Genetic characterization of infectious bursal disease viruses associated with gumboro outbreaks in commercial broilers from asyut province, egypt.

Ten infectious bursal disease virus (IBDV) field strains were isolated from 15 broiler flocks located in various parts of Asyut, Egypt. Seven strains were subjected to comparative sequencing and phylogenetic analyses to help provide optimal control program for protection against IBDV infection. Sequence analysis of a 530 bp hypervariable region in the VP2 gene revealed that the rate of identity and homology was around 95.6~99.1%. Sequence characterization revealed the 7 strains identified as vvIBDV with the four amino acids residues typical of vvIBDV (242I, 256I, 294I, 299S). The BURSA-VAC vaccine was the nearest vaccine in sequence similarity to the local examined IBDV strains followed by CEVACIBDL then Bursine plus and Nobilis Gumboro indicating its probable success in the face of incoming outbreaks when using these vaccines. Phylogenetic analysis revealed that the presence of three clusters for the examined strains and are grouped with reference very virulent IBDVs of European and Asian origin (Japanese and Hong Kong) strains suggesting the different ancestors of our isolates. The antigenic index showed a number of changes on the major and minor hydrophilic antigenic peaks of the virus surface structures indicating a new genetic evolution of the surface structure epitopes that may lead to vaccination failure and reemergence of the disease.

Guanidinium chloride-induced spectral perturbations of 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid confound interpretation of data on molten globule states.

We describe limitations in the use of 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid (bis-ANS) to examine unfolding intermediates associated with guanidinium chloride (GuHCl)-induced protein denaturation. Several studies have used alterations in fluorescence emission of bis-ANS to quantify the population of "molten globule" states. Our findings indicate that the observed changes in bis-ANS spectroscopic properties could originate from the interactions of bis-ANS and GuHCl and the aggregation of the dye at higher GuHCl concentrations. We posit that in the absence of additional complementary structural or spectroscopic measurements, the use of bis-ANS emission alone to monitor protein conformations can be misleading.

Ovarian steroids modulate gonadotropin-releasing hormone-induced biphasic luteinizing hormone secretory responses and inositol phosphate accumulation in rat anterior pituitary cells and alpha T3-1 gonadotrophs.

The ovarian steroids estradiol and progesterone act as important modulators of GnRH-induced luteinizing hormone (LH) secretion from anterior pituitary cells. Recently, we demonstrated that the steroids are able to influence GnRH-stimulated Ca2+ mobilization from extra- and intracellular sources. Here we investigated the actions of estradiol and progesterone on GnRH-induced biphasic LH secretory responses in the model of perifused female rat pituitary cells. A 20 min GnRH stimulus elicited biphasic LH responses composed of an initial peak followed by a prolonged plateau phase. Both phases were equally enhanced by long-term (48 h) estradiol treatment. This action was facilitated by subsequent short-term progesterone treatment. In contrast, combined treatment with estradiol and progesterone for 48 h led to inhibited LH secretory profiles. To determine the steroid actions on the extracellular Ca2+ independent component of LH secretion we performed experiments using cells that were perifused with Ca2+ deficient medium. Under these conditions the cells responded exclusively with a single peak phase of LH secretion, which was augmented or inhibited by estradiol and progesterone treatment as described above. To test the hypothesis that an effect of estradiol and progesterone on GnRH-induced polyphophoinositide hydrolysis is responsible for their modulatory actions on Ca2+ signals and LH secretion we measured inositol phosphate (IP) accumulation after different steroid treatment paradigms in rat pituitary cells and alpha T3-1 immortalized gonadotrophs. GnRH-induced IP production was enhanced by long-term estradiol treatment. Short-term exposure of estradiol-primed cells to progesterone did not lead to significant changes of IP production. The long-term progesterone treatment paradigm enhanced GnRH-induced IP formation, while it decreased Ca2+ signals and LH secretion. Alpha T3-1 cells were used to perform more detailed analysis of IP formation. The actions of estradiol and progesterone on the production of inositol mono-, bis-, and trisphosphates were similar to those observed in the mixed cell population. It is concluded that estradiol and progesterone modulate both peak and plateau phases of GnRH-stimulated LH secretory responses, effects which are associated with their impact on Ca2+ signals. Our findings argue against a role of IP modulation in the mechanism of progesterone actions on Ca2+ signaling and LH secretion in gonadotrophs. Such a mechanism might be involved in the positive effects of estradiol in these cells.