Host-specific genetic variation of highly pathogenic avian influenza viruses (H5N1)

The complete genome sequences of two isolates A/chicken/Egypt/CL6/07 (CL6/07) and A/duck/Egypt/D2br10/07 (D2br10/07) of highly pathogenic avian influenza virus (HPAI) H5N1 isolated at the beginning of 2007 outbreak in Egypt were determined and compared with all Egyptian HPAI H5N1 sequences available in the GenBank. Sequence analysis utilizing the RNA from the original tissue homogenate showed amino acid substitutions in seven of the viral segments in both samples. Interestingly, these changes were different between the CL6/07 and D2br10/07 when compared to other Egyptian isolates. Moreover, phylogenetic analysis showed independent sub-clustering of the two viruses within the Egyptian sequences signifying a possible differential adaptation in the two hosts. Further, pre-amplification analysis of H5N1 might be necessary for accurate data interpretation and identification of distinct factor(s) influencing the evolution of the virus in different poultry species

Body image disturbance and surgical decision making in egyptian post menopausal breast cancer patients

<p>Abstract</p> <p>Background</p> <p>In most developing countries, as in Egypt; postmenopausal breast cancer cases are offered a radical form of surgery relying on their unawareness of the subsequent body image disturbance. This study aimed at evaluating the effect of breast cancer surgical choice; Breast Conservative Therapy (BCT) versus Modified Radical Mastectomy (MRM); on body image perception among Egyptian postmenopausal cases.</p> <p>Methods</p> <p>One hundred postmenopausal women with breast cancer were divided into 2 groups, one group underwent BCT and the other underwent MRM. Pre- and post-operative assessments of body image distress were done using four scales; Breast Impact of Treatment Scale (BITS), Impact of Event Scale (IES), Situational Discomfort Scale (SDS), and Body Satisfaction Scale (BSS).</p> <p>Results</p> <p>Preoperative assessment showed no statistical significant difference regarding cognitive, affective, behavioral and evaluative components of body image between both studied groups. While in postoperative assessment, women in MRM group showed higher levels of body image distress among cognitive, affective and behavioral aspects.</p> <p>Conclusion</p> <p>Body image is an important factor for postmenopausal women with breast cancer in developing countries where that concept is widely ignored. We should not deprive those cases from their right of less mutilating option of treatment as BCT.</p

Acquisition of Human-Type Receptor Binding Specificity by New H5N1 Influenza Virus Sublineages during Their Emergence in Birds in Egypt

Highly pathogenic avian influenza A virus subtype H5N1 is currently widespread in Asia, Europe, and Africa, with 60% mortality in humans. In particular, since 2009 Egypt has unexpectedly had the highest number of human cases of H5N1 virus infection, with more than 50% of the cases worldwide, but the basis for this high incidence has not been elucidated. A change in receptor binding affinity of the viral hemagglutinin (HA) from α2,3- to α2,6-linked sialic acid (SA) is thought to be necessary for H5N1 virus to become pandemic. In this study, we conducted a phylogenetic analysis of H5N1 viruses isolated between 2006 and 2009 in Egypt. The phylogenetic results showed that recent human isolates clustered disproportionally into several new H5 sublineages suggesting that their HAs have changed their receptor specificity. Using reverse genetics, we found that these H5 sublineages have acquired an enhanced binding affinity for α2,6 SA in combination with residual affinity for α2,3 SA, and identified the amino acid mutations that produced this new receptor specificity. Recombinant H5N1 viruses with a single mutation at HA residue 192 or a double mutation at HA residues 129 and 151 had increased attachment to and infectivity in the human lower respiratory tract but not in the larynx. These findings correlated with enhanced virulence of the mutant viruses in mice. Interestingly, these H5 viruses, with increased affinity to α2,6 SA, emerged during viral diversification in bird populations and subsequently spread to humans. Our findings suggested that emergence of new H5 sublineages with α2,6 SA specificity caused a subsequent increase in human H5N1 influenza virus infections in Egypt, and provided data for understanding the virus's pandemic potential

Effects of antibiotic resistance, drug target attainment, bacterial pathogenicity and virulence, and antibiotic access and affordability on outcomes in neonatal sepsis: an international microbiology and drug evaluation prospective substudy (BARNARDS)

Background Sepsis is a major contributor to neonatal mortality, particularly in low-income and middle-income countries (LMICs). WHO advocates ampicillin–gentamicin as first-line therapy for the management of neonatal sepsis. In the BARNARDS observational cohort study of neonatal sepsis and antimicrobial resistance in LMICs, common sepsis pathogens were characterised via whole genome sequencing (WGS) and antimicrobial resistance profiles. In this substudy of BARNARDS, we aimed to assess the use and efficacy of empirical antibiotic therapies commonly used in LMICs for neonatal sepsis. Methods In BARNARDS, consenting mother–neonates aged 0–60 days dyads were enrolled on delivery or neonatal presentation with suspected sepsis at 12 BARNARDS clinical sites in Bangladesh, Ethiopia, India, Pakistan, Nigeria, Rwanda, and South Africa. Stillborn babies were excluded from the study. Blood samples were collected from neonates presenting with clinical signs of sepsis, and WGS and minimum inhibitory concentrations for antibiotic treatment were determined for bacterial isolates from culture-confirmed sepsis. Neonatal outcome data were collected following enrolment until 60 days of life. Antibiotic usage and neonatal outcome data were assessed. Survival analyses were adjusted to take into account potential clinical confounding variables related to the birth and pathogen. Additionally, resistance profiles, pharmacokinetic–pharmacodynamic probability of target attainment, and frequency of resistance (ie, resistance defined by in-vitro growth of isolates when challenged by antibiotics) were assessed. Questionnaires on health structures and antibiotic costs evaluated accessibility and affordability. Findings Between Nov 12, 2015, and Feb 1, 2018, 36 285 neonates were enrolled into the main BARNARDS study, of whom 9874 had clinically diagnosed sepsis and 5749 had available antibiotic data. The four most commonly prescribed antibiotic combinations given to 4451 neonates (77·42%) of 5749 were ampicillin–gentamicin, ceftazidime–amikacin, piperacillin–tazobactam–amikacin, and amoxicillin clavulanate–amikacin. This dataset assessed 476 prescriptions for 442 neonates treated with one of these antibiotic combinations with WGS data (all BARNARDS countries were represented in this subset except India). Multiple pathogens were isolated, totalling 457 isolates. Reported mortality was lower for neonates treated with ceftazidime–amikacin than for neonates treated with ampicillin–gentamicin (hazard ratio [adjusted for clinical variables considered potential confounders to outcomes] 0·32, 95% CI 0·14–0·72; p=0·0060). Of 390 Gram-negative isolates, 379 (97·2%) were resistant to ampicillin and 274 (70·3%) were resistant to gentamicin. Susceptibility of Gram-negative isolates to at least one antibiotic in a treatment combination was noted in 111 (28·5%) to ampicillin–gentamicin; 286 (73·3%) to amoxicillin clavulanate–amikacin; 301 (77·2%) to ceftazidime–amikacin; and 312 (80·0%) to piperacillin–tazobactam–amikacin. A probability of target attainment of 80% or more was noted in 26 neonates (33·7% [SD 0·59]) of 78 with ampicillin–gentamicin; 15 (68·0% [3·84]) of 27 with amoxicillin clavulanate–amikacin; 93 (92·7% [0·24]) of 109 with ceftazidime–amikacin; and 70 (85·3% [0·47]) of 76 with piperacillin–tazobactam–amikacin. However, antibiotic and country effects could not be distinguished. Frequency of resistance was recorded most frequently with fosfomycin (in 78 isolates [68·4%] of 114), followed by colistin (55 isolates [57·3%] of 96), and gentamicin (62 isolates [53·0%] of 117). Sites in six of the seven countries (excluding South Africa) stated that the cost of antibiotics would influence treatment of neonatal sepsis

Prevalence and Antibiogram of Pseudomonas aeruginosa Among Nile Tilapia and Smoked Herring, with an Emphasis on their Antibiotic Resistance Genes (blaTEM, blaSHV, blaOXA-1 and ampC) and Virulence Determinant (oprL and toxA)

Bacterial diseases are one of the most challenging issues facing aquaculture sector. Pseudomonas aeruginosa (P. aeruginosa) has been regarded as one of the most significant threats to the fishing industry, which also affects public health. We aimed to elucidate the occurrence and antibiogram profile of P. aeruginosa recovered from Nile tilapia (Oreochromis niloticus) and smoked herring (Clupea harengus) with emphasis on their antibiotic resistance genes (blaTEM, blaSHV, blaOXA-1 and ampC) and virulence determinant genes (oprL and toxA). A total of 150 fish samples (110 diseased Nile tilapia, and 40 smoked herring) were collected randomly from retails of Gharbia Governorate, Egypt. The retrieved isolates were phenotypically characterized using standard methods of culturing and biochemical tests. Then, verified using molecular assay, 16S rRNA gene was detected in 100% of the tested isolates. The overall incidence of P. aeruginosa was 33.3%, out of which 45% from smoked herring and 29% from Nile Tilapia. The occurrence of P. aeruginosa in various infected organs of O. niloticus showed that the gills were the most obviously infected organ followed by kidney, liver, and spleen, respectively. A significant difference (P&lt; 0.05) was noticed in the distribution of P. aeruginosa among O. niloticus internal organs. The phenotypic susceptibility to nine commonly used antimicrobial agents was detected using disc diffusion assay. The tested strains were extremely susceptible to ciprofloxacin, amikacin, and imipenem, whereas exhibited remarkable resistance to oxacillin, cefpodoxime, amoxicillin with clavulanic acid, ceftriaxone, and nalidixic acid. Interestingly, 100% of P. aeruginosa isolates were multiple antimicrobial resistant (MAR). Three resistance phenotypes profiles were identified with MAR index ranged from 0.4-0.5. Screening for antibiotic resistance genes revealed a diversity of β-lactamases in P. aeruginosa isolates, with blaTEM being the most dominant gene (100%), followed by blaSHV, blaOXA-1 and ampC with a total prevalence of 66.6% to all of them. The identified antimicrobial resistance phenotypes and genotypes were found to be significantly correlated. Subsequently, the distribution of virulence determinants in these strains was identified. These isolates had 100% prevalence of oprL and toxA virulence genes. In conclusion, the emergence of MDR P. aeruginosa in fish particularly ESBL and AmpC beta-lactamases producers could pose a potential health hazard to consumers. Thus, antimicrobial susceptibility must be continuously monitored to assess potential risks to human health. Ciprofloxacin, amikacin, and imipenem were the most efficient antibiotics for treatment of the identified P. aeruginosa, ESBL and AmpC beta-lactamases producers

Mechanical behavior and corrosion properties of Ti-7Mo-8Nb alloy for biomedical applications

The present study investigates the microstructural, mechanical and corrosion properties of Ti-7Mo-8Nb alloy manufactured through powder metallurgy. The performance of the developed alloy is benchmarked against cast Ti-6Al-4V. Microstructure examination of Ti-7Mo-8Nb revealed a Widmanstätten structure containing equiaxed β grains along with acicular α phase. In regards to the mechanical properties, Ti-7Mo-8Nb possessed higher compressive yield strength, higher hardness but lower elastic modulus than Ti-6Al-4V. The elastic modulus of Ti-7Mo-8Nb was almost 44.9 GPa, approaching the usually desired value of 30 GPa for cortical bone. Wear test revealed also a lower wear rate for Ti-7Mo-8Nb. Potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) experiments were carried out for both Ti-7Mo-8Nb and Ti-6Al-4V immersed in Hank’s solution as a simulated body fluid at a temperature of 37 °C. Both experiments revealed higher corrosion resistance for Ti-7Mo-8Nb manifested by lower corrosion and passivation current densities, higher negative phase angle, higher impedance modulus and larger Nyquist semicircle diameter as compared to Ti-6Al-4V alloy. The superior corrosion properties of Ti-7Mo-8Nb are indicative of the development of a more stable passive layer on the surface. The fitting of EIS data into an equivalent circuit suggested the formation of a double oxide layer consisting of an inner compact base passive film along with an external porous layer. The presented combination of high strength, high corrosion resistance along with low elastic modulus puts forward the Ti-7Mo-8Nb alloy as a good candidate for orthopedic biomedical applications

Exploring Antimicrobial Potential of Cinnamon, Clove, Peppermint and Black Cumin Essential Oils against Fish Bacterial Pathogens with an Emphasis on the Dietary Supplementation Effects of Cinnamon Oil on Striped Catfish (Pangasianodon hypophthalmus)

Emergence of antimicrobial resistance (AMR) among fish bacterial pathogens is one of the major global public threats. Attempts are being made to develop novel alternatives as a promising approach to combat multidrug resistance disease-causing bacteria. Natural antimicrobials such as essential oils (EOs) are a potential unique strategy to treat bacterial infections with a reduced risk of resistance developing. This study aimed to evaluate the antimicrobial activity of some essential oils (EOs) namely, cinnamon (Cinnamomum zeylanicum), clove (Syzygium aromaticum), Peppermint (Mentha piperita) , and black cumin (Nigella sativa) against some fish pathogens implicated with aquaculture disease outbreaks like Aeromonas hydrophilia, Pseudomonas fluorescence, Photobacterium damselae and Streptococcus agalactiae using agar well diffusion assay. We found significant differences on the antibacterial activity depending on the type of essential oils and bacterial strain. Among all the tested EOs, cinnamon essential oil (CEO) was shown to be the most effective with minimum bactericidal concentration (MBC) ranged from 0.0156-0.125 ml/ml. As a result, it was selected for our in vivo investigations. We next aimed to investigate the effects of dietary CEO on growth performance, disease resistance and immune response of fish. A total of 150 striped catfish (Pangasianodon hypophthalmus) were fed with different levels of CEO (0, 1.50, 2.0, 2.50, and 3.0 mL/kg diets) (assigned as control, Diet 1, Diet 2, Diet 3 and Diet 4) for 60 days. Compared to control, fish fed with graded levels of dietary CEO showed significant (P &lt; 0.05) increase in final body weight, weight gain %, and specific growth rate particularly at fish group fed diet 3. Of interest, there were no significant differences (P &gt; 0.05) in feed conversion ratio and survival rates among control and CEO-supplemented groups. Moreover, we found significant (P &lt; 0.05) increases in plasma lysozyme activity and total IgM levels in a dose dependent manner with dietary CEO supplementation. After feeding trials, we investigated their potential to defend striped catfish against A. hydrophila challenge. Fish fed control diet had the highest mortality rates; in contrast, fish fed diets supplemented with CEO had higher levels of resistance to the bacterial infection, with the lowest mortality rates in the fish group fed diet 3. Overall, these findings showed that EOs exhibit a great potential to be used as antimicrobial agents against fish pathogens. Moreover, dietary administration of CEO, particularly at 2.5 ml/kg feed, can be regarded as a promising component for improving growth, immunological responses and potential alternatives to conventional antimicrobials for control of microbial infections in fish

The Vpu Protein of Human Immunodeficiency Virus Type 1 Plays a Protective Role against Virus-Induced Apoptosis in Primary CD4(+) T Lymphocytes

Previous data revealed that primary cultures of peripheral blood mononuclear cells (PBMCs) were killed by apoptosis at higher rates after infection with two CRF01_AE primary isolates of human immunodeficiency virus type 1 (HIV-1) than after infection with five other CRF01_AE primary isolates, five subtype B primary isolates, and two subtype B laboratory strains. Here, we show evidence that mutations at the vpu gene which were exclusively identified only in the two CRF01_AE isolates mentioned above are involved in their abilities to induce massive apoptosis in primary CD4(+) T lymphocytes. The rates of virus production by these two isolates in the culture media of infected PBMCs were lower (the same as those of the other CRF01_AE isolates) than those of the subtype B isolates. To confirm the correlation between the higher apoptosis-inducing abilities and the mutations at the vpu gene, infectious molecular clone pNL4-3-based vpu mutants were constructed and examined for their apoptosis induction levels. The apoptosis induction levels after introduction of the vpu mutations were greatly increased in primary CD4(+) T lymphocytes. In contrast, the apoptosis induction abilities of these vpu mutants were lower in human T-cell line MT-4. Thus, the Vpu protein of HIV-1 could play a protective role against virus-induced apoptosis in primary CD4(+) T lymphocytes

Highly Pathogenic H5N1 Avian Influenza Virus Induces Extracellular Ca2+ Influx, Leading to Apoptosis in Avian Cells ▿ †

In this study, we show that the highly pathogenic H5N1 avian influenza virus (AIV) (A/crow/Kyoto/53/04 and A/chicken/Egypt/CL6/07) induced apoptosis in duck embryonic fibroblasts (DEF). In contrast, apoptosis was reduced among cells infected with low-pathogenic AIVs (A/duck/HK/342/78 [H5N2], A/duck/HK/820/80 [H5N3], A/wigeon/Osaka/1/01 [H7N7], and A/turkey/Wisconsin/1/66 [H9N2]). Thus, we investigated the molecular mechanisms of apoptosis induced by H5N1-AIV infection. Caspase-dependent and -independent pathways contributed to the cytopathic effects. We further showed that, in the induction of apoptosis, the hemagglutinin of H5N1-AIV played a major role and its cleavage sequence was not critical. We also observed outer membrane permeabilization and loss of the transmembrane potential of the mitochondria of infected DEF, indicating that mitochondrial dysfunction was caused by the H5N1-AIV infection. We then analyzed Ca2+ dynamics in the infected cells and demonstrated an increase in the concentration of Ca2+ in the cytosol ([Ca2+]i) and mitochondria ([Ca2+]m) after H5N1-AIV infection. Regardless, gene expression important for regulating Ca2+ efflux from the endoplasmic reticulum did not significantly change after H5N1-AIV infection. These results suggest that extracellular Ca2+ may enter H5N1-AIV-infected cells. Indeed, EGTA, which chelates extracellular free Ca2+, significantly reduced the [Ca2+]i, [Ca2+]m, and apoptosis induced by H5N1-AIV infection. In conclusion, we identified a novel mechanism for influenza A virus-mediated cell death, which involved the acceleration of extracellular Ca2+ influx, leading to mitochondrial dysfunction and apoptosis. These findings may be useful for understanding the pathogenesis of H5N1-AIV in avian species as well as the impact of Ca2+ homeostasis on influenza A virus infection