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ABSTRACTA carbapenem-resistant sequence type 512 (ST512) Klebsiella pneumoniaecarbapenemase 3 (KPC-3)-producing K. pneumoniaestrain showing a novel variant plasmid content was isolated in Palermo, Italy, in 2014. ST512 is a worldwide successful clone associated with the spread of blaKPCgenes located on the IncFIIk pKpQIL plasmid. In our ST512 strain, the blaKPC-3gene was unusually located on an IncX3 plasmid, whose complete sequence was determined. Two copies of blaKPC-3::Tn4401acaused by intramolecular transposition events were detected in the plasmid.

AbstractBackground:Globally, antimicrobial drug–resistant Escherichia coliis among the most common etiological agents of invasive disease in humans. In Europe, increasing proportions of infections due to third-generation cephalosporins and/or fluoroquinolone-resistant extraintestinal pathogenic E. coli(ExPEC) strains are reported. E. colifrom poultry are those more closely linked to human E. coli, but lack of reliable data makes it difficult to assess the attributable risk of different food sources. In the present study, our objective was to investigate the antimicrobial resistance profile, phylogenetic background, and virulence factors of E. coliisolates from broiler chicken meat sold at retail in Palermo, Italy.Materials and Methods:Isolation of multidrug resistant (MDR) E. coliwas performed during April–December 2013 on a total of 163 chicken meat samples. Susceptibility to a panel of nine antimicrobial agents was determined. PCR assays were carried out to detect extended-spectrum β-lactamase (ESBL), plasmid-mediated AmpC β-lactamase, and plasmid-mediated quinolone resistance (PMQR) genes, phylogenetic group, and ExPEC-associated traits. A single nucleotide polymorphism (SNP) PCR was done to detect E. colisequence type (ST)131.Results:One hundred thirty-four isolates from 109 meat samples were MDR. B1 was the most prevalent phylogenetic group (47.8%), followed by groups D (25.4%), A (22.3%), and B2 (4.5%). ESBLs and AmpC β-lactamases were detected by PCR in 132 (98.5%) and 15 (11.2%) isolates. PMQR determinants were detected in 122 (91%) isolates. Twenty-two MDR isolates met the molecular definition of ExPEC. SNP-PCR results confirmed that four B2 isolates were ST131. Enterobacterial Repetitive Intergenic Consensus sequence–PCR analysis showed a large heterogeneity with 55 unique profiles and 31 clusters including 2–4 isolates.Conclusions:An alarmingly high prevalence of MDR E. colifrom broiler chicken meat is evident in our geographic area. The ongoing use of antimicrobial drugs in livestock should be urgently restricted, particularly in the poultry sector.

In hepatitis B virus (HBV) cirrhosis patients on long-term lamivudine (LAM), the relationships between HBV suppression, development of viral resistance and disease outcome are unclear. We analysed the dynamic of serum HBV-DNA and its relationship with the clinical course of 59 patients (52 males, mean age 51.4 ±8.4 years, 12 HBeAg positive and 47 HBeAg negative, and 57 genotype D and two genotype A) with cirrhosis (45 in Child-Turcotte-Pugh class A) and high levels of serum HBV-DNA (median 14.7x107genomes/ml) treated with LAM [median (range): 44 (15–78) months]. A total of 50 patients (84.7%) achieved a virological response (serum HBV-DNA negative by PCR) during the first 6 months of therapy, and nine (13.3%) achieved a reduction in viral load of >3 log10. Mutations in the YMDD motif of HBV polymerase were documented in 26 patients [median (range) 18: (7–42) months]. At the time of the emergence of mutants, 22 patients had HBV-DNA <105genomes/ml and normal alanine aminotransferase (ALT) levels. The appearance of virological resistance was followed by an increase of HBV-DNA to =105genomes/ml and of ALT values in 19 out of 26 patients [median (range): 8 (3–19) months]. Event-free survival was significantly longer (P=0.001) in patients who maintained virological suppression than in those who did not have a complete virological response or suffered a breakthrough. Patients with advanced cirrhosis were more likely to develop liver failure after the emergence of YMDD mutants. The risk of development of hepatocellular carcinoma in patients with compensated cirrhosis and YMDD mutations was maintained, regardless of HBV-DNA serum levels.Profound and maintained HBV-DNA suppression correlates with a better outcome. Early identification of LAM resistance mutations allows switching to other antivirals before liver decompensation or hepatocellular carcinoma development.

In chronic hepatitis C the rate of relapse afteran end-of-treatment response to interferon may exceed50%. The usefulness of retreatment of relapsers withinterferon in obtaining a complete sustained response and the role of clinical, virological andimmunological features in determining long-term efficacyof retreatment are unclear. We aimed to assess theefficacy of interferon retreatment in obtaining acomplete sustained response, to evaluate whetherincreasing the dose may enhance responsiveness, and toidentify possible predictors of sustained response. Weenrolled 42 patients with biopsy-proven chronichepatitis C without cirrhosis who had previouslyresponded to a six-month course ofInterferon-α2b(total dose: group A, 22patients, 234 MU; group B, 20 patients, 468 MU) and thenrelapsed. All, except one, were HCV-RNA negative at the end of first cycle ofinterferon; most (31/42, 74%) were infected by HCV 1b.Subjects were randomly allocated to receive anothercycle of interferon either at the original dose (group A1: 234 MU, 11 patients; groupB 468 MU, 10 patient) or twice the original dose (groupA2: 468 MU, 11 patients; group B : 936 MU, 10patients). At the end of the second cycle of interferon,24 subjects (57%) had normal ALT and were HCV-RNAnegative, and 16 (39%) had normal ALT, but were HCV-RNApositive. A complete sustained response was obtained ineight patients (19%), at a similar rate in all treatment groups. Complete sustainedresponders were different from the other patients interms of age (35.9 ± 10.4 vs 44.1 ± 8.8,P = 0.027), rate of infection with non-1b HCV (6/8 vs5/34, P = 0.0005), serum HCV-RNA (74,016 vs 321,428median copies/ml, P = 0.037) and serum levels of90K/MAC-2 BP (5.76 ± 3.01 vs 10.25 ± 5.16units/ml, P = 0.02), an N-glycoprotein implicated incellular defense functions. Multivariate logistic analysisvalidated age and HCV genotype as independent predictorsof CSR. Among noncirrhotic relapsers who received atotal interferon dose ≥234 MU in the first cycle,retreatment usually induced end-of-treatment response. Acomplete sustained response was obtained in only one ofevery five subjects. Increasing the dose of interferonabove that of the first cycle did not enhance the rate of sustained response. In conclusionwe might assert that young subjects infected by non-1bHCV and with low levels of HCV-RNA and of 90K/MAC-2 BPare the best candidates for retreatment.

Transcriptional regulators are thought to play a key role in cell fate determination and territorial specification in sea urchin. Our goals are to clone transcription factors for studying embryonic development. One approach has been to use promoter binding and gene transfer technology to investigate the mechanisms of transcriptional activation and repression of the early H2A histone gene. By this analysis we identified a transcriptional activator, the MBF-1, that binds to the modulator element of the H2A gene and enhances the activity of the H2A promoter. However, the enhancer activity of the modulator and its interaction with MBF-1 also occurs at the gastrula stage when the early histone genes are shut off. Therefore, the silencing of the early H2A histone gene at late stages of development requires the inactivation of the modulator function. To search for antimodulator sequence elements, we took advantage of our previous work showing the presence of phased nucleosomes specifically positioned on the 3'-spacer and in the modulator of the repressed H2A gene. Evidence is described indicating that a 3'-spacer DNA fragment cloned between the modulator and the basal promoter behaves as an antimodulator element. However, this element does not confer temporal capability to the modulator in its function, suggesting that other elements have to be involved in the regulation of the early H2A expression. The second approach relied on the cloning of genes controlling development using probes of regulators known to be involved in regional specification, such as the homeobox genes, with the aim to understand their possible role through the study of their temporal and territorial expression and through the analysis of the mechanism of regulation of their expression. We present evidence that PIHbox 12, a homeodomain encoding gene, is transiently expressed during the early/mid cleavage stages. The abundance of the transcripts reach their maximum in embryos at the 64/128-cell stage concomitantly with the segregation of the specified embryonic territories. Expression of PIHbox l2 is drastically reduced in the absence of cell contacts and/or Ca ions, suggesting that this gene is transcriptionally activated by signal transduction mechanisms. Whole mount in situ hybridization showed that PIHbox 12 transcripts are asymmetrically distributed along the A-V axis, being spatially localized in the blastomeres of the ectodermal lineage. We suggest that PIHbox l2 might be involved in the initial events that lead to the specification of the ectodermal territories.

Here we describe carbapenem resistance determinants in two Klebsiella pneumoniaeisolates recovered from two hospitalised patients in the same intensive care unit of a cancer hospital in Cairo, Egypt. PCR and sequencing were used to detect and characterise β-lactamase genes. Clonal relationships between the isolates were analysed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The first K. pneumoniaeisolate carried the blaNDM-1gene and the second isolate carried the blaOXA-163gene. Both isolates co-expressed the extended-spectrum β-lactamase CTX-M-15. The two isolates belonged to different sequence types (STs), ST11 and ST16, respectively. No history of travel was established for the two patients. The first identification of NDM-1-producing K. pneumoniaein Egypt adds further evidence to the spread of NDM-1-producing Gram-negative micro-organisms in North Africa. The additional detection of blaOXA-163in a K. pneumoniaeisolate confirms its endemic presence in a critical healthcare setting of this geographic area.

ABSTRACTTwo genetically unrelated OXA-163-carrying Klebsiella pneumoniaestrains were identified from two infection cases in June 2009 and May 2010 in Cairo, Egypt. OXA-163-producing Enterobacteriaceaehad been previously reported in Argentina only. Both patients had no history of travel abroad. The emergence of this newly recognized OXA-48-related ß-lactamase able to hydrolyze cephalosporins and carbapenems is especially worrying in a geographic area where OXA-48 is endemic and effective surveillance for antibiotic resistance is largely unaffordable.

Coagulase negative staphylococci are increasingly recognized as leading pathogens in bacteremia, with incidence peaking in intensive care units. Interpretation of blood cultures that are positive for CoNS is often doubtful. We describe a fatal case of bacteremia by a newly recognized species of CoNS, Staphylococcus pettenkoferi, in an ICU patient.