Relation between leptin and estradiol levels in Egyptian lactating Arab mares during foaling heat

Sixteen Arab lactating mares belonging to Al-Zahraa Arab Horse Stud underwent two ultrasound examinations at 3 weeks interval starting from the day of demonstration of foaling heat. In addition, daily blood samples were collected from parturition until after exhibiting first postpartum estrus (day 11) with daily observation of estrous signs. Both leptin and estradiol hormones were assayed. Mean day of foaling heat was 8.9 ± 0.9 day. Most mares came in foaling heat during days 9 and 10 had high conception rate compared to those who came in estrus earlier or later. Estradiol levels were high after day of foaling then decrease after expression of foaling heat. But leptin levels increase from day 8 to day 10 compared to other days before and after the first ovulation. A significant positive correlation was found between estradiol and leptin (r = 0.58, p < 0.025). The positive correlation between leptin and estradiol led us to suggest that leptin hormone plays an important role in ovulation of the first postpartum estrus in mares.     

PYPAF7, a novel PYRIN-containing Apaf1-like protein that regulates activation of NF-kappa B and caspase-1-dependent cytokine processing

PYRIN-containing Apaf1-like proteins (PYPAFs) are members of the nucleotide-binding site/leucine-rich repeat (NBS/LRR) family of signal transduction proteins. We report here that PYPAF7 is a novel PYPAF protein that activates inflammatory signaling pathways. The expression of PYPAF7 is highly restricted to immune cells, and its gene maps to chromosome 19q13.4, a locus that contains a cluster of genes encoding numerous PYPAF family members. Co-expression of PYPAF7 with ASC results in the recruitment of PYPAF7 to distinct cytoplasmic loci and a potent synergistic activation of NF-kappa B. To identify other proteins involved in PYPAF7 and ASC signaling pathways, we performed a mammalian two-hybrid screen and identified pro-caspase-1 as a binding partner of ASC. Co-expression of PYPAF7 and ASC results in the synergistic activation of caspase-1 and a corresponding increase in secretion of interleukin-1 beta. In addition, PYPAF1 induces caspase-1-dependent cytokine processing when co-expressed with ASC. These findings indicate that PYPAF family members participate in inflammatory signaling by regulating the activation of NF-kappa B and cytokine processing.     

Effect of dietary fats on hepatic lipid metabolism in the growing turkey

The influence of dietary fatty acids on hepatic capacity of lipid synthesis and secretion was investigated in 7-week-old male turkeys. They were fed 10% of either lard (rich in saturated and monounsaturated fatty acids) or linseed oil (rich in polyunsaturated fatty acids, especially 18:3n-3). Fattening was identical with both diets (0.15-0.20% of abdominal adipose tissue), but the proportion of muscle Pectoralis major was lower with linseed oil (6.6 vs. 7.4%). Specific activities of lipogenic enzymes (ME, G6PDH, ACX, and Delta9-desaturase) were not influenced by the diet, however, FAS activity was lower with linseed oil (14.3 vs. 25.4 nM NADPH fixed/min). Fasting concentrations of lipoproteins synthesized and secreted by the liver, VLDL and HDL, were also lower with linseed oil, as well as plasma concentrations of phospholipids and cholesteryl esters. However, when VLDL catabolism was inhibited by injection of an antiserum against LPL, VLDL concentration was identical in both groups (100-120 mg/l), whereas that of phospholipids and cholesteryl esters, that are transported by HDL mainly, remained lower with linseed oil. Thus, in the growing turkeys, and contrary to mammals and the chicken, feeding n-3 polyunsaturated fatty acids did not decrease hepatic triglyceride synthesis and secretion, nor fattening. By contrast, in this species, n-3 polyunsaturated fatty acids appear to influence mostly HDL metabolism, with a negative impact on muscular growth.     

Reconstitution and characterization of the human DNA polymerase delta four-subunit holoenzyme

Mammalian DNA polymerase delta was originally characterized as a tightly associated heterodimer consisting of the catalytic subunit, p125, and the p50 subunit. Recently, two additional subunits, the third (p68) and fourth subunits (p12), have been identified. The heterotetrameric human pol delta complex was reconstituted by overexpression of the four subunits in Sf9 cells, followed by purification to near-homogeneity using FPLC chromatography. The properties of the four-subunit enzyme were shown to be functionally indistinguishable from those of pol delta isolated from calf thymus. The physicochemical properties of both the reconstituted heterotetramer and the heterodimer of the p125 and p50 subunits were examined by gel filtration and glycerol gradient ultracentrifugation. These studies show quite clearly that the heterodimer and heterotetramer complexes do not behave in solution as dimeric structures. This issue is of significance because several studies of the yeast pol delta complexes have indicated that the third subunit is able to bring about the dimerization of the pol delta complex. The heterodimer is only weakly stimulated by PCNA, whereas the heterotetramer is strongly stimulated to a level with a specific activity comparable to that of the calf thymus enzyme. These results resolve earlier, conflicting reports on the response of the heterodimer to PCNA. Nevertheless, the heterodimer does have some ability to interact functionally with PCNA, consistent with evidence that the p125 subunit itself has an ability to interact with PCNA. The functional interaction of PCNA with the pol delta complex may likely involve multiple contacts.     

The structural determination of a new steroidal metabolite from the brown alga Sargassum asperifolium

An investigation of the natural products chemistry of the brown alga Sargassum asperifolium from the red sea yielded a new steroidal metabolite, 24-vinyl cholest-4-ene-24-ol-3-one, saringosterone (3), a known steroidal metabolite, 24-vinyl cholest-5-ene-3b,24-diol, saringosterol (4), and known diterpene with a hydroazulene skeleton, dictyone (1). The identification of the isolated metabolites was established mainly by spectral methods and chemical transformation of the dictyone (1) to its acetate (2).     

Plant biodiversity in the Saint Catherine area of the Sinai peninsula, Egypt

This study focuses on plant species richness, taxonomic diversity, and endemism of the Sinai peninsula as a whole and Saint Catherine area in particular. Beta diversity was also measured using _T to represent the biotic change between different landforms in the St. Catherine area. The peninsula supports about 1285 species including the infraspecific taxa of which about 800 species (also including the infraspecific taxa) are recorded in the southern Sinai south of El-Tih Desert. The present study reveals that southern Sinai is more diverse as compared with the Sinai peninsula as a whole. Southern Sinai also supports more than the expected numbers of rare and very rare species (about 62%) and about 4.3% endemic species. Beta diversity between different landforms in the St. Catherine area reflect a large biotic change between slopes and terraces on the one hand and between terraces and ridges on the other. Finally, the study recommends appropriate management of the peninsula and more research for the management requirements of the numerous endemic and rare species in the Sinai peninsula.     

Proteomic Profiling of the Outer Membrane Fraction of the Obligate Intracellular Bacterial Pathogen Ehrlichia ruminantium

The outer membrane proteins (OMPs) of Gram-negative bacteria play a crucial role in virulence and pathogenesis. Identification of these proteins represents an important goal for bacterial proteomics, because it aids in vaccine development. Here, we have developed such an approach for Ehrlichia ruminantium, the obligate intracellular bacterium that causes heartwater. A preliminary whole proteome analysis of elementary bodies, the extracellular infectious form of the bacterium, had been performed previously, but information is limited about OMPs in this organism and about their role in the protective immune response. Identification of OMPs is also essential for understanding Ehrlichia’s OM architecture, and how the bacterium interacts with the host cell environment. First, we developed an OMP extraction method using the ionic detergent sarkosyl, which enriched the OM fraction. Second, proteins were separated via one-dimensional electrophoresis, and digested peptides were analyzed via nano-liquid chromatographic separation coupled with mass spectrometry (LC-MALDI-TOF/TOF). Of 46 unique proteins identified in the OM fraction, 18 (39%) were OMPs, including 8 proteins involved in cell structure and biogenesis, 4 in transport/virulence, 1 porin, and 5 proteins of unknown function. These experimental data were compared to the predicted subcellular localization of the entire E. ruminantium proteome, using three different algorithms. This work represents the most complete proteome characterization of the OM fraction in Ehrlichia spp. The study indicates that suitable subcellular fractionation experiments combined with straightforward computational analysis approaches are powerful for determining the predominant subcellular localization of the experimentally observed proteins. We identified proteins potentially involved in E. ruminantium pathogenesis, which are good novel targets for candidate vaccines. Thus, combining bioinformatics and proteomics, we discovered new OMPs for E. ruminantium that are valuable data for those investigating new vaccines against this organism. In summary, we provide both pioneering data and novel insights into the pathogenesis of this obligate intracellular bacterium.     

Hybridization properties of long nucleic acid probes for detection of variable target sequences,and development of a hybridization prediction algorithm

One of the main problems in nucleic acid-based techniques for detection of infectious agents, such as influenza viruses, is that of nucleic acid sequence variation. DNA probes, 70-nt long, some including the nucleotide analog deoxyribose-Inosine (dInosine), were analyzed for hybridization tolerance to different amounts and distributions of mismatching bases, e.g. synonymous mutations, in target DNA. Microsphere-linked 70-mer probes were hybridized in 3M TMAC buffer to biotinylated single-stranded (ss) DNA for subsequent analysis in a Luminex® system. When mismatches interrupted contiguous matching stretches of 6 nt or longer, it had a strong impact on hybridization. Contiguous matching stretches are more important than the same number of matching nucleotides separated by mismatches into several regions. dInosine, but not 5-nitroindole, substitutions at mismatching positions stabilized hybridization remarkably well, comparable to N (4-fold) wobbles in the same positions. In contrast to shorter probes, 70-nt probes with judiciously placed dInosine substitutions and/or wobble positions were remarkably mismatch tolerant, with preserved specificity. An algorithm, NucZip, was constructed to model the nucleation and zipping phases of hybridization, integrating both local and distant binding contributions. It predicted hybridization more exactly than previous algorithms, and has the potential to guide the design of variation-tolerant yet specific probes.