Ascaris spp. and Capillaria caudinflata infections in captive-bred crested ibis (Nipponia nippon) in China.

Crested ibis (Nipponia nippon), an endan gered native bird, was called the "precious stone" of oriental birds. N. nippon was considered a critically endangered species in the IUCN Red List of Threatened Species and a first-class national protected animal in China. The Chinese government had exerted considerable effort to protect the N. nippon population. An effective approach to increase the number of these birds was captive breeding. However, several pathogens, including parasites, could jeopardize the health of this species. The present study used the fecal flotation method to determine prevalence of intestinal parasites in fresh stool samples by wet mount smearing and iodine staining. Samples were obtained from 63 randomly selected crested ibis bred in Shaanxi Rare Wildlife Rescuing and Breeding Research Center in Zhouzhi County, Xi'an City, Shaanxi Province, China. In the 63 captive individuals, 38 were found positive for intestinal parasites (60.3%, 38/63). Of positive birds, high prevalence of Ascaris spp. (84.2%, 32/38) and Capillaria caudinflata (50.0%, 19/38) were detected. Coccidea (7.8%, 3/38), Fasciolidae (23.7%, 9/38), Blastocystis spp. (15.8%, 6/38), and Entamoeba histolytica (7.8%, 3/38) showed relatively low prevalence rates. This study focuses on the morphological identification of Ascaris spp. and C. caudinflata and their transmission in the N. nippon population. We introduce strategies to improve the breeding management of the birds, enhance their health, and stimulate population productivity.

Encystation--survival of Blastocystis hominis in immunocompetent mice abdomen cavity.

Human Blastocystis hominis were isolated from diarrhea patients' feces and cultured in vitro. Then the cultures were inoculated intraperitoneally to laboratory mice. The B. hominis in living mice were collected and inoculated again to healthy mice. The B. hominis showed dose-dependent pathogenicity in the primary inoculation. No pathogenicity was observed in the secondary inoculation. The protozoan existed in the living mice abdomen cavity for more than 6 months and the cyst was the only form. These results showed that encystation enable the parasite to avoid the immune attack in competent host and simultaneously decrease the pathogenicity to host. Intraperitoneal inoculation to laboratory mice is a good method to maintain and propagate B. hominis. This is also a good model to study the interaction of B. hominis and immune system.

Broad profiling of DNA-binding transcription factor activities improves regulatory network construction in adult mouse tissues.

Molecular systematics involves the description of the regulatory networks formed by the interconnections between active transcription factors and their target expressed genes. Here, we have determined the activities of 200 different transcription factors in six mouse tissues using an advanced mouse oligonucleotide array-based transcription factor assay (MOUSE OATFA). The transcription factor signatures from MOUSE OATFA were combined with public mRNA expression profiles to construct experimental transcriptional regulatory networks in each tissue. SRF-centered regulatory networks constructed for lung and skeletal muscle with OATFA data were confirmed by ChIP assays, and revealed examples of novel networks of expressed genes coregulated by sets of transcription factors. The combination of MOUSE OATFA with bioinformatics analysis of expressed genes provides a new paradigm for the comprehensive prediction of the transcriptional systems and their regulatory pathways in mouse.

Novel high-throughput profiling of human transcription factors and its use for systematic pathway mapping.

Transcription factors (TFs) are crucial components of regulatory networks that control gene transcription. Current TF assays are limited to the analysis of a single TF or require TF-specific antibodies. Here we report the Single Primer Amplification assisted Oligonucleotide Array-based Transcription Factor Assay (SPA-OATFA) which can directly analyze the binding activities of 240 human TFs simultaneously. Examining early events during serum-stimulation of HeLa cells as a model, we demonstrated the utility of SPA-OATFA combined with whole genome gene expression to systematically map the temporal activation of signaling pathways. Both TFs known to function in this stimulation response such as EGR1 and AP1 and new TFs such as HSF1 were identified. This information, combined with mRNA profiling, provided novel insights into the activities of regulatory pathways, and illustrates the potential of SPA-OATFA in detailed systems biology analysis of cell responses.

[Study on the biological characteristic of Blastocystis hominis: morphology, mode of reproduction and the relation to bacteria].

OBJECTIVE: To observe the reproductive modes of Blastocystis hominis and study the relation between this protozoa and bacteria. METHODS: Using the Iodine and Haematoxylin staining, the morphology of B. h from patients and RPMI 1640 medium were observed. The B. h positive mucous diarrheal specimens were cultured and identified any possible known pathogenic intestinal bacteria. B. h and colibacillus were co-cultured to observe the interaction between them. RESULTS: Four modes of reproduction for B. h were confirmed: binary fission, endodyogeny, multiple fission and budding. The fact that there was no other intestinal pathogens in half of the B. h positive specimens suggested B. h may cause disease independently. B. h and colibacillus were restrained each other. CONCLUSION: B. h reproduces in at least four modes. B. h could be a pathogen and its pathogenesis may be related to micro-ecological changes.

[Vitro culture of blastocystis hominis in medium DMEM].

OBJECTIVE: To establish the vitro culture of Blastocystis hominis (B. h) in medium DMEM for the further research on diagnosis, life cycle and pathogenicity of this intestinal protoza. METHODS: The growth, reproduction and relevant factors of B . h under different culture conditions including sorts and concentrations of serum, pHs and number of inoculation were compared. RESULTS: Conditions for the continuously anaerobic culture of B. h in medium DMEM were as follows: the number of inoculation were no less than 10(5) cells per tube, pHs ranged 7.0 - 8.0, concentrations of calf serum (or human serum and horse serum) ranged 10% - 30% , antibiotics and Amphotericin B should be added, subculturing could be choose at the each peaking-day 3,6 or 5 at 37 degrees C. CONCLUSION: The medium DMEM could be used in diagnosis and continuously vitro culture for B. h.

Parallel profiling of active transcription factors using an oligonucleotide array-based transcription factor assay (OATFA).

Cell/tissue-specific gene expression are tightly regulated by various combinations of multiple transcription factors (TFs). Here, we present an oligonucleotide array-based transcription factor assay (OATFA), which allows the simultaneous assay of multiple active TFs. In this proof-of-principle work, both purified TFs and cell extracts were analyzed using OATFA and further antibody-based validation confirmed the chip data. This method could simplify the assay of multiple TFs and may facilitate high-throughput profiling of large numbers of TFs.

[Experimental infection of mice with Blastocystis hominis].

OBJECTIVE: To seek a better pathway and proper number of parasites for Blastocystis hominis (B.h) infection in normal and immunocompromised ICR mice. METHODS: (1) 10(4), 10(5) and 10(6) B.h, cultured in RPMI 1640 medium from 3 generations were used to infect mice through oral and rectum; (2) 10(6) B.h were used to infect immunocompromised mice through rectum. The reproduction of B.h in gastrointestinal tract and the pathologic changes in the tissues were observed. RESULTS: Mice were infected by B.h through either oral or rectum. The infected immunocompromised mice showed slow locomotion, depressed, lethargy, and descended body weight. Some infected mice discharged mucus feces, a few of them died during the experiment. Parasites were found in the whole gastrointestinal tract. Severe edema, hyperemia and congestion were observed in the tissues of jejunum, ileum, cecum and colon. The epithelia of small intestine and colonic mucous membrane showed exfoliation, inflammatory cell infiltration in submucosa, and structural changes in glands. CONCLUSION: Mice were more susceptible to Blastocystis hominis infection through rectum than orally. The parasites can be found in the whole gastrointestinal tract of mice, and can breed rapidly and cause significant pathological change in the gastrointestinal mucosa in immunocompromised mice.

[Study on morphology of Blastocystis hominis in culture and from diarrhea patients].

OBJECTIVE: To study morphology of different stage Blastocystis hominis (B. h) for establishing a base in the research of life cycle and pathogenicity of B. h and providing information for clinical laboratory. METHODS: B. h from diarrheal patients was continuously cultured in LES medium, and morphology of B. h was studied with iodine and iron hematoxylin staining under light microscope. RESULTS: The vacuolar, granular, amoeboid and cyst forms of B. h and transformation among the forms were observed microscopically. CONCLUSION: Among different forms, the vacuolar and granular forms were often seen clinically and the vacuolar form can transform to cysts.