In Week 0 and Week 16, intake was below 2/3 of the RDA in 42.9% of the participants [29]. Mean carbohydrate intake was below the RDA [28] at all time points, whereas fat and protein Selonsertib clinical trial intakes were above 100% of the RDA [28]. Table 3 Recommended daily allowance covered for energy, macronutrients and folic acid at three time points Nutrient ≤ 2/3 RDA > 2/3 RDA ≤ RDA > RDA Macronutrients (%) Protein Week 0 – - 100 Week 8 – - 100 Week 16 – - 100 Carbohydrate Week 0 35.7 64.3 – Week 8 – 92.9 7.1 Week 16 – 100 – Fat Week

0 – - 100 Week 8 – - 100 Week 16 – - 100 Vitamins (%) Folic acid CH5183284 Week 0 42.9 42.9 14.3 Week 8 – - 100 Week 16 42.9 50.0 7.1 RDA, recommended daily allowance. Training profile The results in Figure 1 show the training loads recorded during the study period. Training load is reported here as training time, RPE and distribution among three levels of intensity during the intervention (STp) and post-intervention periods (NSTp). There were no statistically significant differences in training time

between STp and NSTp. Figure 1 Comparison of training variables throughout the experimental trial. *Statistically significant difference (P < 0.05) STp vs NSTp. Overall Ivacaftor cost RPE during STp was significantly lower (P < 0.05) than during NSTp. With regard to the durations of different RHR levels (training intensity), a significant difference (P < 0.05) was found for the 60%–80% range, which accounted for 30.35% of the total training time during STp, and for 35.87% of the training time during the NSTp. There were no significant differences for training intensity levels in the <60% range or the >80% range. Bivariate analysis to calculate Pearson’s correlation coefficient detected statistically significant correlations (P < 0.01) between overall RPE and training intensity levels of 60%–80% RHR (r = 0.64) and >80% RHR (r = 0.76). Biochemical assays The results

of biochemical analyses are shown in Table 4. There were no significant changes in plasma folic acid at any time point, and all values were within the normal crotamiton range for the healthy population. However, plasma concentrations of Hcy increased significantly (P < 0.05) to above the normal range of values during the Week 8 and Week 16 periods compared to baseline values in Week 0. Regarding the relationship between plasma concentrations of Hcy and folic acid and training intensity, we found that both plasma concentrations showed a significant negative correlation (r = −0.75) (P < 0.01) with the level of intensity of <60% RHR. Bivariate analysis disclosed a significant negative correlation (P < 0.01) between Hcy and folic acid concentrations (r = −0.84) in Week 8. Table 4 Biochemical values of clinical and nutritional parameters at three time points N = 14 Study period Biochemical parameters Reference value Week 0 Week 8 Week 16     Mean SD Mean SD Mean SD Transferrin (mg/dl) 200 – 360 261.21 27.82 261.71 33.00 265.50 28.67 Prealbumin (mg/dl) 20 – 40 26.76 3.53 27.

Proc Natl Acad find more Sci USA 2005,102(46):16819–16824.CrossRefPubMed 12. Boles BR, Thoendel M, Singh PK: Self-generated diversity produces

“”insurance effects”" in biofilm communities. Proc Natl Acad Sci USA 2004,101(47):16630–16635.CrossRefPubMed 13. Rice SA, Koh KS, Queck SY, Labbate M, Lam KW, Kjelleberg S: Biofilm formation and sloughing in Serratia marcescens are controlled by quorum sensing and nutrient cues. J Bacteriol 2005,187(10):3477–3485.CrossRefPubMed 14. Coetzee JN, Deklerk HC: Effect Of Temperature On Flagellation, Motility And Swarming Of Proteus. Nature 1964, 202:211–212.CrossRefPubMed 15. Kearns DB, Losick R: Swarming motility in undomesticated Bacillus subtilis. Mol Microbiol 2003,49(3):581–590.CrossRefPubMed 16. Givskov M, Ostling J, Eberl L, Lindum PW, Christensen AB, Christiansen G, Molin S, Kjelleberg S: Two separate regulatory systems participate in control of swarming motility of Serratia liquefaciens MG1. J Bacteriol 1998,180(3):742–745.PubMed 17. Overhage J, Lewenza S, Marr AK, AZD1390 in vitro Hancock RE: Identification of genes involved in swarming motility using a Pseudomonas aeruginosa PAO1 mini-Tn5-lux mutant library. J Bacteriol 2007,189(5):2164–2169.CrossRefPubMed 18. Kaiser D: Bacterial swarming: a re-examination of cell-movement patterns. Curr Biol 2007,17(14):561–570.CrossRef 19. Wang Q, Frye JG, McClelland M, Harshey RM: Gene expression patterns during swarming

in Salmonella typhimurium: genes specific to surface growth and putative new motility and pathogeniCity genes. Mol Microbiol 2004,52(1):169–187.CrossRefPubMed 20. Raf inhibitor Connelly MB, Young GM, Sloma A: Extracellular proteolytic activity plays a central role in swarming motility in Bacillus subtilis. J Bacteriol 2004,186(13):4159–4167.CrossRefPubMed 21. Kim W, Surette MG: Prevalence of surface swarming behavior in Salmonella. J Bacteriol 2005,187(18):6580–6583.CrossRefPubMed

next 22. Kohler T, Curty LK, Barja F, van Delden C, Pechere JC: Swarming of Pseudomonas aeruginosa is dependent on cell-to-cell signaling and requires flagella and pili. J Bacteriol 2000,182(21):5990–5996.CrossRefPubMed 23. Shrout JD, Chopp DL, Just CL, Hentzer M, Givskov M, Parsek MR: The impact of quorum sensing and swarming motility on Pseudomonas aeruginosa biofilm formation is nutritionally conditional. Mol Microbiol 2006,62(5):1264–1277.CrossRefPubMed 24. Steil L, Hoffmann T, Budde I, Volker U, Bremer E: Genome-wide transcriptional profiling analysis of adaptation of Bacillus subtilis to high salinity. J Bacteriol 2003,185(21):6358–6370.CrossRefPubMed 25. Wang Q, Suzuki A, Mariconda S, Porwollik S, Harshey RM: Sensing wetness: a new role for the bacterial flagellum. Embo J 2005,24(11):2034–2042.CrossRefPubMed 26. Hall-Stoodley L, Costerton JW, Stoodley P: Bacterial biofilms: from the natural environment to infectious diseases.

It is exceedingly apparent that caffeine is not effective for non-trained individuals participating in high-intensity exercise. This may be due to the high variability in performance that is typical for untrained subjects. Results, however, are strikingly different for highly-trained athletes consuming moderate doses of caffeine. Collomp et al. [46] examined the use of 250 mg of caffeine (4.3 mg/kg) in trained and untrained swimmers. Swimmers participated in two maximal 100 m freestyle swims; significant increases in swim velocity were only recorded for the trained swimmers. Similar results were reported by MacIntosh and Wright [74] in a study

that examined the effects of caffeine in trained swimmers, but the caffeine treatment was provided at a higher dose (6 mg/kg) and the protocol involved a 1,500-meter Selleckchem GNS-1480 swim. Results indicated a significant improvement in swim times for those subjects who consumed caffeine, as compared to placebo. Moreover, time was measured at 500-m splits, which resulted in significantly faster times for each of the three splits for the caffeine condition [74]. As suggested

by Collomp et al., [29] it is possible that specific physiologic adaptations present in highly trained anaerobic athletes, such as PKC inhibitor enhanced regulation of acid-base balance (i.e., intracellular buffering of H+), is intrinsic for caffeine to exert an ergogenic effect [29]. Participants in a study published by Woolf et al. [30] were highly trained anaerobic athletes, and results of that investigation demonstrated a significant increase AZD8931 molecular weight in peak power with a moderate dose

of caffeine (5 mg/kg) as compared to placebo [30]. Wiles et al. [44] reported a 3.1% improvement in performance time for a 1-kilometer time trial (71.1s for caffeine; 73.4s for placebo) at a caffeine dose of 5 mg/kg, and results also included a significant increase in both mean and peak power [44]. Wiles et al. [44] indicated that subjects in the study reported regular interval sprint training, which may support the theory that caffeine is most beneficial in trained athletes who possess physiological adaptations to specific high-intensity training Bay 11-7085 [44]. A recent study published by Glaister et al. [31] examined a 5 mg/kg dose of caffeine on sprint interval performance. Subjects were defined as physically active trained men and performed 12 × 30 m sprints at 35 s intervals. Results indicated a significant improvement in sprint time for the first three sprints, with a consequential increase in fatigue for the caffeine condition [31]. The authors suggested that the increase in fatigue was due to the enhanced ergogenic response of the caffeine in the beginning stages of the protocol and, therefore, was not meant to be interpreted as a potential negative response to the supplement [31]. Bruce et al. [32] tested two doses of caffeine (6 mg/kg, 9 mg/kg) on 2000 m rowing performance in competitively trained oarsmen.

Actually, it has been shown that Salmonella expands its population in the liver by increasing the number of infection foci rather than undergoing massive intracellular growth in individual host cells, where the bacterial spreading from the initial infection foci to nearby cells may be facilitated by inducing cytotoxic effects in the infected cells [47, 48]. How sseJ STM reduces the cytotoxicity in S. Typhi is not clear. It is known that the lipid imbalance associated to the presence of lipid DNA Damage inhibitor alcohols, fatty acid and sterols is related to cytotoxicity and apoptosis [49, 50]. Any

process that limits the accumulation of these species is likely to be cytoprotective [50]. One such process involves the presence of different acyltransferase gene Selleck Baf-A1 families that generate neutral lipids or steryl esters from these lipid alcohols [50]. SseJ, that presents glycerophospholipid: cholesterol acyltransferase (GCAT) activity in eukaryotic cells [51], might plausibly contribute to the reduction of the lipid-associated cytoxicity. The precise mechanisms underlying this process is unknown, but one possibility is that the presence of sseJ STM in S. Typhi is affecting the lipid remodelling in the infected cells, in turn reducing the cytotoxicity.

All our results together suggest that the loss of the sseJ gene in S. Typhi contributed to the adaptation to the systemic infection by increasing the bacterial-induced cytotoxicity and by decreasing the retention/proliferation inside the epithelial cells. Conclusions Based on our results we conclude that the mutation that inactivate the sseJ gene in S. Typhi resulted in evident changes in the behaviour of bacteria in contact with eukaryotic cells, plausibly contributing to the S. Typhi adaptation to the systemic

infection in humans. Methods Bacterial strains, media and growth conditions The S. Typhi and S. Typhimurium strains used in this study are described in Table 2. Strains were routinely grown in Luria-Bertani (LB) selleck chemicals llc medium (Bacto Tryptone 10 g × l-1; Thiamet G Bacto Yeast Extract 5 g × l-1, NaCl 5 g × l-1) at 37°C, with vigorous shaking, or anaerobically by adding an overlay of 500 μl of sterile mineral oil as a barrier to oxygen prior to invasion assays with cultured human cells. When required, the medium was supplemented with antibiotics at the following concentrations: chloramphenicol 20 μg × ml-1, ampicillin 100 μg × ml-1 and kanamycin 50 μg × ml-1. Media were solidified by the addition of agar (15 g × l-1 Bacto agar). Table 2 Bacteria strains and plasmids used in this study Strain or plasmid Relevant characteristic Reference or Source Strains     Serovar Typhimurium     ATCC14028s Wild-type strain, virulent ATCC LT2 Wild-type strain S.

016 474 AAC → AAT –         498 GCG → GCT –         502 GTA → GTG –         518 ACA → ACG – ST5- MRSA-I (5) C (1)/t045 (1) Cape Town, RSA ≥ 256 481 CAT → TAT H481Y         498 GCG → GCT –         630 AAT → AAC –         658 GGT → GGA – ST612- MRSA-IV (8) AR-13324 supplier D (2), E (5), sporadic isolates (2)/t064 (3), t1443 (5), t1257 (1) Cape Town, RSA ≥ 256 481 CAT → AAT H481N         498 GCG → GCT –         512 CGT → CGC –         527 ATT → ATG I527M ST612- MRSA-IV (8) ND6 (2)/t064

(2) RSA (N83; N84) ≥ 256 481 CAT → AAT H481N         498 GCG → GCT –         512 CGT → CGC –         527 ATT → ATG I527M ST612- MRSA-IV (8) ND (1)/t064 (1) Australia (04-17052) ≥ 256 481 CAT → AAT H481N         498 GCG → GCT –         512 CGT → CGC –         527 ATT → ATG I527M ST612- MRSA-IV (8) ND (1)/t7571 (1) Australia (09-15534) ≥ 256 481 CAT → AAT H481N         498

GCG → GCT –         512 CGT → CGC –         527 ATT→ATG I527M         579 AAA→AGA K579R 1 Clonal types are indicated using the current international nomenclature (sequence type (ST) – antimicrobial phenotype – staphylococcal cassette chromosome mec (SCCmec) type) 2 PFGE, pulsed-field gel electrophoresis 3 As determined by E-test 4 S. aureus co-ordinates 5 RSA, Republic of South Africa 6 ND, not determined In addition to the mutations associated with amino acid substitutions in RpoB, silent single nucleotide polymorphisms (SNPs) were detected in the rpoB sequences of all 16 isolates (Table 2). Based on a comparison with the corresponding sequence eFT508 supplier of the rifampicin-susceptible S. aureus strain RN4220, all isolates shared a common SNP at amino acid 498 (GCG → GCT), as shown in Table 2. Otherwise between one and three additional SNPs particular to each clonal type were identified. Of note is the conserved SNP at amino acid 512 (CGT → CGC), which was detected in Adenylyl cyclase all 13 ST612-MRSA-IV isolates (Table 2). Discussion A number

of factors drive the emergence and spread of antibiotic resistance, including antibiotic usage, infection control Capmatinib order practices and the organism’s genetics [1]. Previous studies carried out in South Africa have reported large proportions of rifampicin-resistant MRSA isolates [2–5], and this study is no exception with the prevalence of rifampicin-resistance among MRSA isolates ranging from 39.7% to 46.4% (Figure 1). It is likely that the frequent use of rifampicin to treat tuberculosis in South Africa has driven the high prevalence of rifampicin-resistance among local MRSA. Support for this suggestion comes from the work of Sekiguchi et al. [14] who reported a significantly higher prevalence of rifampicin-resistant MRSA in tuberculosis wards compared to non-tuberculosis wards in two hospitals in Japan. A previous study showed that ST612-MRSA-IV was the dominant clone circulating in public hospitals in Cape Town. The 44 isolates corresponding to this clonal type were uniformly resistant to rifampicin.

KEO assisted in the design of the study, acquired funding

for the project, and provided critical analysis of the manuscript.”
“Background The LAB represents a group of organisms that are functionally related by their general ability to produce lactic acid during homo- or hetro-fermentative Tideglusib manufacturer metabolism. They are predominantly Gram-positive, non-sporulating facultative anaerobic bacteria and have been isolated from sources as diverse as plants, animals and humans (for recent reviews on LAB see [3–7]). LAB can be sub-classified into 7 phylogenetic clades:Lactococcus, Lactobacillus, Enterococcus, Pediococcus, Streptococcus, Leuconostoc and Oenococcus [8]. They represent the single most exploited group of bacteria in the food industry, playing crucial roles in the fermentation of dairy products, meat and vegetables, as well as in the production of wine, coffee, cocoa and sourdough. This is reflected in the fact that to date (July 2008), 65 LAB genomes are either completely sequenced or in progress (source http://​www.​ncbi.​nlm.​nih.​gov). Some LAB, such as Lb. rhamnosus ATCC 53013 and Lb. acidophilus NCFM have been shown to be probiotic, which is defined by the World Health Organisation as: ‘Live microorganisms which when administered in adequate amounts confer a health benefit on the host’. [9] LAB are also a reservoir for antimicrobial peptides, such as bacteriocins. There are numerous examples ATPase inhibitor of bacteriocin producing LAB -one

of the most recent being Lb. salivarius UCC118, which was shown to be effective in reducing L. monocytogenes infections in mice [10]. However, members of the LAB can also be important pathogens, e.g. several Streptococcus and Enterococcus species. Such species are commonly found in the human and animal GI tract of and can occasionally cause disease. Diseases caused by colonisation of this website pathogenic LAB include urinary tract infections,

bacteremia, bacterial endocarditis, diverticulitis, and meningitis. Members of the LAB group have close phylogenetic relationships largely due to their sharing relatively small, AT-rich genomes (~2.4 Mb) and common metabolic pathways [8]. Despite their phylogenetic closeness, the LAB occupy a diverse set of ecological niches including fermenting plants, milk, wine, sour-dough, the human and animal GI tract and the oral cavities of vertebrates. Such niche diversity among closely-related species suggests considerable genetic adaptation during their evolution. The recently sequenced dairy culture Lb. helveticus DPC4571 [1], has 98.4% 16s ribosomal RNA identity to the gut organism Lb. acidophilus NCFM [2]. This gave us a unique opportunity to investigate two very similar organisms occupying extremely different niches and led us to investigate if we could define a specific gene set which is associated with niche adaptation in LAB. Phylogenetically, both Lb. helveticus and Lb. acidophilus branch together with other gut bacteria.

8 9 18.8 3 20.0 Perception of the response by family and friends  Adequate and helpful 7 33.3 35 71.4 7 46.7  Inadequate or nonexistent 14 66.7 14 28.6 8 53.3 Perception of the colleagues’ response  Adequate and helpful 11 52.4 26 54.2 6 40.0  Inadequate or nonexistent 10 47.6 16 33.3 8 53.3  No colleagues – – 6 12.5 1 6.7 References Barling

J, Dupré KE, Kelloway EK (2009) Predicting workplace aggression and violence. Annu Rev Psychol 60:671–692CrossRef Bowling NA, Beehr TA (2006) Workplace harassment from the victim’s perspective: a theoretical model and meta-analysis. J Appl Psychol 91(5):S63845 998CrossRef Buckley P, Cookson H, Pakham C (2010) Violence PCI-34051 mouse at work: findings from the 2008/09 British Crime Survey. Health and Safety Executive, London Cole LL, Grubb PL, Sauter SL, Swanson NG, Lawless P (1997) Psychosocial correlates of harassment, threats and fear of violence in the workplace. Scan J Work Environ Health 23:450–457CrossRef De Puy J, Romain-Glassey N, Gut M, Wild P, Dell’Eva A-S, Asal V (2012) Rapport final présenté à la Suva (groupe Progrès). Etude portant sur les victimes d’agressions au travail ayant consulté l’Unité de médecine des violences entre 2007 et 2010 et sur les ressources de prévention dans le canton de Vaud. Institut universitaire romand de santé au travail

et Centre universitaire romand de médecine légale. Lausanne Dillon BL (2012) Workplace violence: GSK2118436 datasheet impact, causes, and prevention. Work 42(1):15–20 European Foundation for the Improvement of Living and Working Conditions (2007) 4th European working conditions survey EWCS. Office

for official publications of the european communities, Luxembourg European Foundation for the Improvement of Living and Working Conditions (2010) Foundation findings: physical and psychological violence at the workplace. Eurofound, Dublin Gates DM (2004) The epidemic of violence against health care workers. Occup Environ Med 61:649–650CrossRef Gillespie GL, Gates DM, Miller M, Howard PK (2010) Workplace violence in healthcare settings: PRKD3 risk factors and protective strategies. Rehabil Nurs 35(5):177–184CrossRef Graf M, Pekruhl U, Korn K, Krieger R, Mücke A, Zölch M (2007) Quatrième enquête européenne sur les conditions de travail en 2005. Résultats choisis du point de vue de la Suisse. SECO/Fachhochschule Nordwestschweiz, Berne et Brugg Hansen ÅM, Hogh A, Persson R, Karlson B, Garde AH, Ørbæk P (2006) Bullying at work, health outcomes, and physiological stress response. J Psychosom Res 60(1):63–72CrossRef Hogh A, Viitasara E (2005) A systematic review of longitudinal studies of nonfatal workplace violence. Eur J Work Org Psychol 14(3):291–313CrossRef Kowalenko T, Cunningham R, Sachs CJ, Gore R, Barata IA, Gates D, Kerr HD (2012) Workplace violence in emergency medicine: current knowledge and future directions. J Emerg Med 43(3):523–531CrossRef LeBlanc MM, Kelloway EK (2002) Predictors and outcomes of workplace violence and aggression.

In this study we described six NDM-4-producing EPZ004777 purchase E.coli isolates obtained from two patients admitted to an Italian hospital. We also present data on the localization and the genetic environment of the bla NDM-4 gene. Methods Bacterial strains Six E.coli isolated from urine samples of two inpatients at the San Martino-IST University Hospital (Genoa, Italy)

were studied. Isolates were taken as part of standard patient care and informed consent for the use of clinical data has been obtained by both patients. Strain identification, CRT0066101 antibiotic susceptibility testing and phenotypic screening for MBL production Routine identification and antibiotic susceptibility testing were carried out using the Vitek-2 automated system (BioMérieux, Marcy-L’etoile, France). In vitro activity of carbapenems, aztreonam, fosfomycin and nitrofurantoin was further determined by the broth microdilution method and interpreted according to the of European Committee on Antimicrobial Susceptibility Testing (EUCAST ) guidelines (Version 4.0, 2014) [6]. To detect metallo-β-lactamase (MBL) production,

a synergy test using imipenem and EDTA discs was used [7]. Pulsed-field gel electrophoresis (PFGE) Genomic DNA was prepared, digested with XbaI (New England Biolabs Inc., MA, USA) and subjected to PFGE with the CHEF DRII device (Bio-rad, Milan, Italy), as described previously [8]. Fingerprinting pattern was interpreted Momelotinib concentration according to the method of Tenover et al. [9]. Multilocus sequence typing (MLST) MLST was carried out using protocols and conditions described on the E.coli MLST website (http://​mlst.​warwick.​ac.​uk/​mlst/​dbs/​Ecoli/​documents/​primersColi_​html).

Sequence types were assigned using the website interface. Molecular analysis techniques Polymerase chain reaction (PCR) amplification of the bla NDM gene and direct sequencing of the PCR products was performed as previously described [10]. Screening for resistance genes was carried out using primers and conditions previously described [11–13]. Phylogenetic analysis using multiplex PCR method as described previously [14] was used. PCR experiments were performed to identify the upstream- and downstream-located regions of the bla NDM-4 gene [15]. Mapping of the variable region of class 1 integron was performed by PCR as described previously [16]. The genetic environment of bla NDM-4 was studied by PCR mapping and sequencing Amylase as described previously [13]. Conjugation assay and plasmid study Plasmid transfer was attempted by conjugation, using E.coli J53 as the recipient, as described previously [17]. Plasmid DNA, isolated from E.coli, was obtained by the alkaline lysis method and was used as a template in PCR analysis with primers that are specific for bla NDM and bla CTX-M[18]. To rule out chromosomal DNA contamination the template was used to amplify an internal fragment of the house-keeping recA gene. A PCR-based replicon typing method was used to identify the incompatibility group [19].

0) CT computed tomography aActual osmolality bNot approved for intravascular administration Invasive diagnostic imaging including cardiac angiography or percutaneous catheter intervention Does CKD increase the risk for developing CIN after CAG? Answer: 1. It is highly likely that CKD (GFR <60 mL/min/1.73 m2) increases the risk for developing CIN after CAG.

The risk for developing CIN increases find more as kidney function decreases.   2. We recommend that physicians explain CIN to patients with an eGFR of <60 mL/min/1.73 m2 who are going to undergo CAG, and that they take appropriate preventive measures such as fluid therapy before and after CAG.   Recently, CAG and catheter-based revascularization have become common procedures,

and the use of contrast media has increased substantially. It has been reported that in patients with CKD the risk of CIN increases as kidney function (GFR) decreases (Fig. 1) [8]. In 2001, Shiraki et al. [73] reported that 61 of 1,920 patients (3.2 %) who underwent CAG developed CIN, and 1 of them (0.05 %) required hemodialysis. In Idasanutlin in vitro another study, Fujisaki et al. [74] reported that CIN SAHA purchase developed in 12 of 267 patients (4.5 %) who underwent CAG, and hemodialysis was required in 2 patients (0.7 %). In a report from the Mayo Clinic in 2002, CIN developed in 254 of 7,586 (3.3 %) patients who underwent CAG, and 20 (7.9 %) of these required hemodialysis [4]. Mortality at 1 and 5 years were 12.1 and 44.6 %, respectively, in patients with CIN, which were significantly higher than those in patients without CIN (3.7 and 14.5 %, respectively). Montelukast Sodium In a study reported in 2009, Abe et al. [75] reported that the incidence of CIN within 5 days after

CAG was 4.0 % in 1,157 consecutive patients who underwent CAG, and risk factors for CIN included a baseline SCr level of ≥1.2 mg/dL and the use of a large volume (≥200 mL) of contrast media. In the earlier-mentioned studies, CIN was defined as an increase in SCr levels by ≥0.5 mg/dL. The risk of CIN after CAG was 3.0–5.0 %, and CIN developed mainly in high-risk patients such as those with diabetes, anemia, dehydration, or an underlying kidney diseases, and/or those who were elderly or were receiving nephrotoxic agents [50]. It is recommended that patients with CKD should receive appropriate preventive treatment such as fluid therapy and be closely monitored for kidney function after CAG. Fig. 1 Risk for developing CIN according to baseline kidney function. The incidence of CIN is higher in patients with lower baseline eGFR, and is higher in patients with diabetes than in those without diabetes. CIN contrast-induced nephropathy, eGFR estimated glomerular filtration rate. Adapted from J Am Coll Cardiol. 2008;51:1419–1428 [8], with permission from Elsevier Inc.

This requirement seriously hampers epidemiological investigations, particularly at international scales [21, 23].

Typing procedures based on DNA sequences overcome these limitations, Selleck PND-1186 since sequence data may easily be exchanged and stored in databases that are accessible via the internet. Accordingly, a scheme for multilocus sequence typing (MLST) of C. difficile was developed recently that is based on sequences from seven housekeeping gene fragments [31]. While MLST to date has been applied to a limited number of isolates, available data allowed a first glimpse at the largely clonal genetic population structure of C. difficile [23, 31, 32]. In clonal bacteria, novel genotypes in the course of evolution are generated primarily AZD0530 order through mutations, which in slowly evolving housekeeping genes are rare. Hence, it is this very clonality of C. difficile and the associated linkage disequilibrium that causes MLST to provide poor discriminatory power, which is exemplified by the fact that relevant epidemic strains are not resolved [31]. In addition, MLST remains too Tanespimycin expensive to be applied for routine typing aside from dedicated research

projects. More variable genomic regions may provide improved discrimination ability. In contrast to MLST, it may even suffice to sequence a single locus or very few genetic loci that are sufficiently variable, since – analysing a clonal population – phylogenetic inferences will rarely be confounded through homologous genetic recombination. Sequence-based typing schemes relying on one or several highly discriminatory markers have previously been established for a number of pathogens, including Staphylococcus aureus (spa gene) [33], Campylobacter jejuni (flaA) [34, 35], Streptococcus pyogenes (emm) [36] and Neisseria meningitidis (porA, fetA) [37–39]. The surface layer protein

gene slpA has recently been proposed as a promising target for sequence-based typing of C. difficile [40]. The limited data available suggests extremely high sequence variation among isolates and, correspondingly, excellent discriminatory power [23, 40]. To date, however, slpA sequencing reportedly has been applied to a total of only 11 different ribotypes, and it is not clear if the method is universally applicable why [23, 40]. It is anticipated that the requirement for degenerate oligonucleotide primers may restrict the general utility of the current protocol [39]. The method has as yet not been successfully transferred to any other laboratory [23, 40]. This present report describes the development and application of a new assay for genotyping C. difficile that is based on sequence analysis of two stretches of repetitive DNA. Investigating a panel of 154 diverse C. difficile isolates, we demonstrate extensive sequence variation in these genomic regions, resulting in high discriminatory power, and excellent concordance with PCR ribotyping.