Non-vertebral anti-fracture reduction is 20 to 30%, less than half the vertebral fracture risk reduction reported in most trials [44]. One explanation may be the differing access of drugs to intracortical remodeling sites initiated upon Haversian canals within the large cortical matrix volume [4] and [34]. Risedronate has a lower

mineral binding affinity than alendronate and penetrates deeper into cortical bone [4] and [34]. Risedronate reduced non-vertebral fracture rates selleck compound in two of the three main trials [45], [46] and [47], while alendronate did not [48] and [49]. Nakamura et al. reported that in a 24-month study of 1194 postmenopausal Japanese women and men (placebo, n = 480; denosumab 60 mg every 6 months, n = 472; or open-label alendronate 35 mg weekly, n = 242) [50], new or worsening vertebral fractures occurred Sotrastaurin in 8.5%, 2.4%, and 5.0% of women, respectively (p = 0.0001 denosumab versus placebo). Major non-vertebral fractures occurred in 3.9%, 1.7%, and 2.3% of women, respectively (p = 0.057, denosumab versus placebo). Thus, numerically, fewer fractures occurred in the denosumab than alendronate group but statistical analyses comparing the two antiresorptives was not reported. Moreover, women treated with denosumab in the pivotal phase 3 trial, although a placebo comparator arm was not available in the 4th and 5th years, had a low reported non-vertebral fracture rate, an observation not

reported for alendronate or zoledronic acid, the latter also having high affinity for bone mineral [51]. This study has limitations. StrAx1.0 analysis does not quantify pore size and number so that the relative contribution of reductions in pore number versus pore size to the reduction in porosity cannot be determined at this time. Measures of porosity using StrAx1.0 are more sensitive than measures of density to motion artifacts and this resulted in loss of some images. In summary, this is the first randomized double-blind, placebo controlled trial comparing the effect of two remodeling suppressant therapies on intracortical porosity in vivo. Denosumab reduced Unoprostone remodeling more rapidly, more completely and decreased porosity more than alendronate.

Given the exponential relationship between porosity and bone stiffness, partly reversing cortical porosity is likely to contribute to reductions in fracture risk. Whether this greater reduction in porosity translates into better anti-fracture efficacy will require additional comparator trials. This study was funded by Amgen Inc. RM Zebaze has received grant and/or research support from Amgen and speaker fees from Servier. RM Zebaze is one of the inventors of the StrAx1.0 algorithm and a director of Straxcorp. C Libanati is an employee of Amgen and has received Amgen stock or stock options. M Austin is an employee of Amgen and has received Amgen stock or stock options. A Ghasem-Zadeh is one of the inventors of the StrAx1.0 algorithm.

The incidence of

diagnosed VTE during residence in the current study was higher than reported in 3 earlier nursing home studies16, 17 and 18 but equivalent to that of a second of 2 databases in one of these studies.16 Compared with the current study finding of 3.68 cases per 100 PY, VTE incidence rates in nursing home studies were 1.2 to 1.5 (MCMRP data/Minnesota),16 3.6 (Rochester Epidemiology Project data/Minnesota),16 1.3 (MDS and Medicare data/Kansas),17 and 1.4 to 1.6 (medical chart data/Israel)18 per 100 PY. The high incidence rate found in our study may be a consequence of differences in the pool of nursing homes studied Lapatinib supplier (eg, a potentially greater number of residents receiving subacute care) or in the methods used, or it may be due to the later time period (2007–2009) than the earlier studies (1988–2001). The effect of changes in resident case-mix or a historic trend in the incidence of VTE remain unknown given the

lack of details in the current and earlier studies regarding levels of resident acuity and changes in criteria to diagnose VTE. Findings buy Rapamycin from the Rochester Epidemiology Project16 would suggest that the MDS might be undercounting the incidence of fatal VTE, especially because residents who die in the hospital after nursing home discharge are less likely to have VTE recorded in the final MDS assessment. PE events may be especially undercounted. In a recent national study25 of hospitalizations with a diagnosis of VTE, the ratio of DVT to PE was much lower than our findings: crude estimated average annual rates in that study were 0.152 (DVT) and 0.121 (PE) per 100 hospitalizations, respectively; the relative proportion due to PE declined with advancing age, although in an earlier community study,6 the inverse

relationship was observed. The high incidence rate observed in our study Acetophenone might also be a consequence of the growth in associated risk factors among hospitalized patients admitted to nursing homes in recent years with high disease acuity, short hospital stays, and increased use of surgical and other interventional procedures. Improved diagnostics for recognizing asymptomatic VTE may be a key factor, although we have no means of describing how newer diagnostics, such as portable Doppler ultrasound, have affected incidence rates over time. Stein et al26 found that the incidence of DVT in hospitalized patients increased from 0.8% to 1.3% of all hospital admissions over the period 1979 to 1999, yet the incidence of PE remained unchanged at 0.4%. These authors hypothesized that increased use of venous ultrasound may have increased DVT incidence, and early diagnosis and treatment of DVT may have prevented a concurrent rise in PE.26 Our study found a 1:5 ratio of PE cases to DVT cases during residence.

Darüber hinaus wird Quecksilber in einem breiten Spektrum von Produkten und Verfahren verwendet: von der Saatgutaufbereitung über Konsumprodukte und Zahnfüllungen bis hin zu Konservierungsstoffen für Impfstoffe. Daher sind wir alle Quecksilber in irgendeiner Form oder Konzentration ausgesetzt. Diese die

Umwelt und die öffentliche Gesundheit betreffenden Aspekte des Quecksilbers sind von Clarkson 2002 in einem Übersichtsartikel ausführlich behandelt worden [1]. Der Leser sei auf diesen Review mit dem Titel „The three modern faces of mercury” verwiesen, in dem auf hervorragende Weise alle wichtigen Themen im Zusammenhang mit der Exposition von Menschen gegenüber Quecksilber und die besonderen Charakteristika der Chemie des Quecksilbers diskutiert werden. Es liegen zahlreiche INK 128 solubility dmso Übersichtsartikel vor, die sich mit der Toxizität von Quecksilber und seinen Verbindungen bei Säugetieren und Menschen befassen. Einen besonders umfassenden und gründlichen Review haben Clarkson und Magos publiziert [2]. In diesem Übersichtsartikel wird die Toxikologie sowohl von anorganischem als auch von

organischem Quecksilber behandelt. Er beleuchtet insbesondere einige „Rätsel”, die immer noch unsere wissenschaftliche Neugier herausfordern. ALK inhibitor Ein solcher Aspekt ist das verzögerte Auftreten von Symptomen nach einer Exposition gegenüber Alkylquecksilberverbindungen. Die Latenzphase nach der Exposition kann einige Wochen bis mehrere Monate dauern, wobei die Symptome, wenn sie erst einmal eingesetzt haben, sich rasch verschlimmern. Die Latenzphase verkürzt sich nicht mit steigender Dosis und der Mechanismus, der die Latenzphase bewirkt, ist immer noch unbekannt. Noch nicht einmal der Schlüsselmechanismus ist bekannt, der der Neurotoxizität von Alkylquecksilberverbindungen zugrunde liegt, wobei ein vorgeschlagener Hauptmechanismus sowohl die Natur der Latenzphase mafosfamide als auch die Zellspezifität der Schäden erklären sollte. Im Folgenden präsentieren wir eine kurze Übersicht über die allgemeine

Toxikologie des Quecksilbers. Im Hauptteil des Artikels befassen wir uns jedoch mit der Toxikologie von Alkylquecksilber und machen den Versuch einer Bewertung der möglichen Mechanismen, die bei Überlegungen im Hinblick auf ein sicheres Ausmaß der Quecksilber-Exposition, z. B. durch den Verzehr von Fisch, von praktischer Bedeutung sind. Elementares Quecksilber (Hg0) ist bei Raumtemperatur flüchtig und die Dämpfe können für den Menschen gefährlich sein. Eine Exposition gegenüber Quecksilber kann in Labors, an Arbeitsplätzen sowie in häuslicher Umgebung stattfinden. In privaten Haushalten werden u. U. beschädigte Quecksilberthermometer zu einer Expositionsquelle, da es sehr schwierig sein kann, ausgelaufenes Quecksilber aufzusammeln. In vielen Ländern ist die Verwendung von Quecksilber in Thermometern inzwischen verboten, um das Risiko für die Verbraucher und die Gefahr einer Freisetzung von Quecksilber in die Umwelt zu verringern.

This article evaluates patient-related and procedure-related risk factors for ERCP-related adverse events, and discusses strategies for the prevention, diagnosis and management of these events. Tarun Rustagi and Priya A. Jamidar Post–endoscopic retrograde cholangiopancreatography pancreatitis (PEP) is the most common complication GSK458 manufacturer of endoscopic retrograde cholangiopancreatography (ERCP), and not uncommonly is the reason behind ERCP-related lawsuits. Patients at high risk for PEP include young women with abdominal pain, normal liver tests, and unremarkable imaging. Procedure-related factors include traumatic and persistent cannulation attempts, multiple injections of the pancreatic duct, pancreatic

sphincterotomy, and, possibly, use of precut sphincterotomy. GDC-0449 purchase Aggressive hydration, use of rectal indomethacin, and prophylactic pancreatic stenting can diminish the risk (and likely severity) of PEP. Though hugely beneficial, these measures do not supersede careful patient selection and technique. Nayantara Coelho Prabhu and Louis M.

Wong Kee Song Videos demonstrating endoscopic hemostasis accompany this article Acute gastrointestinal bleeding is a common cause for hospitalization. Endoscopic hemostasis plays a central role in the management of lesions with active bleeding or high-risk stigmata for rebleeding. The efficacy and safety of endoscopic hemostasis rely on the identification of lesions suitable for endoscopic therapy, selection of the appropriate hemostatic devices, attention to technique, and prompt recognition and management of procedure-related adverse events. In this article, practical applications of hemostatic devices and pitfalls related to endoscopic hemostasis are discussed. John J. Vargo II Defining the risk of procedural sedation for gastrointestinal endoscopic procedures remains a vexing challenge. The definitions as to what constitutes a cardiopulmonary unplanned event are beginning to take focus but the existing literature is an amalgam of various definitions and subjective outcomes,

providing a challenge to patient, practitioner, and researcher. Gastrointestinal endoscopy when undertaken by trained personnel after the appropriate Phosphatidylethanolamine N-methyltransferase preprocedural evaluation and in the right setting is a safe experience. However, significant challenges exist in further quantifying the sedation risks to patients, optimizing physiologic monitoring, and sublimating the pharmacoeconomic and regulatory embroglios that limit the scope of practice and the quality of services delivered to patients. Nikhil A. Kumta, Christine Boumitri, and Michel Kahaleh Increasingly invasive therapeutic endoscopic and laparoscopic procedures have resulted in endoscopists more frequently encountering complications including perforations, fistulas, and anastomotic leakages, for which nonsurgical closure is desired. Devices and techniques are available and in development for endoscopic closure of gastrointestinal wall defects.

The study was approved by learn more the University of Cape Town’s Faculty of Health Sciences Research Ethics Committee and informed written consent was obtained from all volunteers before the study was initiated. Cervical cytobrush samples were collected according to the protocol described by Nkwanyana et al. (2009). Briefly, cervical immune cells were collected from all women under speculum examination by inserting a Digene cervical sampler into the endocervical os, rotating 360° and immediately placing the cytobrush in 3 ml R10 [RPMI

1640 (GibcoTM) supplemented with 5 mM glutamine, fungazone, penicillin, streptomycin and 10% FCS (Delta Bioproducts)]. Cytobrush samples with visible blood contamination (11/215; 5%) or excessive mucous contamination (21/215; 10%) were excluded from further analysis. Phenotypic and functional assessments of cytobrush-derived T cells were conducted in the remaining 183 samples. Samples were transported between the clinic and laboratory

in temperature-controlled CH5424802 benchtop coolers. Upon arrival in the laboratory (≤ 4 h of collection), the cytobrushes were flushed ~ 30 times with the same 3 ml transport media using a sterile plastic disposable Pasteur pipette and 25 ul of the suspension was removed for ex vivo CD3+ T cell enumeration using a Guava automated cell counter. The samples were divided into four groups to evaluate alternative processing conditions. Group 1 cytobrushes (n = 113) were processed immediately and used for flow cytometry analysis of immune subsets by intracellular cytokine staining (function, n = 98, Group 1a) and surface staining (viability, n = 15; Group 1b; ex vivo cytobrushes). Group 2 cytobrushes (n = 27) were not processed immediately but incubated at 37 °C for 24 h prior to flushing cells off the brush

and analysed for Vasopressin Receptor phenotype and function. Similarly, processing of cytobrushes from Groups 3 (n = 5) and 4 (n = 25) was delayed for 24 h and during this time, cytobrushes were maintained at 4 °C (to mimic cold overnight transport) or room temperature (~ 20 °C; to mimic overnight transport without refrigeration). After removing cervical cells off the cytobrush by gentle flushing, cells were washed once in R10, counted, phenotyped, and functionally evaluated using a Guava cell counter or FACS Calibur flow cytometer (BD Biosciences, San Jose, CA), respectively. Cervical cytobrush cells were counted using an automated Guava cell counter according to the method described by Nkwanyana et al. (2009). CD3-PE (T cells; Guava technologies) was used to label T cells in each cytobrush samples which were then counted using a Guava Automated Cell counter. Briefly, 25 μl cytobrush cells were stained with pre-titrated CD3-PE monoclonal antibodies and incubated at 4 °C for 30 min. Cells were washed with 1 ml wash buffer (1% FCS PBS) and centrifuged at 1500 rpm (437 ×g) for 5 min.

This study also assessed the impact of treatment duration on SVR and the regimen showed efficacy with only 12 weeks of treatment. This therapeutic duration is consistent with publication of viral kinetic modeling data suggesting 10 weeks of treatment with a potent antiviral regimen may be needed to clear infected hepatocytes.29 In contrast, current treatment for GT 1-infected patients includes peginterferon, ribavirin, and the NS3 protease inhibitors telaprevir or boceprevir Selleckchem Z VAD FMK for up to 48 weeks.30 Shorter treatment durations are preferable because they may improve patient compliance. In this study, treatment periods of

both 12 and 24 weeks yielded high SVR rates, suggesting no advantage for extending treatment duration to 24 weeks. The study using ABT-450 boosted with ritonavir, ABT-333, ABT-267, and ribavirin in GT 1 treatment-naive patients also showed high rates of SVR with only 12 weeks of therapy.27 Similarly, regimens including sofosbuvir and daclatasvir with or without ribavirin also showed high rates of SVR with 12 weeks of treatment.22 and 31 In our study, virologic failure was uncommon and observed in only 3 patients in the 150 mg twice-daily dosing groups: 2 patients with viral breakthrough, and 1 patient with virologic relapse. The reasons for treatment failure in these patients remain unclear but could

include baseline virus polymorphisms, host immune status, and/or reduced drug exposure or adherence. Two of these patients were infected with HCV GT 1a. One patient had a pre-existing NS5A variant with increased resistance to daclatasvir (L31M, U0126 250-fold change in the EC50 of daclatasvir in vitro). The second patient had baseline resistance-associated polymorphisms to daclatasvir and asunaprevir (NS5A-H58P and NS3-V36M, respectively), which alone do not appear to alter the EC50 value of the direct-acting antivirals in vitro. It is possible that these variants acted

in a compensatory manner by enhancing the fitness of the emergent variants. The emergent linked NS5A substitution M28A-Q30R confers high-level resistance to daclatasvir in vitro (>200,000-fold resistance). NS3-V36M enhances resistance by approximately 3-fold against asunaprevir when combined with NS3-R155K in vitro. PRKD3 The HCV-RNA sequences of the third patient with HCV GT 1b reported at screening have not been amplified successfully despite numerous attempts, thus the HCV GT remains unconfirmed and the presence of baseline and emergent variants is unknown. The relationship between pre-existing polymorphisms and treatment failure of interferon-free regimens is unclear because patients in this and other studies with similar findings have achieved a sustained response. Thus, larger studies are needed to clarify the impact of pre-existing polymorphisms on efficacy.

Para dificultar ainda mais, existe um espectro de doenças auto-imunes, cujas características clinicas e laboratoriais se sobrepõem à HAI e que podem coexistir no mesmo doente, como os síndromes de sobreposição3 and 4. Pela necessidade de comparar grupos de doentes, foi criado, em 1993, um score diagnóstico numa tentativa de homogeneizar os critérios diagnósticos de hepatite auto-imune, pelo International Autoimmune Hepatitis Group 5. Em 1999, a revisão desses Epacadostat order critérios (chamados critérios clássicos) de diagnóstico tornou-os mais específicos para a exclusão de outras patologias auto-imunes

e passou a incluir PD0332991 solubility dmso também a resposta à terapêutica 6. Por serem complexos e difíceis de usar na prática clinica diária, foi publicado, em 2008, um score mais simplificado que inclui apenas 4 itens: título de auto-anticorpos, níveis de IgG, histologia hepática e exclusão de

hepatite vírica 7. Estes critérios de diagnóstico simplificados, embora não validados em estudos prospectivos mostraram uma elevada sensibilidade e especificidade para o diagnóstico de hepatite auto-imune 2, 7 and 8. Assim, os critérios diagnósticos clássicos foram criados para comparar grupos diferentes de doentes em cenário de investigação clínica, excluem os síndromes de sobreposição e por terem múltiplos itens e múltiplas associações são difíceis de aplicar na prática MYO10 clínica. Como são usados na identificação dos doentes com hepatite auto-imune terão, por definição, sensibilidade de 100%1, 2, 3, 9 and 10. Os critérios de diagnóstico simplificados são menos sensíveis (sensibilidade de 80 a 88%)mas mais específicos (97 a 99%) pois foram concebidos para serem aplicados na prática clínica1, 3 and 9. Desde então têm-se tentado comparar estes 2 sistemas de critérios diagnósticos numa comparação que

é ingrata e quase impossível. Se não há um gold standard diagnóstico qual vai ser a base da comparação? Como comparar 2 sistemas de critérios criados para fins diferentes? Como comparar 2 sistemas em que a selecção dos doentes é feita a partir de um deles? Os vários estudos existentes, apesar maioritariamente retrospectivos e com pequenas amostras, confirmam a aplicabilidade e fiabilidade dos critérios simplificados em várias populações distintas. O grau de concordância entre os 2 sistemas de critérios tem sido descrito mesmo em indivíduos com coexistência de outras hepatopatias crónicas com hepatite auto-imune de apresentação aguda8, 10, 11, 12, 13 and 14. Do mesmo modo, o estudo de Correia L. et al 15 compara estes 2 sistemas de classificação numa população portuguesa.

, 2002), and ADHD patients show greater frontal and parietal alpha EEG power (8–10 Hz) during a sustained attention task (Loo et al., 2009). Considering

that bright light plays a therapeutic role in ADHD (Gruber et al., 2007) and schizophrenia (Aichhorn et al., 2007), selleck chemical lighting conditions seem to modulate human attentional processing. Therefore, it is important to understand how illumination influences attentional processing and cognitive performance. Among the various aspects of attention, we selected sustained attention to assess under specific combinations of illumination parameters. We supposed that a stationary illumination condition might affect a sustained mental state, so sustained attention was considered to be one of the appropriate targets to investigate possible influences by background illumination. Sustained attention, the http://www.selleckchem.com/products/PD-98059.html capability to maintain the focus of attention over time (Mirsky et al., 1991), can be generally assessed, using the continuous performance test (CPT; Riccio et al., 2001), which is featured by a rapid presentation of continuously changing stimuli with an infrequently occurring target stimulus. Several studies have evaluated different aspects of EEG activity recorded during sustained attention tasks.

Since ongoing tonic alpha activity has been reported to be associated with the sustained attention processing (Dockree et al., 2007 and Orekhova Astemizole et al., 2001), we focused on the analysis of EEG alpha activity related to sustained attention task-performance under different illumination conditions. Because lower prestimulus alpha power facilitates task-performance (Ergenoglu et al., 2004 and Hanslmayr et al., 2007), we investigated whether background illumination conditions can affect the prestimulus alpha activity level, which reflects prestimulus preparatory mental states for the sustained and selective allocation of neural processing

resources to target information. In particular, neural activity related to sustained attentional processing has been reported in the parietal brain region (Lee et al., 2013 and Thakral and Slotnick, 2009). Therefore, we hypothesized that light conditions would modulate alpha activity in the parietal region during a sustained attention task. For example, it was suggested that parietal alpha synchronization reflects an active inhibition of certain parietal networks involved in maintaining attention to peripheral visual field (Orekhova et al., 2001), and that parietal alpha activity ipsilateral to the attended hemi-field was enhanced relative to the control condition when attention was shifted away from fixation (Cosmelli et al., 2011). Moreover, we hypothesized that the background illumination condition would affect selective sensory gain control in the visual pathways.

The salt influxes were concentrated in the deep portion of the channels at 0–6 km and 14.8–15.2 km, rather than in the shoal region at the Cape Henry cross-section. The baroclinic component of the tidally averaged salt flux excluding QfS0 was also calculated, and the magnitude is about half of the total PI3K inhibitor flux,

as shown in the bottom panel. It is concluded that both barotropic and baroclinic components contributed to oceanic saltwater influxes during the first stages of the hurricanes. Local winds that exert stress on the surface of the water can cause direct wind mixing, and reduce the stratification, but a moderate down-estuary wind can also induce a wind-straining effect, which under certain conditions increases stratification

(Scully et al., 2005). Due to their tracks, Hurricanes Floyd and Isabel produced distinctly different local wind stresses, a down-estuary and an up-estuary stress. This difference provides a natural test bed for examining how the direction of the axial wind affects the vertical stratification and the salt transport. find more In order to reasonably compare the wind-induced mixing process between the two hurricanes, a controlled experiment is required to ensure that the local and remote winds are separated, that different pre- and post-hurricane conditions are equalized, and that the background conditions are uniform. To start with, the background state of the estuarine system is required to be in a quasi-steady state prior to the hurricane. Upon the passage of the hurricane, the estuarine system will experience the hurricane’s wind forcing, and then eventually return to the quasi-steady state when all of

the external perturbations Sorafenib mouse are removed. Table 6 shows seven experiments that were performed to examine the mixing process induced by the local and remote meteorological external forcing during the two hurricanes, Floyd (FL) and Isabel (IS). Four types of wind forcing were considered: no wind (NW), local (L), remote (R), and combined (C). Fig. 15 shows wind and pressure fields selected from the real hurricane conditions for the controlled experiment. The base run used only the M2 tidal constituent and a constant river discharge of 550 m3 s−1, which characterizes the summer average flow in the Bay. The use of a single semi-diurnal tidal constituent precludes investigation of the effect of spring–neap tides on salinity. A constant ambient current of 10 cm s−1 was specified at the cross-shore open boundaries in the continental shelf, based on the work of Cho (2009). To obtain the initial salinity condition in an equilibrium state, the model was spun up for 180 days without meteorological forcing from a cold start, such that salinity had a linear variation horizontally from the Bay head (0 ppt) to the open ocean (34–35 ppt) with no stratification in the vertical direction.

Toxin

encoding DNA was amplified in the first PCR step (E-PCR1) using gene-specific primers listed in Table 1 (PCR-conditions: 10× Fermentas PCR-buffer, dNTPs 0.2 mM RG7422 supplier each, forward and reverse primer 0.5 μM each, 0.05 U/μl Taq DNA polymerase, 2 ng DNA, ad MilliQ H2O to a final volume of 50 μl. Initial denaturation at 95 °C for 10 min, denaturation at 94 °C for 30 s, primer annealing at 54 °C for 30 s, primer extension at 72 °C for 45 s, final extension at 72 °C for 5 min; number of cycles: 30) ( Table 2). 100 ng PCR product from the first PCR step was directly applied to the second PCR amplification (E-PCR2) procedure. In E-PCR2 adapter primers were used to add tag-encoding sequences and regulatory sequences at the 5′- and 3′-end of the final PCR-product for cell-free expression (Suppl. Table S1). Amplification was performed according to the manufacturers recommendations (EasyXpress Linear Template Kit PLUS, Qiagen, Hilden, Germany). E-PCR2 was performed in a final volume of 25 μl (PCR-conditions: 5 μl 5× High Fidelity PCR MAPK inhibitor buffer, 2.5 μl adapter primer each, High Fidelity DNA Polymerase 0.05 U/μl, initial denaturation at 95 °C for 5 min, denaturation at 94 °C for 60 s, primer annealing at 50 °C for 60 s, primer extension at 72 °C for

45 s, final extension at 72 °C for 10 min; number of cycles: 30). All E-PCR2 products were analyzed by agarose (1%) gel electrophoresis to determine quality and concentration by comparison with a known DNA marker. A 9 μl aliquot of the individual linear E-PCR2 products was directly used in the cell-free prokaryotic system without any further purification. Genomic DNA extraction from V. parahaemolyticus ifenprodil O3:K6 strain was performed with the RTP Bacteria DNA Kit from Stratec Molecular, Berlin, Germany. Primers used for the amplification

of the tdh2 gene for the construction of an E. coli recombinant plasmid were VparaF (5′-CAA AGC CTC ATA GAG TTG TAA G-3′) and VparaR (5′-GAA GCG AAT AAA TAG CGT G-3′) amplifying an 972 bp fragment of the genomic DNA of the O3:K6 strain PMA1.6 containing the complete coding sequence of the tdh2 gene ( Suppl. Fig. S3). PCR reaction was performed with DreamTaqTM DNA Polymerase (Fermentas, St. Leon-Rot, Germany) according to the manufacturers recommendations. The PCR product was inserted into the multiple cloning site of the vector pJET2 (Fermentas, St. Leon-Rot, Germany). Finally, the plasmid pJET2-TDH2 was introduced into E. coli DH5α. Sequencing of plasmids and PCR products was carried out by QIAGEN sequencing services (Hilden, Germany). The obtained sequences were analyzed using the Lasergene program “SeqMan” (DNASTAR, Inc., Madison, USA). Sequence translations were performed using the program Accelrys (DS-) gene (Accelrys Inc., San Diego, USA).