In addition, the iron chelator 2, 2′-dipyridyl was able to kill t

In addition, the iron chelator 2, 2′-dipyridyl was able to kill the mioC mutant strain (Fig. 1b). Subsequently, bacterial sensitivities were tested with three different metals: As, Zn and Cu (Fig. 1c). Consistent with the PM assay, the mutant was notably sensitive to As and Zn. Although Cu was not used in the PM assay, we performed the sensitivity test using Cu because it is known to promote cell death. However, the sensitivity of the mutant to Cu was not different from that of the

other two strains. To summarize, we confirmed the results observed with the Biolog PM system using sensitivity tests. The mioC mutant strain displayed significant reductions in biofilm formation during static aerobic growth (Fig. 2a). Therefore, we thought that the mutant might be able to reduce BMN 673 manufacturer cell aggregation of P. aeruginosa under biofilm conditions. Interestingly, aggregation of the mutant cell was reduced during

static Idasanutlin ic50 aerobic growth (Supporting Information, Fig. S1). Under iron excess condition, biofilm formations of the mutant and over-expressed complementation strains were reduced compared with that of the wild type (Fig. 2a and Fig. S2). Thus, the balance of the mioC gene product may be important for maintaining biofilm formation ability under iron excess condition. Interestingly, biofilm formation of the mutant was significantly induced by the iron chelator 2,2′-dipyridyl compared with the other two strains (Fig. 2a and Fig. S2). The growth mutant appeared to be slower under the iron chelator than was the wild type (Fig. 1b), whereas biofilm formation ability was enhanced by

0.5 mM dipyridyl (Fig. 2a and Fig. S2). No biofilm formation occurred in the absence of dipyridyl, but robust biofilm formation occurred in the presence of dipyridyl, which clearly demonstrated that dipyridyl Glycogen branching enzyme treatment increased biofilm formation of the mutant (Fig. S2). In addition, biofilm formation was increased in the mioC mutant cell under Zn and As stresses (Fig. 2b). Consistent with sensitivity data, biofilm formation under Cu stress was similar to that under normal conditions (Fig. 2b). Subsequently, the colony morphology test was performed using Congo red and Brilliant blue (Fig. 2c–e). Congo red and Brilliant blue, a constituent of the agar used in the experiments, are known to bind the glucose-rich exopolysaccharide pellicle and proteins, respectively (Dietrich et al., 2008). Interestingly, red color formation was not observed in the mioC mutant strain, compared with the wild type under iron-rich conditions (Fig. 2d). Red color was recovered in the mioC over-expressed complementation strain under iron excess (Fig. 2d). However, this pellicle appeared in the mutant but disappeared in the other two strains under iron depletion (Fig. 2e). We also performed motility tests (Fig. S3). Interestingly, the swarming motility of the mioC mutant strain had a branch form.

We also assayed the strains for the presence of mutations in the

We also assayed the strains for the presence of mutations in the quinolone resistance–determining regions (QRDRs) of gyrA gene encoding GyrA subunit of DNA gyrase and parC gene encoding ParC subunit of topoisomerase IV. We prospectively collected 121 consecutive single-patient MDR A. baumannii clinical strains during 2006 and 2007 at Cedars-Sinai Medical Center. We considered

a strain as MDR if it was resistant to two or more antibiotic classes that included anti-pseudomonal penicillin and its combination with β-lactamase inhibitor (e.g. piperacillin/tazobactam), anti-pseudomonal cephalosporins (e.g. ceftazidime or cefepime), carbapenems (e.g. IMP), aminoglycosides [e.g. tobramycin or amikacin (AN)], and fluoroquinolones (e.g. ciprofloxacin or levofloxacin) find more based on VITEK® Cabozantinib 2 (bioMérieux, Inc.). All 121 strains were analyzed by repetitive PCR (rep-PCR) amplification using the DiversiLab®Acinetobacter Fingerprinting Kit according to manufacturer’s instructions

(bioMérieux, Inc.). Briefly, bacterial DNA was extracted using UltraClean™ Microbial DNA Isolation Kit (MO BIO Laboratories, Inc.). Amplification reactions were performed in the GeneAmp® PCR System 9700 under the following conditions: 2 min at 94 °C, 35 cycles of denaturation (30 s at 94 °C), annealing (30 s at 50 °C) and extension (90 s at 70 °C), and a final extension before of 3 min at 70 °C. Rep-PCR products were separated by electrophoresis using the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc.). Band patterns for each strain

were aligned and interpreted with web-based DiversiLab software provided by the manufacturer (bioMérieux, Inc.). Strains were grouped by ≥ 95% similarity. Medical record review identified an incident episode of nosocomial acquisition according to Centers for Disease Control surveillance definitions (Horan et al., 2008). Accordingly, 19 strains from patients with evidence of infection or colonization with A. baumannii prior to or at the time of admission to our institution during the study period were considered as having either a repeat episode or non-nosocomial A. baumannii infection, respectively, and their clinical strains were excluded from this study. Of the remaining strains, those belonging to the two prevalent clones, A and B, were selected for further analyses. Etest (bioMérieux, Inc.) was performed on 33 representative strains that were resistant to at least three classes of antibiotics (26 of clone A and seven of clone B) for susceptibility to IMP, COL, AN, DOX, tigecycline (TGC), RIF, and azithromycin (AZT) as per manufacturer’s recommendations.

fumigatus, has been reported to support an aspergilloma (Lee, 201

fumigatus, has been reported to support an aspergilloma (Lee, 2010; Muller et al., 2011). One such recent case study described an Aspergillus flavus aspergilloma in a neonate who had urinary catheters placed for genitourinary complications (Martinez-Pajares et al., 2010). Aspergillus species of industrial importance can also be problematic. For example, adhesion of Aspergillus niger spores may cause surface deterioration on different substrates, and has

also been associated with colonization of contact lenses (Marques-Calvo, high throughput screening 2002). However, many of the characteristics associated Aspergillus biofilms are beneficial with respect to industrial processes. Various organic acids have been produced by Aspergillus biofilms using different supports and bioreactors. In one of the oldest publications, A. niger was grown attached to the vertical discs of a rotating disc reactor (Blain et al., 1979), producing fourfold higher citric acid titres than in stirred tank reactor (Anderson et al., 1980). It was also found that find more A. niger immobilized on polyurethane foam (biofilms) in a bubble reactor for citric acid production performed better than free-living pellets (Lee et al., 1989). Other organic acids have been produced by Aspergillus biofilms. For example, Aspergillus

terreus grown attached on polyurethane foam used for itaconic acid production (Kautola et al., 1989), gluconic acid has also been produced by passively immobilized A. niger (Vassilev et al., 1993; Fiedurek, 2001). Moreover, several enzymes have been produced by Aspergillus biofilm systems, such as the production of glucose oxidase, inulinase, amylase and cellulases by A. niger (Fiedurek & Ilczuk, 1991; Murado et al., 1994; Skowronek & Fiedurek, 2006; Gamarra et al., 2010), production

of β-frutofuranosidase by Aspergillus japonicas (Mussatto et al., 2009) and production of xylanases by A. terreus and A. niger (Gawande & Kamat, 2000). Aspergillus foetidus biofilms have been shown to degrade some plastics under growth (Upreti & Srivastava, 2003). Also, Aspergillus versicolor has been found to form biofilms on perlite particles in a packed column reactor, and in this condition, it could degrade n-alkanes, aromatic hydrocarbons and carbazoles of petroleum samples (Sanchez et al., 2006). Removal of heavy metals (copper, Loperamide chromium, iron and nickel) by biosorption of either A. niger or A. terreus biofilms formed on polyurethane, has also been reported to be a highly efficient method of metal removal (Tsekova & Ilieva, 2001; Dias et al., 2002). Clearly, Aspergillus biofilms are important in many industrial processes, particularly because they are much more productive than in the classical submerged fermentation with free-living mycelia. Filamentous growth is a fundamental feature of fungal biofilms and is an important morphological characteristic of A. fumigatus required during the development of an aspergilloma (Beauvais et al., 2007; Ramage et al., 2009; Loussert et al.

Transmitter domains consist of a dimerization and histidine phosp

Transmitter domains consist of a dimerization and histidine phosphorylation domain (DHp), and a catalytic and ATPase domain (CA). The CA domain belongs to the GHKL (gyrase, Ridaforolimus Hsp90, HK, MutL) family of ATPases (Dutta & Inouye, 2000). GHKL ATPases contain a distinctive ATP-binding pocket known as a Bergerat fold, which is an α/β sandwich composed of a mixed β sheet and an α helix bundle (Bergerat et al., 1997). Based on the sequences of their transmitter domains, HKs have been grouped into 12 families (Grebe & Stock, 1999; Karniol & Vierstra, 2004). The M. xanthus genome encodes 131 HKs that fall into one of these 12 families (Goldman et al., 2006). Many of the 131 HKs have been

linked to the development of spore-filled fruiting bodies through expression profiling (Shi et al., 2008) and/or mutational analyses (Shi et al., 2008; Whitworth & Cock, 2008). One M. xanthus gene codes for a putative HK (Nla6S) that cannot be placed in any of the 12 classical HK families; it is predicted to have a typical DHp find more domain, but lacks a recognizable CA domain. Here, we show that Nla6S is indeed a HK and is the prototype

for a new family of HKs found to date only in the fruiting members of the Cystobacterineae suborder of the myxobacteria. All strains and plasmids used in this study are listed in Supporting information, Table S1. All primers used in this study are listed in Table S2. Myxococcus xanthus strains were grown at 32 °C in CTTYE broth or on CTTYE agar plates (Caberoy et al., 2003). CTTYE broth and CTTYE agar plates were old supplemented with 50 μg mL−1 of kanamycin as needed. Fruiting body development was carried out at 32 °C in six-well microtiter plates containing MC7 buffer (Søgaard-Andersen et al., 1996). Escherichia coli strains were grown in Luria–Bertani (LB) broth or on LB agar plates. For protein expression and purification, E. coli strains were grown in 2XYT broth

(1.6% tryptone, 1% yeast extract, 0.5% NaCl). LB broth, 2XYT broth, and LB agar plates were supplemented with 100 μg mL−1 of ampicillin or 50 μg mL−1 of kanamycin as needed. The Jpred 3 secondary structure prediction server (Cole et al., 2008) was used to predict the secondary structure of Nla6S. The TopPred topology of membrane protein server (von Heijne, 1992; Claros & von Heijne, 1994) was used to identify potential membrane-spanning regions in proteins. Sequence alignments for phylogenetic analysis were generated with clustalw2 (Larkin et al., 2007) using the predicted transmitter domain of the HKs. The phylogenetic tree was constructed using the maximum-likelihood method with PhyML-aLRT (Guindon et al., 2010). The nla6S gene was codon optimized for expression in E. coli (Table S3) (DNA2.0). The 609-bp region of the codon optimized nla6S gene, which encodes the 203 amino acid C-terminal transmitter domain of Nla6S (Nla6S-TD), was cloned into the pET28b vector (EMD Biosciences).

We are very grateful to Sir David Hopwood for critical reading of

We are very grateful to Sir David Hopwood for critical reading of and useful suggestions and corrections on the manuscript. We thank Huarong Tan for kindly providing a cosmid containing the entire nikkomycin biosynthetic gene cluster. This work was supported by grants from National ‘973’ project (2011CBA00801), National Nature Science Foundation of China (31121001), and the Chinese Academy of Sciences project (KSCX2-EW-G-13) to Z.Q. M.Z., X.J., and P.X. contributed equally to this work. Please note: Wiley-Blackwell is not responsible

for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Microbiology, St. Joseph’s click here Health Centre, Toronto, ON, Canada Pseudomonas aeruginosa PA96 is a clinical isolate from Guangzhou, China, that is multiresistant to antibiotics. We previously described the 500-kb IncP-2 plasmid, pOZ176 that encodes many resistance genes including the IMP-9 carbapenemase. BMS-777607 nmr Whole-genome sequencing of PA96 enabled characterization of its genomic islands, virulence factors, and chromosomal resistance genes. We filled gaps using PCR and used optical mapping to confirm the correct contig order. We automatically annotated the core genome and manually annotated the genomic islands.

The genome is 6 444 091 bp and encodes 5853 ORFs. From the whole-genome sequence, we constructed a physical map and constructed Teicoplanin a phylogenetic tree for comparison with sequenced P. aeruginosa strains. Analysis of known core genome virulence factors and resistance genes revealed few differences with other strains, but the major virulence island is closer to that of DK2 than to PA14. PA96 most closely resembles the environmental strain M18, and notably shares a common

serotype, pyoverdin type, flagellar operon, type IV pilin, and several genomic islands with M18. “
“Salmonella enterica serovar Typhimurium is an animal and zoonotic pathogen of worldwide importance. Intestinal colonization, induction of enteritis and systemic translocation by this bacterium requires type III protein secretion. Strategies that target this process have the potential to control infection, pathology and transmission. We defined the global transcriptional response of S. Typhimurium to INP0403, a member of a family of salicylidene acylhydrazides that inhibit type III secretion (T3S). INP0403 treatment was associated with reduced transcription of genes involved in T3S, but also increased transcription of genes associated with iron acquisition. We show that INP0403 restricts iron availability to Salmonella, and that inhibition of T3S system-1 by INP0403 is, at least in part, reversible by exogenous iron and independent of the iron response regulator Fur.

We are very grateful to Sir David Hopwood for critical reading of

We are very grateful to Sir David Hopwood for critical reading of and useful suggestions and corrections on the manuscript. We thank Huarong Tan for kindly providing a cosmid containing the entire nikkomycin biosynthetic gene cluster. This work was supported by grants from National ‘973’ project (2011CBA00801), National Nature Science Foundation of China (31121001), and the Chinese Academy of Sciences project (KSCX2-EW-G-13) to Z.Q. M.Z., X.J., and P.X. contributed equally to this work. Please note: Wiley-Blackwell is not responsible

for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Microbiology, St. Joseph’s RG-7204 Health Centre, Toronto, ON, Canada Pseudomonas aeruginosa PA96 is a clinical isolate from Guangzhou, China, that is multiresistant to antibiotics. We previously described the 500-kb IncP-2 plasmid, pOZ176 that encodes many resistance genes including the IMP-9 carbapenemase. BAY 80-6946 mouse Whole-genome sequencing of PA96 enabled characterization of its genomic islands, virulence factors, and chromosomal resistance genes. We filled gaps using PCR and used optical mapping to confirm the correct contig order. We automatically annotated the core genome and manually annotated the genomic islands.

The genome is 6 444 091 bp and encodes 5853 ORFs. From the whole-genome sequence, we constructed a physical map and constructed Astemizole a phylogenetic tree for comparison with sequenced P. aeruginosa strains. Analysis of known core genome virulence factors and resistance genes revealed few differences with other strains, but the major virulence island is closer to that of DK2 than to PA14. PA96 most closely resembles the environmental strain M18, and notably shares a common

serotype, pyoverdin type, flagellar operon, type IV pilin, and several genomic islands with M18. “
“Salmonella enterica serovar Typhimurium is an animal and zoonotic pathogen of worldwide importance. Intestinal colonization, induction of enteritis and systemic translocation by this bacterium requires type III protein secretion. Strategies that target this process have the potential to control infection, pathology and transmission. We defined the global transcriptional response of S. Typhimurium to INP0403, a member of a family of salicylidene acylhydrazides that inhibit type III secretion (T3S). INP0403 treatment was associated with reduced transcription of genes involved in T3S, but also increased transcription of genes associated with iron acquisition. We show that INP0403 restricts iron availability to Salmonella, and that inhibition of T3S system-1 by INP0403 is, at least in part, reversible by exogenous iron and independent of the iron response regulator Fur.

Between 2001 and 2008, almost all infants born to HIV-infected wo

Between 2001 and 2008, almost all infants born to HIV-infected women in the UK and Ireland received antiretroviral PEP, mostly with one drug. Use of triple PEP increased over time, particularly for infants whose mothers were untreated or viraemic despite HAART, in line with current guidelines. Post-exposure antiretroviral prophylaxis for infants

born to HIV-infected women is an important component of the standard package of interventions used for prevention of mother-to-child transmission (MTCT) of HIV in resource-rich and resource-poor countries [1–3]. The Pediatric AIDS Clinical Trial Group first demonstrated in a randomized trial in 1994 that the administration of zidovudine in pregnancy, during labour and to the infant reduced the risk selleck screening library of transmission

to the child by two-thirds [4]. The independent contribution of neonatal post-exposure prophylaxis (PEP) has since been shown in a number of clinical trials and observational studies [5–7]. The British HIV Association (BHIVA) recommends single-drug PEP for most infants from birth [3]. In addition, consideration of combination PI3K inhibitor prophylaxis is recommended for infants born to women who (i) have an unplanned delivery before starting antiretroviral therapy, (ii) present late, with no information on HIV parameters, or (iii) are diagnosed after Demeclocycline delivery [3,8,9]. Since 2005, British guidelines have recommended that combination PEP should also be considered for infants born to women with persistent viraemia despite combination antiretroviral therapy in pregnancy. However, sick or very premature infants may be unable to receive oral medication, leaving intravenous zidovudine as the only option [3]. Although neonatal PEP continues to be recommended, a decline in use and duration was reported in the European Collaborative Study [10]. Conversely, an Italian study showed use of neonatal prophylaxis increasing in recent years, including combined prophylaxis with two

or more antiretroviral drugs, in a cohort of over 3500 infants [11]. Our aims were to review the use of neonatal PEP in the United Kingdom (UK) and Ireland using national surveillance data and to investigate factors associated with the use of combination prophylaxis in the context of changes in national guidelines. Active population-based surveillance of obstetric and paediatric HIV infection in the UK and Ireland is carried out through the National Study of HIV in Pregnancy and Childhood (NSHPC) [12]. Information on maternal demographic and pregnancy characteristics, antiretroviral therapy, neonatal prophylaxis and infection status of the child is routinely collected.

Governmental actions including increasing awareness of the import

Governmental actions including increasing awareness of the importance of vitamin D and guidelines on how to obtain it

are necessary. Creating areas where women, particularly those of lower socio-economic status, can enjoy sun exposure as well as fortifying more foods would go some way towards tackling this problem. “
“Behçet’s disease (BD) is a systemic vasculitis disease with oral and genital aphthous ulceration, uveitis, skin manifestations, arthritis and neurological involvement. Many investigators have published articles on BD in the last two decades since introduction of diagnosis criteria by the International Study Group for Behçet’s Disease in 1990. However, there is no scientometric analysis available for this increasing amount of literature. A scientometric analysis I-BET-762 order method was used

to achieve a view of scientific articles about BD which were published between 1990 and 2010, by data retrieving from ISI Web of Science. The specific features such as publication year, language of article, geographical distribution, main journal in this field, institutional affiliation and citation characteristics were retrieved and analyzed. International collaboration was analyzed using Intcoll and Pajek softwares. There was a growing trend in the number of BD articles from 1990 to 2010. The number of citations to BD literature also increased around 5.5-fold in this period. The countries found to have the highest output were Turkey, Japan, the USA and England; the first two universities R788 were from Turkey. Most of the top 10 journals publishing BD articles were in the field of rheumatology, consistent with the subject areas of the articles. There was a correlation between the citations per paper and the impact factor of the publishing journal. This

is the first scientometric analysis of BD, showing the scientometric characteristics of ISI publications on BD. “
“The historical significance Megestrol Acetate of the Medici family of Florence is widely recognised, but the diseases which afflicted leading members of this family have only been scientifically studied in recent decades. Paleopathological findings on exhumed skeletons, supplemented by medical descriptions in historical documents, have permitted a retrospective diagnosis of a triple pathological syndrome in the senior branch of the Medici family. Peripheral joint and spinal conditions, with the presence of skin disease, are identified in several generations of the family in the 15th century and are presented as the ‘Medici syndrome’. Radiological findings are compared with macro- and microscopical descriptions in the diagnosis of the peripheral joint disease and spinal ankylosis/stenosis within the syndrome. “
“Objective:  To investigate the effect of Kashin-Beck disease (KBD)-affected feed and T-2 toxin on the bone development of Wistar rats.

, 1999) They are widespread throughout the photic regions of the

, 1999). They are widespread throughout the photic regions of the world’s oceans between 40°S and 50°N, with cell densities of up to 105 cells mL−1 in the central oligotrophic gyres (Partensky et al., 1999). They are principally distinguished

into two taxonomic GSK1120212 in vivo clades due to physiological niche adaptation to light intensity: high light- and low light-adapted ecotypes (Moore et al., 1998; West & Scanlan, 1999; Rocap et al., 2003). A great deal of interest has arisen around Prochlorococcus due to its small size and specifically its near-minimal genome. Indeed, the chromosomes of most Prochlorococcus strains demonstrate significant genomic reduction, revealing a central conserved core set of essential genes and a variable shell, which is hypothesized to reflect each individual strain’s evolutionary adaptation to a specific environmental niche (Kettler et al., 2007; Shi & Falkowski, 2008). Closer inspection of Prochlorococcus genomes reveals that Selleckchem FK228 the majority of these strain-specific genes (74% in the case of Prochlorococcus strain MED4) are located in highly variable ‘genomic islands’, suggesting a mosaic structure that continually undergoes genomic rearrangement (Coleman et al., 2006). A suggested source of pressure for these organisms to reduce genome as well as cell size is thought to be reduced nutrient

availability (Raven, 1998), which is a characteristic of subtropical oceans,

particularly phosphate (P). Indeed, P concentrations are hypothesized to have affected domain shifts from a eukaryotic to a prokaryotic life in these oligotrophic regions (Karl et al., 1995, 2001). Also, recent studies have found that phytoplanktonic species within nutrient-poor oceanic biomes substitute phospholipids with sulpholipids in order to conserve Farnesyltransferase ambient phosphorous for more essential metabolic use in the face of competition from heterotrophic bacteria (Van Mooy et al., 2006, 2009). A recent study of MED4 showed that a unique suite of genes was upregulated under P stress (Martiny et al., 2006). Most of these genes are orthologues of Escherichia coli genes located in and around the phoB operon, but another set are located within a variable genomic island, ‘Island 5’, and unique to MED4. The function of these genes is as yet uncharacterized; however, some putative annotations are available at GenBank (http://www.ncbi.nlm.nih.gov/). It is clear that the availability and ambient concentration of inorganic P within oligotrophic regions is a crucial factor determining the success of MED4 within those environments. Therefore, this study seeks to ascertain the global quantitative proteomic response of MED4 to longer term P starvation, and thereby providing further insight into how this organism responds to P stress.

With regard to our primary endpoint, neither total bilirubin nor

With regard to our primary endpoint, neither total bilirubin nor ATV status was significantly associated with FMD of the brachial artery. This is consistent with both the study by Flammer et al. and the SABAR (Switch to Atazanavir and Brachial Artery Reactivity) study by Murphy et al., which showed that switching to ATV from another PI, whether boosted with ritonavir or not, did not improve endothelial function measured using FMD after 24 weeks, despite significant improvement in lipid levels [13, 14]. It is conceivable that the effect of a modest increase in bilirubin in this population is masked by the ongoing heightened

inflammation resulting from chronic HIV infection. Indeed, with potent ART, inflammation and endothelial dysfunction do improve [18, 19], but not to SB431542 research buy normal levels, when compared with HIV-uninfected individuals [2, 19]. Further, participants included in this study did not have extremely elevated total bilirubin levels (median 1.8 mg/dL; IQR 1.1–2.6 mg/dL; minimum 0.3 mg/dL and maximum 5 mg/dL). Although seeing an effect with extreme hyperbilirubinaemia would be mechanistically intriguing, this would have uncertain clinical relevance. Another consideration is that the antioxidant effect of elevated bilirubin was outweighed by the oxidative stress induced by ART [20]. Although a different method for measuring endothelial function

was used, our results are Staurosporine mouse incongruent with the study by Dekker et al., in which ATV-induced hyperbilirubinaemia did improve endothelium-dependent vasodilation measured using forearm blood flow response to acetylcholine in participants with type II diabetes mellitus after 3 days [11]. In our study, perhaps an effect would have been seen if FMD had been measured earlier after ATV was initiated [median (IQR) duration on ATV in our study was 28.5 (16.8–47.7) months]. The clinical implication of a solely transient acute effect would also be questionable, however. Another consideration is that endothelial dysfunction is more pronounced in subjects with diabetes mellitus

than in our Benzatropine HIV-infected population and is why an effect was seen in this potentially higher risk group. To better assess whether the degree of endothelial dysfunction played a role in the association between total bilirubin level and FMD in our study, the correlation between total bilirubin level and FMD in those with the lowest FMD (FMD less than the median FMD of 3.3%) was determined. Total bilirubin was not correlated with FMD in this subgroup (Spearman correlation coefficient = 0.13; P = 0.38). With regard to our secondary endpoints, neither total bilirubin nor ATV status was associated with markers of inflammation, coagulation or oxidation, with the exception of fibrinogen. Fibrinogen levels were higher among participants taking ATV. This result is consistent with a study by Madden et al., where PI use was associated with elevated fibrinogen levels.