The camera faced the whole cage and allowed

monkeys’ latencies to take food buy Dasatinib rewards placed at the back of the Perspex box to be measured and general behaviour and facial expressions to be recorded for later analysis. Four macaque monkeys (Macaca mulatta) were tested on a social valuation task (Rudebeck et al., 2006) before and after mOFC lesions. Briefly, animals were tested in the WGTA (Fig. 3A) and on every trial the monkey retrieved a small food item that was placed in a fixed central position on the top of a transparent plastic box. Two different emotive toy snakes (static and moving) were used to investigate fearfulness (experiment 1a). The five short films of other macaques (detailed above) were used to investigate social valuation in experiment 1b. Responsiveness to videos of humans staring was also assessed in experiment 1c. Finally, responsiveness to neutral control objects was also assessed in order to provide a baseline against which to compare any changes in fearfulness and social valuation (experiment 1d). On each trial, stimuli were placed in the Perspex box or displayed on a screen behind the box. The animal had 30 s to retrieve the food item or else an opaque moveable screen was lowered in between the animal and the box for the duration of a 30-s Ipilimumab ic50 intertrial interval. The latency to reach for the piece of food indexed the

macaques’ assessments of the value of obtaining additional information about the stimulus before reaching, and reflected their relative valuation of the stimulus in contrast very to the incentive value of the food. On each day, animals were exposed to ten different stimuli of possible social or emotional importance and 20 neutral objects. The test was repeated

over four sessions (with a day of rest in between sessions) and the median reaching latency for each stimulus per animal was calculated. Each stimulus either in the box or on the screen was presented once per day. Objects in the box and images on the screen were presented in a pseudorandom order. The constraints enforced on order were that neutral object trials always followed trials in which potentially fear-inducing or social stimuli (snake or monitor stimuli) were presented. In experiment 1 two animals (mO1 and mO2) had acted as unoperated controls in a previous experiment (Rudebeck et al., 2006). The other two animals (mO3 and mO4) were tested only in this study pre- and postoperatively. The data from all four animals were considered together because there was no discernable difference between animals’ performances in relation to the time of testing. Animals were first habituated to the testing environment and then trained to take food from the top of the Perspex box while it was empty. The food reward was located at the centre of the back edge of the box nearest the PC monitor so that during the actual test the animal would have to reach over anything in the box or as close as possible to the monitor.

He suffered from diabetes mellitus type 2, but was otherwise healthy. In the previous years he had complained of intermittent abdominal pain, but both an ultrasound and X-ray performed the previous year were normal. He did not return to Sri Lanka or visit other tropical areas in the period of 2005 to 2007. At admission his blood samples showed white blood cell count (WBC) of 10.7 × 109 L−1 and the C-reactive protein (CRP) level was 100 mg/L. Abdominal computed tomography (CT) scan demonstrated a splenic abscess (Figure

1A), and he was transferred to the regional hospital for further treatment. The abscess was drained, and treatment with antibiotics was started. A fistula between the spleen and colon was eventually diagnosed, and a splenectomy was performed. Histological examination of biopsies from colon and spleen demonstrated subacute inflammation,

fibrosis, and necrosis. One week after surgery he developed a subphrenic abscess JQ1 price that was drained successfully. Five days after admission there was growth in blood culture of a nonfermentative, oxidase-positive, gram-negative rod with bipolar staining. The bacteria grew on blood and lactose agar. After some days of culture, the colonies appeared large and dry with a typical wrinkled surface. The bacteria isolated from blood were identified as Burkholderia pseudomallei by the Vitek 2 system with Ganetespib manufacturer 96.4% probability. Sequencing of the 16S rRNA gene demonstrated Etomidate DNA sequences identical to sequences of B pseudomallei in GenBank. Later, the bacteria were isolated from both the splenic and the subphrenic abscesses. The commercial biochemical test API 20 NE (BioMérieux, Marcy l’Etoile, France) supported the identification. The rod grew at 42°C, which is in contrast to the characteristics of Burkholderia mallei. The minimum inhibitory concentration (MIC) values obtained from the E-tests (AB Biodisk, BioMérieux) performed on the isolates are summarized in Table 1. The patient was treated with antibiotics intravenously for a total of 6 weeks.

At the admission to hospital he initially received cefuroxime and metronidazole, but because of lack of clinical response this was changed to meropenem after a few days. For the last couple of weeks of the treatment he received piperacillin-tazobactam according to susceptibility data, available before the bacteria were identified. Although piperacillin-tazobactam appears to be effective in vitro, there is little clinical experience on which to recommend their use.1 However, the clinical condition of the patient improved during this period, so there is reason to believe that also in vivo susceptibility existed for this antibiotic. He was thereafter transferred back to his local hospital and received eradication therapy with trimethoprim-sulfamethoxazole (TMP-SMX) and doxycycline for a total of 20 weeks with gradual improvement of his clinical condition.

g., transgenic reporter mice (Jonsson et al., 2009) or pluripotent stem cells (Takahashi

& Yamanaka, 2006; Tabar et al., 2008; Lindvall & Kokaia, 2009). We thank Anneli Josefsson and Ulla Jarl for expert technical assistance and Dr Eilís Dowd for valuable guidance in adapting the corridor task to mice. The study was supported by grant from the Swedish Research Council (04X-3874) and, in part, also from the EU 7th Framework Programme, NeuroStemcell (222943). Abbreviations 6-OHDA 6-hydroxydopamine CPu caudate–putamen unit DA dopamine DAergic dopaminergic KPBS potassium selleck screening library phosphate-buffered saline MFB medial forebrain bundle MPTP 1-methyl-1,2,3,4-tetrahydropyridine NAc nucleus accumbens PD Parkinson’s disease SN substantia nigra TH tyrosine hydroxylase VTA ventral tegmental area Fig. S1. Correlation of behavioural impairments and degeneration of the nigrostriatal pathway. Fig. S2. Correlation of behavioural impairments and degeneration of the mesolimbocortical pathway. As a service to our authors and readers, this journal provides supporting information supplied by

the authors. Such materials are peer-reviewed and may be re-organized for online delivery, but are not copy-edited or typeset by Wiley-Blackwell. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. “
“Serotonin (5-hydroxytryptamine; 5-HT) is a physiological signal that translates both internal and external information about behavioral context into changes in sensory processing through a diverse array see more of receptors. The details of this process, particularly how receptors interact to shape sensory encoding, are poorly understood. In the inferior colliculus, a midbrain auditory nucleus, 5-HT1A receptors have suppressive and 5-HT1B receptors have facilitatory effects on evoked

responses of neurons. We explored how these two receptor classes interact by testing three hypotheses: that they (i) affect separate neuron populations; (ii) affect different response properties; Lenvatinib manufacturer or (iii) have different endogenous patterns of activation. The first two hypotheses were tested by iontophoretic application of 5-HT1A and 5-HT1B receptor agonists individually and together to neurons in vivo. 5-HT1A and 5-HT1B agonists affected overlapping populations of neurons. During co-application, 5-HT1A and 5-HT1B agonists influenced spike rate and frequency bandwidth additively, with each moderating the effect of the other. In contrast, although both agonists individually influenced latencies and interspike intervals, the 5-HT1A agonist dominated these measurements during co-application. The third hypothesis was tested by applying antagonists of the 5-HT1A and 5-HT1B receptors. Blocking 5-HT1B receptors was complementary to activation of the receptor, but blocking 5-HT1A receptors was not, suggesting the endogenous activation of additional receptor types.

, 2009). Various structures of Mt-DapD have been obtained, both in native form and in complex

with succinyl-CoA see more (Schuldt et al., 2008, 2009). A ribbon model of Mt-DapD is shown in Fig. 2. Mt-DapD forms a biologically relevant homotrimer, and each monomer is composed of three distinct domains – an N-terminal α/β-globular domain, a left- handed parallel β helix and a small C-terminal domain (Schuldt et al., 2008, 2009). The amino acid residues Glu 199 and Gly 222 of Mt-DapD are important for enzymatic activity. Mt-DapD is activated by Mg2+, Ca2+ and Mn2+ and inhibited by Co2+ and Zn2+ (Schuldt et al., 2009). The sixth step in this pathway is catalysed by Mt-DapC (Rv0858c), which transfers an amino group

from l-glutamate Olaparib molecular weight and converts the substrate N-succinyl-2-amino-6-ketopimelate to N-succinyl diaminopimelate by the use of a pyridoxal phosphate (PLP) cofactor (Weyand et al., 2006, 2007). Mt-DapC belongs to the aminotransferase family of class I PLP-binding proteins. Mt-dapC has been heterologously expressed, purified and crystallized in two related crystal forms that arise from a pH difference between the crystallization conditions (Weyand et al., 2006). In the tetragonal crystal form, a monomer was present in the asymmetric unit, whereas in the orthorhombic crystal form, a dimer was present in the asymmetric unit (Weyand et al., 2006). Because of the presence of PLP in the crystal, both crystal forms appeared as pale yellow (Weyand et al., 2006). The three-dimensional structure of Mt-DapC was refined to a resolution of 2.0 Å (Weyand et al., 2007) and displayed the characteristic S-shape of class I PLP-binding proteins. Distinct from other class I PLP structures, Mt-DapC has an eighth β-strand inserted between strands three and four (Weyand et al., 2007). A ribbon diagram of Mt-DapC is shown in Fig. 2. 6-phosphogluconolactonase Mt-dapE (Rv1202) encodes the N-succinyl-l,l-diaminopimelic

acid desuccinylase. DapE catalyses the hydrolysis of N-succinyl-l,l-diaminopimelic acid (SDAP) to l,l-diaminopimelic acid and succinate (Born et al., 1998; Davis et al., 2006). The enzyme is a metal-dependent peptidase (MEROPS family M28) catalysing the hydrolysis of substrate by water with the help of one or two metal ions located in the active site (Born et al., 1998; Nocek et al., 2010). DapEs have been over-expressed and purified from Helicobacter pylori, E. coli, Haemophilus influenzae and Neisseria meningitidis (Bouvier et al., 1992; Karita et al., 1997; Born et al., 1998; Bienvenue et al., 2003; Badger et al., 2005). DapEs from E. coli and H. influenzae are small proteins (approximately 42 kDa) requiring two Zn2+ ions per mole of polypeptide for their activity (Bouvier et al., 1992; Born & Blanchard, 1999; Bienvenue et al., 2003).

ART success was defined as VL < 400 copies/mL or stable/rising CD4 counts or both. Data on demographics,

adherence, CD4 counts, weights, and post-travel VL were compared between the two groups, between those who had or did not have ART failure and where appropriate before and after travels. t-Test, Wilcoxon-rank-sum (z), Fisher’s exact, and Chi-square (χ2) tests and measures of effect were used for comparison between groups as appropriate, with two-sided p-value < 0.05 regarded as significant. A nested case-controlled analysis was done to determine the role of Hajj in ART failure. Analysis was done using STATA (version 10.0) (College Station, TX, USA). A total of 32 HP on ART performed the Hajj in 2008 to 2009 whereas selleck screening library 32 NP patients Selleck Caspase inhibitor were recruited in the study. One participant each among HP and NP had both high pre-travel and post-travel VL (> 400 copies/mL) and were excluded from analyses. Eventually, 31 HP and 27 NP had the required data and their characteristics are presented in Table 1. The HP spent [median (range)] 36 days (28–43 days) whereas the NP spent 84 days (28–84 days) away before their follow-up appointments (Wilcoxon-rank-sum, z = − 4.09; p < 0.0001). The two groups were broadly on similar three-drug ART regimens. They were on two-drug back bone regimens of Zidovudine/Lamivudine (30), Stavudine/Lamivudine (15) and Tenofovir/Emtricitabine,

or Lamivudine (13) coupled with a non-nucleoside reverse transcriptase inhibitor (NNRTI), either Nevirapine (47) or Efavirenz (7), or the ritonavir-boosted Protease Inhibitor Lopinavir–ritonavir (4); all the latter four were HP patients. The daily dosing frequencies were similar between the two groups with majority on twice daily regimens 27/31 (87%) and 27/27 (100%), respectively (Fisher’s exact; p-value = 0.116). The risk ratio (RR) (95% confidence interval [CI]) of missing at least one ART dose among HP compared with NP in the month preceding their journey was Tolmetin 2.18 (0.46–10.33)

(Table 1). The proportion who missed at least one ART dose among HP and NP while away was 16/31 (51.6%) and 5/27 (18.5%), respectively with RR (95% CI) 2.79 (1.18–6.60). Among HP, the proportion who missed at least one dose during Hajj (16/31 [51.6%]) compared with the month before (5/31 [16.1%]) was with a significantly higher RR (95% CI) 3.20 (1.34–7.65). In addition, the proportion among HP who missed a dose after returning from HP was 9.7%, significantly lower than the proportion who missed a dose during the Hajj (p = 0.0003). In contrast, there was no statistical difference in these proportions among the NP before, during, and after travels. Of the 16 HP who missed a dose during Hajj, 14 did not take ART for a median of 34.5 days (range 1–50 days). Five patients were unable or were not allowed passage with ART medications at airports of departure (1) and arrival (4); all discarded their ART supplies.

Magnetotactic bacteria (MTB) are ubiquitous in aquatic environments, for example marines and lakes. They can form intracellular nanosized magnetite or greigite crystals, known as magnetosomes, which are membrane bound and are generally organized into one or more chains (Schüler, 2008). The net magnetic moment of magnetosome chains can interact with the Earth’s magnetic field and thus navigate MTB along local geomagnetic fields (magnetotaxis) (Faivre & Schüler, 2008). It is widely believed that the magnetotaxis

in conjunction with aerotaxis and other chemotaxis can help MTB to efficiently locate and maintain the most optimal position in vertically stratified sediments or water columns (Frankel et al., 1997; Pan et al., 2009b). All currently known MTB belong to the Proteobacteria and Nitrospira phyla based on the comparison of 16S rRNA genes (Amann et al., 2006). MTB can play important roles in mediating some geochemical selleck products processes, for example iron and sulfur cycling (Simmons & Edwards, 2006). Moreover, fossil magnetosomes preserved in sediments are important natural remanent magnetization carriers (Chang http://www.selleckchem.com/products/Dasatinib.html & Kirschvink, 1989; Moskowitz et al., 1993; Pan et al., 2005a, b; Kopp & Kirschvink, 2008), and can serve as a potential proxy for paleoenvironmental reconstruction

(Snowball et al., 1999; Snowball et al., 2002; Paasche et al., 2004; Kopp & Kirschvink, 2008). Therefore, understanding the patterns of MTB communities in environments is of great importance. A handful of studies have examined the

diversity and vertical distribution of MTB in a single location and have shown that the majority of MTB are usually close to the oxic–anoxic transition zone in chemically P-type ATPase stratified aquatic habitats (Spring et al., 1992, 1993; Bazylinski et al., 1995; Bazylinski & Frankel, 2004; Simmons et al., 2004; Flies et al., 2005a; Pan et al., 2008; Lin & Pan, 2009; Lin et al., 2009). However, due to the lack of detailed studies, the distribution of MTB communities between different locations and their temporal variations remain unclear (Spring et al., 1994; Flies et al., 2005b). Our previous studies revealed that large amounts of MTB (up to 106 cells mL−1) existed in sediments from Lake Miyun near Beijing, China, where the enriched MTB affiliated within both Proteobacteria and Nitrospira phyla (Lin et al., 2008, 2009). In the present study, we used a combination of a cultivation-independent approach and unifrac analysis to investigate the temporal variations of MTB in two freshwater sediment microcosms, which were collected from two separate sites in Lake Miyun, Beijing. The diversity and variation of MTB communities in two microcosms were also compared. The MTB-bearing sediment samples used in this study were collected from two separate sites (MY8 and MY11) in the southern margin of Lake Miyun near Beijing, China (Fig. 1).

Similar results were obtained with rPHY expressed from the AOX1 promoter (data not shown). The N-glycans of rPHY expressed from GAP and AOX1 promoters were separated by HPLC on an NH2P-50 column to investigate the presence of negatively charged

mannose residues (Fig. 3). It was clearly shown that N-glycans of rPHY from both expression systems exhibited different oligosaccharide structures. N-glycans of rPHY produced from the AOX1 promoter were separated into three distinct peaks. The first group of peaks detected at 10–20 min corresponded to neutral glycan, whereas the other two possibly represent mono and di-mannosylphosphorylated glycans (Fig. 3b, retention PD0332991 order time 20–50 min; Wang et al., 1997). On the other hand, rPHY produced from the GAP promoter contained neutral glycans as a major fraction with small populations of negatively charged mannans (Fig. 3a). To confirm that these negatively charged glycans were of the mannosylphosphorylated

type, the N-glycans extracted from rPHY were treated with mild acid and subsequent alkaline phosphatase, which converts phosphorylated glycans to neutral oligosaccharides. The peaks corresponding to negatively selleck compound charged N-glycans detected at 20–50 min retention time from both rPHYs were completely shifted to 10 min retention time after treatment, indicating that these samples were phosphorylated oligosaccharides (Fig. 3). Pichia thermomethanolica BCC16875 is a thermotolerant yeast that

can grow at temperatures from 10 °C up to 40 °C (data not shown). Different growth temperatures might affect the structure of oligosaccharides produced in the host cells. Therefore, we next investigated the N-linked sugar chain structures of cell wall mannoproteins from P. thermomethanolica BCC16875 grown at various temperatures (Fig. 4). Yeast grown at 20 and 30 °C exhibited a similar pattern of N-glycan structures, in which there were comparable ratios of long- and short-chain N-linked mannoproteins, whereas cell wall mannoproteins from the 37 °C culture tended to produce more short-chain N-linked glycans (Fig. 4). Pichia thermomethanolica BCC16875 (recently renamed Ogataea thermomethanolica), was isolated MG132 from soil in southern Thailand (Limtong et al., 2005, 2008). Since this strain is methylotrophic, we reasoned that P. pastoris expression vectors would be functional. Recombinant plasmid vectors with P. pastoris GAP and AOX1 promoters driving expression of recombinant phytase were integrated into the P. thermomethanolica genome and the proteins were secreted as functional enzymes, although the level of protein expression was not as high as when expressed in P. pastoris (Promdonkoy et al., 2009). Pichia thermomethanolica BCC16875 has not been characterized genetically and so the degree of conservation in promoter function and gene regulatory mechanism with P. pastoris is unknown.

In 6 months, the probability of remaining in state 1 is about 0.20. The chances of being in the other states as HPV status changes and/or VL decreases are: 0.08 for HPV positive

and VL > 400 copies/mL (state 2); 0.60 for HPV negative and VL < 400 copies/mL (state 3) and 0.12 for HPV positive and VL < 400 copies/mL (state 4). The probabilities stabilize in about 2.5 years to around 0.11, 0.08, 0.58 and 0.23 for the four states, respectively. They suggest that, whereas the probabilities of being HPV positive and HPV negative are similar when VL > 400 copies/mL (at around 0.10), the probability of being HPV positive is about 0.4 times the probability of being HPV negative when VL ≤ 400 copies/mL. There were 145 subjects included in this analysis, because two of the subjects did http://www.selleckchem.com/products/PLX-4032.html not provide CD4 cell counts. Of the 140 subjects with both HPV and CD4 results at baseline, 107 subjects (76%) started with a CD4 count <350 cells/μL, and HPV was detected in 86 subjects (61%). Data availability trends over time were similar to those for the VL model above. In the CD4 model (Fig. 1b), similar conclusions were drawn for the two HPV sets (set 2 results are shown). Comparison between γ21 and γ43 suggested that a woman with a current CD4 count >350 cells/μL was more likely to clear HPV than a woman with a CD4 count ≤ 350 cells/μL

(hazard ratio γ43/γ21 = 2.65; P = 0.018). The statistical tests on other comparisons were not significant. There was no evidence that HPV detection rates differed between Veliparib subjects with CD4 counts ≤350 and >350 cells/μL (γ12/γ34 = 1.03; P = 0.94), and HPV status did not seem to affect CD4 state transition rates (γ13/γ24 = 0.920; P = 0.78; γ31/γ42 = 0.408; P = 0.18). Figure 2b presents the model-based probability curves over 5 years for a HAART-initiating woman,

starting as HPV negative and with a CD4 count ≤350 cells/μL (state 1). The probabilities stabilize in about 3.5 years to around 0.12 in state 1, 0.11 in state 2 (HPV positive and CD4 count ≤350 cells/μL), 0.56 in state 3 (HPV negative and CD4 count >350 cells/μL) and 0.21 in state 4 (HPV positive and CD4 count >350 cells/μL). The probability of being HPV positive is about 0.4 times the probability Lck of being HPV negative when the CD4 count is >350 cells/μL. Studies have shown varying effects of HAART on HPV infection and HPV-related cervical dysplasia. Several studies have shown a higher HPV prevalence in women with a lower CD4 cell count and higher likelihood of clearance of HPV with improved CD4 cell count [9, 13, 14]. Research on the association between HIV VL and HPV detection has been limited. One study showed that HIV VL and CD4 cell count in combination appeared to be associated with HPV detection, with varying effects of HIV RNA level on HPV prevalence and incident detection depending on the CD4 cell count stratum [4].

coli isolates from diseased piglets in Guangdong Province, China. It also describes the association between AMR and VGs, and between resistance and phylogenetic background. Other such studies describing associations between resistance and virulence traits have invariably investigated a limited number of antimicrobials (principally ampicillin, tetracycline, chloramphenicol, streptomycin, and sulfonamides), whereas we have extended our observations to include doxycycline, florfenicol, apramycin, and amikacin. Such studies, reporting an association between the resistance of this range of antimicrobials and VGs among E. coli strains from diseased swine in South China, are not available at

present.

The results from this study showed alarming frequencies of resistance to many antimicrobial agents ALK mutation commonly used in China. In agreement with previous reports (White et al., 2000; Lanz et al., 2003; Maynard et al., 2003), most E. coli isolates from swine were resistant to sulfamethoxazole, tetracycline, streptomycin, and chloramphenicol. Multidrug resistance phenotypes of E. coli isolates from animals have been reported worldwide (Lanz et al., 2003; Maynard et al., 2003; Yang et al., 2004), and in accordance with this, >50% of E. coli strains in our study were resistant to 8–10 antimicrobials tested. Doxcycline and florfenicol have been approved selleck compound for use in food-producing animals in China, and are now used Protirelin extensively with livestock, resulting in the emergence of resistance to both drugs. Many E. coli isolates showed high resistance or reduced susceptibility to doxycycline as well as to florfenicol in this study, which is similar to previous studies (Bischoff et al., 2002; Dai et al., 2008). The likely reasons for the high resistance rates are the inappropriate use of these antimicrobials in veterinary practice and cross-resistance among antibiotics of the same class, such as tetracycline and chloramphenicol, although chloramphenicol has been prohibited for use in food animals in China. Similarly high resistance rates to ciprofloxacin seen in this study have

also been observed in other studies in China among E. coli isolates from swine and chickens (Yang et al., 2004; Liu et al., 2007; Dai et al., 2008), which suggests that this agent has become ineffective in veterinary medicine in China (Xu, 2001). Ceftiofur is the only cephalosporin approved for systemic use in food-producing animals since 2002 in China, and it is highly effective against E. coli isolates. The rate of resistance to ceftiofur was higher in our study than in previous studies (Yang et al., 2004; Liu et al., 2007), presumably as a consequence of the increasing use of cephalosporins on animal farms. Prudent use of antimicrobials in veterinary practice is therefore fundamental to the reduction of resistance development.

pneumoniae isolates were identified after the recovery of the C-NS isolates. In each case, the C-S isolate GPCR & G Protein inhibitor was closely related to

the C-NS from the same patient, including the PFGE subtype and the β-lactamase content. It is again difficult to explain these identifications of the C-S isolates. They might have resulted from long-term colonization in other body sites where the C-S organisms could have survived antimicrobial treatment or from recurrent acquisitions from unidentified sources, specific for each of these patients. Although the material collected was not complete, it is possible that at least in some of the patients, the C-NS K. pneumoniae strain variants evolved during the course of colonization or infection, selected during the carbapenem treatment. PD-0332991 mw Several cases of such an evolution of ESBL- or AmpC-producing Enterobacteriaceae strains have been documented in other studies (Bidet et al., 2005; Thiolas et al., 2005; Livermore & Woodford, 2006; Cuzon et al., 2010). Numerous reports on clinical and laboratory strains demonstrated that reduced susceptibility to carbapenems may emerge in organisms expressing various types of ESBLs or AmpCs (Jacoby et al., 2004; Bidet et al., 2005; Kaczmarek et al., 2006; Martínez-Martínez, 2008; Doumith

et al., 2009), including the SHV-5 and DHA-1 enzymes identified in this work (Jacoby et al., 2004; Lee et al., 2007; Cuzon et al., 2010). Interestingly, Idoxuridine the blaDHA-1 gene was found in the integron variant like in the K. pneumoniae RBDHA isolate from France from 2002 (Verdet et al., 2006), suggesting wider spread of this particular resistance determinant in Enterobacteriaceae across Europe. Based on the porin analysis data, it may be proposed that the major alteration of porin profiles affecting carbapenem susceptibility in the isolates studied was the loss of OmpK36, similar to a number

of other reports (Martínez-Martínez, 2008; Gröbner et al., 2009; Wang et al., 2009). None of the C-NS isolates produced OmpK36 that would be detectable by SDS-PAGE and Western blot, and the failure of ompK36 amplification with the ‘external’ pair of primers (Kaczmarek et al., 2006) indicated significant DNA rearrangements at this locus. The different behavior of the C-NS isolates with the ‘internal’ PCR primers demonstrated that different types of ompK36 alterations occurred in particular PFGE types. In one of these (type A), the 5′ part of the gene was detected; however, the frame-shift resulting from tetranucleotide insertion prevented it from producing a functional protein. Previous studies demonstrated a variety of events leading to the inactivation of the major porin genes in Enterobacteriaceae, including nonsense or frame-shift mutations at multiple positions, insertions of IS elements, or gene deletions (Hernández-Allés et al., 1999; Kaczmarek et al., 2006; Doumith et al., 2009).