bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of Sporozoite Surface Proteins and Antigens of Eimeria nieschulzi (Apicomplexa)
Authors: TILLEY MICHAEL , UPTON STEVE J. 
Published: May 2007

ABSTRACTSodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting, lectin binding, and 125I surface labeling of sporozoites were used to probe sporozoites of the rat coccidian, Eimeria nieschulzi. Analysis of silver stained gels revealed >50 bands. Surface iodination revealed about 14 well labeled, and about 10 weakly labeled but potential, surface proteins. The most heavily labeled surface proteins had molecular masses of 60, 53–54, 45, 28, 23–24, 17, 15, 14, 13, and 12 kD. Following electrophoresis and Western blotting, 2 of the 12 125I labeled lectin probes bound to two bands on the blots, which collectively indicated that two bands were glycosylated. Concanavalin A (ConA) specifically recognized a band at 53 kD, which may represent a surface glycoprotein, and a lectin derived from Osage orange (MPA) bound to a single band at 82–88 kD, that may also be a surface molecule. Immunoblotting using sera collected from rats inoculated orally with oocysts, as well as sera from mice hyperimmunized with sporozoites, revealed that many surface molecules appear to be immunogenic.
Nolleria pulicis N. Gen., N. Sp. (Microsporida: Chytridiopsidae), a Microsporidian Parasite of the Cat Flea, Ctenocephalides felis (Siphonaptera: Pulicidae)
Authors: BEARD CHARLES B. , BUTLER JERRY F. , BECNEL JAMES J. 
Published: May 2007

ABSTRACTA new species of microsporidium, Nolleria pulicis, is described and named here from the cat flea, Ctenocephalides felis. The genus Nolleria is created and placed within the family Chytridiopsidae. The family is slightly modified to accommodate certain features of intracellular development seen in N. pulicis, which is otherwise very similar to other species in the family Chytridiopsidae. Sporulation is described from ultrastructural analysis of infected midgut epithelial cells of adult C. felis. The term “multiple division by vacuolation” is proposed for describing sporogony as it occurs in this species and certain related species of microsporidia. The probable mode of transmission and apparent absence of merogony are discussed.
Trypanosoma acomys (Wenyon, 1909): Continuous Culturing with a Mouse Fibroblast Cell‐Line (A9)
Authors: ABDALLAH MONA A. , ABDEL‐HAFEZ SAMI K. , AL‐YAMAN FADWA M. 
Published: May 2007

ABSTRACTThe continuous culturing of Trypanosoma acomys in the presence of a murine areolar‐adipose cell line (A9) was possible for the 1st time. The trypanosomes were cultured at 37° C with A9 in DMEM supplemented with 20% heat inactivated fetal bovine serum, using an initial inoculum from primary cultures of lung or blood clots from infected spiny mice. The cultures were maintained for 115 days and underwent 15 passages before termination and cryopreservation. Using this culture system T. acomys subcultures were initiated from 3 different initial inocula (3 × 104, 1.5 × 105 and 7.4 × 105 parasites/ml) and growth curves revealed that the lowest inoculum gave the best growth pattern. This inoculum yielded a population doubling time of less than 12 h for 4 days, a high peak density of 7 × 106 parasites/ml and the most gradual decline compared to the other 2 inocula. Rosetting epimastigotes and nests of amastigotes were observed in close association with the feeder layer cells. Epimastigotes were the most predominant form in culture supernatants but other morphological forms observed included trypomastigotes and sphaeromastigotes.
Ultrastructural Study of the Development of Vairimorpha necatrix (Kramer, 1965) (Protozoa, Microsporida) in Larvae of the Corn Earworm, Heliothis zea (Boddie) (Lepidoptera, Noctuidae) with Emphasis on Sporogony
Authors: MITCHELL MAUREEN J. , CALI ANN 
Published: May 2007

ABSTRACTThe ultrastructure of the life cycle of Vairimorpha necatrix (Kramer) was studied in Heliothis (Helicoverpa) zea larvae raised at temperatures ranging from 15° C to 30° C. In contrast to the description of the type species, both patterns of development (Nosema‐like and Thelohania‐like) that characterize the genus Vairimorpha were present at all temperatures examined. No temperature‐related differences in the ultrastructure of Vairimorpha necatrix were observed; however, the effects of temperature variation were apparent in 1) the time between inoculation with V. necatrix and the initiation of the parasite life cycle and 2) the duration (or length) of the parasite life cycle. An additional form of the parasite, that of a diplokaryon in a sporophorous vesicle, was present during early sporogony. The nuclei of the diplokaryon separated followed by cytokinesis, forming two monokaryotic parasites, and eventually two plasmodia, within the sporophorous vesicle. This pattern of development is characteristic neither of Nosema nor Thelohania, nor has it been described for Vairimorpha.
Production and Characterization of Monoclonal Antibodies Against a Highly Immunogenic Fraction of Entamoeba histolytica (NIH:200) and Their Application in the Detection of Current Amoebic Infection
Authors: SENGUPTA KRISANU , DAS PRADEEP , JOHNSON TONY M. , CHAUDHURI PARTHA P. , DAS DEBASISH , NAIR G. BALAKRISH 
Published: May 2007

ABSTRACTSix monoclonal antibodies (MAbs) were produced against a highly immunogenic fraction derived by the chromatographic separation of the soluble preparation of axenic Entamoeba histolytica (strain NIH:200) trophozoites. Isotype characterization of the six MAbs revealed that four belonged to the IgM class and one each to the IgG1 and the IgG2a subclasses. The immunoreactivity patterns and the specificity of the MAbs with homologous and heterologous antigens were analyzed by the enzyme‐linked immunotransfer blot technique and by the enzyme‐linked immunosorbent assay. The MAbs reacted intensely with isolates of E. histolytica (strain NIH:200 as well as a local isolate MX1) but showed no reactivity with Entamoeba coli, Iodamoeba butschlii, Endolimax nana, Entamoeba hartmanni, free‐living amoeba (Acanthamoeba harticolus) and other enteric parasites. Using the IgG1 MAb as a detecting antibody, a polyclonal‐monoclonal antibody‐based enzyme‐linked immunosorbent assay was developed for the detection of E. histolytica antigens in stool samples of infected patients. The detection limit of the assay was 8 ng of amoebic antigen. This test was found to be specific and sensitive and yielded 100% positive results in cases with amoebiasis but did not react with controls included in the evaluation. The MAb‐based enzyme‐linked immunosorbent assay developed in this study will be an important test for the diagnosis of E. histolytica in the feces of infected humans; however, the limitation of the test is the inability to discriminate the pathogenic status of the amoeba detected in the stool.
Purification and Characterization of Phospholipase C Preferentially Hydrolysing Phosphatidylcholine in Tetrahymena Membranes
Authors: ALAM SHARIFUL , BANNO YOSHIKO , NOZAWA YOSHINORI 
Published: May 2007

ABSTRACTA phospholipase C (PLC) activity that preferentially hydrolyses phosphatidylcholine to diacylglycerol and phosphorylcholine was found to be present in Tetrahymena pyriformis, strain W and most of its activity was recovered in the membrane fraction. This enzyme was extracted with 1% Triton X‐100 from the membrane fraction and purified to apparent homogeneity by sequential chromatographies on Fast Q‐Sepharose, hydroxyapatite HCA‐100S, Mono Q and Superose 12 gel filtration columns. The purified enzyme had specific activity of 2083 nmol of diacylglycerol released/mg of protein/min for dipalmitoylphosphatidylcholine hydrolysis. Its apparent molecular mass was 128 kDa as determined by SDS‐polyacrylamide gel electrophoresis and was 127 kDa by gel filtration chromatography, indicating that the enzyme is present in a monomeric form. The enzyme exhibited an optimum pH 7.0 and the apparent Km value was determined to be 166 μM for dipalmitoylphosphatidylcholine. A marked increase was observed in phosphatidylcholine hydrolytic activity in the presence of 0.05% (1.2 mM) deoxycholate. Ca2+ but not Mg2+ enhanced the activity at a concentration of 2 mM. This purified phospholipase C exhibited a preferential hydrolytic activity for phosphatidylcholine but much less activity was observed for phosphatidylinositol (∼ 9%) and phosphatidylethanolamine (∼ 2%).
Pharmacological Characterization of an Opioid Receptor in the Ciliate Tetrahymena
Authors: CHIESA RICARDO , SILVA WALTER I. , RENAUD FERNANDO L. 
Published: May 2007

ABSTRACTA pharmacological characterization has been performed of the opioid receptor involved in modulation of phagocytosis in the protozoan ciliate Tetrahymena. Studies on inhibition of phagocytosis by mammalian prototypic opioid agonists revealed that morphine and β‐endorphin have the highest intrinsic activity, whereas all the other opioids tested can only be considered partial agonists. However, morphine (a mu‐receptor agonist) is twice as potent as β‐endorphin (a delta‐receptor agonist). Furthermore, the sensitivity for the opioid antagonist naloxone, determined in the presence of morphine and β‐endorphin, is very similar to the sensitivity exhibited by mammalian tissues rich in mu‐opioid receptors. We suggest that the opioid receptor coupled to phagocytosis in Tetrahymena is mulike in some of its pharmacological characteristics and may serve as a model system for studies on opioid receptor function and evolution.
Trypanosoma brucei brucei and T. b. gambiense: Stumpy Bloodstream Forms Express More CB1 Epitope in Endosomes and Lysosomes than Slender Forms
Authors: BRICKMAN MARLA J. , BALBER ANDREW E. 
Published: May 2007

ABSTRACTCB1‐glycoprotein is a component of flagellar pocket, endosome, and lysosome membranes of long, slender bloodstream forms of the Trypanosoma brucei subgroup of African trypanosomes. We have used immunoblotting, immunofluorescence, and cryoimmunoelectron microscopy to study CB1‐glycoprotein expression as long, slender bloodstream forms of pleomorphic T. b. brucei and T. b. gambiense transform through intermediate stages into short, stumpy forms. Intermediate and stumpy forms express more CB1‐glycoprotein than long, slender forms. These results, coupled with previous work showing that procyclic forms do not express CB1‐glycoprotein, show that the expression of lysosomal membrane glycoproteins is regulated coordinately with other aspects of lysosome and endosome function as these trypanosomes go through their life cycle.
Artificial Infections of Pneumocystis carinii in the SCID Mouse and their Use in the In Vivo Evaluation of Antipneumocystis Drugs
Authors: COMLEY JOHN C. W. , STERLING ALETHEA M. 
Published: May 2007

ABSTRACTA model for the in vivo evaluation of antipneumocystis drugs has been developed in SCID mice infected intratracheally with cryopreserved mouse‐derived Pneumocystis carinii. The development of a highly reproducible fatal P. carinii pneumonia occured within 10 weeks (mean survival time ± SEM = 72.2 ± 1.2 days). Continuous administration of dexamethasone (2 mg/liter in the drinking water) exacerbated the rate of onset of severe P. carinii pneumonia (mean survival time ± SEM = 63 ± 1.3 days) in SCID mice. The number of cysts per g of lung homogenate (homogenate counts) were maximal with an inoculum of 20,000 cysts at 6 weeks post infection. Homogenate counts correlated with infection scores (graded assessments of immunofluorescent cysts on lung impression smears) suggesting that infection scoring accurately and rapidly reflects the severity of P. carinii pneumonia in SCID mice. These studies led to the development of a drug screening protocol in which Pneumocystis‐free female SCID mice (20–25 g) were started on dexamethasone 7 days prior to IT inoculation with a single dose of 20,000 cysts. Drugs were evaluated either for: a) prophylaxis (continuously from day 1 post infection) or b) treatment (from day 21 post infection) until day 42 post infection, when all mice were killed and infection scores determined. Co‐trimoxazole (at 250 mg sulfamethoxazole + 50 mg trimethoprim/kg/day) given in the drinking water was found to be highly effective in both the prophylaxis and treatment of mouse P. carinii pneumonia. Co‐trimoxazole remained very effective in the prophylaxis P. carinii pneumonia in the SCID mouse at 125 mg sulfamethoxazole + 25 mg trimethoprim/kg/day p.o. and showed some enhancement of efficacy over sulfamethoxazole alone at 125 mg/kg/day p.o., suggesting limited synergy between sulfamethoxazole and trimethoprim. The results presented provide confirmation of the usefulness and predictability of the model.
Molecular Characterization of a Cytoplasmic Dynein from Dictyostelium
Authors: KOONCE MICHAEL P. , GRISSOM PAULA M. , LYON MARY , POPE THERESA , McINTOSH J. RICHARD 
Published: May 2007

ABSTRACTCytoplasmic dynein is a high molecular weight, microtubule‐based mechanochemical ATPase that is believed to provide motive force for a number of intracellular motilities, including transport of membrane‐bound organelles. Cytoplasmic dynein also localizes to the mitotic spindles of some organisms and to the kinetochore regions of some condensed chromosomes, where it may play an active role in spindle assembly, spindle position, and/or chromosome movement during cell division. Despite active research efforts from a number of laboratories, little detail is yet available about dynein‐based cellular activities. This paper describes our efforts to characterize cytoplasmic dynein from Dictyostelium and to use this protist as a molecular genetic factory to probe structure‐function relationships of this molecule.
Effect and Localization of Trifluralin in Plasmodium falciparum Gametocytes: An Electron Microscopic Study1
Authors: KAIDOH TOSHIYUKI , NATH JAYASREE , FUJIOKA HISASHI , OKOYE VINCENT , AIKAWA MASAMICHI 
Published: May 2007

ABSTRACTTrifluralin, a herbicide which is known to bind to plant and algal tubulin, induced ultrastructural changes in the microtubules of the mature Plasmodium falciparum gametocytes in vitro. Trifluralin treatment led to disassembly of the well ordered subpellicular microtubules, whereas it had no effect on microtubules of human platelets or of rat neuronal cells in vitro. The disassembled microtubules showed fragmented large tubular structures, which frequently were associated with the pellicular membranes. Electron microscopic autoradiography showed radioactive trifluralin associated with the microtubule fragments. These results provide evidence that trifluralin selectively binds to microtubules in malaria parasites and causes disruption of their structure.
Molecular Evolutionary Analyses of Nuclear‐Encoded Small Subunit Ribosomal RNA Identify an Independent Rhizopod Lineage Containing the Euglyphina and the Chlorarachniophyta
Authors: BHATTACHARYA DEBASHISH , HELMCHEN THOMAS , MELKONIAN MICHAEL 
Published: May 2007

ABSTRACTThe Rhizopoda comprise a diverse assemblage of protists which depend on lobose or filose pseudopodia for locomotion. The biochemical and morphological diversity of rhizopods has led to an uncertain taxonomy. Ribosomal RNA sequence comparisons offer a measure of evolutionary relatedness that is independent of morphology and has been used to demonstrate a polyphyletic origin of the Lobosea. We sequenced complete small subunit ribosomal RNA coding regions from the filose amoebae, Euglypha rotunda and Paulinella chromatophora (Euglyphina) to position these taxa in the eukaryote phylogeny. The neighbor‐joining analyses show that E. rotunda and P. chromatophora share a monophyletic origin and are not closely related to any lobose amoebae in our analyses. Instead, the Euglyphina form a robust sister group to the Chlorarachniophyta. These results provide further evidence for the polyphyly of the Rhizopoda and support the creation of a new amoeboid lineage which includes the Euglyphina and the chlorarachniophyte algae; taxa with tubular mitochondrial cristae and filose or reticulate pseudopodia.
DNA Polymorphism Revealed by Arbitrary Primers Polymerase Chain Reaction Among Blastocystis Strains Isolated from Humans, a Chicken, and a Reptile
Authors: YOSHIKAWA HISAO , NAGONO ISAO , YAP EU HIAN , SINGH MULKIT , TAKAHASHI YUZO 
Published: May 2007

ABSTRACTDNA polymorphisms of different strains of Blastocystis isolated from humans, a chicken, and a reptile were examined by an arbitrary primer PCR method. Two strains of Blastocystis hominis isolated from humans in the USA and Japan yielded nearly identical PCR products. However, one strain of B. hominis (isolated from a human in Singapore) yielded quite different PCR products. Blastocystis sp. isolated from a chicken yielded PCR products similar to those of the former two strains, while Blastocystis lapemi, isolated from a reptile, shared no bands with any of the other isolates. These results indicate the possibility that our isolate from the chicken is a zoonotic strain, and that there is intraspecific variation of Blastocystis hominis.
Diagnosis of PCP by ITSs Nested PCR on Notinvasive Oropharingeal Samples
Authors: ATZORI CHIARA , AGOSTONI FIORENZA , GUBERTINI GUIDO , CARGNEL ANTONIETTA 
Published: May 2007

SUMMARY Pneumocystis carinii pneumonia is an opportunistic disease usually affecting immunocompromised hosts. Diagnosis is based on the microscopical detection of microrganism on BAL, an invasive sample which requires pt compliance for collection and skilled health care personnel for examination. In order to verify the possible diagnostic utility of noninvasive specimens collected in the upper respiratory tract we examined by the Internal Transcribed Spacers (ITSs) nested PCR, 39 oropharingeal samples (garglings) collected from 20 HIVAb positive pts and from 15 healthy controls.
Algorithmic Analysis of in Vitro Chemotherapy Against Pneumocystis carinii
Authors: ATZORI CHIARA , AGOSTONI FIORENZA , ANGELI ELENA , CARGNEL ANTONIETTA , DRAGO LORENZO , GISMONDO MARIA R. 
Published: May 2007

SUMMARYThe development of in vitro experimental chemotherapy against P.carinii has been hindered by the lack of efficient methods for continous cultivation of the microorganism. Various short‐term systems, allowing the production of infectious forms of Pneumocystis, can be employed for in vitro experimental chemotherapy [1,2,3]. The purpose of the present study is to describe a statistically relevant algorithmic model for the evaluation of P.carinii in short‐term in vitro culture and drug screening.
In Vitro Cultivation of Cryptosporidium parvum in the Non‐Adherent Human Monocytic THP‐1 Cell Line
Authors: LAWTON PHILIPPE , NACIRI MURIEL , MANCASSOLA ROSELYNE , PETAVY ANNE‐FRANCOISE 
Published: May 2007

SUMMARYThis study shows that the human monocytic cell line THP‐1 supports the growth of C. parvum.. Immunofluorescence controls showed that only source oocytsts remained after infection and disappeared within the first 24 h of culture. A continuous asexual life cycle proceeded throughout the experiments, with at least 15‐d cultures. This model provides a useful tool for studies on the biologyof C. parvum in cells involved in its transport in immunocompromised host.
The Feeding Ecology of Actinophrys sol (Sarcodina: Heliozoa) in Chesapeake Bay
Authors: PIERCE RICHARD W. , COATS D. WAYNE 
Published: May 2007

ABSTRACTThe occurrence of Actinophrys sol, a planktonic heliozoan, in Chesapeake Bay was monitored over a four‐year period (1988–1991). Actinophrys sol was widely‐distributed throughout Chesapeake Bay and could exceed densities of 5,000 cells liter −1. It was most abundant during the warmer months. Feeding experiments were conducted with field populations of heliozoa using 1‐μm fluorescent microspheres to label ciliate prey. Two ciliates, a small Strobilidium sp. (30 μ in diameter) and a Pleuronema sp. (45 μ length), were the primary ciliate‐prey items in the water column when the experiments were conducted, although a wide range of ciliate taxa was ingested. Two other ciliates not present in situ, a Cyclidium sp. (20 μ length) and a Uronema sp. (40 μ length), were also labeled and added at various concentrations to field populations of plankton containing A. sol. Heliozoan ingestion rates on in situ prey at concentrations of 30 Strohilidium and one Pleuronema ml−1 were 0.2 to 0.3 prey heliozoan−1 hour−1. Ingestion rates increased to a maximum of 1.2 prey heliozoan −1 hour−1 with additions of 100 Uronema ml−1. A mean clearance rate of 0.15 ml heliozoan−1 day−1 did not change with increasing prey abundance. The abundance and distribution of A. sol suggests that these sarcodines may exert strong grazing pressure on the planktonic ciliate populations of Chesapeake Bay at certain times of the year, and may be important in shaping the ciliate community composition and distribution.
Construction of a Normalized cDNA Library for the Trypanosoma cruzi Genome Project
Authors: ÜRMÉNYI TURÁN P. , BONALDO MARIA F. , SOARES MARCELO B. , RONDINELLI EDSON 
Published: May 2007

ABSTRACTSequencing of the Trypanosoma cruzi genome is underway. Expressed sequence tags, obtained from cDNA libraries, facilitate mapping and gene discovery. The efficiency of large‐scale generation of such tags is increased when using normalized cDNA libraries, where the frequency of individual clones is brought within a narrow range. Repetitive sequencing of abundant clones is therefore minimized. We constructed a normalized cDNA library from epimastigotes of clone CL Brener, and the efficiency of normalization of representative clones was assessed and shown to be adequate. The normalized cDNA library has been distributed to several groups and large‐scale sequencing is currently in progress.
Relationship between periodontal findings and Behçet's disease: a controlled study
Authors: Akman Ayse , Kacaroglu Hasan , Donmez Levent , Bacanli Ali , Alpsoy Erkan 
Published: May 2007

AbstractBackground: Behçet's disease (BD) is a chronic, relapsing, systemic vasculitis of unknown aetiology. The involvement of oral mucosal surfaces represents the onset feature of the disease in the majority of patients.Objective: The aim of this study was to evaluate the periodontal status of BD patients and then compare with recurrent aphthous stomatitis (RAS) patients and healthy controls. We also determined the relationship between the periodontal condition and the clinical severity of the disease in BD patients.Methods: Eighty‐six patients with BD, 63 patients with RAS and 82 healthy subjects were included in the study. The periodontal status of all subjects was evaluated according to the community periodontal index of treatment needs (CPITN). BD patients were also assessed for clinical severity score (CSS) as described previously.Results: The mean CPITN were observed to be higher in BD patients (1.79 ± 0.96) compared with RAS patients (1.22 ± 0.87) and healthy controls (1.18 ± 0.98) (p<0.001). There was a positive association between CSS and CPITN (p=0.017) in BD patients.Conclusion: Our results showed that periodondal status is worse in BD patients and associated with disease severity. We can speculate that periodontitis may induce a systemic inflammatory process that may contribute to the development and/or progression of BD.