymnaea stagnalis heart muscle cells were shown to conduct Na+ in the absence of extracellular Ca2+. They did not appear to be 'voltage-gated' as they were not activated by voltage. Also, they remained active over a wide range of membrane potentials. However, they were weakly 'voltage-sensitive' as their activity usually tended to increase with depolarization. The weak voltage-sensitivity and similarity to other non-voltagegated Ca2+ channels suggested that one or both of these channels may be receptor-operated Ca2+ channels. 2. One of the two channels had a slope conductance of 15 pS. The other appeared to have at least two subconductance states with slope conductances of 50 and 72pS. Both these conductance states had very similar open dwell-time constants and identical reversal potentials. The open dwell-time constants of both conductance states were not affected by voltage, suggesting that the channels' weak voltage-sensitivity was mediated by one of the closed states. 3. With divalent cations in the patch pipette, non-voltage-gated Ba2+ and Ca2+ currents were also detected. The Ba2+ conductance (12pS) was similar to the Ca2+ conductance (11pS).

Animals↗

X-ray analysis of Lymnaea stagnalis muscle fibres does not suggest that the sarcolemma is permeable to lanthanum ions.

The cross-striated muscle from the heart ventricle and the smooth penis retractor muscle of the freshwater snail Lymnaea stagnalis have been investigated by X-ray microanalysis to establish whether lanthanum can cross the plasma membrane, as has been reported by other investigators. Tissues were incubated in 1 mM ionic lanthanum before fixation in phosphate- or cacodylate-buffered fixative. X-ray mapping for emissions in the lanthanum energy range indicates a concentration of emissions that coincided only with the network of sub-surface transverse tubules formed by the invagination of the plasma membrane and with the plasma membrane/extracellular space interface. X-ray energy spectra were collected from various cell compartments; peak-to-background ratios were obtained and analysed statistically. Cacodylate buffer is less effective than phosphate buffer in precipitating lanthanum, but no evidence to suggest the redistribution of lanthanum in cacodylate-buffered preparations was found. Lanthanum is precipitated only in the sub-surface transverse tubules and at the plasma membrane/extracellular space interface in both heart ventricle muscle and penis retractor muscle, fixed in either phosphate or cacodylate buffer. There was no evidence of lanthanum precipitation in the background cytoplasm or on any cytoplasmic organelle. These results confirm our hypothesis that lanthanum does not cross the plasma membranes in these molluscan tissues.

Animals↗

The sites of action of praziquantel in a smooth muscle of Lymnaea stagnalis.

Praziquantel (1.6 X 10(-6) M), a new anthelmintic drug, was shown to produce a sustained contracture in the penis retractor muscle of the freshwater snail Lymnaea stagnalis. Calcium-free saline greatly reduced the magnitude and duration of the praziquantel contractures if the preparation had been previously exposed to praziquantel. The praziquantel contracture in zero-calcium saline was abolished if the preparation was pretreated with successive applications of 2 X 10(-2) M caffeine. Lanthanum (1.0 mM) in zero-calcium saline prevented the contractures normally produced by 30 mM potassium saline, but did not prevent the praziquantel contracture. In normal saline, however, 1.0 mM lanthanum greatly reduced the magnitude of a praziquantel-induced contracture. We conclude that praziquantel increases sarcolemmal calcium permeability and releases calcium from intracellular stores in Lymnaea stagnalis smooth muscle.

Animals↗

Insecticidal and neuroexcitant actions of DDT analogs on the cockroach, Periplaneta americana.

The American cockroach (Periplaneta americana) was used to test 14 DDT analogs for their ability to cause a) death and b) repetitive firing in the abdominal nerve cord. P,p' -DDT, p,p' -DDD, p,p' -methoxychlor, and o,p -DDT were all found to induce repetitive firing in the nerve cord after a twice threshold stimulus, but only p,p' -DDT, p,p' -DDD, and p,p' -methoxychlor produced mortality within 4 days from injection of the pesticides. Also, cockroaches exposed to sublethal doses of p,p' -DDT and p,p' -methoxychlor were found to exhibit repetitive firing in the abdominal nerve cord three weeks after injection of the pesticide. The 'cause and effect' relationship between neuroexcitation and lethality is therefore questioned, at least in the case of cockroaches.

Action Potentials↗

Frescon: neurophysiological action of a molluscicide.

The molluscidice N-trityl morpholine (Frescon) has an unusual effect on the central nervous system of a freshwater snail. Nerve impulses become grouped into spontaneous "bursts", with many cells firing synchronously. This may result from interference with inhibitory processes.

Animals↗

Temperature-sensitive neurons in the brain of brook trout.

The brain temperature of trout (Salvelinus fontinalis) was altered by perfusion of the gills with warm or cool water. Neuronal activity was recorded with microelectrodes. Twelve neurons responded to an increase in temperature with increased activity, and five neurons responded to a decrease in temperature with an increase in activity.

Animals↗