Anatomical spatial distribution of Influenza virus receptors in some poultry species raised in Egypt

Background: Avian influenza H5N1 has been distressing not only the poultry industry but also humans causing fatal infections in Egypt. Understanding the initial steps in the viral infection was proposed by many to be a key for solving the entire problem. Domestic healthy chicken, Pekin duck, Egyptian goose, Japanese quail, pigeon and turkey were purchased; three adult birds per each species. Lectin histochemistry was performed using fluorescein isothiocyanate labelled Sambucusnigra agglutinin specific for SAα2,6-gal receptors, and FITC labelled Maackiaamurensis agglutinins specific for SAα2,3-gal receptors. Methods: From each bird, three specimens per each trachea, lung, duodenum, colon, liver and brain were used. In chicken, duck, goose, Japanese quail, domestic pigeon and turkey, both SAα2,3-gal and SAα2,6-gal receptors were expressed in at least one segment of respiratory and intestinal tracts except in pigeons where SAα2,3-gal receptors were not expressed in the respiratory tract while in ducks were not expressed in lower respiratory tract and in turkey not expressed in small intestine. The human type receptors were not expressed in the lower trachea of goose, large intestine of chicken and intestinal tract and liver of turkey and pigeons. Results: The widespread detection of both SAα2,6-gal and SAα2,3-gal receptors in different tissues from each species suggests that these birds’ organs may be potential targets for both avian and human influenza viruses, and can act as adaptive host for avian influenza viruses to change receptor specificity. This may indicate that different native bird species in Egypt could have participated equally or variably in the generation of H5N1 viruses that were able to extensively infect humans. All experimental procedures were approved by Damanhour university ethics committee. The widespread detection of both SAα2,6-gal and SAα2,3-gal receptors in different tissues from each species suggests that these birds’ organs may be potential targets for both avian and human influenza viruses, and can act as adaptive host for avian influenza viruses to change receptor specificity. Conclusion: This may indicate that different native bird species in Egypt could have participated equally or variably in the generation of H5N1 viruses that were able to extensively infect humans.

Immunohistochemical Expression of Synaptophysin in the Adult and Developing Cervical and Lumbar Enlargements of the Spinal Cord of Rabbit

Background: The principle findings of synaptophysin immunoreactivity (SynpIR) during the ontogeny of rabbit spinal cord are: At E14, SynpIR precedes in the entire marginal layer especially at the entrance zone of dorsal root and motor neurite outgrowth emerged from the basal plate. At E21, SynpIR is expressed in the motoneurons of ventral and lateral horns of mantle layer growing into the ventrolateral columns of marginal layer. Methods: We found intensely stained thick tracts and diffuse axons among proliferating neuroblasts of mantle layer. The peripheral parts of ventral horns were occupied with closely packed multipolar neurons from which long dendrites departed toward the surface of marginal layer. Results: At E28, pronounced SynpIR presented in the ventral grey horn while the white matter was faintly stained., meanwhile the dorsal horn was more cellular than ventral and lateral horns. Few intensively SynpIR fibers cross the dorsal and ventral commissures. In adult, profuse SynpIR appeared in the entire grey matter, and stained dendrites departed from neurons in the lateral laminae into the adjacent funiculi as finger-like projections. These projections did not reach the surface, so that the outer one-third to onefourth of the funiculi contained little or no SynpIR. In the periphery of ventral horns, we found large multipolar neurons with faintly stained cytoplasm. The white matter and the neuroepithelial cells surrounding the central canal were almost unstained. Conclusion: Synaptophysinis a reliable marker for fiber outgrowth and synapse formation in therabbit spinal cord, and its differential expression levels is specific and almost completed before birth.

Bovine conceptus of Bos indicus produced by somatic cell nuclear transfer and parthenogenesis present morphological variations since the blastocyst stage / Conceptos bovinos (Bos indicus) produzidos por transferência nuclear de células somáticas e partenogênese apresentam variações morfológicas desde o estágio de blastocisto

In cattle, embryo development is characterized by the appearance of two distinct cell layers, the trophectoderm and the inner cell mass. The latter will undergo differentiation to form the embryonic disc consisting of the epiblast and hypoblast. The aim of this study was to ultrastructurally characterize the bovine embryo from different in vitro production techniques, with emphasis on trophectoderm and inner cell mass cells. Bovine embryos on day 7 (conception = D1) of pregnancy, derived via in vitro production techniques, were fixed for light and transmission electron microscopy processing. Results suggested that embryos produced by nuclear transfer of somatic cells and parthenogenesis showed significant changes in macroscopic and microscopic structure. Size was reduced, and the inner cell mass had no defined shape. Furthermore, organelles responsible for the absorption processes, communication, growth, and cellular metabolism were fewer and had changes in shape, when compared to results in embryos produced by in vitrofertilization. We concluded that embryos produced by parthenogenesis and SCNT exhibit morphological differences when compared with IVF embryos, such as undeveloped blastocoel, poorly defined distribution of ICM, and morphological differences in organelles.

Em bovinos, o desenvolvimento embrionário é caracterizado pelo surgimento de duas camadas distintas, o trofectoderma e a massa celular interna. Este último irá sofrer diferenciação para formar o disco embrionário, o qual consiste em epiblasto e hipoblasto. O objetivo deste estudo foi caracterizar ultraestruturalmente o embrião bovino proveniente de diferentes técnicas de produção in vitro, com ênfase no trofectoderma e na massa celular interna. Embriões bovinos com sete dias de gestação (fecundação = D1), derivados de técnicas de produção in vitro, foram fixados para processamento de microscopia de luz e eletrônica de transmissão. Os resultados sugerem que os embriões produzidos por transferência nuclear de células somáticas e partenogênese apresentaram alterações significativas em suas estruturas macro e microscópica. O tamanho foi reduzido, e a massa celular interna não tinha uma forma definida. Além disso, organelas responsáveis por processos de absorção, comunicação, crescimento e metabolismo celular estavam em menor número e tinham alterações na forma quando comparadas aos resultados em embriões produzidos por fertilização in vitro. Conclui-se que os embriões produzidos por SCNT e partenogênese apresentam diferenças morfológicas quando comparados aos embriões de fertilização in vitro, tais como blastocele pouco desenvolvida, massa celular interna pouco definida e diferenças morfológicas nas organelas.

Development and optimization of a fluorescent differential display PCR system for studying bovine embryo development in vitro

Differential display is a widely used methodology to identify genes that are differentially expressed in biological samples. We developed a new protocol for the amplification and recovery of differentially expressed genes from extremely small initial amounts of RNA (10 to 25 pg mRNA) from preimplantation bovine embryos. The cDNAs generated with an anchor primer, associated with a universal sequence, were amplified with an arbitrary primer and a single fluorescently labeled primer. Amplification products were easily visualized with a fluorescence scanner, without the need for radioisotopes. Nineteen isolated fragments were cloned and sequenced, confirming the expected primer sequences and allowing the recognition and identification of gene transcripts involved in bovine embryonic physiology.

A probable case of Leishmaniasis (Espundia) acquired in Japan.

A case of mucocutaneous leishmaniasis is described in a Japanese patient who had never been abroad, suggesting either contact with an infected person from Brazil or the presence in Japan of indigenous cutaneous leishmaniasis. This case illustrates the value of electron microscopic examination in such infections, since it provided the definitive diagnosis.