Isolation and mutation trend analysis of influenza A virus subtype H9N2 in Egypt

ABSTRACT: BACKGROUND: Avian influenza virus H9N2 is a panzootic pathogen that affects poultry causing mild to moderate respiratory distress but has been associated with high morbidity and considerable mortality. Interspecies transmission of H9N2 from avian species to mammalian hosts does occur. The virus possesses human virus-like receptor specificity and it can infect humans producing flu-like illness. METHODS: Recently, mild influenza like symptoms were detected in H5N1 vaccinated flocks. Influenza A subtype H9N2 was isolated from the infected flock. The virus evolution was investigated by sequencing the viral genes to screen the possible virus recombination. The viral amino acid sequences from the isolated H9N2 strains were compared to other related sequences from the flu data base that were used to assess the robustness of the mutation trend. Changes in the species-associated amino acid residues or those that enabled virulence to mammals were allocated. RESULTS: Phylogenetic analyses of haemagglutinin and neuraminidase genes showed that the recently isolated Egyptian strain belonged to the H9N2 sub-lineage that prevails in Israel. The six internal segments of the isolated virus were found to be derived from the same sub-lineage with no new evidence of reassortment. The results demonstrated conserved genetic and biological constitution of H9N2 viruses in the Middle East. The recently isolated H9N2 virus from chicken in Egypt possessed amino acids that could enable the virus to replicate in mammals and caused severe disease in domestic chickens. CONCLUSION: The study highlights the importance of continuous monitoring of the mutations evolved in avian influenza viruses and its impact on virulence to avian species in addition to its importance in the emergence of new strains with the capacity to be a pandemic candidate.     

A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

Novel Comparative Efficiency of Ozone and Gamma Sterilization on Fungal Deterioration of Archeological Painted Coffin,Saqqara Excavation,Egypt

An archeological wooden painted coffin was excavated in Tety tomb from Saqqara excavation. It belonged to the Ministry of Antiquities. This coffin was discovered in a bad state of conservation with many destroyed big and small pieces in Saqqara stores. Analyses and investigation study were performed on the ground layer of the coffin by X-ray diffraction (XRD), Energy dispersive X ray analysis (EDX) equipped with environmental scanning electron microscopy (ESEM) and Fourier transform infrared spectroscopy (FTIR). Results confirmed that the degradation factors affecting the wooden painted coffin are essentially attributed to direct effects of microbial phenomena, which have lead to many deterioration forms as: macro- and microcracks, hydrated salts, flaking, coloration, scaling and defoliation microbiological spots. Nine deteriorating fungal species were isolated from the painted and ground layers of the tested coffin. Fusarium moniliforme followed by Aspergillus flavus able to significantly solublize calcium salts as major components of the ground layer of archeological wooden coffin. Effect of ozone and Gamma sterilization on growth; lipid, tryptophan oxidation and protein, nucleic acid leakage in the most dominant toxigenic deteriorated fungal species were detected. No mycelial growth was observed at 4 ppm of ozone at all exposure times. As Gamma radiation dose increased over 250 Gy, the growth parameter gradually decreased to reach the lethal dose at 2000 Gy. The production of mycotoxins by the tested toxigenic fungi was completely disappeared under the exposure to 3 ppm and 90 min to ozone.     

The use of adjunctive GPIIb/IIIa inhibitors in patients with unstable angina/non-Q-wave MI undergoing percutaneous coronary intervention

Glycoprotein IIb/IIIa receptor inhibitors represent a relatively new therapeutic approach in the field of antiplatelet therapy. Following the development of abciximab a number of small molecule GPIIb/IIIa inhibitors have been introduced such as tirofiban and eptifibatide. In this fast-moving field the interventional cardiologist needs a framework to guide decision-making for the individual patient. This review covers the efficacy and safety data from the clinical trials of GPIIb/IIIa inhibitors in the context of patients undergoing percutaneous coronary intervention for unstable angina/non-Q-wave myocardial infarction. There is an increasing body of evidence to support the efficacy of GPIIb/IIIa inhibitors in reducing the risk of adverse ischemic events in high and low risk patients undergoing percutaneous coronary intervention. A number of unresolved efficacy and safety issues remain, including the duration of treatment before and after intervention; whether a reduction in the heparin dose would further decrease the risk of hemorrhage without affecting the periprocedural thrombotic rate in patients undergoing PTCA with adjunctive GPIIb/IIIa inhibitors; and the cost-effectiveness of this therapy. When a thorough analysis of cost-effectiveness has been made, it will be easier to advocate the widespread use of these agents in all patients undergoing coronary intervention.     

Microbial transformation of zearalenone to a zearalenone sulfate.

The conversion of zearalenone by various microorganisms was studied. A new polar metabolite was formed in addition to alpha- and beta-zearalenols. The structure of the new metabolite was determined as zearalenone-4-O-sulfate conjugate on the basis of enzymatic and acid hydrolysis, followed by mass spectrometry, nuclear magnetic resonance, and infrared spectroscopic analysis. The results obtained demonstrate that Rhizopus arrhizus catalyzes sulfation of zearalenone at the C-4 hydroxyl group.     

A high-throughput cloning system for reverse genetics in <Emphasis Type="Italic">Trypanosoma cruzi</Emphasis>

Background  

The three trypanosomatids pathogenic to men, Trypanosoma cruzi, Trypanosoma brucei and Leishmania major, are etiological agents of Chagas disease, African sleeping sickness and cutaneous leishmaniasis, respectively. The complete sequencing of these trypanosomatid genomes represented a breakthrough in the understanding of these organisms. Genome sequencing is a step towards solving the parasite biology puzzle, as there are a high percentage of genes encoding proteins without functional annotation. Also, technical limitations in protein expression in heterologous systems reinforce the evident need for the development of a high-throughput reverse genetics platform. Ideally, such platform would lead to efficient cloning and compatibility with various approaches. Thus, we aimed to construct a highly efficient cloning platform compatible with plasmid vectors that are suitable for various approaches.     

Concurrent Demonstration of Desoxyribose nucleic Acid and 1,2-Glycols. 2. Periodate Oxidation

Formalin-fixed sections of rat intestine were treated in a solution of 0.5% periodic acid in 50% aqueous phosphoric acid for the concurrent demonstration of 1,2-glycols and DNA. The effects of varying the concentration of reagents, time of exposure and temperature on the results showed that, at 28°C, good reactions were obtained in 10-15 min when the concentration of the phosphoric was between 40 and 60% and that of the periodic acid between 0.05 and 0.50%.