Hemoglobin catabolism and the killing of intraerythrocytic<Emphasis Type="Italic">Plasmodium falciparum</Emphasis> by chloroquine

To evaluate how chloroquine kills malaria parasites, hemoglobin catabolism was studied at the various stages of intraerythrocytic parasite development. We found that hemoglobin catabolism is switched off whenPlasmodium falciparum parasites mature to the late trophozoite or early schizont stages and is switched on again during the ring stage. When hemoglobin catabolism is switched off, the parasites are resistant to the morphologic effects of chloroquine. Although the ring stage parasites failed to mature in the presence of chloroquine, some of them switched on hemoglobin ingestion and became stuffed with hemoglobin-filled vesicles, indicating a distal block in catabolism. In fact, we demonstrated a high-grade block in hemozoin production during a 22 h incubation of synchronized ring forms; ferriprotoporphyrin IX (FP) incorporation into the β-hematin of hemozoin decreased from 900 to 50 pmol/10⁶ parasitized erythrocytes. We propose that the primary effect of chloroquine on hemoglobin catabolism is to block FP polymerization to β-hematin. Secondarily, toxic FP and FP-chloroquine complexes accumulate and are available to exert their several toxicities, which include inhibition of hemoglobindegrading proteases and membrane damage. As a consequence, maturation is arrested and eventually the parasites die and lyse.     

Hemoglobin catabolism and the killing of intraerythrocyticPlasmodium falciparum by chloroquine

To evaluate how chloroquine kills malaria parasites, hemoglobin catabolism was studied at the various stages of intraerythrocytic parasite development. We found that hemoglobin catabolism is switched off whenPlasmodium falciparum parasites mature to the late trophozoite or early schizont stages and is switched on again during the ring stage. When hemoglobin catabolism is switched off, the parasites are resistant to the morphologic effects of chloroquine. Although the ring stage parasites failed to mature in the presence of chloroquine, some of them switched on hemoglobin ingestion and became stuffed with hemoglobin-filled vesicles, indicating a distal block in catabolism. In fact, we demonstrated a high-grade block in hemozoin production during a 22 h incubation of synchronized ring forms; ferriprotoporphyrin IX (FP) incorporation into the -hematin of hemozoin decreased from 900 to 50 pmol/10⁶ parasitized erythrocytes. We propose that the primary effect of chloroquine on hemoglobin catabolism is to block FP polymerization to -hematin. Secondarily, toxic FP and FP-chloroquine complexes accumulate and are available to exert their several toxicities, which include inhibition of hemoglobindegrading proteases and membrane damage. As a consequence, maturation is arrested and eventually the parasites die and lyse.     

The effect of chloroquine upon the developing lens

Applied to the developing lens of the 14-day-old chick embryo, in organ culture conditions, chloroquine prevented the elongation of the primary lens fibres, destroyed the equatorial ones and provoked vacuolisation and/or destruction in the epithelial cells.     

Studies on correlations between chloroquine-induced tissue damage and serum enzyme changes in the rat

The administration of chloroquine to rats resulted in a significant elevation of serum enzymes and a corresponding decrease of these enzymes in the tissues. The changes in serum and kidney enzymes were most marked, thus indicating a primary renal dysfunction.     

The effect of monensin and chloroquine on the endocytosis and toxicity of chimeric toxins

The toxicity of two conjugates containing ribosome-inactivating proteins (RIPs, i.e. saporin and ricin-A chain x-linked to transferrin) has been measured on a prostatic cancer line (PC3) naturally overexpressing the transferrin receptor, in the presence of monensin and chloroquine. This paper investigates whether the increased toxicity of Tf-RIPs induced by monensin and chloroquine may be due to alterations of the normal endocytotic pathway of the complexes mediated by the transferrin receptor. Monensin, besides inducing alkalinization of normally acid intracellular compartments, causes an accumulation of the receptor-bound Tf-RIP in a perinuclear region contiguous to the cisternae of the trans-Golgi network. Chloroquine, though increasing the intracellular pH, seems not to modify the endocytotic pathway of these chimeric molecules. We believe that the enhanced toxicity of the Tf-RIPs may be related to intracellular alkalinization (i.e. endosomal or lysosomal pH) rather than to the effects on the recycling of transferrin receptor-bound toxins. We conclude that the efficacy of chimeric toxins may be modulated not only by the carrier used for their engineering but also by addition of drugs able to influence the stability and activation of the toxins inside the cell. Received 22 December 1997; received after revision 30 March 1998; accepted 15 May 1998     

Malaria prophylaxis in travellers: The current position

Summary Malaria prevention is a main challenge for physicians, nurses, health officers and tour operators. The attack rate of malaria in travellers is 1–10/10,000 departures, and the case fatality rate of imported malaria is around 0.5/100. Travellers should be informed about the risk they are going to take, how to protect against mosquito bites, about the antimalarials they will have to take and about what to do when a malaria breakthrough should occur.The 4-aminoquinolines (chloroquine, amodiaquine) remain the drug of choice for the prevention ofPlasmodium vivax and of sensitiveP. falciparum infections. The problem is to find an effective and safe drug combination for travellers to areas whereP. falciparum is either resistant to chloroquine, to Fansidar (the combination of pyrimethamine plus sulfadoxine) or to both. These travellers will probably best be protected by an individually tailored drug combination, which includes amodiaquine or mefloquine as baseline drugs, and a supplementation with Fansidar, Maloprim (the combination of pyrimethamine with dapsone), paludrine or an antibiotic.     

Effect of intercalating mutagens on crossing-over inDrosophila melanogaster females

Summary In order to evaluate the effect of several intercalating compounds on crossing-over inDrosophila melanogaster females, acridine orange, acriflavine, chloroquine, ethidium bromide and quinacrine were fed separately to larvae ofy ct f/+++ genotype. Our results show that acridine orange, acriflavine and ethidium bromide increase significantly the recombination frequency at thect-f region and support the view that, for intercalating agents, there is a relationship between clastogenic activity and female recombination induction.     

Role of lysosomes, microtubules and microfilaments in the mechanism of the lactogenic action of prolactin in the rabbit mammary gland]

The lysosomotropic agents, ammonium chloride and chloroquine, added to the culture medium of pseudopregnant Rabbit mammary gland, did not inhibit the initiation of casein synthesis by prolactin. By contrast, they considerably reduced the down-regulation of prolacting receptors. Converse-y, colchicine totally blocked the lactogenic action of prolactin without altering the down-regulation of the receptor. Cytochalasin B inhibited only partly the lactogenic action of prolactin while it has no effect on the down-regulation of the receptor. These data suggest that the degradation of the prolactin-receptor complex in lysosome is not a compulsory step in the mechanism of prolactin action. The integrity of microtubules but not of microfilaments is required for prolactin to initiate casein synthesis. These elements of the cytoskeleton are not strictly involved in the down-regulation of the receptor.     

Know your enemy: understanding the role of PfCRT in drug resistance could lead to new antimalarial tactics

The prevention and treatment of malaria is heavily dependent on antimalarial drugs. However, beginning with the emergence of chloroquine (CQ)-resistant Plasmodium falciparum parasites 50 years ago, efforts to control the disease have been thwarted by failed or failing drugs. Mutations in the parasite's 'chloroquine resistance transporter' (PfCRT) are the primary cause of CQ resistance. Furthermore, changes in PfCRT (and in several other transport proteins) are associated with decreases or increases in the parasite's susceptibility to a number of other antimalarial drugs. Here, we review recent advances in our understanding of CQ resistance and discuss these in the broader context of the parasite's susceptibilities to other quinolines and related drugs. We suggest that PfCRT can be viewed both as a 'multidrug-resistance carrier' and as a drug target, and that the quinoline-resistance mechanism is a potential 'Achilles' heel' of the parasite. We examine a number of the antimalarial strategies currently undergoing development that are designed to exploit the resistance mechanism, including relatively simple measures, such as alternative CQ dosages, as well as new drugs that either circumvent the resistance mechanism or target it directly.     

Antimalarial drugs: recent advances in molecular determinants of resistance and their clinical significance

Molecular determinants of antimalarial drug resistance are useful and informative tools that complement phenotypic assays for drug resistance. They also guide the design of strategies to circumvent such resistance once it has reached levels of clinical significance. Established resistance to arylaminoalcohols such as mefloquine and lumefantrine in SE Asia is mediated primarily by gene amplification of the P. falciparum drug transporter, pfmdr1. Single nucleotide polymorphisms in pfmdr1, whether assessed in field isolates or transfection experiments, are associated with changes in IC₅₀ values (to arylaminoalcohols and chloroquine), but not of such magnitude as to influence clinical treatment outcomes. Recently described emerging in vitro resistance to artemisinins in certain areas correlates with mutations in the SERCA-like sequence PfATP6 and supports PfATP6 as a key target for artemisinins. Received 13 February 2006; revised after revision 7 March 2006; accepted 29 March 2006     

Evidence for the metabolism of phospholipids and triacylglycerols in humanAscaris lumbricoides

Summary HumanAscaris lumbricoides has the necessary mechanism for the biosynthesis and degradation of phospholipids and triacylglycerols, as in most other species. Piperazine decreases the level of triacylglycerols of this parasite by stimulating the activity of lipase and partially inhibiting the activity of phosphatidate phosphatase.Acknowledgments. Thanks are due to the University of Kerala for facilities provided, Dr T. Ramasarma of the I.I. Sc., Bangalore for help in the labelling studies, Dr. G. D. Cain of the University of Iowa for valuable comments and Dr P. Vijayagopal of ANU for the gift of CDP-choline.     

Protein content of various developmental stages of three strains of the pink bollworm,Pectinophora gossypiella

Summary Comparison of the total body protein of male and female larvae, pupae, and adults of a diapause and 2 nondiapause strains of the pink bollwormPectinophora gossypiella indicated that the protein content of larvae of the diapause strains was significantly higher than of the non-diapause strain.I thank Drs D.R. Nelson and R.A. Bell of the U.S.D.A. Metabolism and Radiation Research Laboratory, Fargo, for facilities and guidance.     

Cerebellar decussation of fibres from the nucleus reticularis tegmenti pontis in the brain of the albino rat

Summary The existence of a cerebellar decussation of fibres from the medial portion of each nucleus reticularis tegmenti pontis (Rtp) of the albino rat is indicated. Definite cell loss in the medial aspect, of the most rostral third of Rtp is detectable after cerebellar hemisection involving parts or the entire depth of sublobule VIb. Cell loss in the medial aspect of the caudal half of Rtp is evident as a consequence of experimental lesions which damage both sublobules IIb and III.This work was supported by the Medical Research Council of New Zealand.Acknowledgments. A. special acknowledgment to. Mrs Heather Reid for her excellent preparation of the histological specimens. Grateful thanks to Mrs Maureen Owen for her careful typing assistance.     

Transparency in organisms

Summary The occurrence in animal phyla of species having a relatively transparent body is noted and measurements of the transmittance of medusae made in a spectrophotometer are reported, but the approximate nature of the results obtained with a commercial instrument and the importance of the correct physical design of the measuring apparatus are emphasized. The application to invertebrates of the structural explanation of the predominant transmission of incident light by the vertebrate cornea is discussed and the role of other factors considered. Destructive interference of the scattered rays, sufficient to account for the transparency of the cornea, has been shown not to demand a completely regular arrangement of collagen fibres. The small diameter and regularity of the fibrillar components in the muscles ofSagitta may be adequate to account for their transparency.I am grateful to Dr.D. M. Maurice for the encouragement of this interest, to Dr.E. G. Jordan for electron micrographs ofSagitta and to Mr.G. Ross (Department of Physics, Queen Elizabeth College) for helpful and critical discussion.     

Age-related response of citrate synthase to hydrocortisone in the liver and brain of male rats

Summary The activity of citrate synthase of the liver and brain of rats shows a gradual increase as a function of age. Adrenalectomy causes no significant change in the activity of citrate synthase in either of these tissues in young, adult or old rats. Administration of hydrocortisone to adrenalectomized rats depresses the activity of this enzyme maximally in the liver and brain of young rats. Administration of actinomycin D tends to normalize, the depressed level of this enzyme.Acknowledgments. The authors are indebted to Professor M.S. Kanungo of the Department of Zoology of Banaras Hindu University for helpful advice and facilities. A part of this project was presented at the 12th International Congress of Biochemistry, Australia, 1982.     

Origin of the differences of superficial potentials in Rana esculenta]

The spatial distributions of superficial D.C. potentials on the skin of Rana esculenta have been compared to those of the intensity of short-circuit current (S.C.C.) expressing the transcutaneous active transport of sodium ions. It has been observed that the sites of maximum D.C. potentials coincide with the localisations of maximum S.C.C. values. Moreover, superficial D.C. potentials and S.C.C. are similarly modified by the depression of metabolic activity due to lowered temperature or poisoning by dinitrophenol (DNP). It is thus proposed that the spatial distribution of the transcutaneous active transport system for sodium ions is the origin of the electric generator of superficial D.C. potentials.     

Iron-sulfur proteins: Recent developments in the field

Summary Iron-sulfur clusters in proteins are now recognized as among the main types of electron-transferring groups in biological systems, besides heme and flavins. Recent developments have brought forth a better understanding about the ways the protein environment modulates the potential of the cluster by placing the cluster in a more or less hydrophobic surrounding. Refinement in models, extensive studies on the kinetics of electron transfer (e.g. by measurement of the electronic spin lattice relaxation time) and the introduction of novel spectroscopic methods (EXAFS, magnetic CD and others) in the elucidation of structures in various systems are among the main developments. Other advances include EPR studies of the spatial orientation of Fe−S centers in complex membraneous systems (e.g. in mitochondria) and the recent elucidation of the nature of center X in photosystem I by M?ssbauer-spectroscopy. M?ssbauer studies have also been described on a number of Fe−S proteins (nitrogenase, aconitase, some ferredoxins, etc.) and revealed the existence of novel structures that enlarged the number of known basic units of Fe−S centers. These advances include: 1. the discovery of a novel non-heme Fe-protein (called desulforedoxin) of the rebredoxin type, 2. the elucidation of the nitrogenase Fe−S centers and the nitrogenase cofactor and 3. the discovery of a three-iron cluster in several enzymes and some ferredoxins. The latter 3-Fe cluster seems capable of being converted into a classical 4-Fe cluster under appropriate conditions, a phenomenon that plays a role in activation-deactivation of some enzymes (e.g. aconitase). It is now recognized that some iron-sulfur clusters may be involved in systems devoided of any oxydation-reduction reaction and may act as sensors of the surrounding redox potential, triggering the activation/deactivation of an enzyme (cf. e.g. aconitase).     

The quantification and differentiation of the drug receptor theory, c. 1910-1960

While historians have dealt with the origins of the concept of drug receptors in the work of Paul Ehrlich (1854-1915) and John N. Langley (1852-1925) as well as with some of its applications in modern pharmaceutical research, the history of the receptor theory as such has been neglected. Discussing major developments and conceptual changes in receptor theory between about 1910 and 1960 (including relevant contributions by A. V. Hill, A. J. Clark, J. H. Gaddum, E. J. Ari?ns and others), this paper attempts to fill this gap in historiography. It provides a case study of the unfolding of research under a new paradigm, but it considers also contemporary criticism and scepticism. By the early 1960s, quantitative investigations of drug action and interpretations of the experimental findings in terms of the receptor concept had become constitutive of the emerging field of 'molecular pharmacology'. Even then, however, receptors were still hypothetical entities.     

Induction of microsomal drug-metabolizing enzymes caused by hexobarbital

Summary Hexobarbital was given to anaesthetized mice for a period of 7 h by repeated i. p. injection, first of 100 mg/ kg, then several times of 50 mg/kg. A high level of hexobarbital was maintained in the liver. The activity of microsomal drug-metabolizing enzymes was induced by this treatment with hexobarbital. 30 min after a single i. p. injection of 100 mg/kg of hexobarbital, there was a significant inhibition of aminopyrine N-demethylase but none of cytochrome c and neotetrazolium reductases.—Hexobarbital in vitro inhibits aminopyrine N-demethylase but not cytochrome c reductase.     

Sterilization by irradiation of Cadra coutella (Wlk.) (Lepidoptera, Pyralidae) males increased by female sex pheromone

Sterilization by irradiation was studied in 0-24 hour old males of Cadra cautella (Ephestia cautella) exposed to 40 Krad of gamma radiation from a 60 cobalt source at a rate of 2250-2200 rad/minute in the presence of female sex pheromone. This radiation dose failed to induce the desired degree of sterility in the absence of pheromone. Pheromone was extraced from the virgin females and applied to filter paper at doses of .1, .5, and 1 female equivalent which was then introduced into containers containing 15-18 males 3 minutes prior to irradiation. Excitation and increased activity of the insects was observed when compared to the controls without the pheromone. The proportion of low percentage hatch increased with increased pheromone presence. The difference between absence and presence of pheromone was marked. It is concluded that these results are evidence of the value of this approach of preconditioning to irradiation when induction of sterility is the objective.