A total of 10.140 Anopheles mosquitoes were collected during the period 1985-1995 in 5 study sites. ELISA(+) reactions were detected in 229 mosquito samples from 10 Anopheles species of An.minimus, An.dirus, An.vagns, An.maculatus, An.subpictus, An.campestris, An.tessellatus, An.jeyporiensis, An.aconitus, An.spl. In Bai Tranh (Thanh Hoa province), Khanh Phu and Khanh Nam (Khanh Hoa province), 7 species were found to be infected with malaria sporozoites as An.minimus, An.dirus, An.vagus, An.maculatus. An.aconitus, An.tessellatus, An.jeyporiensis. In the brackish water commune Phong Phu (Binh Chanh district, HCM city) and in Can Giuoc district (Long An province), 7 species were found to be infected with malaria sporozoites as An. subpictus, An.sinensis, An.sundaicus, An.campestris, An.vagus, An.tessellatuss, An.spl. The positive infection rate was An.dirus (4,5% - 8,5%), An.minimus (8,3%) in Khanh Phu and Khanh Nam (Khánh Hòa) and An.sinensis (4,5% - 11%). An.campestris (6,6% - 6,7%), An.vagus (4,6%) in Phong Phu (HCM city) and Can Giuoc (Long An)
malaria ; diagnosis ; malaria, falciparum ; mosquitoes ; Sporozoites ; Enzyme-Linked Immunosorbent Assay ; malaria ; malaria, falciparum ; mosquitoes ; Sporozoites ; malaria, vivax

A prerequisite for the development of a cancer cell selective targeting adenovirus is the generation of adenoviral vectors that lack native receptor binding ability and additionally contain domains redirecting the vector to cancer cell specific receptors. Towards this goal, we have generated an E1B 55kDa-deleted oncolytic and coxoackie and adenovirus receptor (CAR)-binding ablated adenovirus, YKL-K420A. This newly engineered adenovirus resulted in a dramatic reduction of transduction efficiency compared to the control adenovirus, YKL-1, in all of the cell lines tested. The malaria circumsporozoite (CS) protein interacts with glycosaminoglycans (GAG) present on the liver cell surface, and plays a prominent role in sporozoite attachment and invasion into hepatocytes. To redirect the CAR binding ablated adenovirus YKL-K420A to hepatocytes, CS protein epitope (EWSPCSVTCGNGIQVRIK) was incorporated onto the C-terminus of the YKL-K420A fiber protein, generating an YKL-K420A-hepa. The In vitro efficacy and specificity of YKL-K420A-hepa was then evaluated by comparing the cytopathic effect in hepatoma and other cancer cells from different origins. In hepatoma cells, YKL-K420A-hepa exerted upto 20-fold higher cytolytic ability compared to the control adenovirus, YKL-1, in hepatoma cell lines. Treatment with YKL-K420A-hepa also significantly suppressed tumor growth in a hepatoma xenograft tumor model when compared to YKL-1. Taken together, these studies demonstrate that the strategy to alter adenovirus tropism may greatly improve adenoviral utilities in gene therapy applications.
Adenoviridae* ; Carcinoma, Hepatocellular ; Cell Line ; Genetic Therapy ; Glycosaminoglycans ; Hepatocytes ; Heterografts ; Liver ; Malaria ; Sensitivity and Specificity ; Sporozoites ; Tropism

The aim of this study was to identify antigens for a vaccine or drug target to control rabbit coccidiosis. A combination of 2-dimensional electrophoresis, immunoblotting, and mass spectrometric analysis were used to identify novel antigens from the sporozoites of Eimeria stiedae. Protein spots were recognized by the sera of New Zealand rabbits infected artificially with E. stiedae. The proteins were characterized by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF/TOF-MS) analysis in combination with bioinformatics. Approximately 868 protein spots were detected by silver-staining, and a total of 41 immunoreactive protein spots were recognized by anti-E. stiedae sera. Finally, 23 protein spots were successfully identified. The proteins such as heat shock protein 70 and aspartyl protease may have potential as immunodiagnostic or vaccine antigens. The immunoreactive proteins were found to possess a wide range of biological functions. This study is the first to report the proteins recognized by sera of infected rabbits with E. stiedae, which might be helpful in identifying potential targets for vaccine development to control rabbit coccidiosis.
Coccidiosis* ; Computational Biology ; Eimeria ; Electrophoresis ; HSP70 Heat-Shock Proteins ; Immunoblotting ; Mass Spectrometry ; Rabbits ; Sporozoites

The cytological and ultrastructural findings of Pneumocystis carinii(PC) obtained from rats by bronchoalveolar lavage(BAL) are described. All developmental forms of the PC organisms were obtained in the lavage fluid. Ultrastructurally, the cysts were almost circular in shape, and were nearly devoid of surface tubular extensions. The wall of the cyst was composed of an unit membrane, and intermediate electron lucent layer and an external electron dense layer. The cysts frequently contained intracystic bodies, so called sporozoites. Occasionally empty or collapsed cysts with no intracystic bodies, and precysts were found. Trophozoites were variable in size and shape with abundant tubular extensions along the single electron dense pellicle. BAL is a useful method for concentrating the various morphologic forms of PC organisms, and is a rapid diagnostic method for PC pneumonia.
Animals ; Bronchoalveolar Lavage* ; Membranes ; Pneumocystis carinii* ; Pneumocystis* ; Pneumonia ; Pneumonia, Pneumocystis* ; Rats ; Sporozoites ; Therapeutic Irrigation ; Trophozoites

In Tansan and Wondang m 1960, and m Guidandong and Yongju-eup in 1961, routine entomological work was carried out according to the plan of operation for the ma1aria pre-eradication survey. During the present work, six anopheline mosquito species were recorded as follows: 1. Anopheles (Anopheles) sinensis Wiedmann, 1828. 2. Anopheles (Anopheles) sineroides Yamada, 1925. 3. Anopheles (Anopheles) lesteri Baisas and Hu, 1936. 4. Anopheles (Anopheles) koreicus koreicus Yamada and Watanabe, 1918. 5. Anopheles (Anopheles) koreicus edwardsi Yamada, 1925, and, 6. Anopheles (Anopheles) lindesayijaponicus, Yamada, 1918. A. sinensis is the most predominant species, although A. koreicus koreicus was also found to be predominant after A. sinensis in Guidandong (a mountainous area) A. sineroides is the next most predominant species after A. sinensis. Anopheline mosquitos begin to appear from late April or May and disappear in October each year. The resting places for the anopheline mosquitos are mainly cow sheds and outdoors. The population density of A. sinensis in cow sheds shows a peak either in June or in July in most places with a second small peak in late August or in September. Night biting habits appear to be active throughout the whole night but are more active from sunset to midnight. Most of the anophelines caught appeared to be zoophilic; however, the results of precipitin tests for A. sinensis showed a likelihood that these are facul-tative anthropophilic. Dissection of salivary glands in the present study of 2736 female A. sinensis mosquitos failed to show or to prove the presence of sporozoites, although sinensis is suspected as a potential of malaria. The body weight, moisture and fat content in A. sinensis appeared to decrease in July from a high peak in June and then to increase again m September. Insecticide susceptibility tests proved that the species was susceptible to DDT and Dieldrin in Guidandong and Yoju. The bionomics of A. sineroides, A. koreicus koreicus, A. koreicus edwardsi, A. lesteri and A. Iindesayi ja-ponicus was discussed; the latter two species are probably the first to be recorded in Korea. The mosquitos caught in hibernating places were found to be nulliparous and to have sperms in the spermathecae during the winter months. Anopheline hibernated probably in the adult stage.
Adult ; Anopheles* ; Body Weight ; Culicidae* ; DDT ; Dieldrin ; Ecology ; Female ; Humans ; Korea* ; Malaria ; Population Density ; Precipitin Tests ; Salivary Glands ; Spermatozoa ; Sporozoites

OBJECTIVETo identify the biological forms, sporozoite rate and molecular characterization of the Anopheles stephensi (An. stephensi) in Hormozgan and Sistan-Baluchistan provinces, the most important malarious areas in Iran.

METHODSWild live An. stephensi samples were collected from different malarious areas in southern Iran. The biological forms were identified based on number of egg-ridges. Molecular characterization of biological forms was verified by analysis of the mitochondrial cytochrome oxidase subunit I and II (mtDNA-COI/COII). The Plasmodium infection was examined in the wild female specimens by species-specific nested-PCR method.

RESULTSResults showed that all three biological forms including mysorensis, intermediate and type are present in the study areas. Molecular investigations revealed no genetic variation between mtDNA COI/COII sequences of the biological forms and no Plasmodium parasites was detected in the collected mosquito samples.

CONCLUSIONSPresence of three biological forms with identical sequences showed that the known biological forms belong to a single taxon and the various vectorial capacities reported for these forms are more likely corresponded to other epidemiological factors than to the morphotype of the populations. Lack of malaria parasite infection in An. stephensi, the most important vector of malaria, may be partly due to the success and achievement of ongoing active malaria control program in the region.

Animals ; Anopheles ; genetics ; parasitology ; DNA, Mitochondrial ; genetics ; DNA, Protozoan ; genetics ; Eggs ; classification ; parasitology ; Female ; Iran ; Male ; Parasite Load ; Plasmodium ; genetics ; isolation & purification ; Polymerase Chain Reaction ; Sporozoites

BACKGROUND: More than 200 million people suffer from malaria worldwide. In Korea this infectious disease had been on a decreasing trend since 1930s and was considered to be eradicated from 1985. However, since 1993 when a case of malaria was reported, its incidence is progressively increased. Recent efforts have been focused on the development of vaccine against the infective sporozoite stage of Plasmodium vivax. It has been found that sporozoites are complicated with genetic variation within the circumsporozoite gene and phenotypic heterogeneity in the protein it encodes. So, we investigated the distribution of circumsporozoite gene of Plasmodium vivax in Korea. METHODS: Polymerase chain reactions (PCR) were performed on samples confirmed with microscopic examination for P. vivax and negative samples with clinical and microscopic examination. The amplified products were analyzed by dot hybridization with oligonucleotide probes VK210 and VK247 which are respectively complementary to the predominant and variant strains of the circumsporozoite gene of P. vivax. RESULTS: The incidence of isolation in VK210 and VK247 strains were 96.3%, respectively and individuals who were infected with both strains were 92.7%. Compared to the microscopic examination, the results of PCR showed 82% in sensitivity, 100% in specificity. CONCLUSIONS: The present study suggests that a single-epitope vaccine based on either one circumsporozoite domain is unlikely to be protective because both VK210 and VK247 strains of P. vivax were found widely in Korea. The PCR method appears not to be feasible as a screening, but suitable as a confirmatory test for the identification of Plasmodium species.
Communicable Diseases ; Genetic Variation ; Genotype* ; Incidence ; Korea* ; Malaria ; Mass Screening ; Oligonucleotide Probes ; Plasmodium vivax* ; Plasmodium* ; Polymerase Chain Reaction ; Population Characteristics ; Sensitivity and Specificity ; Sporozoites

Despite the development of new technologies, new challenges still remain for large scale proteomic profiling when dealing with complex biological mixtures. Fractionation prior to liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis is usually the preferred method to reduce the complexity of any biological sample. In this study, a gel LC-MS/MS approach was used to explore the stage specific proteome of Cryptosporidium (C.) parvum. To accomplish this, the sporozoite protein of C. parvum was first fractionated using SDS-PAGE with subsequent LC-MS/MS analysis. A total of 135 protein hits were recorded from 20 gel slices (from same gel lane), with many hits occurring in more than one band. Excluding all non-Cryptosporidium entries and proteins with multiple hits, 33 separate C. parvum entries were identified during the study. The overall goal of this study was to reduce sample complexity by protein fractionation and increase the possibility of detecting proteins present in lower abundance in a complex protein mixture.
Chemical Fractionation/methods ; Chromatography, Liquid/methods/veterinary ; Cryptosporidium parvum/*chemistry/growth & development/metabolism ; Electrophoresis, Polyacrylamide Gel/methods/veterinary ; Gene Expression Profiling/*methods/veterinary ; Proteome/analysis ; Proteomics/*methods ; Protozoan Proteins/*analysis ; Sporozoites/chemistry/metabolism ; Tandem Mass Spectrometry/methods/veterinary

Malaria is an infectious disease affecting humans, which is transmitted by the bite of Anopheles mosquitoes harboring sporozoites of parasitic protozoans belonging to the genus Plasmodium. Despite past achievements to control the protozoan disease, malaria still remains a significant health threat up to now. In this study, we cloned and characterized the full-unit Plasmodium yoelii genes encoding merozoite surface protein 1 (MSP1), circumsporozoite protein (CSP), and Duffy-binding protein (DBP), each of which can be applied for investigations to obtain potent protective vaccines in the rodent malaria model, due to their specific expression patterns during the parasite life cycle. Recombinant fragments corresponding to the middle and C-terminal regions of PyMSP1 and PyCSP, respectively, displayed strong reactivity against P. yoelii-infected mice sera. Specific native antigens invoking strong humoral immune response during the primary and secondary infections of P. yoelii were also abundantly detected in experimental ICR mice. The low or negligible parasitemia observed in the secondary infected mice was likely to result from the neutralizing action of the protective antibodies. Identification of these antigenic proteins might provide the necessary information and means to characterize additional vaccine candidate antigens, selected solely on their ability to produce the protective antibodies.
Animals ; Anopheles ; Antibodies ; Clone Cells ; Coinfection ; Communicable Diseases ; Culicidae ; Humans ; Immunity, Humoral ; Life Cycle Stages ; Malaria ; Merozoite Surface Protein 1* ; Mice ; Mice, Inbred ICR ; Parasitemia ; Parasites ; Plasmodium yoelii* ; Plasmodium* ; Rodentia ; Sporozoites ; Vaccines

The antigen location of Cryptosporidium parvum, which stimulates antibody formation in humans and animals, was investigated using infected human sera. Immuno-electron microscopy revealed that antigenicity-inducing humoral immunity was located at various developmental stages of parasites, including asexual, sexual stages, and oocysts. The amount of antigen-stimulating IgG antibodies was particularly high on the oocyst wall. The sporozoite surface was shown to give stimulation on IgG and IgM antibody formation. Trophozoites implicated the lowest antigenicity to humoral immunity, both IgG and IgM, by showing the least amount of gold labeling. Immunogold labeling also provided clues that antigens were presented to the host-cell cytoplasm via feeder organelles and host-parasite junctions.
Animals ; Antibodies, Protozoan/*immunology ; Antigens, Protozoan/*analysis ; Cryptosporidium parvum/*chemistry/*immunology/ultrastructure ; Female ; Humans ; Immunoglobulin G/immunology ; Immunoglobulin M/immunology ; Mice ; Microscopy, Immunoelectron ; Sporozoites/chemistry/immunology/ultrastructure ; Staining and Labeling/methods ; Trophozoites/chemistry/immunology/ultrastructure