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Background Rift Valley Fever Virus (RVFV) is an arbovirus, a zoonotic disease that resurfaces as a potential hazard beyond geographic boundaries. Fever that can proceed to encephalitis, retinitis, hemorrhagic fever, and death is the main manifestation observed in human infections. RVFV has no authorized medication. The RNA interference (RNAi) gene silencing pathway is extremely well conserved. By targeting specific genes, small interfering RNA (siRNA) can be used to suppress viral replication. The aim of this study was to design specific siRNAs against RVFV and evaluate their prophylactic and antiviral effects on the Vero cells. Methods and results Various siRNAs were designed using different bioinformatics tools. Three unique candidates were tested against an Egyptian sheep cell culture-adapted strain BSL-2 that suppressed RVFV N mRNA expression. SiRNAs were transfected a day before RVFV infection (pre-transfection), and 1 h after the viral infection (post-transfection), and were evaluated to detect the silencing activity and gene expression decrease using real-time PCR and a TCID50 endpoint test. The degree of N protein expression was determined by western blot 48 h after viral infection. D2 which targets the (488–506 nucleotides), the middle region of RVFV N mRNA was the most effective siRNA at 30 nM concentration, it almost eliminates N mRNA expression when utilized as antiviral or preventive therapy. siRNAs had a stronger antiviral silencing impact when they were post-transfected into Vero cells. Conclusion Pre and post-transfection of siRNAs significantly reduced RVFV titer in cell lines, offering novel and potentially effective anti-RVFV epidemics and epizootics therapy.

Microorganisms associated with marine invertebrates consider an important source of bioactive products. This study aimed to screen for antimicrobial and anticancer activity of crude extracts of bacteria associated with Red sea nudibranchs and molecular identification of the bioactive isolates using 16Sr RNA sequencing, in addition to whole-genome sequencing of one of the most bioactive bacteria. This study showed that bacteria associated with Red sea nudibranchs are highly bioactive and 16Sr RNA sequencing results showed that two isolates belonged to Firmicutes, and two isolates belonged to Proteobacteria, and Actinobacteria. The whole genome sequence data of the isolated Nocardiopsis RACA4 isolate has an estimated genome length of 6,721,839 bp and the taxonomy showed it belongs to the bacteria Nocardiopsis dassonvillei. The De novo assembly of RACA-4 paired reads using Unicycler v0.4.8 initially yielded 97 contigs with an N50 value of 214,568 bp and L50 value of 10, The resulting assembly was further mapped to the reference genome Nocardiopsis dassonvillei strain NCTC10488 using RagTag software v.2.1.0 and a final genome assembly resulted in 39 contigs and N50 value of 6,726,007 and L50 of 1. Genome mining using anti-smash showed around 9.1% of the genome occupied with genes related to secondary metabolites biosynthesis. A wide variety of secondary metabolites belonging to Polyketides, Terpenes, and nonribosomal peptides were predicted with high degree of similarity to known compounds. Non-characterized clusters were also found which suggest new natural compounds discovered by further studies.

Staphylococcus aureus (S. aureus) is a versatile bacterium which exhibits multiple antibiotic resistances. To ameliorate the undesirable diseases causing potential, there is a need to design a protective vaccine capable of stimulating immune response against this pathogen. In a similar study in our laboratory, reverse vaccinology approach was used to nominate potential vaccine candidate genes against S. aureus. Zinc Metalloproteinase Aureolysin (aur) gene was one of the nominated genes based on that previously published in-silico study. The objective of this study is the cloning, expression, purification of aur gene and testing the aur protein reactivity with serum antibodies collected from groups of human patients with confirmed Staphylococcal disease. Cloning was done in pH6HTN His6HaloTag® vector and it was expressed in E. coli BL21 (DE3) using these conditions; 0.5 mM Isopropyl β-D-1-thiogalactopyranoside (IPTG) for 4 h at 37°C. Purification was carried out by Immobilized Metal Affinity Chromatography (IMAC). The his-tag aur protein was detected at ~86 KDa as a single band after western blot assay and was successfully reacted with antibodies obtained from humans infected with S. aureus. The results encourage further testing of aur protein as a potential vaccine candidate for S. aureus. Key words: Staphylococcus aureus, Zinc Metalloproteinase Aureolysin (aur), cloning, expression.

In 1869, the Suez Canal was opened, which brought the waters of the Mediterranean and the Red Sea into direct contact. Notably, the Suez Canal was constructed for navigation purposes without focusing on the ecological impacts. The Suez Canal paved the way for species migration from the Red Sea to the Mediterranean Sea through Lessepsian migration, named after Ferdinand de Lesseps, while the migration from the Mediterranean Sea to the Red Sea is called the anti-Lessepsian migration. It has been argued in the past that the migrating species had negative consequences for the host environment as well as of humans. Few studies to date have attempted to map the microorganism migration problem because the traditional ways of measuring the community's richness and dissimilarities failed to provide enough detection of the migrating taxa. We collected 22 seawater samples from different locations in Egypt, in relationship to the migration across and to/from the Suez Canal. The V3-V4 regions of 16s genes were amplified and sequenced by the next generation Illumina MiSeq sequencer. Bioinformatics analysis revealed 15 taxa that migrated from the Mediterranean Sea to the Red Sea (i.e., anti-Lessepsian migration) such as the genera Fluvicola, HTCC2207, and Persicirhabdus. The family OCS155 is the only one that migrated from the Red Sea to the Mediterranean Sea (Lessepsian migration). Seven anti-Lessepsian migrants colonized the Suez Canal more than the Mediterranean Sea such as the genera Marinobacter and Halomonas. These findings collectively suggest that the anti-Lessepsian migration is more predominant than the Lessepsian migration in the bacterial community. This study paves the way for future research questions as well. For example, why is the anti-Lessepsian migration more common than the Lessepsian route in bacteria? Why do certain taxa stop migration at the Suez Canal, and why do certain taxa present in higher frequencies in the Suez Canal? Which taxa continue migration to the Indian Ocean and the Atlantic Ocean, and what is the impact of the anti-Lessepsian migration on the bacterial community? Understanding microbial diversity in a context of microorganism migration across seas and oceans remains a prime topic in biodiversity research and systems science.

To screen for a variety of marine bacteria with anti-quorum sensing and anti-biofilm activities. Among 188 bacterial isolates from water, sediment, and corals in the Red Sea region, approximately 35% (65 isolates) of the isolates displayed a significant degradation in the purple pigment of the bioreporter strain without affecting cell growth. The quorum quenching bacteria obtained from coral-associated bacteria were 66.2% out of the total isolates. The PCR amplification results revealed that the recorded Acyl Homoserine lactone (AHL) inhibition by 91% of the anti-QS marine bacteria was not due to lactonase activity. On the other hand, lactonase genes were recorded only in the remaining 9% (6 isolates) and those were belonging to genus Bacillus, Nocardiopsis, and Enterobacter based on 16S rRNA gene sequences. The results also showed that marine bacteria with anti-QS activity inhibited 67% of the biofilm formed by Aeromonas hydrophila, Pseudomonas aeruginosa, and Vibrio alginolyticus. The computational profiling analysis confirmed the presence of the functional region in the detected genes. Coral microbial communities are rich sources for pharmacologically important natural products with anti-quorum sensing and anti-biofilm activities.

The Erythraean species of mantis shrimp, Erugosquilla massavensis, has been reported as the first Lessepsian migrant stomatopod to the Mediterranean Sea in 1933, off Alexandria, Egypt via the Suez Canal. Currently, it extended to the middle and western Mediterranean competing with the native spot-tail mantis shrimp Squilla mantis (Linnaeus, 1758). The continuous increase of invasive species to the Mediterranean claimed the necessity to update and revise the identification of those early migrants. The present study aimed to confirm the morphological identification of E. massavensis from the coast of the Egyptian Mediterranean Sea using the DNA barcoding gene cytochrome oxidase subunit I (COI) and conducting a phylogenetic analysis. Genomic DNA of samples collected from the Mediterranean Sea at Port Said in 2016 was extracted and COI was amplified and sequenced. Our phylogenetic analysis revealed that E. massavensis (MH447072 and MH447073) constitutes a single monophyletic clade and appeared more linked with E. woodmasoni than the Japanese mantis shrimp Oratosquilla oratoria (divergence value < 3%). The phylogenetic relationship between E. massavensis, E. woodmasoni, and O. oratoria was confirmed by the Neighbor-Joining tree. The present study confirmed the morphological identification and document, for the first time in GenBank/EMBL/ DDBJ genetic databases, the genetic status of E. massavensis from the Mediterranean of Egypt using COI.

The novelty of the present work looks in the synthesis of aqueous dispersed selenium nanoparticles (Se NPs) using gamma rays with the aid of various natural macromolecules such as citrus pectin (CP), sodium alginate (Alg), chitosan (CS) and aqueous extract of fermented fenugreek powder (AEFFP) using Pleurotus ostreatus for investigating their impact in vitro toward carcinoma cell. The synthesized Se NPs were characterized by XRD, UV–Vis., DLS, HRTEM, SEM, EDX and FTIR. Nucleation and growth mechanisms were also discussed. The factorial design was applied to examine the importance of multiple parameters on Se NPs production with a special focus on temperature and gamma rays influences. FTIR spectrum exhibited the existence of several functional groups in Se NPs-capping macromolecules. Results revealed that Se NPs' size was dramatically-influenced by the type of stabilizer, precursors concentration, pH and the absorbed gamma rays dose. The current research reported the promising antitumor application of Se NPs against Ehrlich Ascites Carcinoma (EAC) and human Colon Adenocarcinoma (CACO) in vitro. The proliferation of EAC was significantly-hindered by Se NPs-CS (38.0 μg/ml) at 60 kGy (IC50 = 23.12%) and Se NPs-AEFFP (19.00 μg/ml) at 15 kGy (IC50 = 7.21%). Also, Se NPs control the generation of CACO cells, IC50 was recorded as 25.32% for Se NPs-CS (38.0 μg/ml) and 8.57% for Se NPs-AEFFP (19.00 μg/ml).

An effective vaccine against Staphylococcus aureus infection is a major planetary heath priority, particularly with increasing antibiotic resistance worldwide. Previous efforts for a highly effective S. aureus vaccine were largely unsuccessful, in part, because the vaccine designs have tended to target mainly the B cell immunity and development of opsonic antibodies. In contrast, recent observations suggest that cell mediated immunity may be critical for protection against S. aureus. In addition, the S. aureus surface proteins are among the key immunodominant antigens because they are the first molecules to interact with the host organism cells and tissues. We report here an original vaccinomics study in which we used a reverse vaccinology and immunoinformatics in silico strategy integrated with genomics. After analyzing 2767 proteins, we defined 16 proteins of S. aureus as promising subunit vaccine candidates. Phosphatidylinositol phosphodiesterase (Plc) is secreted by extracellular pathogens such as S. aureus. We mapped the B and T cell epitopes for the Plc protein, tested the reactivity of the synthesized epitopes by Western blotting, and verified our findings in a pilot study of 10 patients with S. aureus infection. The peptides were then tested for their protective effect in groups of mice challenged with pathogenic S. aureus strain, which showed high protection level. These findings warrant further translational research for development of novel vaccines against S. aureus infection. Reverse vaccinology is an advanced approach that can be applied to identify new vaccine candidates against a host of microorganisms, including S. aureus.

Microbial communities residing in the nose play important roles in human health and disease. We report marked differences in nasal microbiota between a rural community and an industrial setting located near a major urban city. Nasal samples were collected from 19 healthy male subjects: 9 samples from persons living in a rural village, and 10 samples from ceramic factory workers in a major industrial Egyptian city. The nasal microbiota in the rural sample had higher and distinct diversity compared with industrial samples from workers exposed to pollution daily. Taxonomic analysis of the sequences revealed five major phyla; among these phyla were Actinobacteria, Proteobacteria, Bacteroidetes, and Fusobacteria, revealing significant abundance variation by geographical location. For example, the rural group had a significant increase in representation of Actinobacteria and Bacteroidetes (

DNA extraction with high quality is critical to all molecular genetic analyses. However, obtaining DNA from microbes associated with animals is challenging. Despite the availability of various DNA extraction kits in the markets, no studies were conducted to date to evaluate their potential for the invertebrates such as nudibranch, one of Mollusca. This study compared the Quick—DNA Fungal/Bacterial Miniprep Kit and CTAB methods and tested them across four samples of Chromodoris quadricolor gut and skin tissues. The universal bacterial primers 331f and 797r and the animal-specific primers LCO1490-JJ and HCO2198-JJ were used to amplify the 16S rRNA gene and partial mitochondrial cytochrome oxidase I gene using extracted DNA as a template. The DNA and PCR products’ quality and concentration were verified with agarose gel and Nanodrop, respectively. The two methods' quality assessed using the deep pyrosequencing of the 16S rRNA gene in capturing a more diverse microbiome. The highest yield and purity (over 1000 ng / µL) were obtained with the CTAB method, while it was not exceeded 260 ng/µL with Quick DNA kit and display high purity. Also, 16S rRNA community amplicon sequencing revealed that the CTAB way could catch more diverse bacterial groups. The most efficient method of DNA extraction was CTAB, as it achieved both high concentration and purity

Bacterial vaginosis (BV) is highly common, adversely affecting the health of millions of women. New therapeutic targets and diagnostics are urgently needed for BV. Microbiome research offers new prospects in this context. We report here original findings on changes in the vaginal microbiome in pregnant and nonpregnant women with BV. Reproductive age women were recruited for this study after a clinical examination. The total sample (

An effective vaccine against

Red Sea marine sponges (phylum Porifera) and associated microorganisms harbor a wide range of microorganisms, which are considered an essential source of bioactive products. In this study, we screened both the crude extracts of Red Sea marine sponges and their associated bacterial extract for antimicrobial activity and antiviral activity. Molecular characterization of bioactive producers was performed using16S rRNA sequencing, in addition to metagenomic analysis of three representative sponges utilizing the 16S rRNA gene V3-V4 region sequencing in two different seasons. Twelve samples were collected from five different sponge species by scuba diving, and all the crude extracts of sponges showed antimicrobial activity except Negombata corticata. Moreover, 84 out of 110 bacterial isolates extracts demonstrated antimicrobial activity against at least one tested microorganism. These results revealed the bioactivity and biodiversity of the Red Sea marine invertebrates-associated bacteria. It was found that the bioactive isolates belong to several bacterial groups. The bacterial communities of Negombata magnifica, Negombata corticata, and Siphonochalina siphonella were shown with great diversity and differences in the bacterial percentage, diversity, and unique community composition at different seasons in each sponge species. Unique microenvironment for each sponge species may be linked to the production of specific bioactive product.

Measles virus considers an important cause of child morbidity and mortality in some areas as Africa. Ribavirin's activity as a nucleoside analog can disclose the surprisingly broad spectrum action against several RNA viruses under laboratory cell culture conditions. The Current study aimed to investigate the antiviral activity of ribavirin Nano gold particles (AuNPs) against measles virus on vero cell line. Ribavirin- AuNPs was prepared, characterization and the cytotoxicity of ribavirin, AuNPs and ribavirin -AuNPs were tested on vero cells using MTT assay. Antiviral activiry of ribavirin, AuNPs and ribavirin- AuNPswere determined on vero cells using simultaneous, pre-infection and post-infection protocols. Results indicated safety of ribavirin and ribavirin-AuNPs on vero cells, there was a reduction by 78.1% when vero cells treated with ribavirin -AuNPs at 99.5µg/ml while, the viral reduction was 25.4% when ribavirin 500 µg /ml was used for the same viral concentration. Our results concluded that ribavirin - AuNPs had a higher antiviral activity with lower dose than ribavirin alone and the maximal activity showed when it used after the virus infection.

Pseudomonas aeruginosa is a common and major opportunistic human pathogen, its causes many and dangerous infectious diseases like cystic fbrosis, wounds, burns inflammation, urinary tract infection, other many infections otitis external, nosocomial infection and causes bacteremia. A main problem in P. aeruginosa infection that exhibits a high degree of resistance to a broad spectrum of antibiotics. A total of 147 patients of Pseudomonas aeruginosa were isolated from Ismailia hospitals. The clinical isolates were collected from wound, sputum and urine. Carbapenem sensitivity was performed per Kirby-Bauer disk diffusion method and minimum inhibitory concentration (MIC) was preformed towards meropenem and imipenem resistant isolates by agar dilution methods. Imipenem and meropenem sensitivity result were 91% and 70%. MICs were carried out for carbapenem resistant isolates. Furthermore P. aeruginosa screening should be managed and antibiotic abuse should be monitored.

Fungal infections caused by dermatophytes recently became more common. Available antifungal drugs are limited because of emergence of resistant strains due to prophylaxis with them, so there is an urgent need for novel antifungals. This study is aimed to detect the antifungal activity of silver nanoparticles (SNPs) on Fluconazole resistant dermatophytes isolated from primary school children clinically suffering from tinea capitis and attending El-Sheikh Zaid Dermatology Center in Ismailia. The study was done on 112 clinical cases. Examination with potassium hydroxide(KOH) of hair samples was done, followed by routine identification using culturing, macroscopical and microscopical examination and biochemical tests, finally molecular identification using Polymerase Chain Reaction (PCR) with (GACA) 4 was done. Fluconazole resistance of these dermatophytes was detected by different methods including agar disc diffusion method and broth microdilution susceptibility testing. Silver nanoparticles susceptibility testing was carried out on these Fluconazole resistant dermatophytes. The Ubiquitin 1 (Ub 1) gene was detected in samples which were Fluconazole resistant but SNPs susceptible. In this study dermatophytes were found only in 70 samples (62.5%). They were belonged to 3 species: Trichophyton violaceum, Microsporum gypseum and Microsporum canis. Fluconazole resistance was found in 58 samples (82.85%). Both M. canis and M. gypseum were resistant to all used concentrations of SNPs, while T. violaceum was susceptible to 50 µg/ml SNPs solution. The Ub1 gene was detected in 1 sample (4.8%). Therefore SNPs can be used for treatment of T. violaceum, while they can't be used for treatment of M. canis or M. gypseum.

Bacteria regulate their social activities and physiological processes through a quorum sensing (QS) mechanism. Many bacteria inhabit the mesohyl matrix of sponges. This symbiotic relationship is believed to contribute to the health and nutrition of sponges by production of protective antibiotics, the acquisition of limiting nutrients and nitrogen fixation. Marine isolates were analysed by bioassays using two different acylated homoserine lactone (AHL) reporter bacteria Chromobacterium violaceum 026 (CV026) and Agrobacterium tumefaciens KYC55. Thirty four marine isolates activated the A. tumefaciens and produce blue color. Using degenerate LuxS primers, partial LuxS gene sequences were detected in both Gram-negative and Gram-positive species including Bacillus species, Proteus species, uncultured Bacterium species, Micrococcus species and Bacterium NLAE-zl-H221. Using degenerate LuxR primers, partial LuxR gene sequences were detected in Proteus spp., Providencia species and Bacterium NLAE-zl-H221. Partial LuxS gene sequences were detected in both Gram-negative and Gram-positive. The results thus assure the fact that LuxS gene has been proposed to be a universal interspecies signaling molecule. This study showed that, Proteus mirabilis strain SZH18, Proteus vulgaris and Bacterium NLAE-zl-H221 have both luxS and luxR genes, thus it was claimed that these isolates have the potential to use two different QS systems, for intraspecies communication and communication with other species. Key words: Quorum sensing, autoinducers, biofilm, double layer method, quorum quenching, N-acyl homoserine lactones.

Atopic dermatitis (AD) is a relapsing, chronic, and inflammatory skin disorder. Its causes remain unclear. Here, we reported the first proteome study of the bacterial community in AD patients. Bacterial community in 7 patients and 1 healthy control using bottom-up proteomics were examined starting with in-solution digestion followed by purification steps with subsequent analysis using LC-MS/MS and ended with data processing and bioinformatic analysis. Overall, great bacterial changes between patient samples and healthy one were noticed with the presence of Staphylococcus aureus, Aeromonas hydrophila, and Shewanella species, and others that were present uniquely in patient samples suggesting their role in AD. Additionally, detection of some important proteins that trigger bacterial pathogenesis and the immune system such as enolase, glyceraldehyde-3-phosphate, Chaperone proteins DnaK and HtpG beside protein pathways needed for bacterial growth and pathogenesis like chaperones and folding catalysts; and Energy metabolism. These new findings of the microbiome and detected proteins could start a new era of proteomics to study the bacterial community as a whole and detect the way it interacts with each other and with the host. SIGNIFICANCE: This paper would represent a reference work for investigations on microbiota that present on AD, from both a microbiological and a functional proteomic point of view. We focused on analysisng bacteria community and proteins produced and its role in the disease, highlighting some functional characteristics of certain proteins and discussing its potential role in AD.

Obesity and diabetes are reaching epidemic levels globally. Metagenomics and microbiome science have recently emerged as new tools for studying common complex human diseases. We report in this study notable differences in gut microbiome in adult patients with obesity and diabetes in Egypt. The experimental design was based on comparisons of four study groups: (1) Controls (C) with a normal body mass index, without obesity or diabetes, (2) Obese adults (O) without diabetes, (3) adults with diabetes (D) who are not obese, and (4) Adults who are both obese and diabetic (OD). In a total study sample of 60 participants, we sequenced the 16S ribosomal RNA (rRNA) gene using the Illumina MiSeq platform. Alpha diversity analysis revealed greater diversity in bacterial communities of (D) than controls. Phylum-level analysis identified a trend for overrepresentation of

Atopic dermatitis (AD) is a complex, multifactorial, chronic pruritic inflammatory skin disease. We report the first microbiome study and new insights on the relationship between skin microbiota variation and AD susceptibility in a population sample from Egypt. We characterized the skin microbiome in 75 patients with AD and 20 healthy controls using Illumina MiSeq sequencing of 16S rRNA gene. Overall, bacterial diversity of skin microbiome in patients with AD was less than those of the healthy subjects. Genus level analysis revealed significant abundance variations by age, disease severity, locality, or immune response. Among these genera, Streptococcus, Cutibacterium, and Corynebacterium appeared to be specific signatures for AD in children, adolescents, and adults, respectively, while Staphylococcus was noted as a potential biomarker candidate for AD. Additionally, functional potential of metagenomes shifted the overall metabolic pathways to participate in the exacerbation of disease. Total immunoglobulin E (IgE) levels were positively correlated with relative enrichment of certain Staphylococcus aureus subspecies. Finally, AD-related differences in skin bacterial diversity appeared to be in part linked to the serum IgE level. These new observations attest to the promise of microbiome science and metagenomic analysis in AD specifically, and clinical dermatology broadly.

The aim of this study was to characterize the beta lactamases genes of bacteria isolated from urinary tract infection (UTI) in Assiut, Egypt. Results revealed that one hundred fifty nine [31.8%] out from 500 urine samples were culture-positive. Escherichia coli was the most common UTI pathogen [61%] followed by Klebsiella pneumoniae [23.3%], Proteus mirabilis [8.2%] and Pseudomonas aeruginosa [7.5%]. Sensitivity of isolates to ampicillin was [15%], amoxicillin/clavulanic acid [43.5%], ceftriaxone [24%], imipenem [95.6%], amikacin [75%], ciprofloxacin [21.4%] and trimethoprim /sulfamethoxazole [37%]. Confirmatory phenotypic detection of extended-spectrum β-lactamases [ESBLs] by ESBL E-test method resulted in [42.7%] isolates were ESBLs producers. Genotypic characterization of ESBLs genes in phenotypically positive isolates resulted in [91.2%] were ESBL producers. The presence of CTX-M type ESBL was [75%] followed by TEM [37%], OXA [24%] and SHV [21%]. Sequencing of ESBLs genes showed that CTX-M-15, OXA [1,116], TEM-1 and SHV [1, 11,111,115] as new ESBL types. Multiple sequence alignment of sequenced genes showed mutation in L31R in SHV-11[Novel SHV-115], E29Q in SHV-1[Novel SHV-111], and P65R in TEM-1 and I97M in OXA-1 [Novel OXA-116]. This study is one from first studies in Egypt that highlights the presence of multiple mutations in ESBLs. Key words: Uropathogens, extended-spectrum β-lactamases (ESBLs), mutation, Egypt.

Bibliometric analysis of Egyptian literature on HCV provides the intelligence needed for decision makers and gives an insight into research productivity in this area. We propose our database (HCVDBegy) on MS-SQL server by querying PubMed for "HCV and Egypt" with time limit till 31st March 2013. Fifty eight out of the 716 records were excluded and the rest 658 were divided into 22 domains. Analysis used data mining add-ins for Microsoft Excel, including association and regression algorithms. A fluctuation in numbers of papers was noticed from 2004 to 2009 with a steady increase onward. Eighty six percent of publications were the contribution of three or more authors. Top publishing bodies were Cairo and Ain Shams Universities, Faculty of Medicine, National Research Center and National Cancer Institute. Three Egyptian journals came on top, whereas other publishing journals were mainly from the USA. Few controlled clinical trials and meta-analyses were published. HCV epidemiology, review articles and sequence analysis domains were the most cited. Forecasting model showed a breakthrough in numbers of publications on 2013 and 2014 than those forecasted. Dependency network based on association rule model of MeSH topics was also extensively analyzed. Number of publications showed a promising increase which points to the better national awareness of HCV problem. Studying MeSH terms clustering showed some hot topics. We recommend that the PubMed should alarm authors of the challenges of author affiliations. HCVDBegy availability opens the door for more drill down analysis for decision makers.

Vibrio cholerae is a waterborne bacterium inhabiting the aquatic ecosystem. Faecal contamination of water resources, represents modes of dissemination of the bacteria and thus the disease. This study is reporting an association between V. cholerae and chironomids larvae and exuviae, collected from Lake Manzala. The collection sites were chosen to cover the southern sector of the lake that is reportedly characterized by industrial, agricultural and domestic pollution. The only identified species was Chironomus transvaalensis, an East African species. The isolation of V. cholerae was carried out according to the standard microbiological methods, and followed by 16S rRNA sequence analysis. Non-O1/non-O39 V. cholerae was found to be present in all of the examined C. transvaalensis larvae and exuviae. These findings further support the use of chironomids as a monitoring agent of V. cholerae.

The objective of this work was to isolate bacteria from Red Sea invertebrates, determine their antimicrobial activity, and screen for the biosynthetic gene clusters [polyketides (PKs) and nonribosomal peptides (NRPs)] which could be involved in the production of bioactive secondary metabolites. Eleven different samples of marine invertebrates’ were collected from Egypt’s Red Sea (El-Tor-Sharm El-Sheikh and Hurghada) by scuba diving, and a total 80 isolates of the associated microorganisms were obtained from the cultivation on six different cultural medium. Seven isolates of them showed an antimicrobial activity against five pathogenic reference strains, while the most active antimicrobial agent was isolate number HFF-8 which was 99% identical to Bacillus amyloliquefaciens. HFF-8’s extract showed positive results against Gram negative bacteria, Gram positive bacteria and yeast. Moreover, the isolates gave positive bands when screened for the presence of PK synthase (PKS) I and II and NRP synthetase (NRPS) I and II biosynthetic genes, those biosynthetic fragments when cloned and sequenced were primitively predicted as biosynthetic fragments for kirromycin and leinamycin production by NaPDoS program with 56 and 55%, respectively. The Red Sea can provide a sustainable solution to combat bacterial resistance. The contribution of this work is that B. amyloliquefaciens was isolated from Heteroxenia fuscescens, Red Sea, Egypt. Moreover, the bacterial extract showed a broad spectrum with a potent antimicrobial activity.

Aqueous dispersed zinc nanoparticles (ZnNPs) were synthesized using natural polysaccherides (chitosan (CS), citrus pectin (CP) and alginate (Alg)) using aqueous fermented fenugreek powder (FFP) by Pleurotus ostreatus as reducing and stabilizing agent or using gamma irradiation. The synthesized ZnNPs are characterized by ultra violect spectroscopy (UV), Transmission electronmicroscopy (TEM), Dynamic light scattering (DLS), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FT-IR). XRD analysis of the ZnNPS confirmed the formation of metallic nanoparticles. The nucleation and growth mechanism of ZnNPs is also discussed. TEM showed that the average diameter of ZnNPs was in the range of 46nm. The size of nanoparticles is influenced by certain parameters such as the choice of stabilizer, pH during synthesis and absorbed dose. Evaluating the antioxidant and anticancer activities of ZnNPs was performed. The results indicating the ZnNPs synthesized by aqueous fermented fenugreek extract have high activity and the average size is 46nm. The results explored that ZnNPs show anticancer activity against Ehrlich Ascites Carcinoma (EAC) and human colon adenocarcinoma (CACO) and the IC50% was 47.5μg/ml and 65μg/ml respectively. Also, ZnNPs had excellent bactericidal activity against gram positive and negative bacteria and yeast.

Measles virus considers an important cause of child morbidity and mortality in some areas as Africa. Ribavirin's activity as a nucleoside analog can disclose the surprisingly broad spectrum action against several RNA viruses under laboratory cell culture conditions. The Current study aimed to investigate the antiviral activity of ribavirin Nano gold particles (AuNPs) against measles virus on vero cell line. Ribavirin- AuNPs was prepared, characterization and the cytotoxicity of ribavirin, AuNPs and ribavirin -AuNPs were tested on vero cells using MTT assay. Antiviral activiry of ribavirin, AuNPs and ribavirin- AuNPswere determined on vero cells using simultaneous, pre-infection and post-infection protocols. Results indicated safety of ribavirin and ribavirin-AuNPs on vero cells, there was a reduction by 78.1% when vero cells treated with ribavirin -AuNPs at 99.5µg/ml while, the viral reduction was 25.4% when ribavirin 500 µg /ml was used for the same viral concentration. Our results concluded that ribavirin - AuNPs had a higher antiviral activity with lower dose than ribavirin alone and the maximal activity showed when it used after the virus infection.

The double-stranded RNA dependent protein kinase (PKR) plays a vital role in the immune system. During HCV infection, PKR has antiviral effect by inhibition of protein synthesis of the HCV. The functional single nucleotide polymorphisms (SNPs) in PKR promoter region might have a relation to HCV disease outcome and response to treatment. The objective of the present work was threefold. First, it proposed an optimized protocol for PCR amplification of PKR promoter. Second, it screened the promoter region of PKR gene in HCV Egyptian patients to detect the possible SNPs' function. Third, to study the association between the detected SNPs and the response to treatment.The functional SNPs in PKR promoter region were detected using DNA sequencing in 40 HCV infected patients; 20 sustained virologic response (SVR) patients and 20 nonresponse (NR) patients after combined interferon/ribavirin therapy. Twenty healthy subjects were included as a control.Two functional SNPs were detected: rs62133148TG and rs12992188CT within our target PKR promoter region. In rs62133148 polymorphism, there is a significant difference between patients and control subjects for TT and TG genotypes (p 0.0001). In addition, the G allele is more predominant in HCV patients. In rs12992188 polymorphism, the CC genotype is significantly different between patients and healthy control subjects (OR/95% CI: 0.033/0.006-0.172, p 0.0001). The presence of C allele was significantly associated with the NR patients (OR/95%CI: 0.25/0.097-0.643, p = 0.006). The TT genotype is significantly different between SVR and NR (OR/95%CI: 8.5/1.54-46.871, p = 0.014).This study is a pioneer clinical study on these two functional SNPs (rs62133148TG and rs12992188 CT). The rs62133148 polymorphism does not show any association with response to treatment. The TT genotype in rs12992188 polymorphism shows association with response to treatment. Therefore, patients with TT genotypes were more likely to achieve SVR.

Background: Resistant Pseudomonas aeruginosa is a serious concern for antimicrobial therapy, as the common isolates exhibit variable grades of resistance, involving beta-lactamase enzymes, beside native defense mechanisms. Objectives: The present study was designed to determine the occurrence of Metallo-β- Lactamases (MBL) and Amp C harboring P. aeruginosa isolates from Suez Canal university hospital in Ismailia, Egypt. Methods: A total of 147 P. aeruginosa isolates, recovered from 311 patients during a 10-month period, were collected between May 2013 and February 2014; the isolates were collected from urine, wound and sputum. Minimum inhibitory concentration (MIC) determined by agar dilution methods was ≥2 μg/mL for meropenem and imipenem. Identification of P. aeruginosa was confirmed using API 20NE. Metallo-β- Lactamases and Amp C were detected based on different phenotypic methods. Results: Overall, 26.5% of P. aeruginosa isolates (39/147) were carbapenem resistant isolates. Furthermore, 64.1% (25/39) were MBL producers, these isolates were screened by the combined disc and disc diffusion methods to determine the ability of MBL production. Both MBL and Amp C harbored P. aeruginosa isolates were 28% (7/25). Sixty-four percent of P. aeruginosa isolates were multidrug resistant (MDR) (16/25). The sensitivity toward polymyxin, imipenem, norfloxacin, piperacillin-tazobactam and gentamicin was 99%, 91%, 88%, 82% and 78%, respectively. The resistance rate towards cefotaxime, ceftazidime, cefepime, aztreonam and meropenem was 98.6%, 86%, 71.4%, 34% and 30%, respectively. Conclusions: Multidrug resistance was significantly associated with MBL production in P. aeruginosa. Early detection of MBL-producing P. aeruginosa and hospital antibiotic policy prescription helps proper antimicrobial therapy and avoidance of dissemination of these multidrug resistance isolates.

This study was designed to investigate the prevalence of metallo-β-lactamase (MBL) in Pseudomonas aeruginosa isolates collected from Suez Canal University Hospital in Ismailia, Egypt. Antibiotic susceptibility testing and phenotypic and genotypic screening for MBLs were performed on 147 isolates of P. aeruginosa. MICs were determined by agar dilution method for carbapenem that was ≥2 μg/mL for meropenem. MBL genes were detected by multiplex and monoplex PCR for P. aeruginosa-harbored plasmids. Mutation profile of sequenced MBL genes was screened using online software Clustal Omega. Out of 147 P. aeruginosa, 39 (26.5%) were carbapenem-resistant isolates and 25 (64%) were confirmed to be positive for MBLs. The susceptibility rate of P. aeruginosa toward polymyxin B and norfloxacin was 99% and 88%, respectively. Identification of collected isolates by API analysis and constructed phylogenetic tree of 16S rRNA showed that the isolates were related to P. aeruginosa species. The frequency of blaGIM-1, blaSIM-1, and blaSPM-1 was 52%, 48%, and 24%, respectively. BlaVIM and blaIMP-like genes were 20% and 4% and the sequences confirm the isolate to be blaVIM-1, blaVIM-2, blaVIM-4, and blaIMP-1. Three mutations were identified in blaVIM-4 gene. Our study emphasizes the high occurrence of multidrug-resistant P. aeruginosa-producing MBL enzymes.

A successful 17 β estradiol (E2) degrading bacterium was isolated from waste water of Saft Elhena drain in Egypt. Molecular identification of the isolated bacteria was carried out using DNA (RAPD), the amplification was made by restriction analysis of amplified 16s rDNA and sequencing. Alos, the bacterium was incubated in saline media under different levels of 17 β-estradiol anaerobically. Levels of CO2 production were examined by GC and residual 17 β-estradiol was tested by HPLC to measure biodegradation capacity. Identification of the genetic element responsible of the gene resistance in the bacteria was tested. The sequence analysis of 16S rRNA gene for 17 β estradiol degrader bacterium showed that its affiliations to phylum Enterobacteriaceae, and it belonged to the genera Klebsiella sp. Results revealed that Klebsilla was cabable of degrading estradiol and that the responsible resistance gene was mostly the chromosomal gene. However, further studies are still going on to locate and examine the gene responsible for E2 resistance.

This study was initiated to provide the first recor d of monitoring of 17 �- estradiol (E2) residues in some Egyptian aquatic ec osystems. Samples of water were collected from three water bodies located in the Su ez Canal region. Samples were extracted, filtered and examined by HPLC on a C 18 column using Florescence detectors. Results provided evidence of the presence of estra diol in the studied area. Interestingly, marine lakes contained significant l evels of 17 �-estradiol (P