Characterization of Acylated Anthocyanins in Callus Induced From Storage Root of Purple-Fleshed Sweet Potato, Ipomoea batatas L

Four anthocyanins were isolated from a highly pigmented callusinduced from the storage root of purple-fleshed sweet potato(Ipomoea batatas L) cultivar Ayamurasaki. Theanthocyanins were respectively identified as cyanidin3-O-(2-O-(6-O-(E)-caffeoyl-β-D-glucopyranosyl)-β-D-glucopyranoside)-5-O-β-D-glucopyranoside, cyanidin3-O-(2-O-(6-O-(E)-p-coumaroyl-β-D-glucopyranosyl)-6-O-(E)-caffeoyl-β-D-glucopyranoside)-5-O-β-D-glucopyranoside, cyanidin3-O-(2-O-(6-O-(E)-p-coumaroyl-β-D-glucopyranosyl)-6-O-(E)-p-coumaroyl-β-D-glucopyranoside)-5-O-β-D-glucopyranoside, and peonidin3-O-(2-O-(6-O-(E)-p-coumaroyl-β-D-glucopyranosyl)-6-O-(E)-p-coumaroyl-β-D-glucopyranoside)-5-O-β-D-glucopyranoside by chemical and spectroscopic analyses. These anthocyanins wereexamined with respect to the stability in neutral aqueous solutionas well as the radical scavenging activity against the1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. These acylatedanthocyanins exhibited both higher stability and higher DPPHradical scavenging activity than corresponding nonacylatedcyanidin and peonidin 3-O-sophoroside-5-O-glucosides.     

New species and records of the mite genus Prolistrophorus (Acariformes: Listrophoridae) from rodents of the subfamily Sigmodontinae (Rodentia: Cricetidae)

Six fur-mite species of the genus Prolistrophorus Fain, 1970 (Acariformes: Listrophoridae) were recorded from Central and South American rodents of the subfamily Sigmodontinae (Rodentia: Cricetidae). Among them, Prolistrophorus (Aprolistrophorus) parabidentatus sp. nov. from Akodon azarae from Argentina and Prolistrophorus (Aprolistrophorus) tylomys sp. nov. from Tylomys nudicaudus from Guatemala are described as new for science. New hosts are recorded for the following species: Prolistrophorus (Prolistrophorus) grassii (Radford, 1954) from Zygodontomys brevicauda from Colombia, P. (P.) frontalis (Hirst, 1921) from Oligoryzomys sp. from Argentina, P. (P.) argentinus (Hirst, 1921) from Melanomys caliginosus, Akodon affinis from Colombia and Scapteromys aquaticus from Argentina, Prolistrophorus (Beprolistrophorus) hirstianus Fain, 1973 from Scapteromys aquaticus from Argentina.     

Phenotypic and molecular characterization of Escherichia albertii: Further surrogates to avoid potential laboratory misidentification

Escherichia albertii is an emerging gastrointestinal pathogen, related to Escherichia coli, which can be misidentified as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC), due to the presence of the eae gene in E. albertii. The aim of this study was to verify our hypothesis that E. coli cytolethal distending toxin-II (Eccdt-II) gene-positive E. coli is E. albertii and to accumulate the data regarding the bacteriological characteristics of E. albertii. For these purposes, we attempted to detect E. albertii in eae gene-positive bacteria previously identified as E. coli and to examine if re-identified E. albertii contained Eccdt-II-homologous gene and remaining eae gene-positive E. coli did not. A total of 373 eae gene-positive E. coli strains were analyzed by biochemical tests, multilocus sequence analysis and an E. albertii-specific PCR. The strains re-identified as E. albertii were also examined for the presence of cdt genes by using ³²P-labled DNA probes, followed by their toxin-typing. Of the 373 strains, 17 were re-identified as E. albertii by three above-mentioned methods. Furthermore, all the 17 re-identified E. albertii possessed cdt genes highly homologous to Eccdt-II and Eacdt genes. Moreover, Eccdt-I or both Eccdt-I and stx2f genes were detected in two re-identified E. albertii strains. However, the remaining 356 strains did not carry such cdt genes. These data indicate that all re-identified E. albertii isolates specifically carried cdt genes homologous to Eccdt-II and Eacdt genes. We suggest that Eccdt-II gene-positive E. coli may be identical to E. albertii.     

Genomic structure of the whole D–J–C clusters and the upstream region coding V segments of the TRB locus in pig

In the vertebrate immune system, T cells play a central role in host defense against microbial or viral infection. Previous studies suggested that at least two sets of TRBD–J–C clusters are harbored in the porcine genome. In this study, we determined 212,193 bp of a continuous porcine genomic sequence covering the entire TRBC region. EPHB6, TRPV6, TRY, and ten TRBV genes were conserved in the vicinity of the TRBD–J–C clusters. Interestingly, three TRBD–J–C clusters were identified in this sequence; each TRBD–J–C cluster consisted of one TRBD and seven TRBJ segments, with one TRBC region composed of four exons. The distribution of repetitive sequences and phylogenetic analysis indicated that the TRBD–J–C cluster, located at the center of the three clusters identified, had a structure combined with the others. Most of the TRBJ segments were available in public databases, suggesting that all three TRBD–J–C clusters are functional in pigs.     

Strong linkage disequilibria among HLA-Bw50, Bfs1, and HLA-DR3/DR7, and mapping of the Bf locus

Based on genotypic and phenotypic studies we have found strong linkage disequilibria in Caucasians among the genes HLA-Bw50, Bf^s1, and HLA-DR3 and/or -DR7. The relative disequilibria, which are among the highest described in man, are Δ_r(Bf^s1, DR7) = 0.51, Δ_r (Bw50, Bf^s1, DR7) = 0.36, Δ_r (Bw50, DR3 or 7) = 0.72, Δ_r (Bf^s1, DR3 or 7) = 0.91, Δ_r (Bw50, Bf^s1, DR3 or 7) = 0.73. The previously described high Δ_r (Bw50, Bf^s1) andΔ_r (Bw50, DR7) have also been confirmed. A B//DR crossover family is also presented that, together with previously reported recombinant families, confirms that the Bf locus resides between HLA-B and HLA-DR. These data suggest the existence of a supergene complex of Bw50, Bf^s1. DR3/7 (or MB2), and hypotheses to account for the observed disequilibria are discussed.     

Regulation and expression of human Fabs under the control of the Escherichia coli arabinose promoter,PBAD

Background: The l-arabinose operon from E. coli contains an inducible promoter P_BAD which has been extensively studied for the control of gene expression. P_BAD has a number of potential advantages over P_lac, and has been used successfully to promote high level expression of recombinant proteins. Objectives: The aim of this study was to investigate P_BAD as an alternative system to P_lac for the bacterial expression of recombinant Fabs. Study design: The promoter P_BAD from the E. coli arabinose operon araBAD and the gene encoding the regulator of this promoter, were cloned into the phagemid expression vector MCO1. Expression of human recombinant tetanus toxoid (TT) and c-erb B2 Fabs under the control of P_BAD was compared at two induction temperatures with the same Fabs produced under the control of P_lac. Results: Expression of TT and c-erb B2 Fabs under the control of P_BAD was comparable to Fab expression from P_lac. However, highly expressed TT Fab under the control of P_BAD was localised to the soluble periplasmic fraction whereas under the control of P_lac, there was greater leakage of Fab into the culture supernatant. In addition, Fab expression from P_BAD could be more tightly repressed than from P_lac. Conclusion: P_BAD is a useful and cheaply inducible alternative to the more commonly used P_lac for the rapid expression of soluble recombinant human antibody fragments.     

DNA microarray based on arrayed-primer extension technique for identification of pathogenic fungi responsible for invasive and superficial mycoses

An oligonucleotide microarray based on the arrayed-primer extension (APEX) technique has been developed to simultaneously identify pathogenic fungi frequently isolated from invasive and superficial infections. Species-specific oligonucleotide probes complementary to the internal transcribed spacer 1 and 2 (ITS1 and ITS2) region were designed for 24 species belonging to 10 genera, including Candida species (Candida albicans, Candida dubliniensis, Candida famata, Candida glabrata, Candida tropicalis, Candida kefyr, Candida krusei, Candida guilliermondii, Candida lusitaniae, Candida metapsilosis, Candida orthopsilosis, Candida parapsilosis, and Candida pulcherrima), Cryptococcus neoformans, Aspergillus species (Aspergillus fumigatus and Aspergillus terreus), Trichophyton species (Trichophyton rubrum and Trichophyton tonsurans), Trichosporon cutaneum, Epidermophyton floccosum, Fusarium solani, Microsporum canis, Penicillium marneffei, and Saccharomyces cerevisiae. The microarray was tested for its specificity with a panel of reference and blinded clinical isolates. The APEX technique was proven to be highly discriminative, leading to unequivocal identification of each species, including the highly related ones C. parapsilosis, C. orthopsilosis, and C. metapsilosis. Because of the satisfactory basic performance traits obtained, such as reproducibility, specificity, and unambiguous interpretation of the results, this new system represents a reliable method of potential use in clinical laboratories for parallel one-shot detection and identification of the most common pathogenic fungi.     

RAS/RAF mutations and their associations with epigenetic alterations for distinct pathways in Vietnamese colorectal cancer

KRAS, NRAS, and BRAF are potential tumor-driven genes that are involved in the RAS/RAF/MAPK signaling pathway. RAS/RAF mutations importantly contribute to colorectal tumorigenesis since they remain the activated status of downstream pathways without regulation of the upstream EGFR signal. However, it has not been unclear how epigenetic alterations involved in colorectal tumorigenesis mediated by KRAS, NRAS, or BRAF mutations. Therefore, in this study, we investigated the frequency and distribution of KRAS/NRAS/BRAF mutations in Vietnamese colorectal cancer (CRC) and explored the relationship between genetic and epigenetic abnormalities in 156 tumors of CRC. Somatic mutations of KRAS (exon 2, codon 12/13; exon 3, codon 61), NRAS (exon 2, codon 12/13; exon 3, codon 61), and BRAF (exon 15, codon 600) was determined by Cobas® KRAS Mutation Test, Therascreen NRAS Pyro Kit and Cobas® 4800 BRAF V600 Mutation Test, respectively. Methylation status of BRCA1, MLH1, MGMT, p16, RASSF1A, and APC was detected by methylation-specific PCR. Distribution of each abnormality in clinicopathological features was also analyzed. Results showed the mutation rates of KRAS, NRAS, and BRAF were 41.0 %, 9.6 %, 8.3 % respectively, while the methylation rates of BRCA1, MLH1, MGMT, p16, RASSF1A, and APC were 16.7 %, 16.7 %, 32.7 %, 30.1 %, 30.1 %, and 37.2 % respectively. The distribution of KRAS mutation was mutually exclusive against that of NRAS (p < 0.001) and BRAF (p < 0.001) mutations in CRC. RAS/RAF mutations were more common in adenocarcinoma subtype (p = 0.020), whereas RASSF1A methylation was more frequent in mucinous adenocarcinoma subtype (p = 0.007). In addition, the frequency of having KRAS mutations was significantly higher in MGMT (p = 0.035) or RASSF1A (p = 0.043) methylated cases than in those without methylation. BRAF mutations were positively associated with MLH1 hypermethylation (p = 0.028) but were inversely associated with APC hypermethylation (p = 0.032). Overall, our results show specific interactions of genetic and epigenetic alterations and suggest the presence of independent oncogenic pathways in tumorigenesis of CRC.     

Imd pathway is involved in the interaction of Drosophila melanogaster with the entomopathogenic bacteria,Xenorhabdus nematophila and Photorhabdus luminescens

Xenorhabdus nematophila/Steinernema carpocapsae and Photorhabdus luminescens/Heterorhabditis bacteriophora are nemato-bacterial complexes highly pathogenic for insects. Using a syringe as artificial vector, we have analyzed the effects of the two bacteria, X. nematophila and P. luminescens on the genetic tool insect, Drosophila melanogaster. Both bacteria were found to kill adult flies in a dose dependent manner with X. nematophila being the fastest. On the other hand, when an injection of non-pathogenic bacteria, Escherichia coli, is performed 1 day before challenge with the entomopathogenic bacteria, then the survival of Drosophila flies was prolonged by at least 20 h. After injection of entomopathogenic bacteria, Drosophila mutant Dif¹, affected on the Toll pathway, showed a similar phenotype than wild-type flies whereas Drosophila mutant Dredd^D55, affected on the imd pathway, was not protected by a prior injection of E. coli. This suggested that members of the imd pathway might be targets of these entomopathogenic bacteria albeit synthesis of antimicrobial peptides through this signaling pathway was induced by X. nematophila as well as P. luminescens. Finally, P. luminescens phoP mutant, an avirulent mutant in the Lepidopteran insect, Spodoptera littoralis, was found poorly virulent for D. melanogaster. phoP mutant partially protected D. melanogaster flies if injected 1 day before the injection of P. luminescens wild-type TT01 to the same extent than the E. coli-induced protection. However, phoP recovered a level of pathogenicity comparable to P. luminescens wild-type TT01 when injected to Drosophila flies affected on the imd pathway.     

PhoP/Q regulated genes inSalmonella typhi: identification of melittin sensitive mutants

Many of the genes (pags (phoPactivatedgenes) andprgs (phoPrepressedgenes) ) regulated by the PhoP and PhoQ proteins (PhoP/Q) are necessary for survival ofSalmonella typhimuriumin murine macrophages and pathogenesis in mice. Although a great deal is known about theS. typhimuriumphoP/Qregulon, little has been done with the human specific pathogenS. typhi, prompting us to investigateS. typhiphoP/Qregulated genes. IsogenicphoP12(null) andphoP24(constitutive) strains were constructed inS. typhiTy2 andS. typhimuriumC5 strains. Comparison of whole cell proteins from these strains by SDS-PAGE showed differences in both the number and molecular mass of PhoP/Q regulated proteins. This suggested thatS. typhiandS. typhimuriummay have different PhoP/Q regulated proteins and/or that their regulation may be different. A genetic procedure was developed to isolate mutations in PhoP/Q regulated genes. This involved random MudJ transposon mutagenesis of aphoP12mutant, creatinglacZ-gene fusions, and screening for Lac⁺ or Lac^- colonies. A mobilizable plasmid carrying thephoP24mutant gene was conjugated into these insertion mutants. Those that changed from Lac^- to Lac⁺ were inferred to bepag::MudJ insertions and those that changed from Lac⁺ to Lac^- were inferred to beprg::MudJ insertions. Five mutants with PhoP/Q regulated MudJ fusions were found by this scheme. The mutations were termedpqa(PhoPQ activated) andpqr(PhoPQ repressed) to distinguish them from other PhoP/Q regulated genes. Thepqa/pqr::MudJ mutations were transduced intoS. typhiphoP⁺ andphoP24strains by Vi-I phage transduction. Characterization of the mutants (Southern blot analysis, β-galactosidase activity on indicator plates and in liquid cultures) strongly suggested that their MudJ insertion mutations were in five different genes. Further characterization involved determining cationic peptide sensitivity and mouse virulence. Two mutants were found to be sensitive to the antimicrobial peptide melittin.     

Organization of the plasmid cpe Locus in Clostridium perfringens type A isolates

Clostridium perfringens type A isolates causing food poisoning have a chromosomal enterotoxin gene (cpe), while C. perfringens type A isolates responsible for non-food-borne human gastrointestinal diseases carry a plasmid cpe gene. In the present study, the plasmid cpe locus of the type A non-food-borne-disease isolate F4969 was sequenced to design primers and probes for comparative PCR and Southern blot studies of the cpe locus in other type A isolates. Those analyses determined that the region upstream of the plasmid cpe gene is highly conserved among type A isolates carrying a cpe plasmid. The organization of the type A plasmid cpe locus was also found to be unique, as it contains IS1469 sequences located similarly to those in the chromosomal cpe locus but lacks the IS1470 sequences found upstream of IS1469 in the chromosomal cpe locus. Instead of those upstream IS1470 sequences, a partial open reading frame potentially encoding cytosine methylase (dcm) was identified upstream of IS1469 in the plasmid cpe locus of all type A isolates tested. Similar dcm sequences were also detected in several cpe-negative C. perfringens isolates carrying plasmids but not in type A isolates carrying a chromosomal cpe gene. Contrary to previous reports, sequences homologous to IS1470, rather than IS1151, were found downstream of the plasmid cpe gene in most type A isolates tested. Those IS1470-like sequences reside in about the same position but are oppositely oriented and defective relative to the IS1470 sequences found downstream of the chromosomal cpe gene. Collectively, these and previous results suggest that the cpe plasmid of many type A isolates originated from integration of a cpe-containing genetic element near the dcm sequences of a C. perfringens plasmid. The similarity of the plasmid cpe locus in many type A isolates is consistent with horizontal transfer of a common cpe plasmid among C. perfringens type A strains.     

Interspecies Variations in Bordetella Catecholamine Receptor Gene Regulation and Function

Bordetella bronchiseptica can use catecholamines to obtain iron from transferrin and lactoferrin via uptake pathways involving the BfrA, BfrD, and BfrE outer membrane receptor proteins, and although Bordetella pertussis has the bfrD and bfrE genes, the role of these genes in iron uptake has not been demonstrated. In this study, the bfrD and bfrE genes of B. pertussis were shown to be functional in B. bronchiseptica, but neither B. bronchiseptica bfrD nor bfrE imparted catecholamine utilization to B. pertussis. Gene fusion analyses found that expression of B. bronchiseptica bfrA was increased during iron starvation, as is common for iron receptor genes, but that expression of the bfrD and bfrE genes of both species was decreased during iron limitation. As shown previously for B. pertussis, bfrD expression in B. bronchiseptica was also dependent on the BvgAS virulence regulatory system; however, in contrast to the case in B. pertussis, the known modulators nicotinic acid and sulfate, which silence Bvg-activated genes, did not silence expression of bfrD in B. bronchiseptica. Further studies using a B. bronchiseptica bvgAS mutant expressing the B. pertussis bvgAS genes revealed that the interspecies differences in bfrD modulation are partly due to BvgAS differences. Mouse respiratory infection experiments determined that catecholamine utilization contributes to the in vivo fitness of B. bronchiseptica and B. pertussis. Additional evidence of the in vivo importance of the B. pertussis receptors was obtained from serologic studies demonstrating pertussis patient serum reactivity with the B. pertussis BfrD and BfrE proteins.     

MiR-148a inhibits angiogenesis by targeting ERBB3

MicroRNAs (miRNAs) play an important role in carcinogenesis in various solid cancers including breast can-cer. Down-regulation of microRNA-148a (miR-148a) has been reported in certain cancer types. However, the biological role of miR-148a and its related targets in breast cancer are unknown yet. In this study, we showed that the level of miR-148a was lower in MCF7 cells than that in MCF10A cells. V-erb-b2 erythroblastic leukemia viral oncogene homolog 3 (ERBB3) is a direct target of miR-148a in human breast cancer cells through direct binding of miR-148a to ERBB3 3’-UTR region. Overexpression of miR-148a in MCF7 cells inhibited ERBB3 expression, blocked the downstream pathway activation including activation of AKT, ERK1/2, and p70S6K1, and decreased HIF-1α expression. Furthermore, forced expression of miR-148a attenuated tumor angiogenesis in vivo. Our re-sults identify ERBB3 as a direct target of miR-148a, and provide direct evidence that miR-148a inhibits tumor an-giogenesis through ERBB3 and its downstream signaling molecules. This information would be helpful for target-ing the miR-148a/ERBB3 pathway for breast cancer prevention and treatment in the future.     

Occurrence and molecular characterization of Hysterothylacium aduncum (Nematoda: Anisakidae) from Merlangius merlangus euxinus and Trachurus trachurus off the Turkish coast of Black Sea

A total of 286 larval forms of Hysterothylacium aduncum were collected from Merlangius merlangus euxinus and Trachurus trachurus captured at different sites of the Black Sea coast of Turkey. Prevalence of H. aduncum in M. merlangus euxinus and T. trachurus was 37.4 and 29.3 %, respectively. The fourth-stage larvae from M. merlangus euxinus and T. trachurus of H. aduncum were characterized genetically using a molecular approach. The ribosomal DNA (rDNA) internal transcribed spacer (ITS) region (ITS-1, 5.8S subunit, ITS-2) was amplified and sequenced. Two isolates of H. aduncum obtained from M. merlangus euxinus and T. trachurus in Black Sea showed a 100 % nucleotide similarity. Pairwise comparison between the entire ITS fragment including ITS-1, 5.8S, ITS-2 sequences of the H. adumcum isolates of M. merlangus euxinus and T. trachurus from Black Sea (Turkey, JX413596-JX413597) and other H. adumcum isolates from Baltic Sea (Poland, AJ937672), North Sea (Denmark, HM598666), Mediterranean Sea (Tunisia, HQ270427), Japan Sea (Japan, AB277826), Adriatic Sea (Croatia, JQ934878), East Greenland Sea, English Channel, Bay of Biscay, Adriatic Sea, and North Sea showed differences ranging from 0.1 to 0.7 %. With the present study, larvae of H. aduncum infecting M. merlangus euxinus and T. trachurus caught off the Black Sea, Turkey were characterized for the first time by sequencing of the ITS rDNA.     

Detection of mecA- and mecC-Positive Methicillin-Resistant Staphylococcus aureus (MRSA) Isolates by the New Xpert MRSA Gen 3 PCR Assay

An advanced methicillin-resistant Staphylococcus aureus (MRSA) detection PCR approach targeting SCCmec-orfX along with mecA and mecC was evaluated for S. aureus and coagulase-negative staphylococci. The possession of mecA and/or mecC was correctly confirmed in all cases. All methicillin-susceptible S. aureus strains (n = 98; including staphylococcal cassette chromosome mec element [SCCmec] remnants) and 98.1% of the MRSA strains (n = 160, including 10 mecC-positive MRSA) were accurately categorized.     

Distribution of MICA alleles and haplotypes associated with HLA-B in Greek population

IntroductionThe Major Histocompatibility Complex Class I-related chain A gene (MICA) is a highly polymorphic functional gene located close to the HLA-B locus. Certain MICA alleles have been related to inflammatory and autoimmune diseases while MICA antibodies have been implicated in organ allograft rejection or graft-versus-host disease (GVHD).AimThe aim of this study was to identify the frequencies of MICA alleles and MICA ~ HLA-B haplotypes in the Greek population since, as far as we know, these data are still limited.MethodsDNA was obtained from 277 unrelated healthy Greek individuals of Caucasian origin, volunteer donors of blood stem cells. HLA-B* and MICA* genotyping was performed by reverse PCR-SSOP.ResultsA total of 18 MICA alleles were defined in the present study. The five most frequent alleles in the Greek population were MICA*008 (24.6%), MICA*009 (22.36%), MICA*018 (16.03%), MICA*002 (8.02%) and MICA*004 (7.17%) which altogether account for 77.8% of all alleles. The most common MICA ~ HLA-B haplotypes were MICA*018 ~ B*18 (12.5%) and MICA*009 ~ B*51(11.5%).ConclusionsThe five most frequent MICA alleles in the Greek population were *008, *009, *018, *002, *004. In other Caucasian populations, two of these alleles (*008, and *004) were observed in similar frequencies. MICA*002 was observed less frequently (8.02%) in the Greek population compared to other Caucasian groups (frequencies > 15%). Also, MICA*009 and MICA*018 had elevated frequencies (above 15%) whereas in other Caucasian populations they were found around 10% or less. These data may be important for the elucidation of the role that MICA polymorphisms play in organ and stem cell transplantation and to identify the relation of certain MICA with susceptibility to specific diseases.     

Characterization of Enterococcus faecium isolates and first report of vanB phenotype–vanA genotype incongruence in the Middle East

We aimed to characterize the vancomycin genotype/phenotype, carriage of putative virulence genes, and genetic relatedness of Enterococcus faecium isolates in Saudi Arabia. E. faecium isolated from inpatients at our medical center were studied. Sensitivity to ampicillin, linezolid, teicoplanin, quinupristin/dalfopristin, tetracycline, and ciprofloxacin was determined. The presence of van genes and virulence genes for aggregation substance (Asa-1), enterococcal surface proteins (esp), cytolysin (cylA, cylL, cylM), gelatinase (gelE), E. faecium endocarditis antigen (EfaA _fm ), hyaluronidase (hyl), and collagen adhesion (Ace) was assessed. Genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE). Twenty-nine E. faecium isolates were obtained and the majority of isolates (n/N?=?22/29) were from stool specimens. PFGE analysis identified eight pulsotypes (A–H) based on 80?% similarities. Isolates were represented in five major pulsotypes: type A (n?=?5), type B (n?=?3), type D (n?=?6), type E (n?=?5), and type F (n?=?7). All isolates were vanA gene-positive. Thirteen isolates had vanA⁺/vanB⁺ genotype. Of these, ten exhibited a vanB phenotype and three had a vanA phenotype. Eight isolates with vanA⁺/vanB^? genotype exhibited vanB phenotype. Six of these eight isolates belonged to the same pulsotype. All isolates were positive for gelE, esp, and EfaA _fm genes. Five were CylA-positive and 24 had the hyl genes. Of the eight isolates harboring a combination of gelE, esp, EfaA _fm , and hyl genes, five showed vanB phenotype–vanA genotype incongruence. This is the first report of vanB phenotype–vanA genotype incongruent E. faecium in the Middle East region. Molecular typing indicates clonal spread and high occurrence of virulence genes, especially esp genes, associated with epidemic clones.