In comparative study of respiratory metabolism, it was established that the relative proportions ... more In comparative study of respiratory metabolism, it was established that the relative proportions of respiratory end-products (succinic, acetic and lactic acids) differed consistently in two strains of Hymenoiepis diminutu (Toronto and ANU). The ANU strain produced more lactic acid and less succinic acid under aerobic and anaerobic conditions. In the shift from aerobic to anaerobic conditions both strains compensated by increasing their outputs of succinic acid. The ANU strain possessed significantly higher activities of hexokinase, pyruvate kinase, lactate dehydrogenase, cytosolic and mitochondrial malic enzyme and cytosolic a-glycerophosphate dehydrogenase. The Toronto strain had significantly higher activities of fumarase, succinate dehydrogenase, and fumarate reductase. There were no significant differences in the activities of phosphoenolpyruvate carboxykinase and malic dehydrogenase between strains. The fumarase activity in the Toronto strain was 16 times that of the ANU strain, its Km (malate) wds 0.8mM, as opposed to 2.5 mM, and it was less sensitive to inhibition by NAD or ATP. These observations are consistent with the patterns of end-product formation in the two strains. Ratios of end-products and calculations of approximate redox balance suggest that the Toronto strain may have a greater capacity for aerobic metabolism.
Amphiphilic cobalt() cage complexes with bridgehead octyl, dodecyl and hexadecyl hydrocarbon c... more Amphiphilic cobalt() cage complexes with bridgehead octyl, dodecyl and hexadecyl hydrocarbon chain substituents have been synthesized simply by co-condensation of formaldehyde and long chain aliphatic aldehydes with the tripodal cobalt() hexaamine complex, [Co(sen)] 3ϩ {sen = 4,4Ј,4Љ-ethylidynetris(3-azabutan-1-amine)}. The synthetic methodology was also used to prepare a novel chiral surfactant by capping the Λ-(Ϫ) D-[Co(sen)] 3ϩ stereoisomer. The cobalt() cage complexes with octyl to hexadecyl substituents are all surface active and reduce the surface tension of water to levels approaching those of organic solvents. The dodecyl substituted cage complex forms aggregates in aqueous solution with a critical micelle concentration of (1.3 ± 0.1) × 10 Ϫ3 mol dm Ϫ3 at 25.00 ЊC. The surfactant cage complexes are biologically active and are lethal at millimolar levels to the tapeworm Hymenolepis diminuta, and the parasitic eukaryote, Tritrichomonas foetus, in vitro. The biological activity of these surfactants appears to involve insertion of the paraffin tail into the organism's exterior membrane and consequent incorporation of the highly charged head-group, which perturbs the normal membrane potential and leads to disintegration of the membrane and death of the organism. The cobalt() cage head-group of these surfactants also undergoes a chemically reversible one-electron reduction to the corresponding cobalt() cage complex and the construction of oriented films of such redox reagents should be feasible. The reduction potential of the cobalt()/() couple is shifted from Ϫ0.72 to Ϫ0.61 V (vs. saturated calomel electrode) by replacing a bridgehead hydrocarbon chain substituent with an alkoxy substituent. The shift in potential correlates with the electrochemical polar substituent constants of alkyl versus alkoxy chains.
Drosophila possesses the core gene silencing machinery but, like all insects, lacks the canonical... more Drosophila possesses the core gene silencing machinery but, like all insects, lacks the canonical RNAdependent RNA polymerases (RdRps) that in C. elegans either trigger or enhance two major small RNA-dependent gene silencing pathways. Introduction of two different nematode RdRps into Drosophila showed them to be functional, resulting in differing silencing activities. While RRF-1 enhanced transitive dsRNA-dependent silencing, EGO-1 triggered dsRNA-independent silencing, specifically of transgenes. The strain w; da-Gal4; UAST-ego-1, constitutively expressing ego-1, is capable of silencing transgene including dsRNA hairpin upon a single cross, which created a powerful tool for research in Drosophila. In C. elegans, EGO-1 is involved in transcriptional gene silencing (TGS) of chromosome regions that are unpaired during meiosis. There was no opportunity for meiotic interactions involving EGO-1 in Drosophila that would explain the observed transgene silencing. Transgene DNA is, however, unpaired during the pairing of chromosomes in embryonic mitosis that is an unusual characteristic of Diptera, suggesting that in Drosophila, EGO-1 triggers transcriptional silencing of unpaired DNA during embryonic mitosis.
The Gram-negative bacterium Shigella flexneri is the causative agent of shigellosis, a diarrhoeal... more The Gram-negative bacterium Shigella flexneri is the causative agent of shigellosis, a diarrhoeal disease also known as bacillary dysentery. S. flexneri infects the colonic and rectal epithelia of its primate host and induces a cascade of inflammatory responses that culminates in the destruction of the host intestinal lining. Molecular characterization of hostpathogen interactions in this infection has been challenging due to the host specificity of S. flexneri strains, as it strictly infects humans and non-human primates. Recent studies have shown that S. flexneri infects the soil dwelling nematode Caenorhabditis elegans, however, the interactions between S. flexneri and C. elegans at the cellular level and the cause of nematode death are unknown. Here we attempt to gain insight into the complex host-pathogen interactions between S. flexneri and C. elegans. Using transmission electron microscopy, we show that live S. flexneri cells accumulate in the nematode intestinal lumen, produce outer membrane vesicles and invade nematode intestinal cells. Using two-dimensional differential in-gel electrophoresis we identified host proteins that are differentially expressed in response to S. flexneri infection. Four of the identified genes, aco-1, cct-2, daf-19 and hsp-60, were knocked down using RNAi and ACO-1, CCT-2 and DAF-19, which were identified as up-regulated in response to S. flexneri infection, were found to be involved in the infection process. aco-1 RNAi worms were more resistant to S. flexneri infection, suggesting S. flexneri-mediated disruption of host iron homeostasis. cct-2 and daf-19 RNAi worms were more susceptible to infection, suggesting that these genes are induced as a protective mechanism by C. elegans. These observations further our understanding of the processes involved in S. flexneri infection of C. elegans, which is immensely beneficial to the routine use of this new in vivo model to study S. flexneri pathogenesis.
The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolate... more The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolated from the livers of Wistar rats infected with Fasciola hepatica were examined in relation to the aberrant mitochondrial respiration previously reported [Rule, Behm, and Bygrave (1989) Biochem. J. 260, 517-523]. At 2 weeks post-infection, elevated NEFA levels were associated with uncoupling of mitochondrial respiration that was reversible in vitro by the addition of BSA. State IV respiration rates showed a strong correlation with NEFA content. At 3 weeks post-infection, NEFA content had increased further and uncoupled mitochondria no longer showed any response to BSA. 31P-NMR analyses of cholate extracts of mitochondria from infected livers at 3 weeks post-infection revealed a marked loss of several major phospholipid species with a concomitant increase in catabolic products, particularly glycerophosphocholine and glycerophosphoethanolamine. Similar changes were observed in microsomal ex...
One-year old worm free, merino wethers, were each infected with 5000 H. contortus larvae of a str... more One-year old worm free, merino wethers, were each infected with 5000 H. contortus larvae of a strain resistant to mebendazole at a rate of 52 mg/kg body weight of sheep. After 21 days, they were assigned to two trials. The preliminary trial showed that mebendazole and levamisole acted synergistically on the H. contortus infection. In the second trial, sheep were treated with 0.35 mg/kg levamisole (one seventh the minimum effective dose against susceptible worms) or 40 mg/kg mebendazole (40 times the minimum effective dose against susceptible worms). In each case the anthelmintics did not reduce worm burdens, although mebendazole depressed egg production. However, when mebendazole and levamisole, at the above dose rates, were administered simultaneously, total worm counts in sheep were reduced by almost 60%. Similar results were obtained when the levamisole was administered 8 h or 14 h after mebendazole treatment. The implications of these observations for the treatment of benzimidazole-resistant haemonchiasis in sheep are discussed.
Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We creat... more Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We created transgenic strains of Drosophila melanogaster in which the rrf-1 or ego-1 RdRp genes from C. elegans were placed under the control of the yeast GAL4 upstream activation sequence. Activation of the gene was performed by crossing these lines to flies carrying the GAL4 transgene under the control of various Drosophila enhancers. RT-PCR confirmed the successful expression of each RdRp gene. The resulting phenotypes indicated that introduction of the RdRp genes had no effect on D. melanogaster morphological development.
BEHM C. A., BRYANT C. AND JONES A. J. 1987. Studies of glucose metabolism in Hymenolepis diminuta... more BEHM C. A., BRYANT C. AND JONES A. J. 1987. Studies of glucose metabolism in Hymenolepis diminuta using t3C nuclear magnetic resonance.
The respiratory properties of mitochondria isolated from the livers of rats infected with the par... more The respiratory properties of mitochondria isolated from the livers of rats infected with the parasite Fasciola hepatica were examined. Oligomycin-sensitive ATPase activity was also examined during the acute stage (2-4 weeks post-infection). At 2, 4 and 6 weeks post-infection, mitochondrial respiration in vitro (supported by site I and site II substrates) was completely uncoupled. Limited respiratory control had returned by 11 weeks post-infection, but complete recovery was not observed even at 21 weeks post-infection. At 4 weeks post-infection, uncoupled respiration (from all three energy-conserving sites) was also markedly attenuated (to the greatest extent with NADH-linked substrate). Except for pyruvate-supported respiration, this attenuation was not apparent at any other stage of the infection. The attenuation of pyruvate-supported respiration declined, but was still present, at 6 weeks post-infection. In addition to these perturbations in mitochondrial respiratory properties, mitochondrial ATPase activity at 4 weeks post-infection was insensitive to oligomycin, indicating a change in the structural integrity of the ATPase complex.
ABSTRACT Bennet E.-M., Behm C.A. and Bryant C. 1978. Effects of mebendazole and levamisole on tet... more ABSTRACT Bennet E.-M., Behm C.A. and Bryant C. 1978. Effects of mebendazole and levamisole on tetrathyridia of Mesocestoides corti in the mouse. International Journal for Parasitology8: 463–466. Mebendazole, but not levamisole, administered to mice carrying artificial infections of 50 tetrathyridia of Mesocestoides corti, was effective in killing the parasites. However, simultaneous administration of mebendazole and levamisole was still more effective. Treatment with levamisole before infection had no additional effect.Injection of mice with dead larvae offered some protection against a subsequent challenge with 50 live larvae; however, levamisole did not then improve the anthelmintic efficacy of mebendazole. In mice rendered immunoincompetent by radiation mebendazole was less effective than in non-irradiated controls and levamisole again did not enhance the effect of mebendazole. It is concluded that anthelmintic efficacy of mebendazole depends on its anthelmintic activity supplemented by the host's immune response; and that levamisole stimulates the latter.
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1992
Mitochondria isolated from male Wistar rats experimentally infected with the common liver fluke, ... more Mitochondria isolated from male Wistar rats experimentally infected with the common liver fluke, Fasciola hepatica, exhibit loss of respiratory control from 2 weeks post-infection (Rule, et al. (1989) Biochem. J. 260, 517-523). We now report that subcutaneous injections of the anti-inflammatory drug, dexamethasone, during the final week of infection prevented the mitochondrial uncoupling and restored respiratory control almost to the levels of uninfected controls. Further investigations have shown that mitochondria from infected rat livers are unable to synthesize ATP and that abnormal respiration is also evident in hepatocytes isolated from infected rats. These abnormalities were absent when infected rats were treated with dcxamethasone. In addition, liver mitochondrial function in infected, congenitally athymic, nude rats (CBH/R nu/nu) was not significantly different from that in uninfected nude or Wistar controls. These results provide evidence that the mitochondrial dysfunction in fascioliasis is host-mediated and that T lymphocytes in particular may be involved. Abbreviations: CCCP, carbonyl cyanide m-chlorophenylhydrazone; ADP, adenosine diphosphate; ROS, reactive oxygen species; RNI, reactive nitrogen intermediates.
Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1994
Biochi~ic~a et Biophysica A~ta Characterization of the oligomycin-sensitivity properties of the F... more Biochi~ic~a et Biophysica A~ta Characterization of the oligomycin-sensitivity properties of the F1F0-ATPase in mitochondria from rats infected with the liver fluke Fasciola hepatica
Mice deficient in interleukin-5 (IL-5-/- mice) were generated by gene targeting in embryonal stem... more Mice deficient in interleukin-5 (IL-5-/- mice) were generated by gene targeting in embryonal stem cells. Contrary to previous studies, no obligatory role for IL-5 was demonstrated in the regulation of conventional B (B-2) cells, in normal T cell-dependent antibody responses or in cytotoxic T cell development. However, CD5+ B cells (B-1 cells) in the peritoneal cavity were reduced by 50%-80% in 2-week-old IL-5-/- mice, returning to normal by 6-8 weeks of age. The IL-5-/- mice did not develop blood and tissue eosinophilia when infected with the helminth Mesocestoides corti, but basal levels of eosinophils with normal morphology were produced in the absence of IL-5. IL-5 deficiency did not affect the worm burden of infected mice, indicating that increased eosinophils do not play a significant role in the host defence in this parasite model.
The concentration of cytochrome P450, measured spectrophotometrically in microsomal preparations ... more The concentration of cytochrome P450, measured spectrophotometrically in microsomal preparations from the livers of rats infected with 30 metacercariae of Fasciola hepatica, declined by approximately 50% at 3 weeks post-infection. Treatment of infected rats with the anti-inflammatory agent dexamethasone (2 mg/kg at 48 h intervals for 8 days prior to assay) abolished the decline in P450 content. Assay of P450 in infected congenitally athymic (nude) rats showed normal levels. These results demonstrate that the T-cell-dependent inflammatory response in the liver of the host is a necessary factor in the development of the decline in hepatic P450, which is known to compromise the metabolism of certain drugs in infected hosts.
The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolate... more The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolated from the livers of Wistar rats infected with Fasciola hepatica were examined in relation to the aberrant mitochondrial respiration previously reported [Rule, Behm, and Bygrave (1989) Biochem. J. 260, 517-523]. At 2 weeks post-infection, elevated NEFA levels were associated with uncoupling of mitochondrial respiration that was reversible in vitro by the addition of BSA. State IV respiration rates showed a strong correlation with NEFA content. At 3 weeks post-infection, NEFA content had increased further and uncoupled mitochondria no longer showed any response to BSA. 31P-NMR analyses of cholate extracts of mitochondria from infected livers at 3 weeks post-infection revealed a marked loss of several major phospholipid species with a concomitant increase in catabolic products, particularly glycerophosphocholine and glycerophosphoethanolamine. Similar changes were observed in microsomal extracts. The NEFA content and phospholipid composition of mitochondria isolated from infected, athymic nude rats were not significantly different from uninfected, athymic rats. These findings suggest that uncoupling of liver mitochondria during infection with F. hepatica is the result of phospholipase activation mediated by the immune system of the host.
In comparative study of respiratory metabolism, it was established that the relative proportions ... more In comparative study of respiratory metabolism, it was established that the relative proportions of respiratory end-products (succinic, acetic and lactic acids) differed consistently in two strains of Hymenoiepis diminutu (Toronto and ANU). The ANU strain produced more lactic acid and less succinic acid under aerobic and anaerobic conditions. In the shift from aerobic to anaerobic conditions both strains compensated by increasing their outputs of succinic acid. The ANU strain possessed significantly higher activities of hexokinase, pyruvate kinase, lactate dehydrogenase, cytosolic and mitochondrial malic enzyme and cytosolic a-glycerophosphate dehydrogenase. The Toronto strain had significantly higher activities of fumarase, succinate dehydrogenase, and fumarate reductase. There were no significant differences in the activities of phosphoenolpyruvate carboxykinase and malic dehydrogenase between strains. The fumarase activity in the Toronto strain was 16 times that of the ANU strain, its Km (malate) wds 0.8mM, as opposed to 2.5 mM, and it was less sensitive to inhibition by NAD or ATP. These observations are consistent with the patterns of end-product formation in the two strains. Ratios of end-products and calculations of approximate redox balance suggest that the Toronto strain may have a greater capacity for aerobic metabolism.
Amphiphilic cobalt() cage complexes with bridgehead octyl, dodecyl and hexadecyl hydrocarbon c... more Amphiphilic cobalt() cage complexes with bridgehead octyl, dodecyl and hexadecyl hydrocarbon chain substituents have been synthesized simply by co-condensation of formaldehyde and long chain aliphatic aldehydes with the tripodal cobalt() hexaamine complex, [Co(sen)] 3ϩ {sen = 4,4Ј,4Љ-ethylidynetris(3-azabutan-1-amine)}. The synthetic methodology was also used to prepare a novel chiral surfactant by capping the Λ-(Ϫ) D-[Co(sen)] 3ϩ stereoisomer. The cobalt() cage complexes with octyl to hexadecyl substituents are all surface active and reduce the surface tension of water to levels approaching those of organic solvents. The dodecyl substituted cage complex forms aggregates in aqueous solution with a critical micelle concentration of (1.3 ± 0.1) × 10 Ϫ3 mol dm Ϫ3 at 25.00 ЊC. The surfactant cage complexes are biologically active and are lethal at millimolar levels to the tapeworm Hymenolepis diminuta, and the parasitic eukaryote, Tritrichomonas foetus, in vitro. The biological activity of these surfactants appears to involve insertion of the paraffin tail into the organism's exterior membrane and consequent incorporation of the highly charged head-group, which perturbs the normal membrane potential and leads to disintegration of the membrane and death of the organism. The cobalt() cage head-group of these surfactants also undergoes a chemically reversible one-electron reduction to the corresponding cobalt() cage complex and the construction of oriented films of such redox reagents should be feasible. The reduction potential of the cobalt()/() couple is shifted from Ϫ0.72 to Ϫ0.61 V (vs. saturated calomel electrode) by replacing a bridgehead hydrocarbon chain substituent with an alkoxy substituent. The shift in potential correlates with the electrochemical polar substituent constants of alkyl versus alkoxy chains.
Drosophila possesses the core gene silencing machinery but, like all insects, lacks the canonical... more Drosophila possesses the core gene silencing machinery but, like all insects, lacks the canonical RNAdependent RNA polymerases (RdRps) that in C. elegans either trigger or enhance two major small RNA-dependent gene silencing pathways. Introduction of two different nematode RdRps into Drosophila showed them to be functional, resulting in differing silencing activities. While RRF-1 enhanced transitive dsRNA-dependent silencing, EGO-1 triggered dsRNA-independent silencing, specifically of transgenes. The strain w; da-Gal4; UAST-ego-1, constitutively expressing ego-1, is capable of silencing transgene including dsRNA hairpin upon a single cross, which created a powerful tool for research in Drosophila. In C. elegans, EGO-1 is involved in transcriptional gene silencing (TGS) of chromosome regions that are unpaired during meiosis. There was no opportunity for meiotic interactions involving EGO-1 in Drosophila that would explain the observed transgene silencing. Transgene DNA is, however, unpaired during the pairing of chromosomes in embryonic mitosis that is an unusual characteristic of Diptera, suggesting that in Drosophila, EGO-1 triggers transcriptional silencing of unpaired DNA during embryonic mitosis.
The Gram-negative bacterium Shigella flexneri is the causative agent of shigellosis, a diarrhoeal... more The Gram-negative bacterium Shigella flexneri is the causative agent of shigellosis, a diarrhoeal disease also known as bacillary dysentery. S. flexneri infects the colonic and rectal epithelia of its primate host and induces a cascade of inflammatory responses that culminates in the destruction of the host intestinal lining. Molecular characterization of hostpathogen interactions in this infection has been challenging due to the host specificity of S. flexneri strains, as it strictly infects humans and non-human primates. Recent studies have shown that S. flexneri infects the soil dwelling nematode Caenorhabditis elegans, however, the interactions between S. flexneri and C. elegans at the cellular level and the cause of nematode death are unknown. Here we attempt to gain insight into the complex host-pathogen interactions between S. flexneri and C. elegans. Using transmission electron microscopy, we show that live S. flexneri cells accumulate in the nematode intestinal lumen, produce outer membrane vesicles and invade nematode intestinal cells. Using two-dimensional differential in-gel electrophoresis we identified host proteins that are differentially expressed in response to S. flexneri infection. Four of the identified genes, aco-1, cct-2, daf-19 and hsp-60, were knocked down using RNAi and ACO-1, CCT-2 and DAF-19, which were identified as up-regulated in response to S. flexneri infection, were found to be involved in the infection process. aco-1 RNAi worms were more resistant to S. flexneri infection, suggesting S. flexneri-mediated disruption of host iron homeostasis. cct-2 and daf-19 RNAi worms were more susceptible to infection, suggesting that these genes are induced as a protective mechanism by C. elegans. These observations further our understanding of the processes involved in S. flexneri infection of C. elegans, which is immensely beneficial to the routine use of this new in vivo model to study S. flexneri pathogenesis.
The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolate... more The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolated from the livers of Wistar rats infected with Fasciola hepatica were examined in relation to the aberrant mitochondrial respiration previously reported [Rule, Behm, and Bygrave (1989) Biochem. J. 260, 517-523]. At 2 weeks post-infection, elevated NEFA levels were associated with uncoupling of mitochondrial respiration that was reversible in vitro by the addition of BSA. State IV respiration rates showed a strong correlation with NEFA content. At 3 weeks post-infection, NEFA content had increased further and uncoupled mitochondria no longer showed any response to BSA. 31P-NMR analyses of cholate extracts of mitochondria from infected livers at 3 weeks post-infection revealed a marked loss of several major phospholipid species with a concomitant increase in catabolic products, particularly glycerophosphocholine and glycerophosphoethanolamine. Similar changes were observed in microsomal ex...
One-year old worm free, merino wethers, were each infected with 5000 H. contortus larvae of a str... more One-year old worm free, merino wethers, were each infected with 5000 H. contortus larvae of a strain resistant to mebendazole at a rate of 52 mg/kg body weight of sheep. After 21 days, they were assigned to two trials. The preliminary trial showed that mebendazole and levamisole acted synergistically on the H. contortus infection. In the second trial, sheep were treated with 0.35 mg/kg levamisole (one seventh the minimum effective dose against susceptible worms) or 40 mg/kg mebendazole (40 times the minimum effective dose against susceptible worms). In each case the anthelmintics did not reduce worm burdens, although mebendazole depressed egg production. However, when mebendazole and levamisole, at the above dose rates, were administered simultaneously, total worm counts in sheep were reduced by almost 60%. Similar results were obtained when the levamisole was administered 8 h or 14 h after mebendazole treatment. The implications of these observations for the treatment of benzimidazole-resistant haemonchiasis in sheep are discussed.
Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We creat... more Drosophila melanogaster, along with all insects and the vertebrates, lacks an RdRp gene. We created transgenic strains of Drosophila melanogaster in which the rrf-1 or ego-1 RdRp genes from C. elegans were placed under the control of the yeast GAL4 upstream activation sequence. Activation of the gene was performed by crossing these lines to flies carrying the GAL4 transgene under the control of various Drosophila enhancers. RT-PCR confirmed the successful expression of each RdRp gene. The resulting phenotypes indicated that introduction of the RdRp genes had no effect on D. melanogaster morphological development.
BEHM C. A., BRYANT C. AND JONES A. J. 1987. Studies of glucose metabolism in Hymenolepis diminuta... more BEHM C. A., BRYANT C. AND JONES A. J. 1987. Studies of glucose metabolism in Hymenolepis diminuta using t3C nuclear magnetic resonance.
The respiratory properties of mitochondria isolated from the livers of rats infected with the par... more The respiratory properties of mitochondria isolated from the livers of rats infected with the parasite Fasciola hepatica were examined. Oligomycin-sensitive ATPase activity was also examined during the acute stage (2-4 weeks post-infection). At 2, 4 and 6 weeks post-infection, mitochondrial respiration in vitro (supported by site I and site II substrates) was completely uncoupled. Limited respiratory control had returned by 11 weeks post-infection, but complete recovery was not observed even at 21 weeks post-infection. At 4 weeks post-infection, uncoupled respiration (from all three energy-conserving sites) was also markedly attenuated (to the greatest extent with NADH-linked substrate). Except for pyruvate-supported respiration, this attenuation was not apparent at any other stage of the infection. The attenuation of pyruvate-supported respiration declined, but was still present, at 6 weeks post-infection. In addition to these perturbations in mitochondrial respiratory properties, mitochondrial ATPase activity at 4 weeks post-infection was insensitive to oligomycin, indicating a change in the structural integrity of the ATPase complex.
ABSTRACT Bennet E.-M., Behm C.A. and Bryant C. 1978. Effects of mebendazole and levamisole on tet... more ABSTRACT Bennet E.-M., Behm C.A. and Bryant C. 1978. Effects of mebendazole and levamisole on tetrathyridia of Mesocestoides corti in the mouse. International Journal for Parasitology8: 463–466. Mebendazole, but not levamisole, administered to mice carrying artificial infections of 50 tetrathyridia of Mesocestoides corti, was effective in killing the parasites. However, simultaneous administration of mebendazole and levamisole was still more effective. Treatment with levamisole before infection had no additional effect.Injection of mice with dead larvae offered some protection against a subsequent challenge with 50 live larvae; however, levamisole did not then improve the anthelmintic efficacy of mebendazole. In mice rendered immunoincompetent by radiation mebendazole was less effective than in non-irradiated controls and levamisole again did not enhance the effect of mebendazole. It is concluded that anthelmintic efficacy of mebendazole depends on its anthelmintic activity supplemented by the host's immune response; and that levamisole stimulates the latter.
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease, 1992
Mitochondria isolated from male Wistar rats experimentally infected with the common liver fluke, ... more Mitochondria isolated from male Wistar rats experimentally infected with the common liver fluke, Fasciola hepatica, exhibit loss of respiratory control from 2 weeks post-infection (Rule, et al. (1989) Biochem. J. 260, 517-523). We now report that subcutaneous injections of the anti-inflammatory drug, dexamethasone, during the final week of infection prevented the mitochondrial uncoupling and restored respiratory control almost to the levels of uninfected controls. Further investigations have shown that mitochondria from infected rat livers are unable to synthesize ATP and that abnormal respiration is also evident in hepatocytes isolated from infected rats. These abnormalities were absent when infected rats were treated with dcxamethasone. In addition, liver mitochondrial function in infected, congenitally athymic, nude rats (CBH/R nu/nu) was not significantly different from that in uninfected nude or Wistar controls. These results provide evidence that the mitochondrial dysfunction in fascioliasis is host-mediated and that T lymphocytes in particular may be involved. Abbreviations: CCCP, carbonyl cyanide m-chlorophenylhydrazone; ADP, adenosine diphosphate; ROS, reactive oxygen species; RNI, reactive nitrogen intermediates.
Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1994
Biochi~ic~a et Biophysica A~ta Characterization of the oligomycin-sensitivity properties of the F... more Biochi~ic~a et Biophysica A~ta Characterization of the oligomycin-sensitivity properties of the F1F0-ATPase in mitochondria from rats infected with the liver fluke Fasciola hepatica
Mice deficient in interleukin-5 (IL-5-/- mice) were generated by gene targeting in embryonal stem... more Mice deficient in interleukin-5 (IL-5-/- mice) were generated by gene targeting in embryonal stem cells. Contrary to previous studies, no obligatory role for IL-5 was demonstrated in the regulation of conventional B (B-2) cells, in normal T cell-dependent antibody responses or in cytotoxic T cell development. However, CD5+ B cells (B-1 cells) in the peritoneal cavity were reduced by 50%-80% in 2-week-old IL-5-/- mice, returning to normal by 6-8 weeks of age. The IL-5-/- mice did not develop blood and tissue eosinophilia when infected with the helminth Mesocestoides corti, but basal levels of eosinophils with normal morphology were produced in the absence of IL-5. IL-5 deficiency did not affect the worm burden of infected mice, indicating that increased eosinophils do not play a significant role in the host defence in this parasite model.
The concentration of cytochrome P450, measured spectrophotometrically in microsomal preparations ... more The concentration of cytochrome P450, measured spectrophotometrically in microsomal preparations from the livers of rats infected with 30 metacercariae of Fasciola hepatica, declined by approximately 50% at 3 weeks post-infection. Treatment of infected rats with the anti-inflammatory agent dexamethasone (2 mg/kg at 48 h intervals for 8 days prior to assay) abolished the decline in P450 content. Assay of P450 in infected congenitally athymic (nude) rats showed normal levels. These results demonstrate that the T-cell-dependent inflammatory response in the liver of the host is a necessary factor in the development of the decline in hepatic P450, which is known to compromise the metabolism of certain drugs in infected hosts.
The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolate... more The non-esterified fatty acid (NEFA) content and phospholipid composition of mitochondria isolated from the livers of Wistar rats infected with Fasciola hepatica were examined in relation to the aberrant mitochondrial respiration previously reported [Rule, Behm, and Bygrave (1989) Biochem. J. 260, 517-523]. At 2 weeks post-infection, elevated NEFA levels were associated with uncoupling of mitochondrial respiration that was reversible in vitro by the addition of BSA. State IV respiration rates showed a strong correlation with NEFA content. At 3 weeks post-infection, NEFA content had increased further and uncoupled mitochondria no longer showed any response to BSA. 31P-NMR analyses of cholate extracts of mitochondria from infected livers at 3 weeks post-infection revealed a marked loss of several major phospholipid species with a concomitant increase in catabolic products, particularly glycerophosphocholine and glycerophosphoethanolamine. Similar changes were observed in microsomal extracts. The NEFA content and phospholipid composition of mitochondria isolated from infected, athymic nude rats were not significantly different from uninfected, athymic rats. These findings suggest that uncoupling of liver mitochondria during infection with F. hepatica is the result of phospholipase activation mediated by the immune system of the host.
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Papers by Carolyn A Behm