bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of a trypanocidal factor againstTrypanosoma equiperdumin normal human serum
Authors: Verducci G. , Perito S. , Rossi R. , Mannarino E. , Bistoni F. , Marconi P.
Published: June 2009

Normal human serum (HS) contains trypanolytic activity and agglutinins toTrypanosoma equiperdum, while such activities are not found in sera from a range of animals susceptible to infection. HS given toT. equiperdum-infected mice caused a rapid decrease in the number of circulating trypanosomes and protection from lethal infection. Trypanolytic activity of human serum was found to be associated, after DEAE chromatography and Sephadex G-200 gel filtration, with the fraction containing 19S antibodies. Immunofluorescence assays confirmed a binding of human IgM and C1qcomplement component onto the surface ofT. equiperdum. Anti-T. equiperdumactivity of HS was specifically directed toT. equiperdumsurface components and not to some mouse serum components adsorbed on parasites during the growth in the host, because HS adsorbedin vivoin CD-1 mice retained full protective and agglutinating properties. Trypanocidal activity appears in human serum about the 7th month after birth and persists until late in life. On the contrary, human purified high-density lipoprotein had no significantin vitroorin vivotrypanocidal activity. In conclusion, strong natural anti-T. equiperdumactivity in human serum was mainly mediated by natural antibodies of the IgM class. The presence of natural IgM active againstT. equiperdumin HS could represent one of the natural mechanisms of resistance of refractory hosts against trypanosome infections. This phenomenon provides further evidence that host specificity of trypanosomes may be partly conditioned by the presence of natural antibodies.
Aerobic to anaerobic transition in the carbohydrate metabolism of Schistosoma mansoni cercariae during transformation in vitro
Authors: Van Oordt B. E. P. , Tielens A. G. M. , Van Den Bergh S. G.
Published: June 2009

Schistosoma mansoni cercariae in water were shown to possess a largely aerobic energy metabolism, the Krebs cycle being the main terminal of carbohydrate breakdown. A metabolic transition towards a more anaerobic breakdown of carbo-hydrate could be achieved by incubation conditions which also stimulated biological transformation. Incubation of cercariae in a simple salt medium containing 5 mM glucose induced such a metabolic transition: beside carbon dioxide large amounts of lactate and pyruvate were excreted. The results indicate that the production of pyruvate was coupled to electron transfer in the respiratory chain. Some aspects of this unusual pyruvate production are discussed. The observed change in the end-product pattern of carbohydrate breakdown is very rapid: most of the switch occurred within 2 h. Our results show that the metabolic transition was triggered by the biological transformation itself, or by the same event that induces the biological transformation. The metabolic and the biological changes proceeded synchronously.
Detection of peptidases inTrypanosoma cruziepimastigotes using chromogenic and fluorogenic substrates
Authors: Healy N. , Greig S. , Enahoro H. , Roberts H. , Drake L. , Shaw E. , Ashall F.
Published: June 2009

Detergent extracts ofTrypanosoma cruziepimastigotes catalysed the hydrolysis of a range of amino-acyl and peptidyl p-nitro-anilides and aminomethylcoumarins. At least three enzymes were detected that cleave Z–Phe–Arg–MCA. Two of these were optimally active at alkaline pH, the other at pH 4·0. Of the two enzymes with alkaline pH optima, one was a cysteine peptidase and was unable to cleave Bz–Arg–MCA readily, whilst the other cleaved Bz–Arg–MCA and was inhibited by diisopropyl fluorophosphate. The acidic enzyme was similar to cathespin L of other eukayrotes with respect to its pH profile, substrate-specificity and inhibitor-sensitivity. Evidence was presented that epimastigotes contain a cysteine-type dipeptidyl aminopeptidase, one or more aminopeptidases, and a serine peptidase that cleaves Boc–Ala–Ala–pNA. Digitonin solubilization of the activities from cells supports the hypothesis that the cathespin L-like enzyme and the dipeptidyl aminopeptidase are lysosomal, whilst the Bz–Arg–MCA hydrolase, the aminopeptidases and the Boc–Ala–Ala–pNA serine peptidase are cytosolic.
Nematodirus battus: scanning electron microscope studies of the duodenal mucosa of infected lambs
Authors: Martin Jean , Lee D. L.
Published: June 2009

Lambs given 60000 third-stage larvae of Nematodirus battus were killed on days 16, 20, 22, 24 and 32 of the infection and the duodenal mucosa was examined by scanning electron microscopy (SEM). On day 16 of the infection the villi were long and finger-like and, although goblet cells were visible, the surface of the villi was not extensively folded, as is the surface of villi of uninfected control animals. By day 20 of the infection the villi had been reduced to flattened, plate-like structures or had formed low, irregular-shaped ridges. On this day of the infection the nematodes appeared to be enclosed by mucus-like material. As the infection progressed the intestinal damage was repaired, so that, by day 24 of the infection, the surface of the intestine was covered by short, smooth, finger-like projections and, by day 32 of the infection, the morphology of the intestine was somewhat similar to that of the control lamb. The possible role of villus atrophy and of mucus in the rejection of N. battus from the intestine is discussed.
Patency and transmission of Filaroides hirthi infection
Authors: Georgi J. R. , Georgi M. E. , Cleveland D. J.
Published: June 2009

Filaroides hirthi lungworm infection was diagnosed by the recovery of 1st-stage larvae from the faeces of dogs with heavy, artificially induced infections using zinc sulphate flotation. Diagnosis of low-grade natural infections was infrequently achieved. Zinc sulphate flotation was demonstrated to be about 100 times as efficient as the Baermann technique in concentrating F. hirthi larvae from dog faeces. Larvae recovered from faeces proved to be infective when fed to a pup and it was concluded that F. hirthi infection can be transmitted directly and immediately by fresh faecal contamination. Mongrel dogs of diverse ancestry were readily infected by feeding 1st-stage larvae from lung tissue. This, F. hirthi infection was shown not to be limited to the Beagle breed by biological restrictions. The observations that 1st-stage larvae pass through the alimentary tract on their way out of the body and that larvae are found in the mesenteric lymph modes long after a single exposure to infection support the hypothesis that there is an autogenous re-infection of the host by a proportion of these larvae.
Enzyme polymorphism of freshwater fish trypanosomes and its use for strain identification
Authors: Zajíček P.
Published: June 2009

Sixteen strains of trypanosomes isolated from the blood of 10 freshwater fish species (all but one from South Bohemia) were characterized by enzyme analysis using dise electrophoresis in 5% polyacrylamide gel. Six enzymes were selected: ADH, LDH, MDH, ME, G6PDH and GPI. The most variable were ADH and LDH, whereas MDH, G6PDH and GPI showed the same pattern in all strains analysed. Cluster analysis indicated a very high degree of similarity among strains, even those isolated from phylogenetically distant hosts. Moreover, no correlation between enzyme pattern and host species was found.
The transition from an aerobic to an anaerobic energy metabolism in transforming Schistosoma mansoni cercariae occurs exclusively in the head
Authors: Horemans A. M. C. , Tielens A. G. M. , Van Den Bergh S. G.
Published: June 2009

It has been shown that in intact cercariae of Schistosoma mansoni in water, both head and tail had an identical, aerobic energy metabolism. As long as the environment was water, glucose was mainly degraded to carbon dioxide by both head and tail whether or not these two were still connected to each other. Transfer of intact cercariae into a simple salt medium supplemented with glucose resulted in a very rapid transition towards a more anaerobic energy metabolism: the production of lactate and pyruvate increased, whereas the production of carbon dioxide remained more or less constant. A concomitant rise in temperature to 37°C was not essential for this biochemical transition, but made it more pronounced. Experiments on isolated cercarial bodies and tails in a transforming medium demonstrated that the tails oxidized glucose to carbon dioxide, whereas bodies produced mainly pyruvate and lactate. The results showed that the metabolic transition towards a more anaerobic energy metabolism occurred only in the head and not in the tail of the cercariae. Loss of the tail was shown not to be a pre-requisite for this transition, nor did it by itself trigger a metabolic switch in the resulting cercarial body.
Kronbrogia pugettensis sp. nov. (Neorhabdocoela: Fecampiidae), an endoparasitic turbellarian infesting the shrimp Heptacarpus kincaidi (Rathbun), with notes on its life-history
Authors: Shinn G. L. , Christensen A. M.
Published: June 2009

The morphology of Kronborgia pugettensis sp. nov., an endoparasite from the haemocoele of the caridean shrimp Heptacarpus kincaidi is described from specimens collected in the San Juan Island region of the western coast of North America. Members of this species are unisexual as are other members of the genus, but only females were found. An ootype and a type of accessory reproductive gland not known from other Kronborgia spp, are described, but they are probably present in other members of the genus. The life-history is similar to that of other Kronborgia spp. After the female reaches maturity, it emerges from the host and secretes a tubiform cocoon around itself and deposits its egg capsules within it. Embryogenesis lasts about 4 months. A free-swimming larva locates the new host, attaches to its exterior, and secretes a cyst around itself. It then penetrates through the exoskeleton to reach the haemocoele. K. pugettensis is easily distinguished from other members of the genus by the shape of the cocoon, type of host, and geographic distribution. This is the second species of fecampiid described from North America, but several undescribed species are known to exist from their cocoons.
A comparative study of the succinate dehydrogenase–fumarate reductase complex in the genusTrichinella
Authors: Rodriguez–Caabeiro F. , Criado–Fornelio A. , Jimenez–Gonzalez A.
Published: June 2009

Succinate dehydrogenase and fumarate reductase activities from a participate fraction of 4 isolates of the supposed ‘species’ ofTrichinella, T. spiralis, T. pseudospiralis, T. nativaandT. nelsonihave been determined using spectrophotometric methods. The ‘in vitro’ effect of the anthelmintics thiabendazole, mebendazole and ivermectin on succinate dehydrogenase and fumarate reductase activities from the above isolates is also described. The significance of the results is discussed in the context of the controversy that surrounds speciation of the genusTrichinella.
Monosaccharide composition of the surface glycoprotein antigens ofTrypanosoma brucei
Authors: Allsopp B. A. , Njogu A. R.
Published: June 2009

Evidence has been adduced that the surface antigens ofTrypanosoma bruceisubgroup are a group of glycoproteins having D-galactose, D-mannose and D-glucosamine as monosaccharide components.

There was considerable variation in the number of antigen components, and their relative amounts, as revealed by isoelectric focusing on polyacrylamide gel. The relevance of these variations to the adaptation theory of antigenic variation in trypanosomes is discussed.
The distribution of larval Aspiculuris tetraptera Schulz during a primary infection in Mus musculus, Rattus norvegicus and Apodemus sylvaticus
Authors: Behnke Jerzy M.
Published: June 2009

The larvae of Aspiculuris tetraptera were found in the mid-colon of mice within hours of infection. When the colon was divided into 10 equal sections the larvae were mainly found in sections 5, 6 and 7 during the first 6 days of the infection. The worms entered the crypts of Lieberkühn in this region on day 1 and remained there until day 4 or 5. After this time they left the crypts and returned to the lumen of the colon. On day 7 the worms emigrated anteriorly and thereafter were recovered only from the proximal region of the colon (sections 1–4), although in heavier infections a few larvae remained in the mid-colon.

The initial establishment site was the same in both Rattus norvegicus and in Apodemus sylvaticus, but an infection with A. tetraptera in the abnormal hosts, R. norvegicus and A. sylvaticus, was characterized by less than 7% of the inoculum becoming established, a slower rate of growth and a wider distribution centering around the preferred site. The small number of established larvae was lost from the rat before day 12 and from A. sylvaticus before day 8, whereas larvae persisted in laboratory mice for a longer period.
Avian trypanosomes in Simulium and sparrowhawks(Accipiter nisus)
Authors: Dirie M. F. , Ashford R. W. , Mungomba L. M. , Molyneux D. H. , Green E. E.
Published: June 2009

Isolates of avian trypanosomes from nestling sparrowhawks (Accipiter nisus) and from Simulium latipes were compared by isoenzyme electrophoresis with the previously described avian trypanosomes Trypanosoma corvi and T. everetti. Simulium isolates developed into trypomastigotes in semi-defined medium at 37 °C confirming that they belong to the genus Trypanosoma. Cellulose acetate electrophoresis (CAE) and polyacrylamide gel electrophoresis (PAGE) methods were employed. A total of 11 enzymes was examined, of which 8 gave satisfactory results for all lysates (MDH, ICD, PGM, 6PGD, SOD, PEP I, PEP II and PEP D). The zymograms of 3 isolates from Simulium and 4 from A. nisus were similar indicating that they were the same organism and the isoenzyme patterns coincided with those of T. corvi. It is concluded that A. nisus and S. latipes are infected with T. corvi. Flagellate infections were found in the midgut, hindgut and rectum of Simulium where large numbers of epimastigotes were found in the lumen and attached to the cuticular intima of the rectal ampullae by hemidesmosomes at the distal end of an expanded flagellum. Out of 285 nestling sparrowhawks 13 (5%) were infected with trypanosomes. In view of their age and inability to preen themselves it is suggested they became infected via infected prey.
Effects of dietary carbohydrate on the nutritional physiology and blood sugar level of Trichoplusia ni parasitized by the insect parasite, Hyposoter exiguae
Authors: Thompson S. N.
Published: June 2009

The effects of parasitization by Hyposoter exiguae on the nutritional physiology of Trichoplusia ni were influenced by the dietary sucrose level. The results confirmed that host nutrition plays a significant role in the manifestation of parasitic infection during the insect parasite-host association. Increased dietary sucrose level resulted in an increase in haemolymph trehalose concentration in both control and parasitized insects, but the increase was greater in the latter group. However, when the sucrose level was increased without energetic compensation and the dietary casein level was maintained constant, the haemolymph trehalose concentration was affected very little in unparasitized individuals but was markedly elevated in parasitized hosts. This result suggests that the de novo synthesis of carbohydrate in parasitized T. ni may not be regulated by dietary sugar intake.
Schistosoma mansoni: dynamics of migration through the vascular system of the mouse
Authors: Wilson R. A. , Coulson Patricia S.
Published: June 2009

Autoradiography of compressed mouse tissues has been used to estimate the numbers of Se-labelled schistosomula present in different mouse organs. Day 7 parasites extracted from the lungs of donor mice were delivered by injection to the lungs, systemic organs and liver of recipient mice as a discrete pulse. The numbers detected in various locations with time post-injection were then used to analyse the dynamics of intravascular migration. Approximately 98% of cercaria-associated label was lost during the first 14 days of parasite life, two-thirds of this in the first 7 h post-infection. Nevertheless, 99–113% of schistosomula could be detected 30 min post-injection into the locations chosen. The efficiency of the parasite delivery system was 95%. The time required for the number of foci in the lungs to decline to 50%, after injection of parasites via the femoral vein, was 55 h. Adjustment of this data to allow for parasites returning to the lungs after passage round the systemic vasculature gave a value of 30–35 h for the true mean time of lung transit. The distribution of parasites to systemic organs after their exit from the lungs was proportional to the fractional distribution of cardiac output. The probability (P) of a schistosomulum being distributed to splanchnic beds was estimated at 0·32 and itsPof being trapped in the hepatic portal distributaries within the liver as 0·72–0·86. On this basis, the entire hepatic portal population of adult schistosomes would be recruited during 2–3 circuits of parasites around the pulmonary-systemic vasculature. The mean transit time of schistosomula through intestinal capillaries was 6·5 h whilst that through other systemic organs combined (muscles, kidneys, brain, etc) was 16 h, considerably more rapid than lung transit. The time taken for schistosomula to pass between organs, in arterial and venous blood, was shown to be less than 30 min in both cases (probably much less).
Naturally occurring immunoglobulin M antibodies: enhancement of phagocytic and microbicidal activities of human neutrophils againstToxoplasma gondii
Authors: Konishi E. , Nakao M.
Published: June 2009

Naturally occurring immunoglobulin M antibodies toToxoplasma gondiiin human sera were examined for their ability to enhance toxoplasmacidal activities of human neutrophils to show a role of natural antibodies in immunity to the early stage ofToxoplasmainfection in humans. Neutrophils were mixed with tachyzoites at a ratio of 1:5 in the presence or absence of antibodies and were cultured for 1–18 h for microscopical examination. The count of tachyzoites phagocytosed in 200 neutrophils within 1 h was significantly higher in the presence of sera with natural IgM antibody levels of > 0.8 than those of < 0.1, with 2.4- to 2.9-fold differences (P< 0.02). The total tachyzoite counts at 18 h decreased to 26–39% of those at 1 h in the presence of natural IgM antibody levels of > 0.8 (P< 0.01), while the counts at 1 h consistently increased by 18 h in the absence of natural IgM antibodies. These results indicate that relatively high levels of natural IgM antibodies enhanced phagocytic and microbicidal activities of neutrophils againstToxoplasma. The enhancement was dose dependent, and was also weaker than that obtained with the same dilution of sera from individuals with chronic or acute infections. Live and dead tachyzoite counts in infected neutrophils suggested a quicker effect of natural IgM antibodies than IgG antibodies elicited by infection.
Ectoparasite impacts on Gerbillus andersoni allenbyi under natural conditions
Authors: Lehmann T.
Published: June 2009

To assess ectoparasite impact on individuals and populations of Gerbillus andersoni allenbyi under natural conditions, I addressed the following questions. Do ectoparasites affect their host fitness and, if so, how? Do ectoparasites affect host population level? Does this parasite–host interaction support the traditional concept of parasite evolution towards avirulence? For this purpose, host infestation, infection, survival, haematological indices, and physical condition were recorded. A field experiment which included manipulating host infestation while recording host survival was conducted to determine the causal relations between these variables. G. a. allenbyi was infested by 2 fleas (Synosternus cleopatrae and Stenoponia tripectinata), 5 mesostigmatid mites (Androlaelaps centrocarpus, A. hirsti, A. insculptus, A. marshalli and Hirstionyssus carticulatus), 1 tick (Rhipicephalus sanguineus), and 1 louse (Polyplax gerbilli). Ectoparasite burden significantly reduced host survival and red blood cell indices (red cell concentration, haemoglobin concentration and haematocrit). Ectoparasite burden did not significantly affect white blood cell concentration. Gerbils were not infected by haemoparasites or gut helminths which potentially could cause anaemia. The causal relationship between S. cleopatrae burden and host survival was established by manipulation of host infestation. Both ectoparasite removal and initial level of infestation significantly affected host survival. Ectoparasites that caused anaemia were not associated with host physical condition (PC), measured as weight/length3. None of the red blood cell indices was correlated with host PC. These results suggest both that host PC was not affected by ectoparasite burden and that exsanguination leading to anaemia was the main effect of the ectoparasites. Ectoparasite pressure on the host population (based on the ectoparasite effects as estimated by statistical models, combined with dispersion of the infestation within the host population) changed seasonally and was greatest when host density was the highest. A large segment of the gerbil population was affected by ectoparasites during the entire year. An explanation for the evolution of parasite virulence, contrasting parasites that evolve towards increased virulence with parasites that evolve towards avirulence is presented. This classification is primarily based on whether parasite impact is equated with a higher efficiency of host exploitation, or whether it is a ‘side effect’ of parasite biology.
Detection of linkage disequilibrium in Trypanosoma brucei isolated from tsetse flies and characterized by RAPD analysis and isoenzymes
Authors: Stevens J. R. , Tibayrenc M.
Published: June 2009

This study analyses the different populations of Trypanosoma brucei spp. which may coexist within the midgut of wild tsetse flies (Stevens et al. 1994). Cloned trypanosome populations characterized by multilocus enzyme electrophoresis (MLEE) were further analysed by the random amplified polymorphic DNA (RAPD) technique, allowing detection of genetic variation at a finer level than that possible by MLEE. Genetic distance matrices derived from the results of each of the two biochemical methods were calculated and compared using a computer program based on the method of Mantel (1967). The observed correlation was used to investigate the degree of linkage disequilibrium (LD) in the data, association between unrelated polymorphic markers providing a measure of the departure from panmixia. The potential of each biochemical method to detect linkage was evaluated by an extended Mantel test. The MLEE/RAPD correlation test evidenced significant LD within the population, suggesting a predominantly clonal method of reproduction for these West African trypanosomes. Analysis of RAPD data by the extended Mantel test also showed significant LD, while the results with MLEE data were less conclusive, providing an indication of the relative potential of the two techniques to detect fine genetic variation.
203Hg and other gamma-emitting radio-isotopes as labels for Dirofilaria immitis microfilariae
Authors: Lengemann F. W. , Grieve R. B. , Chmielewicz M. , Georgi J. R.
Published: June 2009

The in vitro uptake of gamma-emitting radionuclides by microfilariae of Dirofilaria immitis was investigated. Radionuclides tested were 133Ba, 207Bi, 82Br, 109Cd, 51Cr, 60Co, 59Fe, 203Hg, 125I, 54Mn, 32P, 125Sb, 75Se, and 65Zn. Only 207Bi, 59Fe, 203Hg, 54Mn, and 46Sc showed more than 2% of the available radioactivity to bind to the microfilariae. When tested for retention in vitro only 203Hg showed retention levels approaching 90%. Moreover, when dimethyl suiphoxide was incorporated into the medium at levels of 1% (v/v) the uptake of 203Hg could be increased by 3–5 times; no other radio-isotope tested responded in this manner. The uptake of 203Hg was directly related to temperature and time of incubation. Mercury, as mercuric chloride, was toxic to the microfilariae and respresents an impediment to the incorporation of high levels of 203Hg in microfilariae.
Plasmodium falciparum (human malaria)-induced modifications in human erythrocyte band 3 protein
Authors: Crandall I. , Sherman I. W.
Published: June 2009

A monoclonal antibody, 1C4, was produced which recognizes a 65 kDa protein that is localized to the plasma membrane of human erythrocytes infected with Plasmodium falciparum. By immunofluorescence the antigen was visualized as dots on the surface of the infected cell. The 65 kDa protein was present in 4 strains of diverse geographical origin, and in erythrocytes infected with a knobless strain. The 65 kDa protein was insoluble in non-ionic detergents, but was partly soluble in SDS and some high (1 M) śalt solutions. The 65 kDa protein is recognized by antibodies specific for the cytoplasmic domain and the N-terminal side of the membrane-spanning region of human band 3, but was not recognized by an antibody specific to the C-terminal side of the membrane-spanning region. The results of treatment of the 65 kDa protein with trypsin and chymotrypsin are consistent with the 65 kDa protein being a truncated and covalently modified band 3 molecule which consists of the first 540 amino acids of human band 3.
Specific immunosuppression byTrichinella: fine specificity and effect on lymphocyte functionin vivo
Authors: Baltar P. , Leiro J. , Santamarina M. T. , Sanmartín M. L. , Porto M. C. , Ubeira F. M.
Published: June 2009

Muscle-phaseTrichinellalarvae depress the immune response of mice to the phosphorylcholine (PC)-bearingTrichinellaantigen FCp without affecting responses to other PC-bearing or non-PC antigens. The depressive activity is independent of antigen dose andTrichinellaspecies and, in adoptive cell transfer experiments with lethally irradiated recipient mice, depended on the state of the recipient (infected recipients had a depressed response even a month after their encysted larvae had been killed and regardless of whether the donor had been exposed to FCp) but not on the state of the transferred cells. We conclude that lymphocytes are not permanently altered by the depressive action, that the agent responsible persists in the host at least a month after the death of the encystedTrichinellalarvae, and that the alteration does not eliminate lymphocyte immunological memory.
Trichuris trichiurainfection and cognition in children: results of a randomized clinical trial
Authors: Simeon D. T. , Grantham-McGregor S. M. , Wong M. S.
Published: June 2009

The effects of mild to moderate infections ofTrichuris trichiuraon cognitive functions were investigated in Jamaican children aged 7 to 10 years. In all, 189 infected children and 100 uninfected classmates were studied. The infected children were randomly assigned to receive treatment (albendazole) or a placebo. All children were given cognitive tests on enrolment and 14 weeks later. These included verbal fluency (generation of ideas), digit span (working memory), number choice (speed of processing of visual stimuli), visual search (sustained attention) and a French Vocabulary test (paired-associate learning). At baseline, the infected children had lower scores than the uninfected ones in fluency (P= 0·01), search (P= 0·02) and French (P= 0·01). Treatment effects were examined among infected children and there was no significant treatment effect for any of the tests. However, there was a significant treatment by weight-for-age interaction in fluency (P < 0·05). The children with low weight-for-age (Z-score < – 1) improved with treatment while there was no improvement with treatment among the other children. We concluded that treatment of children with mild to moderateT. trichiurainfections using albendazole produces little benefit in cognition if they are adequately nourished; however, undernourished children are more likely to benefit.
Endotoxins in the blood and cerebrospinal fluid of patients with African sleeping sickness
Authors: Pentreath V. W. , Alafiatayo R. A. , Crawley B. , Doua F. , Oppenheim E. A.
Published: June 2009

Endotoxin levels were measured in the blood and cerebrospinal fluid (CSF) of control individuals and 2 groups of patients with African sleeping sickness. Endotoxin levels were markedly elevated in the blood (infected groups mean endotoxin values 40·2 pg/ml and 53·8 pg/ml, compared to control 11·6 pg/ml, P < 0·0001 for both increases) and CSF (infected groups mean endotoxin values 45·8 pg/ml and 50·1 pg/ml compared to control 6·3 pg/ml, P < 0·0001 for both increases) of the patients. The levels were reduced 6 weeks following different drug treatments in the 2 groups (blood levels to mean 33·8·pg/ml and 28·5 pg/ml; CSF levels to 37·4 pg/ml and 27·0 pg/ml). The blood endotoxin values correlated with the CSF values before treatment (r = 0·74 and 0·57 for the 2 groups; P < 0·0001 for both) and after treatment (r = 0·57 and 0·56 for the 2 groups; P < 0·0001 for both). It is concluded that raised endotoxin equilibrates in the blood and CSF compartments, and may contribute significantly to the pathology of sleeping sickness.
Cytoadherence-related homologous motifs inPlasmodium falciparumantigen Pf155/RESA and erythrocyte band 3 protein
Authors: Iqbal J. , Siddique A. B. , Ahlborg N. , Perlmann P. , Berzins K.
Published: June 2009

Cytoadherence ofPlasmodium falciparum-infected erythrocytes plays an important role in the pathogenesis of cerebral malaria. The identity of cell surface molecules on parasitized erythrocytes involved in cytoadherence is of great interest to understand the molecular basis of this mechanism. Peptide sequences derived from exofacial loops of the erythrocyte antigen band 3 from parasitized erythrocytes have previously been shown to inhibit cytoadherence. We now report that a non-repeated region of Pf155/RESA (residues 213–218) contains a hexapeptide motif being highly homologous to cytoadherence inhibitory sequences from band 3. Synthetic peptides containing the hexapeptide motif of Pf155/RESA inhibited the binding ofP.falciparum-infected erythrocytes to melanoma cellsin vitro. Furthermore, individuals residing in malaria-endemic areas have antibodies reactive with epitopes involving these motifs in band 3 and in Pf155/RESA.
Hymenolepis diminuta-induced fecundity reduction may be caused by changes in hormone binding to Tenebrio molitor ovaries
Authors: Webb T. J. , Hurd H.
Published: June 2009

Aspects of vitellogenesis, known to be controlled by juvenile hormone, are adversely affected by Hymenolepis diminuta infection of Tenebrio molitor, in spite of circulating titres of the hormone remaining unchanged. It has therefore been proposed that juvenile hormone binding is disrupted at the tissue site level. Juvenile hormone III binding sites were located in the nuclear, microsomal and post-microsomal supernatant fractions of the follicle cells of Tenebrio molitor. When JH-III binding was quantified for both control and Hymenolepis diminuta-infected beetles, binding in the nucleus and cytosol were found to be largely unaffected. However, microsomal binding was severely disrupted; on days 3 and 6 post-infection, binding was greatly diminished, on day 9 post-infection, binding was slightly reduced and, by day 15, binding was ‘restored’ to that of control insects. Using follicle cell microsomes at day 3 post-infection, previous Scatchard analysis revealed the presence of at least two JH-III binding sites. The first is of higher affinity, Kd = 5·3 x 10·5 x 10−11 mol/mg protein and the second of lower affinity Kd = 7·7 x 10−7 M, Bmax = 9·75 x 10−11 mol/mg protein. A comparison with microsomal binding parameters of follicle cells from non-infected Tenebrio indicated that although the Bmax values were unchanged, the Kd value of the higher affinity site was increased by approximately 5-fold. These data are indicative of a parasite-induced competitive binding inhibitor.
Gwendolen Rees FRS – fifty-six years (1930 to date) in research
Authors: Williams Harford
Published: June 2009

Professor Gwendolen Rees, or Gwen as she prefers to be known, was educated at the Intermediate School for Girls, Aberdare, between 1918 and 1924. Aberdare, a valley community in South Wales was, in the nineteenth century, world-famous as a centre for the Welsh iron and coal industry. Unlike its neighbouring town, Merthyr, however, it was not totally dominated by industry and it is especially renowned for its spacious and beautiful park near the town centre. During Gwen's early life, Aberdare was one of the largest and most influential centres of literary culture in Wales, and a place where education was of prime importance in the community. As far back as 1867, Aberdare men and women had claimed that ‘what we think today, Wales will think tomorrow’. Could these be the reasons why Aberdare has produced so many distinguished academics? None, however, has achieved greater distinction than Gwendolen Rees who, in 1971, became the first woman academic in Wales to be elected a Fellow of the Royal Society. Since she has often been affectionately referred to as ‘one in a million’ by her former students, the metaphor assumed literal significance in 1971, when the female population of Wales was 1446000. Because of a phenomenal life-long dedication to her chosen career as university teacher, researcher and administrator, together with a genuine lack of desire for personal acclaim, there may have been many people who had never heard of her until 1971.
Identification of isolate-specific proteins on sorbitol-enriched Plasmodium falciparum infected erythrocytes from Gambian patients
Authors: Aley S. B. , Sherwood J. A. , Marsh K. , Eldelman O. , Howard R. J.
Published: June 2009

We have compared the surface radio-iodinated proteins of uninfected and Plasmodium falciparum-infected erythrocytes from natural infections of human patients. Cryopreserved infected blood from Gambian children with falciparum malaria was thawed, cultured to the middle trophozoite stage, and surface radio-iodinated. Trophozoite-infected cells were enriched about 10-fold on a Percoll gradient newly designed to separate cells based on their differential permeability to sorbitol. Infected blood was radio-iodinated and erythrocyes from the fraction enriched in parasitized cells and uninfected erythrocytes from the same sample obtained from the gradient and compared by SDS–PAGE and autoradiography. In each sample, parasitized erythrocytes contained one or more polypeptides of very high molecular weight (Mr 250000–300000) that were not found on uninfected erythrocytes from the same patient. These proteins were isolate-specific in size and number, suggesting that natural isolates contain a variable number of different P. falciparum phenotypes for this surface protein. In addition, these radio-iodinated surface proteins could not be extracted from the host cell membrane by the non-ionic detergent Triton X-100, but were extracted by SDS. The properties of these proteins suggest they are the equivalent for natural infections of the strain-dependent antigen previously described (Leech, Barnwell, Miller & Howard, 1984) on the surface of P. falciparum-infected Aotus erythrocytes. In addition, we observed a second parasite-dependent modification of labelled proteins on infected erythrocytes with the appearance of a new band of Mr 30000. There were also variations in the pattern of radio-isotope labelled proteins on uninfected erythrocytes from different patients.
Sialoglycoproteins and sialic acids ofPlasmodium knowlesischizont-infected erythrocytes and normal rhesus monkey erythrocytes
Authors: Howard R. J. , Reuter G. , Barnwell J. W. , Schauer R.
Published: June 2009

The effects of malaria infection on RBC sialic acids and sialoglycoproteins were studied with asexual blood-stage infections ofPlasmodium knowlesiin rhesus monkeys. Glycoprotein radio-isotope labelling methods were used to compare the sialoglycoproteins of normal RBC andP. knowlesischizont-infected RBC (SI-RBC). Tritiation of glycoproteins from SI-RBC with the standard sialidase + galactose oxidase/NaB3H4method or standard periodate/NaB3H4method was significantly decreased when compared to normal RBC. However, tritium uptake into glycoproteins was normal when SI-RBC were treated with 5-fold higher concentrations of both enzymes in the first labelling method, or with a 5-fold increase in the molar ratio of periodate to sialic acid in the second method. The mobility of tritiated host cell glycoproteins on SDS–polyacrylamide gels was identical for SI-RBC and normal RBC. New bands, possibly glycoproteins, of 230, 160, 90, 52, and 30 kDa were detected after labelling SI-RBC by the modified periodate/NaB3H4method. Sialic acid analysis of normal rhesus monkey RBC (62μg/1010RBC) revealed that 46% of the total sialic acid wasN-glycolylneuraminic acid, 33% wasN-acetyl-9-O-acetylneuraminic acid, and the remainderN-acetylneuraminic acid. SI-RBC collected either directly from infected monkeys or afterin vitroculture of ring-infected RBC in horse serum, had increased total sialic acid (126 or 115μg/1010RBC, respectively). The sialic acid content of infected RBC must increase during parasite development since RBC infected with ring-stageP. knowlesihad the same content as normal RBC. There was no significant difference in the ratio of the three sialic acids of SI-RBC and normal RBC. In contrast, the uninfected RBC from infected blood of different monkeys showed marked variation in sialic acid composition and generally had a lower sialic acid content than normal RBC.
Mouse-to-mouse transmission of infections with Giardia muris
Authors: Belosevic M. , Faubert G.M. , Maclean J. D.
Published: June 2009

The transmission of infection with Giardia muris among CD-I mice was quantified by determining the duration of the latent period and intensity of cyst release during the acute phase of the infection. Housing of experimentally inoculated mice with uninfected mice in the same cage resulted in 100% transmission of infection. The dissemination of the infection was faster if experimentally inoculated hosts were passing high numbers of cysts at the time of housing with uninfected animals. The course of ‘naturally’ acquired and experimentally induced infections was similar. We conclude that G. muris has a high transmission potential and that the number of cysts randomly consumed does not influence the course and eventual outcome of the infection.
Population dynamics in echinococcosis and cysticercosis: biological parameters of Echinococcus granulosus in dogs and sheep
Authors: Gemmell M. A. , Lawson J. R. , Roberts M. G.
Published: June 2009

The numerical distributions of Echinococcus granulosus in an experimental dog population are described. At all dose rates of protoscoleces from 10 to 175000 the distribution of worms was over-dispersed. Host age had no effect. There was a direct proportionality between the infective-stage density and rate of infection, and between the latter and the index of clumping. The worm burdens were significantly higher in the proximal than distal portions of the small intestine. Lengths of the 3- and 4-segmented worms increased from 4 to 10 and 4 to 8 weeks of age, respectively. Thereafter apolysis was asynchronous and could not be determined. Eggs were first detected in the faeces at 6 weeks and the mean age at oogenesis was 7·26 weeks. Retarded growth of the whole population of worms was observed in some dogs. For the first few infections, worm burdens varied widely in the same dog, but by the 6th infection 50% of the dog population had developed a relative insusceptibility to infection. Growth or oogenesis of the worms were not affected. A short-acting immune response was artificially induced in some dogs following the parenteral injection of activated embryos of E. granulosus, Taenia hydatigena, T. ovis, T. multiceps, T. pisiformis and T. serialis. The response affected either the number of worms established, growth or oogenesis or all three parameters. There was a strong positive correlation between numbers and lengths of worms in dogs with acquired and induced immunity, indicating that no ‘crowding’ effects were involved. In sheep populations the mean number of cysts which established was directly proportional to the number of eggs given, implying that there was no negative feedback mechanism operating at this stage of the life-cycle. The distribution of the larval population in sheep was over-dispersed and the index of clumping increased with the size of the egg dose from 25 to 2500 eggs. Protoscoleces were first observed in cysts at 2 years and the proportion producing them increased with age, with an estimate of 50% of cysts containing protoscoleces at 6 years. No deaths were observed in dogs or sheep even when high parasite burdens were present, implying that E. granulosus does not regulate the population of its hosts.
Stereological estimation of gap junction surface area per neuron in the developing nervous system of the invertebrate Mesocestoides corti
Authors: Burns A. J. , Howard C. V. , Allen J. M. , Van Velsen D. , McKerr G.
Published: June 2009

As a major morphological feature in establishing the form of the nervous system, it is recognized that neurons are initially overproduced, then naturally occurring cell death reduces the neuron number to the functional requirement. However, the mechanisms controlling the selective elimination of certain neurons during a general phase of cell death are not fully understood. One event that seems to be pivotal is the establishment of neural connections, the degree of which may be influential regarding the fate of specific neurons. However, little quantitative evidence is available to either support or refute this theory. In this current study, a Stereological measurement of gap junction per neuron was carried out within the invertebrate model system of the tapeworm metacestode Mesocestoides corti, which has previously been shown to overproduce neurons during the asexual reproduction stage of its life-cycle. Novel Stereological estimation methods with ‘ vertical sections’ indicated that prior to asexual division the cerebral ganglion possessed approximately 268 neurons, each with a gap junction surface area of 250 μm2. As division progressed, the neuron number increased to approximately 700, while the total surface area of gap junction remained statistically unchanged. As a result the surface area of gap junction per neuron decreased to 106 μm2, less than half that in the undividing stage. These results provide the first non-biased quantitative data regarding changes in the mean surface area of gap junction per neuron in a developing cerebral ganglion.
The within-host cellular dynamics of bloodstage malaria: theoretical and experimental studies
Authors: Hetzel C. , Anderson R. M.
Published: June 2009

The properties of a mathematical model of bloodstage infection with a single strain of malaria were investigated. Analysing the cell population dynamics in theabsenceof a host immune response we demonstrate a relationship between host and parasite parameters that defines a criterion for the successful invasion and persistence of the parasite. Important parameters are the rates of merozoite production and death and those of erythrocyte production, death and invasion. We present data from experiments designed to evaluate the erythrocyte invasion rate in a rodent malaria system. The model generates patterns of parasitaemia in good qualitative agreement with those seen inPlasmodium bergheiinfections. The sole force behind the rise and fall in parasitaemia in the model without immunity is the density of susceptible erythrocytes, suggesting that resource availability is an important determinant of the initial pattern of infectionin vivo. When we incorporate a simple immune response into the model we find that immunity against the infected cell is much more effective at suppressing parasite abundance than immunity against the merozoite. Simulations reveal oscillating temporal patterns of parasite abundance similar toP. c. chabaudiinfection, challenging the concept that antigenic variation is the sole mechanism behind recrudescing patterns of infection.
Enzyme polymorphism in Endotrypanum and numerical analysis of isoenzyme data
Authors: Franco A. M. R. , Momen H. , Naiff R. D. , Moreira C. F. S. , Deane M. P. , Grimaldi G.
Published: June 2009

In this study, we have analysed enzyme polymorphism among a group of protozoan parasites of the genus Endotrypanum (Kinetoplastida: Trypanosomatidae). Seventeen stocks of Endotrypanum spp. isolated from sloths (Choloepus didactylus and C. juruanus) in the Amazon Region of Brazil were analysed by enzyme electrophoresis, and their electromorphic profiles were compared with reference strains reported previously. The 16 enzymic loci were analysed, and the strains were classified into zymodemes, each representing parasites with unique enzyme profiles. Each zymodeme was considered as an elementary taxon, and using numerical analyses (cladistic, agglomerative hierarchical and ordination techniques) the genus was shown to be monophyletic and the 12 zymodemes characterized could be divided into 3 groups (A, B, C). The heterogeneous population (which may represent a complex of parasite species or strains variants) showed, however, no correlation with the origin (i.e. host species involved or geographic area of isolation) of Endotrypanum stocks. Eight isolates of Endotrypanum sp. from Rondonia State (Brazil) and a parasite strain from Panama were clustered together into a zymodeme, which was phenetically closely related to the E. monterogeii from Costa Rica. The data indicate that E. schaudinni is a species complex.
Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults
Authors: De Graaf D. C. , Berghen P. , Moens L. , De Marez T. M. , Raes S. , Blaxter M. L. , Vercruysse J.
Published: June 2009

Western blot analysis using an anti-globin rabbit serum Rb94 revealed a major band of 17 kDa in extracts of Ostertagia ostertagi adults and 4th-stage larvae. The adult stage globin-like antigen (OoAdGlb) was purified from total worm extracts by liquid chromatography. The protein has an estimated molecular mass of 36 kDa under non-reducing conditions, suggesting a dimeric structure containing 2 non-covalently linked 17 kDa monomers. Tryptic peptides were sequenced and showed strong similarities with the globins of free-living and parasitic nematodes. Immunolocalization revealed the presence of this globin-like antigen in the body wall musculature and/or the cuticle of O. ostertagi adults. An enzyme-linked immunosorbent assay based on the purified OoAdGlb showed no differences in response between calves infected by O. ostertagi and/or Cooperia oncophora and the negative controls.
Malaria parasites enhance blood-feeding of their naturally infected vector Anopheles punctulatus
Authors: Koella J. C. , Packer M. J.
Published: June 2009

We investigated the blood-feeding behaviour of a natural population of the human-feeding mosquito Anopheles punctulatus in Iguruwe, Papua New Guinea. In particular we investigated the relationship between the mosquitoes' blood-feeding behaviour and their infection by the malaria parasites Plasmodium falciparum and P. vivax. Female mosquitoes were caught at 4 times of the night, the amount of blood they had obtained was measured and their status of infection was evaluated. Among uninfected mosquitoes the bloodmeal size steadily increased through the night, possibly because they were progressively less likely to be disturbed by human activity as the night drew on. Infected mosquitoes, on the other hand, tended to feed maximally at all times of the night. This suggests that infected mosquitoes were more tenacious in their blood-feeding behaviour, being either less readily disturbed during a bout of feeding (and thus feeding longer) or more likely to return to continue their feed following disturbance (and thus feeding several times). Either change would increase the parasites' rate of transmission. We conclude that in this natural situation the two species of malaria parasites modified the mosquitoes' behaviour with the effect of increasing their own transmission.