It is worth mentioning that sPBP6, which is the next nearest homolog of DacD, is inactive on pentapeptide substrate (Chowdhury et al., 2010). The crystal structures of sPBP5 and sPBP6 (Nicholas et al., 2003; Chen et al., 2009) show a similar secondary structure with no gross architectural differences. In the absence of crystal structure, CD spectral analysis would be of utmost importance to elucidate the biophysical characteristics of sDacD. It was evident from the CD spectra that purified protein was in a native conformation with characteristics of the molecular

spectra CAL-101 in vitro of alpha and beta structures, indicating the protein was active and stable at room temperature. Unlike sPBP5 and sPBP6 (Chowdhury et al., 2010), more beta-sheets were detected in sDacD (Table 3, Supporting Information, Fig. S1). The occurrence of a larger amount of β-sheet structure in sDacD may cause some structural alteration, which might exert different biological activity than PBP5.

Because DacD shared a high level of aa identity with PBP5, homology modelling (or comparative protein structure modelling) could be applied to generate the three-dimensional conformation of sDacD. For model building, the program modeller 9v1 was used with the pdb coordinate, 3BEC chain A (crystal structure of E. coli PBP5 in complex with a peptide-mimetic cephalosporin; Sauvage et al., 2008) as template. The secondary structure prediction by BI 2536 in vitro predict protein and psipred suggested that sDacD was a αβ protein with a larger amount of β-sheet structure (Table 3 and Fig. S2), which was consistent with the results obtained from CD spectroscopic analyses. The model of lowest energy value had 94.9% residues in the most favoured region in the Ramachandran plot and 98.35% residues had an average Phospholipase D1 3D-1D score above 0.2, as obtained through verify3d profile (Fig. 2), which affirms a well derived model. The model has been

deposited to the PMDB server (ID PM0076504). Like sPBP5, the sDacD model is composed of two Domains placed perpendicular to each other. Domain II is β-sheet-rich, whereas Domain I is composed of both α-helices and β-sheets (Fig. 2a). There is a relative increase in beta-sheet in Domain I of sDacD as compared with sPBP5. Comparison of the calculated secondary structure of the sDacD model generated by stride with that of sPBP5 indicates that residues Gln 38-Arg 39 and His158-Ser159-Ser160 of sDacD create a beta-sheet structure, whereas the respective positions of sPBP5 create coils and turns. Moreover, the Glu 230 and Met 233 of sPBP5 Domain I form turns, whereas the corresponding residues (Gln 229 and Arg 232, respectively) in the sDacD model adopt a beta-conformation. Therefore, both similarities and the differences exist when we take a closer view at the active-site of sDacD and sPBP5.

Calmy, M. Cavassini, C. Cellerai, M. Egger, L. Elzi, J. Fehr, J. Fellay, M. Flepp, P. Francioli (President of the SHCS), H. Furrer (Chairman of the Clinical and Laboratory Committee), C. A. Fux, M. Gorgievski, H. Günthard (Chairman of the Scientific Board), D. Haerry (deputy of ‘Positive Council’), B. Hasse, H. H. Hirsch, B. Hirschel, I. Hösli, C. Kahlert, L. Kaiser, O. Keiser, C. Kind, T. Klimkait, H. Kovari, B. Ledergerber, G. Martinetti, B. Martinez de Tejada, K. Metzner, N. Müller, D. Nadal, G. Pantaleo, A. Rauch, S. Regenass, M. Rickenbach (Head of Data Centre), C. Rudin (Chairman of the Mother & Child find more Substudy), P. Schmid, D. Schultze, F. Schöni-Affolter, J. Schüpbach, R. Speck, P. Taffé,

P. Tarr, A. Telenti, A. Trkola, P. Vernazza,

R. Weber and S. Yerly. “
“The objective was to estimate the utilization of psychotropic drugs in HIV-infected individuals compared with that in the background population. Using data obtained from the Danish HIV Cohort Study and the Danish National Prescription Registry, we analysed aggregated data on redeemed prescription of psychotropic drugs during 1995–2009. We primarily focused our analyses on HIV-infected individuals with no history of injecting drug use (IDU) or hepatitis C virus (HCV) infection. Drug utilization was expressed as defined daily doses per 1000 person-days (DDD/1000PD). The utilization rate ratio (URR) was calculated as utilization in the HIV-infected cohort compared with that in the comparison cohort. We estimated longitudinal trends in utilization and potential www.selleckchem.com/products/cobimetinib-gdc-0973-rg7420.html associations with HIV and exposure to highly active antiretroviral therapy (HAART), especially efavirenz. During 1995–2009, 54.5% of the HIV-infected cohort (3615 non-IDU/non-HCV-infected HIV-infected individuals) and 29.2%

of the comparison cohort (32 535 individuals) had at least one prescription of a psychotropic drug. HIV infection was associated with a URR of 1.13 for antipsychotics, 1.76 check details for anxiolytics, 4.42 for hypnotics and sedatives, and 2.28 for antidepressants. Antidepressants were confined primarily to men who have sex with men (MSM). Older age, more recent calendar time, and increased time after HIV diagnosis were associated with increased drug utilization. However, no association with exposure to HAART or efavirenz was found. HIV-infected individuals had a higher utilization of psychotropic drugs than the background population, which was not confined to individuals with a history of IDU or HCV infection. This emphasizes the need to focus on diagnosis of, and appropriate psychopharmacological interventions for, mental disorders in this population. “
“To assess:1) if HIV screening with rapid tests in neighbourhoods with a substantial African community is feasible and acceptable among GPs and patients; 2) HIV seroprevalence. Multicenter prospective study with 10 trained physicians. Use of HIV standard test and INSTI Ultrarapid test.

AMPA receptors comprise GluA1–GluA4 (GluRA–D or GluR1–4) subunits (Keinänen et al., 1990; Hollmann et al., 1991), and exist mainly as GluA1/GluA2 and GluA2/GluA3 heteromeric channels in brains (Wenthold et al., 1996). Inclusion of GluA2 edited at the ‘Q/R site’ from glutamine to arginine determines the Ca2+ permeability of AMPA receptors (Hollmann et al., 1991; Hume et al., 1991; Verdoorn et al., 1991; Mosbacher et al., 1994).

Moreover, AMPA receptor trafficking and synaptic expression of AMPA receptors are controlled according to the ‘subunit-specific rule’. A long cytoplasmic tail of GluA1 or GluA4 binds to anchoring molecules SAP97 and protein 4.1, Cobimetinib manufacturer whereas a short tail of GluA2 or GluA3 interacts with GRIP1/2 and PICK1 (Jiang et al., 2006–2007). Phosphorylation and dephosphorylation of the C-termini alter the state of interaction with the anchoring molecules, which then regulates endocytosis and insertion of AMPA receptors at synapse in activity-dependent and subunit-dependent manners (Hirai, 2001; Shi et al., learn more 2001; Malinow & Malenka, 2002; Song & Huganir, 2002; Lee et al., 2004). Neuronal AMPA receptors also contain auxiliary subunits termed transmembrane AMPA receptor regulatory proteins

(TARPs). The TARP family comprises six isoforms: four classical (γ-2, γ-3, γ-4 and γ-8) and two atypical (γ-5 and γ-7) TARPs (Kato et al., 2008; Soto et al., 2009). In the brain, their overall expressions are distinct but largely complementary both spatially oxyclozanide and temporally: γ-2 in the cerebellum, γ-3 in the cerebral cortex, γ-4 in

developing brain, γ-7 in the cerebellum and γ-8 in the hippocampus (Tomita et al., 2003; Fukaya et al., 2005; Kato et al., 2007). Ideas about the role of TARPs originally arose from the discovery of the virtual lack of AMPA receptor-mediated excitatory postsynaptic currents at mossy fiber–cerebellar granule cell synapses in the spontaneous mutant mouse stargazer or stg (Hashimoto et al., 1999), which carries an early transposon insertion in intron 2 of the γ-2 or Cacng2 gene (Letts et al., 1998). It is now evident that TARPs promote AMPA receptor expression at synaptic and extrasynaptic membranes (Chen et al., 2000; Tomita et al., 2004; Fukaya et al., 2006) and also modulate AMPA receptor gating both in vitro (Yamazaki et al., 2004; Priel et al., 2005; Tomita et al., 2005; Turetsky et al., 2005; Körber et al., 2007; Kott et al., 2007; Soto et al., 2007) and in vivo (Chen et al., 1999; Hashimoto et al., 1999, Rouach et al., 2005). In the present study, we aimed at elucidating the roles of TARPs in the expression and function of cerebellar AMPA receptors. To this end, we generated mice deficient for γ-2 and γ-7 on the C57BL/6 genetic background, because these are two major TARPs expressed in cerebellar granule cells and Purkinje cells (Fukaya et al., 2005).

The majority of clones in sections 1 and 3 of the phylogenetic tree are likely to be specific to the concentrate diet as are those clones in sections 4–7 to the hay diet. The trend toward a closer phylogenetic relationship of clones retrieved from the specific dietary conditions implies the presence of diet-specific phylotypes of Prevotella. However, more direct evidence is needed in order to link the proposed diet-specific Prevotella lineages to their role in the ruminal fermentation of feed. Our DGGE data further showed a consistently higher number of bands in

samples from hay-fed animals. This finding corresponded with diversity analysis from clone libraries that showed higher diversity values (Chao1 and Shannon index) and a greater number of OTUs for clones generated from the hay diet. These results suggest the possible involvement of more Dabrafenib research buy diverse

members of Prevotella in the degradation of a hay diet than that of concentrate. Selleck VX-770 In conclusion, Prevotella is a major member of the rumen bacterial community, and uncultured Prevotella constitute a large proportion of ruminal Prevotella. The diet-specific association of Prevotella clones observed suggests significant functional diversity of members of this genus in the rumen. This study provides evidence for the potential involvement of diverse groups of Prevotella in the degradation of feed in the rumen, particularly hay. “
“A group of bacterial strains formerly known as CDC group M-5 are opportunistic pathogens to humans. In 1993, a name, Neisseria weaveri, was proposed by two independent studies to harbor CDC group M-5 strains, namely N. weaveri Holmes et al. 1993 and N. weaveri Andersen et al. 1993, with two different ‘type’ strains. However, no study has been conducted on to the relatedness of the two ‘type’ strains,

although the close relationship of the two taxa has long been accepted unofficially. 4��8C Formally, the status of the name N. weaveri Andersen et al. 1993 is illegitimate because it is a later homonym of N. weaveri Holmes et al., 1993; but the name of the strain is still validly published. In this study, we attempt to resolve the confusion caused by the apparent duplication of the species N. weaveri (with different type strains) using whole genome shotgun sequencing. We also sought to gain insight into the genetic characteristics of N. weaveri by conducting comparative genomics. On the basis of genomic similarities revealed through a comparative genomic study, we propose that N. weaveri Andersen et al. 1993 should be re-classified as a later heterotypic synonym of N. weaveri Holmes et al., 1993. The genus Neisseria is composed of commensal bacteria that colonize the mucus membranes of mammals. Neisseria encompasses two important pathogens – Neisseria meningitidis and Neisseria gonorrhoeae – as well as many other opportunistic pathogens (Janda & Knapp, 2003; Han et al., 2006).

To address whether the entire Pet signal peptide functions specifically in the biogenesis of Pet, chimeric constructs were generated with signal peptides representative of the Sec (pMBPssPet and pPhoAssPet) and SRP (pDsbAssPet) targeting pathways (Fig. Trichostatin A nmr 1). pMBPssPet, pPhoAssPet and pDsbAssPet represent the MBP, PhoA and DsbA signal peptides, respectively, fused to Pet lacking its signal peptide (Met55–Phe1295). pPetssPet, a derivative comprising Pet with its signal peptide (Met1–Phe1295), served as a control (Fig. 1). SignalP (Nielsen et al., 1997) analysis predicted that the signal peptide cleavage site of all chimeric ss-pet

constructs was maintained. The plasmids used in the generation of these chimeras contained promoter down mutations (Fig. 3a); pMBPssPet and pPhoAssPet were generated using a plasmid backbone containing a double RXDX-106 cell line point mutation within the −10 promoter region (TATAAT to CATTAT), and a single point mutation within the −35 region (TTGACA to TTTACA). The plasmid backbone used to generate pPetssPet and pDsbAssPet contained only a down mutated −35 promoter region. The ability of cells containing these constructs to express Pet was reliant on an isopropylthiogalactoside-inducible ptrc promoter and monitored by Western blot analysis of supernatant

fractions using anti-Pet passenger domain antibodies. We acknowledge that the use of ptrc promoters with different transcription efficiencies may affect the expression levels of Pet. Although an MBP signal peptide fusion to EspP did not impair the translocation of the passenger domain across the inner membrane, alteration of the native EspP signal peptide caused a significant defect in protein biogenesis (Szabady et al., 2005). Similarly and as hypothesized, in this study, we showed a significant decrease in secretion of the pPhoAssPet and pMBPssPet chimeras (Fig. 3b). In contrast, we found that the pDsbAssPet chimera was released into the culture supernatant almost at wild-type levels (Fig. 3b). Also of note was the finding that the growth

of all cells containing chimeric constructs was comparable to the wild type (data not shown). Overall, although the level of secretion of the Pet chimeras comprising non-native signal peptides was affected, the retained ability Fludarabine to secrete protein indicates that the Pet signal peptide is not specifically required for secretion. As a marker of correct folding of the passenger domain, we determined whether the ESPR Pet deletion mutant and the chimeric Pet proteins displayed cytotpathic activity by performing cytotoxicity assays using HEp-2 epithelial cells. Concentrated supernatants were applied directly to semi-confluent HEp-2 cell monolayers. The results showed that the morphology of the HEp-2 cells was unaltered by treatment with concentrated supernatant from the E.

To address whether the entire Pet signal peptide functions specifically in the biogenesis of Pet, chimeric constructs were generated with signal peptides representative of the Sec (pMBPssPet and pPhoAssPet) and SRP (pDsbAssPet) targeting pathways (Fig. Tipifarnib 1). pMBPssPet, pPhoAssPet and pDsbAssPet represent the MBP, PhoA and DsbA signal peptides, respectively, fused to Pet lacking its signal peptide (Met55–Phe1295). pPetssPet, a derivative comprising Pet with its signal peptide (Met1–Phe1295), served as a control (Fig. 1). SignalP (Nielsen et al., 1997) analysis predicted that the signal peptide cleavage site of all chimeric ss-pet

constructs was maintained. The plasmids used in the generation of these chimeras contained promoter down mutations (Fig. 3a); pMBPssPet and pPhoAssPet were generated using a plasmid backbone containing a double selleck chemicals llc point mutation within the −10 promoter region (TATAAT to CATTAT), and a single point mutation within the −35 region (TTGACA to TTTACA). The plasmid backbone used to generate pPetssPet and pDsbAssPet contained only a down mutated −35 promoter region. The ability of cells containing these constructs to express Pet was reliant on an isopropylthiogalactoside-inducible ptrc promoter and monitored by Western blot analysis of supernatant

fractions using anti-Pet passenger domain antibodies. We acknowledge that the use of ptrc promoters with different transcription efficiencies may affect the expression levels of Pet. Although an MBP signal peptide fusion to EspP did not impair the translocation of the passenger domain across the inner membrane, alteration of the native EspP signal peptide caused a significant defect in protein biogenesis (Szabady et al., 2005). Similarly and as hypothesized, in this study, we showed a significant decrease in secretion of the pPhoAssPet and pMBPssPet chimeras (Fig. 3b). In contrast, we found that the pDsbAssPet chimera was released into the culture supernatant almost at wild-type levels (Fig. 3b). Also of note was the finding that the growth

of all cells containing chimeric constructs was comparable to the wild type (data not shown). Overall, although the level of secretion of the Pet chimeras comprising non-native signal peptides was affected, the retained ability Bcl-w to secrete protein indicates that the Pet signal peptide is not specifically required for secretion. As a marker of correct folding of the passenger domain, we determined whether the ESPR Pet deletion mutant and the chimeric Pet proteins displayed cytotpathic activity by performing cytotoxicity assays using HEp-2 epithelial cells. Concentrated supernatants were applied directly to semi-confluent HEp-2 cell monolayers. The results showed that the morphology of the HEp-2 cells was unaltered by treatment with concentrated supernatant from the E.

A project group with representatives from the five organisations was set up to design a chart with medication safety features. The chart was piloted across the five organisations. The evaluation involved 1) an assessment of the impact on the quality of documentation of the patient’s allergy status and the patient’s venothromboembolus risk assessment; and 2) a user survey

on the chart design and its effect on medication safety. Designated leads at each site prospectively collected documentation data before and after implementation using a proforma. A questionnaire survey (which was administered in person for return via a marked collection point on the ward) was used to gain user views 2 months after implementation. Users were asked to indicate their views on 25 statements relating screening assay to the chart layout, format and booklet design, specialist sections for high risk drugs, and perceived effects of the selleck products changes on safety using

a Likert-like scale. All data was entered onto structured excel data sheets and sent to the lead author for collation and analysis. Statistical significance between documentation rates was assessed using Chi squared (χ2) tests. Ethics approval was not required. A new chart was designed and approved by the relevant Medicines Committees in all five organisations. The safety features included a cut-out section to ensure visibility of the patient demographics and allergy status information; specific sections for prescribing VTE thromboprophylaxis, anti-coagulation and oxygen; dedicated section for medication reconciliation; increased space to reduce the number of concurrent charts required per patient;

use of colour to highlight high risk and specialist areas. The pilot involved 14 wards, 568 patients (255 before; 313 after) and 772 prescription charts (465 before; 307 after). Documentation of essential information improved marginally with the new chart for most parameters (patient name, date of birth and hospital number) except weight where a reduction was seen (from 69/465; 14.8% to 15/307; 4.9%, p < 0.01 χ2 test). Overall allergy status documentation was similar for both charts (95.1% before vs. 95.4% after), but Clomifene for patients with known allergies there was an increase in documentation of the nature of the reaction from 40% to 61.3% (p = 0.02 χ2 test) and allergy severity from 13.1% to 19.4% (not significant). Proportion of patients with a documented VTE risk assessment outcome increased from 17.3% to 24.3% (p = 0.04 χ2 test). Fewer patients required multiple charts following introduction of the new design (30/255; 11.8% compared to 96/313; 30.9%). The survey included responses from 107 users (66 nurses, 23 doctors, 6 pharmacists, 1 pharmacy technician, 4 others and 13 had not stated their profession).

HRQL assessment has become one of the most widely used subjective health evaluations in chronic illness. Life experiences of HIV-infected people are as heterogeneous as the population affected. HRQL assessment in these patients provides valuable information about the effects of ART, disease progression and prognosis, and the factors that influence prognosis; results that clinical analysis is unable to provide. It must be taken into account that the evaluation of HRQL by the patient does RG-7388 not necessarily coincide with the severity of the illness as defined by the patient’s doctor. HRQL provides valuable information for health care managers

and authorities, as it allows evaluation of the efficiency, effectiveness and cost–benefit ratio of health care programmes, and for pharmaceutical companies that gather data on effectiveness, clinical benefit, satisfaction with treatment and treatment adherence [9–11]. The literature shows the importance of factors most closely related to HRQL in HIV-infected people. These factors are psychological aspects and sociodemographic characteristics, clinical indicators unrelated to the infection and the individual illness [6,12–15]. HRQL in the HIV-infected population has not previously been investigated

in our region, and so the aim of this study was to determine the impact of various sociodemographic, clinical and psychological factors on HRQL in an HIV-infected population receiving care at the HIV clinic of a tertiary Spanish selleck hospital, Aurora Kinase and to identify variables that allow us to establish a predictive model to evaluate HRQL in this population and these patients’ overall perception of their health status. A cross-sectional study

was conducted in HIV-infected patients under follow-up at the Río Hortega University Hospital in Valladolid (Spain). The target population comprised individuals with HIV infection who agreed to participate in the study in the period March 2007 to April 2008. Exclusion criteria were: (a) recent diagnosis with HIV infection (less than 6 months ago); (b) age <16 years; (c) the patient not being frequently seen by our specialists; (d) refusal to participate in the study; (e) a physical or mental condition that made interviewing the patient problematic. Nine persons refused to participate in the study (six men and three women) and did not sign the medical consent form; these patients were not a homogeneous group in terms of sociodemographic, epidemiological or clinical characteristics. Following consultation with the Investigation Department, a total of 150 out-patients were consecutively selected after they had signed the medical consent form according to the principles of the Declaration of Helsinki.

Carbapenemase-producing Enterobacteria (CPE) is nowadays a major public health concern worldwide.[1] The link between international spread of antibiotic resistance and travels is well known.[2-4] Carriage

of CPE has been identified in Great Britain in travelers having been hospitalized in Pakistan or India.[5] In France, resistance of Enterobacteria to carbapenems remains uncommon but involves most often patients with a history of hospitalization abroad.[6] The first outbreak of CPE in France, that occurred in 2004 in a hospital of Assistance Publique-Hôpitaux Selleckchem Tanespimycin de Paris (AP-HP), followed the transfer of a patient from a Greek hospital.[7] Following this outbreak, AP-HP launched a long-term program to survey and

control CPE, particularly in patients previously hospitalized in foreign countries. We describe here the emergence of CPE in AP-HP hospitals from 2004 to December 2011 and the link with cross-border exchanges. AP-HP is a public health institution administering 38 teaching hospitals (23 acute care and 15 rehabilitation/long-term care hospitals, spread over Paris, suburbs, and surrounding counties), with a total of 23,000 beds (10% of all public hospital beds in France) and serving 11.6 million inhabitants. AP-HP admits approximately www.selleckchem.com/products/nu7441.html 1 million inpatients per year, employs 19,000 physicians, 18,500 nurses, and 29,800 assistant nurses. Local administrators and medical committees manage AP-HP hospitals, but decisions on large investments and medical developments are taken by the central administration. A local infection control team (LICT) is in charge of prevention and surveillance of hospital-acquired infections in each hospital, but actions of foremost importance for the whole institution, eg, multidrug-resistance (MDR) control program, are coordinated centrally by a multidisciplinary infection control team (head of the infection control team

[CICT], infectious disease physician, bacteriologist, epidemiologist, and nurse).[8] One mafosfamide case was defined as any infected or colonized patient with CPE species. An event was defined as one index case with or without secondary cases. An outbreak was defined as at least two CPE cases (ie, one index case and at least one secondary case) occurring in a given hospital, with a clear epidemiological link (stay during the same period of time in the same unit). In 2004, following the first CPE outbreak in a hospital of AP-HP, every LICT was asked to report quickly every new CPE case to the AP-HP CICT. In 2008, based on the analysis of the CPE events identified during the first 3 years of the survey, LICTs were advised to screen for CPE every patient transferred from a foreign hospital.

At inclusion, a clinical examination was performed and a medical history taken. The duration of HIV infection was defined as the time from the first positive HIV test to inclusion in the study. Blood samples were obtained after an 8-h overnight fast for routine measurement of glucose, total cholesterol, triglycerides, and haematological parameters. In HIV-positive patients, CD4 cell counts (measured using flow cytometry) and HIV-RNA levels [measured using Roche Cobas HIV-1 Monitor and Roche Cobas TaqMan HIV-1, v1.0 (Roche Diagnostics AG, Rotkreuz, Switzerland), with a limit of detection of 50 HIV-1 RNA copies/ml] were determined. Additional samples were processed

by centrifugation, and plasma was learn more stored at –80°C for later analyses. At the end of the study, samples were thawed, and plasma levels of biomarkers were processed simultaneously. For endothelial activation, soluble intercellular adhesion

molecule-1 (sICAM-1), the von Willebrand factor (vWF), and E-selectin were measured [a sandwich enzyme-linked immunosorbent assay (ELISA) technique was used for all three tests; R&D Systems Europe Ltd., Abingdon, UK]. The concentrations of highly sensitive C-reactive protein (hs-CRP; Dade Behring Holdings Inc., Deerfield, Illinois, USA) and p-fibrinogen [chronometric measurement of clot formation (cmcf); STA-R Evolution; Triolab AS, Brøndby, Denmark] were Selleckchem Lumacaftor determined as inflammatory biomarkers, and activation of the coagulation system was assessed using D-dimers (immunoturbidimetric

technique; Tideglusib STA-R Evolution; Triolab), activated partial thromboplastin time (APTT) (cmcf; Triolab) and prothrombin time (PT) (cmcf; Triolab). Endothelial function was assessed noninvasively in the right brachial artery using external ultrasound scanning, as previously described [12, 13]. The artery was scanned longitudinally immediately below the antecubital fossa with a 10-MHz vascular transducer (Acuson Sequoia, Mountain View, CA) after a minimum of 15 min rest in a supine position. The vasodilatory response to reactive hyperaemia [an endothelium-dependent stimulus leading to flow-mediated dilatation (FMD)] was compared with vasodilatation in response to nitroglycerine (NTG; an endothelium-independent stimulus). Vessel diameter was measured four times: (1) at baseline before transient upper arm cuff occlusion (300 mmHg for 4 mins), (2) 45 to 60 s after cuff deflation (reactive hyperaemia), (3) 10 min after cuff deflation (second baseline scan), and finally (4) 3 min after sublingual administration of 400 μg of NTG. Images were recorded on videotape, and a minimum of four cardiac cycles from each scan sequence were analysed by two observers blinded to patient group and the sequence of the scan protocol. FMD and NTG-induced dilation were derived relative to the baseline scan (100%). The mean values obtained by the two observers were used for analysis.