Inference of Antibiotic Resistance and Virulence among Diverse Group A Streptococcus Strains Using emm Sequencing and Multilocus Genotyping Methods

typing (direct sequencing of the genomic segment coding for the antigenic portion of the M protein) or by multilocus genotyping methods. Phenotype analysis, including critical AbR typing, is generally achieved by much slower and more laborious direct culture-based methods. type and the associated AbR and virulence phenotypes. types

The kinase activity of fibroblast growth factor receptor 3 with activation loop mutations affects receptor trafficking and signaling

Amino acid substitutions at the Lys-650 codon within the activation loop kinase domain of fibroblast growth factor receptor 3 (FGFR3) result in graded constitutive phosphorylation of the receptor. Accordingly, the Lys-650 mutants are associated with dwarfisms with graded clinical severity. To assess the importance of the phosphorylation level on FGFR3 maturation along the secretory pathway, hemagglutinin A-tagged derivatives were studied. The highly activated SADDAN (severe achondroplasia with developmental delay and acanthosis nigricans) mutant accumulates in its immature and phosphorylated form in the endoplasmic reticulum (ER), which fails to be degraded. Furthermore, the Janus kinase (Jak)/STAT pathway is activated from the ER by direct recruitment of Jak1. Abolishing the autocatalytic property of the mutated FGFR3 by replacing the critical Tyr-718 reestablishes the receptor full maturation and inhibits signaling. Differently, the low activated hypochondroplasia mutant is present as a mature phosphorylated form on the plasma membrane, although with a delayed transition in the ER, and is completely processed. Signaling does not occur in the presence of brefeldin A; instead, STAT1 is activated when protein secretion is blocked with monensin, suggesting that the hypochondroplasia receptor signals at the exit from the ER. Our results suggest that kinase activity affects FGFR3 trafficking and determines the spatial segregation of signaling pathways. Consequently, the defect in down-regulation of the highly activated receptors results in the increased signaling capacity from the intracellular compartments, and this may determine the severity of the diseases

The Kinase Activity of Fibroblast Growth Factor Receptor 3 with Activation Loop Mutations Affects Receptor Trafficking and Signaling

View of the baths, depicting arch; From the 2nd century AD onwards, gymnasia combining palaestras and baths occupied a large area of the city of Ephesos. Their strong walls and vaults made them expensive to build, and the large quantities of water needed to operate them had to be supplied through reinforced pipes. Surviving buildings include the Vedius Gymnasium, the Theatre Gymnasium, the 'East Gymnasium', the Baths of Varius (renovated in Late Antiquity by a certain Scholastikia) and the Harbour Gymnasium (all mainly 2nd to 3rd century AD). The most usual masonry for walls and vaults in the 1st century AD was rough stonework bonded with lime mortar. In the early 2nd century AD construction in brick began, using square bricks the size of a Roman foot. At the end of the 2nd century AD the bricks became larger, and broken bricks are also found in the opus caementum filling of the great walls of bath buildings. Stone masonry in conjunction with brickwork first occurs in Late Antiquity. [The mosaics in the 40 meter long corridor dates to the 5th century. It has three sections, frigidarium (cold water), tepidarium (warm water) and caldarium (hot water).] Source: Grove Art Online; http://www.oxfordartonline.com/ (accessed 7/13/2008

Isolate properties by PCR/ESI-MS <i>emm</i> group.

<p>Observed rates of antibiotic resistance and scarlet fever exotoxin gene carriage for all identified PCR-ESI/MS emm groups. N+(N) = Number of Positives (Number Tested). Bold indicates 50% or greater rates of resistance. Positives either carry a particular <i>spe</i> gene or show resistance to a particular antibiotic (inclusive of both intermediate and full resistance). ERY, erythromycin; CLI, clindamycin; TET, tetracycline; LEV, levofloxacin; CHL, chloramphenicol; OFX, ofloxacin.^aIndependently significant specific association, positive or negative. ^bSignificant after alpha (Bonferroni) adjustment for multiple measures (highly significant). <i>speA</i>, scarlet fever exotoxin gene A; <i>speC</i>, scarlet fever exotoxin gene C. These following results are not shown in the table: 100% of tested isolates (in all cases N>300) were susceptible to penicillin (PEN), vancomycin (VAN), cefepime (CPM), cefotaxime (CTX), ceftriaxone (CTR), linizolid (LNZ), meropenem (MEM), and gatifloxacin (GAT). 311 were tested with quinupristin-dalfopristim (SYN) and one was resistant, six isolates were tested with trovafloxacin (TVA) and 3 were resistant.</p

Inverse relationships between <i>emm</i> groups and <i>emm</i> types.

<p><i>emm</i> groups (set of equally likely <i>emm</i> types, as identified by T-5000 PCR/ESI-MS analysis) associated with each <i>emm</i> type (identified by <i>emm</i> gene sequence analysis); Contradictory identifications are shown in bold. All identities are shown as “identity (number identified)” for that specific associative category.</p

Rates and significance of nonrandom distribution by sequenced <i>emm</i> type.

<p>Overall rate of antibiotic resistance among all GAS <i>emm</i> types, and the significance of nonrandom association between <i>emm</i> type and antibiotic resistance.^aIncludes both intermediate and fully resistant isolates, as per CLSI definitions. <i>p</i> values represent the probability that the observed distribution of resistance among the types resulted from a random distribution among the real population. Pearson tests were run using Monte Carlo method; N = 10,000.</p><p>ERY, erythromycin; CLI, clindamycin; TET, tetracycline; LEV, levofloxacin; OFX, ofloxacin; CHL, chloramphenicol.</p

Isolate properties by sequenced <i>emm</i> type.

<p>Observed rates of antibiotic resistance and scarlet fever exotoxin gene carriage for all identified sequence-based <i>emm</i> types. N+(N) = Number of Positives (Number Tested). Bold indicates 50% or greater rates of resistance. Positives either carry a particular <i>spe</i> gene or show resistance to a particular antibiotic (inclusive of both intermediate and full resistance). ERY, erythromycin; CLI, clindamycin; TET, tetracycline; LEV, levofloxacin; CHL, chloramphenicol; OFX, ofloxacin.^aIndependently significant specific association, positive or negative. ^bSignificant after alpha (Bonferroni) adjustment for multiple measures (highly significant). <i>speA</i>, scarlet fever exotoxin gene A; <i>speC</i>, scarlet fever exotoxin gene C. These following results are not shown in the table: 100% of tested isolates (in all cases N>300) were susceptible to penicillin (PEN), vancomycin (VAN), cefepime (CPM), cefotaxime (CTX), ceftriaxone (CTR), linizolid (LNZ), meropenem (MEM), and gatifloxacin (GAT). 311 were tested with quinupristin-dalfopristim (SYN) and one was resistant, six isolates were tested with trovafloxacin (TVA) and 3 were resistant.</p

Rates and significance of nonrandom distribution by PCR/ESI-MS <i>emm</i> group.

<p>Overall rate of antibiotic resistance among all GAS <i>emm</i> groups, and the significance of nonrandom association between <i>emm</i> group and antibiotic resistance. ^aIncludes both intermediate and fully resistant isolates, as per CLSI definitions. <i>p</i> values represent the probability that the observed distribution of resistance among the types resulted from a random distribution among the real population. Pearson tests were run using Monte Carlo method; N = 10,000.</p><p>ERY, erythromycin; CLI, clindamycin; TET, tetracycline; LEV, levofloxacin; OFX, ofloxacin; CHL, chloramphenicol.</p

Correlations between <i>emm</i> groups and <i>emm</i> types.

<p><i>emm</i> types (identified by <i>emm</i> gene sequence analysis) associated with each <i>emm</i> group (set of equally likely <i>emm</i> types, as identified by T-5000 PCR/ESI-MS analysis); Contradictory identifications are shown in bold. All identities are shown as “identity (number identified)” for that specific associative category.</p