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J L Bennett

Publications and source records attributed to J L Bennett.

At least 73 records · Page 4Linked to original sources
PubMed1992

Neurobiology of parasitic flatworms: how much "neuro" in the biology?

The nervous systems in the parasitic Platyhelminthes have generally been considered to be degenerate and of marginal significance, but recent studies have shown these systems to be more significant in the biology of these animals than formerly believed. There are many similarities in the construction and apparent neurochemistry of the nervous systems in the parasitic forms as well as in the free-living Turbellaria. In all forms there appears to be a large neurohormonal component. Though the nervous system appears to be important for many aspects of parasitic flatworm biology, little direct or specific information about the physiology of these systems is yet available.

Animals↗
PubMed1991

Impairment of mitochondrial transcription termination by a point mutation associated with the MELAS subgroup of mitochondrial encephalomyopathies.

Defects in mitochondrial DNA (mtDNA) are associated with several different human diseases, including the mitochondrial encephalomyopathies. The mutations include deletions but also duplications and point mutations. Individuals with MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes) carry a common A-to-G substitution in a highly conserved portion of the gene for transfer RNA(Leu(UUR)). Although the MELAS mutation may be comparable to the defect in the tRNA(Lys) gene associated with MERRF (myoclonus epilepsy associated with ragged-red fibres), it is also embedded in the middle of a tridecamer sequence necessary for the formation of the 3' ends of 16S ribosomal RNA in vitro. We found that the MELAS mutation results in severe impairment of 16S rRNA transcription termination, which correlates with a reduced affinity of the partially purified termination protein for the MELAS template. This suggests that the molecular defect in MELAS is the inability to produce the correct type and quantity of rRNA relative to other mitochondrial gene products.

Base Sequence↗
PubMed1991

Oesophageal varices in a region of the Sudan endemic for Schistosoma mansoni.

In a field study of two villages in the Gezira, an area of the Sudan endemic for Schistosoma mansoni, liver ultrasonography was used to detect subjects with Symmers' hepatic periportal fibrosis, some of whom underwent oesophagoscopy to detect oesophageal varices. The prevalence of oesophageal varices in subjects undergoing oesophagoscopy was 54 per cent and 67 per cent respectively, occurring mainly in males aged about 30 years. The varices were usually asymptomatic. Symptomatic varices (with a positive history of haematemesis) occurred in 4 per cent and 3 per cent respectively of subjects with sonographic evidence of liver periportal fibrosis. By detecting oesophageal varices in an asymptomatic phase, hepatic ultrasonography and fibreoptic oesophagoscopy may elucidate the natural history of the varices and their response to periodic anti-schistosomal chemotherapy.

Adult↗
PubMed1991

Purification and characterization of 3-hydroxymethylglutaryl-coenzyme A reductase of Schistosoma mansoni: regulation of parasite enzyme activity differs from mammalian host.

The enzyme 3-hydroxymethylglutaryl-CoA (HMG-CoA) reductase plays a critical role in regulating the production of cholesterol, dolichols, and ubiquinones in mammals. The inhibition of this enzyme in Schistosoma mansoni is accompanied by a cessation of egg production by the female parasite and a reduced ability of the parasite to properly glycoslyate their proteins. Furthermore, we recently demonstrated that mevinolin, if given continuously over a period of 10-14 days, is a potent antischistosomal drug. In this paper, we describe the properties of purified HMG-CoA reductase from S. mansoni. Using affinity chromatography, we were able to obtain a 417-fold purification of the enzyme which had Km values similar to the rat enzyme for HMG-CoA and NADPH. The Ki value for mevinolin, a potent and selective inhibitor of the rat reductase (Ki = 0.6 nM), was significantly higher (Ki = 46 nM) for the schistosome enzyme. SDS-PAGE and HPLC of the purified enzyme resulted in the appearance of a single protein, which had a molecular weight (66,000) in the range reported for the rat enzyme. Parasite reductase activity, unlike that of its host, did not display a circadian rhythm. Furthermore, agents which elevate (cholestyramine) or decrease (cholesterol) mammalian reductase activity had no effect on the parasite enzyme. Our results suggest that the mechanism which regulates production of the parasite's enzyme may differ from its mammalian host.

Acyl Coenzyme A↗
PubMed1991

The schistosomicidal compound Ro 15-5458 causes a reduction in the RNA content of Schistosoma mansoni.

The effect of Ro 15-5458 (10-2-(diethylamino)ethyl-9-acridanone(2-thiazolin- 2-yl)hydrazone) on the steady-state RNA levels of Schistosoma mansoni was studied after dosing the host with 15 mg kg-1 and retrieving parasites. Total RNA content of parasites recovered from the host 12, 72 and 96 h after dosing was reduced by 14, 30 and 41%, respectively. Quantitative filter hybridization of blots of RNA extracted from treated and control parasites with specific probes indicated a decline in actin and superoxide dismutase mRNA as well as rRNA of treated parasites. The decline was observed 12 h after dosing, 48 h before parasites showed drug-induced changes in other vital biological processes. A prominent drug-induced reduction was seen on the 1.9 kb actin mRNA compared to the 1.4 kb. The same dose of the drug did not alter the actin mRNA content of the host liver. Similarly, the administration of the inactive structural analogue Ro 21-6787 (10-2-(diethylamino)ethyl-9-acridanone) was without any effect. We propose that the actions of Ro 15-5458 and/or its products are directed towards the inhibition of the expression of parasite genes.

Acridines↗
PubMed1991

Studies on muscle cells isolated from Schistosoma mansoni: a Ca(2+)-dependent K+ channel.

Muscle cells from adult male Schistosoma mansoni have been isolated using a combination of papain digestions and mechanical dissociation procedures. The muscle fibres isolated in this way lacked nuclei but they did contract and relax in response to high [K+], a response which was blocked in the presence of Co2+. From this we conclude that the isolation procedure yields viable muscle fibres useful for physiological studies. Patch-clamp recordings taken from the isolated fibres show a variety of discrete ionic conductances. In inside-out patches one prominent channel was a Ca(2+)-activated K+ channel with a conductance of 195 pS and a selectivity greater than 10:1 for K+ over Na+, Cs+ or NH4+. Percentage open time was dependent on [Ca2+] at the intracellular face. With [Ca2+] at 1 microM or greater percentage open time was greater than 95%; at 0.1 microM it was less than 2%. No voltage sensitivity could be detected in the voltage range from -50 to -10 mV membrane potential. Ba2+ (10 mM), but neither tetraethylammonium nor 3,4-diaminopyridine blocked the channel from the intracellular face. This Ca(2+)-activated K+ channel in the muscle membrane of this acoelomate animal is similar in most respects to the maxi-K+ channels which have been described in a variety of cells from more highly evolved animals.

Animals↗
PubMed1991

Neurobiology of parasitic platyhelminths: possible solutions to the problems of correlating structure with function.

This paper provides an overview of research on the nervous system of parasitic platyhelminths. We have emphasized studies concerned with the physiological, pharmacological and biochemical nature of the major small molecule neurotransmitters of these parasites. We have attempted to provide a critical review of the work by focusing on important unresolved issues. Finally, we have focused on some recent work in our laboratory, using patch-clamp recording techniques and quantitative fluorescence cytometry, as an example of newer methods that will hopefully resolve some of the unanswered questions concerning the nervous system of these parasites.

Acetylcholine↗
PubMed1991

Nuclear RNase MRP processes RNA at multiple discrete sites: interaction with an upstream G box is required for subsequent downstream cleavages.

RNase MRP is a site-specific endoribonuclease that processes primer RNA from the leading-strand origin of mammalian mitochondrial DNA replication. It is present in active form as isolated from the nucleus, suggesting a bipartite cellular location and function. The relatively high abundance of nucleus-localized RNase MRP has permitted its purification to near homogeneity and, in turn, has led to the identification of protein components of this ribonucleoprotein. Analysis of the mode of RNA cleavage by nuclear RNase MRP revealed the surprising and unprecedented ability of the endonuclease to process RNA at multiple discrete locations. Substrate cleavage is dependent on the presence of a previously described G-rich sequence element adjacent to the primary site of RNA processing. Downstream cleavage occur in a distance- and sequence-specific manner.

Animals↗
PubMed1991

Association of the therapeutic activity of praziquantel with the reversal of Symmers' fibrosis induced by Schistosoma mansoni.

The impact of antischistosomal chemotherapy on the most important complication of intestinal schistosomiasis, Symmers' periportal fibrosis, has not been determined. Since abdominal ultrasonography has proven to be an effective tool in assessing the extent of Symmers' fibrosis in patients, we monitored the effect of chemotherapy, which involved the annual administration of praziquantel, on 48 Sudanese villagers having varying degrees of Symmers' fibrosis. Results indicate no significant differences in the fibrotic status of the 48 patients between 1986 and 1987, but test statistics (both the Wilcoxon signed rank test and Friedman's block/treatment test), indicated a significant decrease between the fibrotic status of the patients in 1986 and their fibrotic status in 1988 and 1989. Thus, after three years of therapy, 12 of the 48 patients no longer had detectable Symmers' fibrosis, while another 16 patients experienced a reduction in the amount of fibrosis in their livers. When coupled with our previous study, which demonstrated that annual treatment of children with praziquantel prevents the appearance of Symmers' fibrosis, it now appears that praziquantel may reverse this schistosomal-induced pathology.

Adolescent↗
PubMed1990

Antischistosomal action of mevinolin: evidence that 3-hydroxy-methylglutaryl-coenzyme a reductase activity in Schistosoma mansoni is vital for parasite survival.

The enzyme 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA reductase) plays an important role in regulating egg production in the human parasitic trematode Schistosoma mansoni. Infected mice treated with a low dose of mevinolin, an inhibitor of this enzyme, stopped egg production by the parasite and blocked the pathology associated with the infection. As an extension of this work, we exposed adult and developing schistosomes to higher doses of mevinolin for an extended period of time and observed the following: administration of 0.2% mevinolin in the diet of infected mice for 14 days resulted in 96-100% elimination of adult parasites. Administration of the same dose for 2 days before, and for 15 days after infection, resulted in 93-96% reduction of adult parasites. Co-administration of mevinolin with 0.5% mevalonate, but not 0.5% cholesterol, blocked the antischistosomal activity of mevinolin. We monitored schistosomal and mouse liver HMG-CoA reductase activity during the course of treatment and observed that HMG-CoA reductase activity was elevated in the liver but significantly reduced in the parasite. In vitro incubation of paired schistosomes with mevinolin, for up to 11 days, resulted in a dose- and time-dependent reduction of parasite motility and lactate production with eventual death. These in vitro effects were no evident when mevalonate was added to the culture system in place of, or in addition to, mevinolin. Collectively, the evidence suggests that mevalonate and/or metabolite not only plays a vital role in schistosome egg production, but is vital for survival of the parasite.

Animals↗
PubMed1990

Schistosoma mansoni: biochemical characteristics of the antischistosomal effects of Ro 15-5458.

We evaluated a variety of biochemical parameters in Schistosoma mansoni isolated from mice up to 4 days after dosing with 15 mg/kg Ro 15-5458. While no drug effect could be demonstrated in the utilization of media glucose, glycogen content, gut pigment, or ATP levels of the parasites, a significant reduction (P less than or equal to 0.05) in parasite weight and protein content was observed. Possible drug actions that may contribute to the loss in parasite protein and perhaps ultimately result in parasite death have been investigated. We noted significant reduction in the incorporation of leucine and thymidine into acid-insoluble fractions of the parasites. The reduction in the incorporation of leucine into parasite proteins was nonspecific and preceded the effect of the drug on the uptake of the amino acid. Parasite and host liver RNA isolated after dosing were translated in vitro in a rabbit reticulocyte system. Drug-treated parasite mRNA, but not that of the host, was less effective than control mRNA in directing the incorporation of [35S]methionine. We propose a hypothesis that attributes the loss in protein content to a defect in the biosynthesis of parasite proteins as a result of a drug-induced reduction in the quantity of mRNA in the parasites This effect of Ro 15-5458 on the parasite may provide the basis for its schistosomicidal action.

Acridines↗
PubMed1990

Studies on intrategumental pH and its regulation in adult male Schistosoma mansoni.

The intrategumental pH in adult male Schistosoma mansoni as measured with pH-sensitive microelectrodes is between 7.0 and 7.2, a value about one pH unit more alkaline than expected for equilibrium. This value is maintained for at least 4 h after isolation in media buffered with Hepes or CO2/HCO3- with or without serum present. CCCP (1 microM) and FCCP (10 microM) cause rapid acidification. DNP (1 mM) and Na-orthovanadate (1 mM) acidify but also cause significant depolarization of the tegument. NH4+ (20 mM) depolarizes and acidifies the tegument with no evidence of transient alkalinization. High K+ (25 mM) accelerates the acidification. Removal of the NH4+ causes a large transient further acidification with recovery requiring as long as 40 min. High K+ (25 mM) delays the early stage of the recovery. Presence of CO2/HCO3- in the incubation medium does not accelerate the recovery rate nor does SITS (100 microM) inhibit the recovery. Intrategumental Na+ is elevated after an acid load and amiloride (3 mM) as well as low Na+ medium interfere with recovery from the acid load indicating that a Na(+)-H+ exchanger may be present in the tegumental membrane.

Animals↗
PubMed1990

The apparent absorption kinetics of Ro 15-5458, a schistosomicidal drug.

The absorption kinetics of Ro 15-5458, a new antischistosomal drug, was studied in rabbits following the administration of a single 50 mg/kg oral dose as an aqueous suspension in 25% glycerol-1% cremophor EL. Ro 15-5458 was absorbed from the gastrointestinal tract rapidly with a lag time of about 6 min and declined without a plateau with a half-life of about 6 h. There was variable extent of drug appearance in plasma after oral administration. Maximum plasma levels ranged from 500 to 1200 ng/ml (1.3-3.1 microM). The plasma concentration-time profile of Ro 15-5458 after a single oral dose appears to follow a one-compartment pharmacokinetic model with zero-order absorption, first-order elimination.

Acridines↗
PubMed1990

Efficient site-specific cleavage by RNase MRP requires interaction with two evolutionarily conserved mitochondrial RNA sequences.

RNase MRP is a site-specific endonuclease that processes primer mitochondrial RNA from the leading-strand origin of mitochondrial DNA replication. Using deletional analysis and saturation mutagenesis, we have determined the substrate requirements for cleavage by mouse mitochondrial RNase MRP. Two regions of sequence homology among vertebrate mitochondrial RNA primers, conserved sequence blocks II and III, were found to be critical for both efficient and accurate cleavage; a third region of sequence homology, conserved sequence block I, was dispensable. Analysis of insertion and deletion mutations within conserved sequence block II demonstrated that the specificity of RNase MRP accommodates the natural sequence heterogeneity of conserved sequence block II in vivo. Heterologous assays with human RNase MRP and mutated mouse mitochondrial RNA substrates indicated that sequences essential for substrate recognition are conserved between mammalian species.

Animals↗
PubMed1989

Glutamine- vs glucose-supported motor activity in Schistosoma mansoni: physiological relevance of aerobic metabolism.

The ability of Schistosoma mansoni to generate energy through aerobic metabolic processes was examined in adult parasites in vitro. Parasite catabolism of radiolabeled glucose, glutamine, and other amino acids to CO2 and Krebs cycle intermediates was measured under a variety of incubation conditions. L-Glutamine was metabolized to CO2 via the intermediates glutamate, alpha-ketoglutaramate, and alpha-ketoglutarate in worms incubated in a balanced salts solution containing this amino acid as the only organic constituent. Of the other amino acids tested, CO2 production was detected from L-glutamate and L-asparagine. The catabolism of L-glutamine to CO2 was reduced by the respiratory inhibitor antimycin A. The motility of schistosomes in culture was maintained for at least 24 hr when L-glutamine was the only carbon source available to the worms. Under these conditions, motility was reduced when parasites were exposed to a respiratory inhibitor such as KCN, antimycin A, rotenone, or oligomycin, but it was completely restored by the addition of glucose to the medium. These results suggest that while the schistosome is capable of limited aerobic energy-generating processes under certain conditions, survival is not contingent upon these processes in the presence of glucose.

Adenine Nucleotides↗
PubMed1989

Comparison of glycolysis and glutaminolysis in Onchocerca volvulus and Brugia pahangi by 13C nuclear magnetic resonance spectroscopy.

Comparison of glycolysis in Brugia pahangi and Onchocerca volvulus by 13C nuclear magnetic resonance (NMR) spectroscopy showed that the former organism is predominantly a lactate fermenter and the latter resembles more closely the metabolism of a mixed acid fermenter producing lactate, succinate, acetate, ethanol, formate and carbon dioxide. Both organisms synthesize glycogen as a storage carbohydrate. Glutaminolysis in both organisms proceeds by the delta-amino-butyrate shunt to produce succinate which is then further metabolized to acetate and carbon dioxide as end-products.

Animals↗
PubMed1989

Physiological role of HMG-CoA reductase in regulating egg production by Schistosoma mansoni.

Pathological lesions observed in humans infected with Schistosoma mansoni are due to the eggs produced by the female parasite. Mevinolin, a potent inhibitor of the enzyme hydroxymethylglutaryl-CoA (HMG-CoA) reductase, blocks egg production by this parasite. In this report, we demonstrate that cholesterol precursors, mevalonate and farnesol, stimulate egg production by the female parasite and that these precursors can reverse the mevinolin-induced inhibition of egg production. Because the parasite cannot synthesize cholesterol, we incubated parasites in a culture media containing radiolabeled acetate with and without mevinolin. We isolated nonsterol lipids from the parasite and observed that mevinolin dramatically reduced the conversion of acetate into the polyisoprenoid (dolichols) lipids of the parasite. Dolichols and other nonsterol lipids did not stimulate egg production. HMG-CoA reductase activity was observed in homogenates of the parasite and was inhibited by mevinolin (Ki = 52 nM), but its activity was tripled when the parasite was chronically exposed to low doses of the drug. Parasites with increased reductase activity produced five to six times more eggs. Lastly, chronic administration of large doses of mevinolin to infected mice resulted in a marked reduction of the pathology associated with the infection. These results suggest that egg production in S. mansoni is associated with the parasite's HMG-CoA reductase activity and that a nonsterol lipid produced in the biochemical pathway regulated by this enzyme stimulates egg production.

Animals↗