Data_Sheet_1_Comparative Genomic Analyses Reveal Potential Factors Responsible for the ST6 Oxacillin-Resistant Staphylococcus lugdunensis Endemic in a Hospital.PDF

Oxacillin-resistant Staphylococcus lugdunensis (ORSL) is considered a life-threatening isolate in healthcare settings. Among ORSL clones, ST6-SCCmec II strains are associated with an endemic spread in hospitals. We analyzed the complete genome of ORSL CGMH-SL118, a representative strain. Results revealed that this strain contained three MGEs (two prophages and one plasmid) other than the SCCmec II element, which showed remarkable differences in genome organization compared to the reference strains from NCBI. Eight multidrug-resistant genes were identified. All but blaZ were carried by MGEs, such as the SCCmec II element [mecA, ant (9)-Ia, and ermA] and the prophage φSPbeta [aac (6')-aph (2'), aph (3')-III, and ant (6)-Ia], indicating that MGEs carrying multidrug-resistant genes may be important for ST6 strains. The prophage φSPbeta contains sasX gene, which was responsible for the pathogenesis of Staphylococcus aureus. A phage-mediated resistant island containing fusB (SlRIfusB-118) was found near φSPbeta, which was highly homologous to type III SeRIfusB-5907 of Staphylococcus epidermidis. In contrast to previous studies, over 20% of ST6 isolates showed a fusidic acid-resistant phenotype, suggesting that phage-mediated intraspecies transmission of resistant islands may become an important issue for ST6 strains. Sixty-eight clinical isolates of ST6 Staphylococcus lugdunensis (50 OSSL, oxacillin-sensitive S. lugdunensis, and 18 ORSL, including CGMH-SL118) collected from various types of specimens in the hospital were studied. Among these isolates in this study, ORSL showed similar drug-resistant genes and phenotypes as CGMH-SL118. The comparative genomic analyses highlight the contribution of MGEs in the development and dissemination of antimicrobial resistance in ST6 strains, suggesting that resistance determinants and virulence factors encoded by MGEs provide a survival advantage for successful colonization and spread in healthcare settings.</p

CI outcomes of the 10 patients with the identified candidate variants.

CI outcomes of the 10 patients with the identified candidate variants.</p

Variants identified in the parents of DE3386 (DE4451 and DE4452).

Variants identified in the parents of DE3386 (DE4451 and DE4452).</p

Pipeline used to identify candidate variants.

Forty-one patients with CI who lacked the known mutations in common deafness-associated genes were subjected to comprehensive genetic analysis by using an Ion Torrent PGM sequencer to target 40 relatively rare deafness genes. Variants were called using plug-in Torrent variant detection algorithms, annotated through wANNOVAR, and initially confirm using the integrative Genome viewer. Annotated variants were filtered by various criteria, including: being located in an exonic region; being non-synonymous; having an allele frequency 85% for homozygotes; and being absent from online databases and 128 ethnically matched normal hearing controls. SIFT, Polyphen 2, and Mutation taster were used to predict the functions of the identified variants; we first filtered for missense variants, and then directly identified indels, splicing site variants, and nonsense variants.</p

Cytotoxic Sesquiterpene Lactones from the Root of <i>Saussurea </i><i>l</i><i>appa</i>

Bioassay-directed fractionation of Saussurea lappa led to the isolation of a novel lappadilactone (1) and seven sesquiterpene lactones (2−8) as cytotoxic principles against selected human cancer cell lines. Lappadilactone (1), dehydrocostuslactone (2), and costunolide (5) exhibited the most potent cytotoxicity with CD50 values in the range 1.6−3.5 μg/mL in dose- and time-dependent manners. The cytotoxicities were not specific and showed similar activities against HepG2, OVCAR-3 and HeLa cell lines. The structure−activity relationship showed that the α-methylene-γ-lactone moiety is necessary for cytotoxicity, and activity is reduced with the presence of a hydroxyl group. In addition, seven noncytotoxic compounds (9−15) were also isolated, including two novel sesquiterpenes, a guaianolide-type with a C17 skeleton, lappalone (13), and 1β,6α-dihydroxycostic acid ethyl ester (14). The structures of the new compounds were elucidated from spectroscopic and/or X-ray data interpretations. Some representative compounds were also tested for antibacterial activity; however, only marginal activities were observed. Therefore, compounds 1−8 are potential cytotoxic agents but without significant antibacterial effect

Families DE3395 and DE3386.

(A) Probands of each family are indicated by arrows. (B) Audiograms of DE3395 (left) and DE3386 (right). Both recipients had bilateral symmetric flat-type audiograms of profound severity. Hearing levels of the right ear and left ear are marked with red and blue lines, respectively.</p

Correlation between vancomycin and daptomycin MICs of 157 MRSA blood isolates determined by E-test; <i>r</i> (Pearson's correlation coefficient) = 0.474.

<p>Correlation between vancomycin and daptomycin MICs of 157 MRSA blood isolates determined by E-test; <i>r</i> (Pearson's correlation coefficient) = 0.474.</p

Thirteen different variants were identified in 10 patients.

Thirteen different variants were identified in 10 patients.</p

Correlations between molecular types and phenotypes of MRSA blood isolates.

<p>*<i>p</i><0.05,</p><p>**<i>p</i><0.01,</p><p>***<i>p</i><0.001.</p><p>Abbreviations: VA, vancomycin; DPC, daptomycin; <i>pvl</i>, Panton-Valentine leucocidin; <i>agr</i>, accessory gene regulator.</p>1<p>Multidrug resistance: resistance to ≥4 classes of antibiotics (disc diffusion test).</p

Candidate variants detected in nine genes of 41 patients.

Each color bar indicates a different variant type, as indicated.</p