6% CI95% [27 6–29 4%] vs 27 7% CI95% [26 5–28 9%] (p = 0 047) fo

6% CI95% [27.6–29.4%] vs. 27.7% CI95% [26.5–28.9%] (p = 0.047) for anti-HBc; 6.4% CI95% [5.6–7.2%] vs. 4.5% CI95% [3.9–5.1%] (p < 10−3) for HBsAg and 3.6% CI95% [3.4–3.7%] vs. 2.4% CI95%

[2.0–2.8%] (p = 0.001) for chronic carriers. Prevalence of anti-HBc and HBsAg increases significantly with age globally for both males and females (p < 10−3). The distribution of HBV markers per governorates and districts is illustrated in Table 1. After standardisation per age significant differences were observed between the two governorates according to anti-HBc prevalence (32.1% CI95% [28.9–32.7%] in Béja and 27.8% CI95% [26.8–28.8%] in Tataouine; p = 0.005) and HBsAg prevalence (4.2% CI95% [3.2–4.8%] in Béja in the north and

selleck compound 5.6% CI95% [5.2–6.2%] in Tataouine in the south; p = 0.001). No significant differences were noted according to chronic carriage prevalence between the two governorates (2.6% CI95% [1.9–3.1%] in Béja vs. 2.8% CI95% [2.6–3.4%] in Tataouine). When the analysis was refined at the subgovernorate level, significant differences were noted between districts according to these three markers (all p values <10−3). Ras el oued and Dhiba (in the south) showed a higher prevalence for all HBV markers than the other districts. If HBV chronic carriage prevalence NU7441 concentration (7.7 and 12.0%, respectively) is considered, these two districts are classified as areas of high endemicity. Khniguet eddhene (in the north) and Rmada est (in the south) show an HBV chronic carriage prevalence of 4.9 and 2.0%, respectively, and can then be classified as areas of intermediate endemicity. All other districts have HBV chronic carriage prevalence less than 2% and are thus classified as areas of low endemicity. Interestingly, the relative proportion of carriers among HBsAg positive subjects differ

significantly ADAMTS5 (p < 10−3) between districts, and ranges from 30 to 90% ( Fig. 1). Not surprisingly, the age-distribution of HBsAg, anti-HBc, and chronic carriage prevalence increased as endemicity decreased. The median age of all HBV infection markers was lower in hyperendemic areas as compared to intermediate and hypo-endemic ones. The median age for anti-HBc positive subjects was 24.3 years, 30.8 years, and 40.0 years (p < 10−3); for HBsAg positive subjects, was 16.9 years, 23.0 years, and 29.9 years (p < 10−3); and for chronic carriers, was 14.7 years, 24.7 years and 29.8 years (p < 10−3) for hyperendemic regions, intermediate endemic regions, and low endemic regions (p < 10−3), respectively. Similarly, the age at which half the population have been infected decreased significantly from low (60 years) to intermediate (40 years) and high endemic regions (10 years) ( Fig. 2a). The age distribution of anti-HBc and chronic carriage showed different patterns according to endemicity ( Fig. 2b). In a hyperendemic area, chronic carriage increased quickly and saturated after the age of 20 years.

These conditions certainly contributed to the rapid loss of the c

These conditions certainly contributed to the rapid loss of the contaminating viruses. Only viruses that are present at very high titers and which grow very rapidly without adaptation would be able to survive such passaging. In a second series of

passages we also monitored more than 50 specimens that did not contain an influenza virus but were positive for other respiratory viruses. In these specimens interference by competing influenza virus growth was excluded. The culture conditions differed, as lower inoculum dilutions were used. Each sample/harvest was diluted 1:100 into the culture, which is the lowest standard dilution applied to recover very low-titred influenza virus. Also under these conditions 54 positive results for 8 different viruses became MEK activation negative after only 2 or 3 passages and this website after a total dilution of the original specimen by a factor of 2 × 10−4 to 2 × 10−5. When similar passages were conducted with adherent Vero cells (“Vero WHO seed”), several positive samples (adenovirus, rhinovirus, enterovirus, metapneumovirus, and bocavirus) remained positive after 2 passages. However, except for adenovirus, the counts did not increase but dropped

(data not shown). These results demonstrate that, under practical conditions as applied to grow influenza viruses, contaminating viruses can be effectively removed by passaging in MDCK 33016PF cells. In combination with their superior isolation efficiency [7] and [28], MDCK cells appear highly suitable to be used as an alternative to embryonated eggs to isolate and propagate candidate vaccine viruses.

The authors would like to thank Knut Schwarz, Marion Wellnitz, MYO10 Veronika Horn and Inge Lettermann for their skillful technical assistance with these studies. We gratefully acknowledge confirmatory PCR test results by independent methods that were partly provided by Marcus Panning, of the Virology Department of the University Clinic in Freiburg, Germany. “
“Dendritic cells (DCs) are key components of the immune system which function by binding and collecting antigens. Following recognition, DCs present the antigen of interest through selective surface markers to T-cells in order to activate a specified immune response [1]. Antigen presentation also stimulates the differentiation of T-cells to B cells which release antibodies specific for the antigen of interest. It is these functions that researchers aim to exploit in the production of vaccines. Non-viral gene delivery to DCs is an attractive approach for DNA vaccination to elicit immune responses towards encoded antigenic sequences [2]. Non-viral techniques often entail delivery of nucleic acids that are bound to a cationic polymer (polycations) resulting in plasmid DNA (pDNA) – polymer products, known as polyplexes [3]. Polycations operate by binding and condensing pDNA into smaller structures thereby facilitating uptake.

Passive surveillance systems are able to identify safety signals,

Passive surveillance systems are able to identify safety signals, but are subject to known limitations, due to underreporting, delayed reporting and a lack of denominator data. Active surveillance in a defined STI571 supplier cohort of vaccines can complement passive surveillance by overcoming problems of delayed and underreporting and enabling calculation of adverse event rates. Recent studies internationally have emphasised the importance

of active surveillance to detect important signals early so that appropriate investigations can be launched and necessary actions taken [8] and [9]. Internationally the usefulness of Patient Reported Outcomes (PROs) utilising available internet tools has been increasingly recognised. There is evidence that in relation to adverse events PROs can identify real-world signals earlier and in higher volume, accurately characterise the signals, allow a focus on specific events

or populations of interest, and permit ongoing efficient safety monitoring [10]. The finding that there was a significantly higher rate of reactions in participants who received IIV in the previous year deserves further investigation as it has not been a consistent finding in previous studies [3]. The initial practice visit by Vaxtracker staff of this pilot phase could be replaced by a brief diagrammatic user guide or online web Protein Tyrosine Kinase inhibitor demonstration to further improve efficiency and reduce the cost of the roll out phase. We estimate that once established the ongoing human resources to operate the system are not great as survey results provide sufficient information for assessment and very few respondents require subsequent telephone clarification of clinical details or support. After the Vaxtracker survey was completed by respondents, case review and data analysis for signal detection quickly take place. The automatic management of survey dispatch and return of completed surveys and email alerts has allowed for the efficient and

prompt review of AEFIs and rapid data analysis and rate calculation. It is essential to reassure the community of vaccine safety and to prompt second early investigation should severe reactions occur or if there is an unexpected increase in the frequency of clinical events [11]. The Vaxtracker active surveillance system achieved encouraging completion rates. These were found to be higher where parents received both mobile phone and email reminders. Feedback and a certificate of appreciation were provided to all General Practice clinics that enrolled participants. Respondents who reported serious AEFI were contacted by telephone to discuss their report, ensure that appropriate clinical management had occurred if required and enquire whether symptoms had resolved. There was no formal feedback to respondents in this pilot but plans are underway to make Vaxtracker safety data available to the public on a website as the programme is expanded.

aureus, Ps acruginosa, P vulgaris, A niger and C albicans as

aureus, Ps. acruginosa, P. vulgaris, A. niger and C. albicans as compare to simple pyrrole. The compounds 2-substituted, Dabrafenib molecular weight 1,2,4-triazole (4a–g), 4-oxadiazole (5a–g) and 4-oxazolidinones (6a–g) have shown good antioxidant activity within the series of compounds synthesized. All authors have none to declare. We are thankful to UGC for providing the financial assistance to carry out the research work (F 12-17, 2004, SR) and also we thank JPR Solutions, Mohali for their partial funding in publishing this research. “
“Quinazolinone derivatives are well-known for their diverse pharmacological (analgesic, anti-allergic, anticonvulsant, anti-depressant, anti-inflammatory, antimalarial, antimicrobial, hypotensive, sedative-hypnotic,

etc) activities. 1 For example, the widely known quinazolinone drug, methaqualone (1) was first synthesized in India in 1951 and was used world-wide as a sedative-hypnotic agent. 2 Also, structural activity relationship studies on 3-phenylsulfonyl-quinazoline-2,4-dione derivatives reveal that the 1-pyridylmethyl and 1-(N-pyridylacetamide) derivatives showed inhibitory concentration (IC50) in the order of 10−8 M as human heart chymase inhibitors. 3 Molecular modeling studies on Pexidartinib the

interaction of one of the derivatives, 7-chloro-3-(4-chlorophenylsulfonyl) quinazoline-2,4(1H, 3H)-dione (2), with the active site of human heart chymase shows good fitting and interaction. 3 The main synthetic pathways to quinazolinone compounds include the condensation of anthranilamide (2-aminobenzamide), (3) with structurally diverse acid

anhydrides, aldehydes or ketones in the presence of various mafosfamide catalysts. 4 and 5 Cycloaddition of anthranilic acid derivatives with amines, imines, iminohalides have also been reported. 6 and 7 There have been reports of microwave-assisted synthesis of quinazolinones from anthranilic acid derivatives and from isatoic anhydride. 8, 9 and 10 Figure options Download full-size image Download as PowerPoint slide The reaction of anthranilamide (3) with phthalic acid anhydride under conventional heating has been reported to give isoindolo[1,2-b]quinazoline-10,12-dione (4).11 This reaction has not been examined under microwave irradiation. In view of our interest in the study of organic reactions under microwave irradiation and construction of nitrogen heterocyclic compounds under such conditions, with simultaneous evaluation of some biological activities of obtained products,12 and 13 we herein report the convenient microwave-assisted access to some quinazolinones, from the reaction of anthranilamide with phthalic anhydride and some other compounds, and their antimicrobial activity. Melting points were determined in open capillary tubes on a Gallenkamp (variable heater) melting point apparatus and are uncorrected. Infrared spectra were recorded (in KBr or Nujol) on a Buck Scientific Spectrometer. Microwave experiments were performed in a domestic oven (24 L oven).

, 1973) It is clear that if ethanol

is taken together wi

, 1973). It is clear that if ethanol

is taken together with food it is diluted and the ethanol absorption is delayed. Human in vivo studies of drug ethanol sensitivity see more would require a combination of high drug doses with ethanol intake and are not ethically feasible. In this study we therefore employed in vitro solubility measurements and in silico absorption simulations to identify compounds potentially sensitive to concomitant ethanol intake. Nine model compounds were included in this study on the basis of their lipophilicity, aqueous solubility (with focus on poorly soluble compounds), and results from a previous study of ethanol sensitivity in FaSSIF (Fig. 2) (Fagerberg et al., 2012). The data set included three acidic compounds (indomethacin, indoprofen and tolfenamic acid), SKI-606 clinical trial three non-ionizable compounds (felodipine, griseofulvin and progesterone), and three weak bases (cinnarizine, dipyridamole and terfenadine); these compounds were selected to cover both charged and non-ionizable compounds with a diversity in physicochemical properties (Table 1). Only compounds available in their free form were included to exclude effects from salt formation. ADMET Predictor (Simulations Plus, CA) was used to calculate lipophilicity

expressed as log P and log DpH2.5, and the total effective permeability (Peff) for the nine compounds. Diffusivity in water was calculated according to the Stoke–Einstein’s equation on the basis of the molecular volume estimated using ACD/Chemsketch 12.0 (Advanced Chemical Development

Inc, Canada). Pharmacokinetic parameters were gathered from the literature. All input data Idoxuridine used in the computational simulations are summarized in Table 2. The composition of FaSSGF was a modification of the gastric medium described by Vertzoni et al. (2005). No pepsin was included and the pH was increased from the suggested 1.6 to 2.5. The latter was done to reflect recent findings regarding the pH of human gastric-fluid aspirates (Kalantzi et al., 2006 and Pedersen et al., 2013) and to avoid unnecessary wear on the stainless-steel fiber-optic dip probes used for concentration determination. A NaCl solution with pH 2.5 (NaClpH2.5) was prepared by dissolving 2 g NaCl in 0.9 L MilliQ water, after which the pH was adjusted to 2.5 by the addition of HCl before adjusting the final volume to 1 L. The resulting NaClpH2.5 was sterile-filtered and stored at 8 °C. NaClpH2.5 with 20% ethanol (NaClpH2.520%Ethanol) was prepared in the same fashion except that 2.5 g NaCl was used and 20% (v/v) ethanol was added to the 1 L volume (final volume 1.2 L). The corresponding biorelevant dissolution media (BDM), i.e. FaSSGF and FaSSGF20%Ethanol, were prepared by dissolving 6 mg SIF powder in 100 mL of each NaCl solutions. Apparent solubility was determined in the four different media using a three-channel μDiss Profiler Plus (pION, MA) described previously (Fagerberg et al.

Clinical outcomes revealed that the majority of

these cas

Clinical outcomes revealed that the majority of

these cases were unrecognized multifetal pregnancies, ongoing or vanishing twins, with a small number of triploid pregnancies also detected. The ability to detect vanishing twin pregnancies is clinically important as it will reduce the number of false-positive results and thereby reduce unnecessary invasive diagnostic procedures. Future longitudinal studies, designed to evaluate the typical SRT1720 time period for which residual fetal cfDNA from vanishing twins remains detectable, may provide greater insight into appropriate clinical care in these patients. “
“LOX-1 is a lectin-like oxidized LDL receptor (also known as oxidized LDL receptor 1—OLR1), which was initially described in endothelial cells by Sawamura et al. [1]. LOX-1 expression has subsequently been described in both smooth muscle cells and macrophage in atherosclerotic plaques [2] as well as Adriamycin in other cell types including adipocytes [3], platelets [4], and chondrocytes [5]. LOX-1 expression can be induced or up-regulated by a number of processes many of which are involved in the atherosclerotic process, including hypertension, sepsis, inflammatory mediators, dyslipidemia, advanced

glycation end products, and fluid shear stress (reviewed in Ref. [6]). LOX-1 performs a number of functions in addition to oxidized LDL (oxLDL) binding, such as binding of apoptotic cell bodies and aged red blood cells [7] and acting as a leukocyte adhesion molecule [8]. Binding of oxLDL to LOX-1 induces endothelial dysfunction and apoptosis, stimulating reactive oxygen species (ROS) production and NFκB activation [9], strongly linking LOX-1 with the process of atherosclerosis through [6] and [10]. Several studies in hyperlipidemic mice have demonstrated a link between LOX-1 and atherosclerosis. Mehta et al. [11] created a LOX-1−/−/LDLR−/− mouse, which on high-fat diet exhibited reduced plaque development in the aorta compared to controls. In addition, the LOX-1−/−/LDLR−/− mice also

demonstrated a number of anti-atherosclerotic features, e.g., increased IL-10 levels and eNOS activity, with a concomitant reduction in MAPK p38 and NFκB activation. Inoue et al. [12] created a bovine LOX-1 transgenic mouse, where LOX-1 was overexpressed in multiple cell types including vascular and cardiac tissue. Among the pathologies displayed in this transgenic mouse was an increase in ox-LDL uptake and atheroma-like lesions in coronary arteries. In addition, Ishigaki et al. [13] used an adenoviral vector to overexpress LOX-1 in the liver, enhancing hepatic uptake of ox-LDL and reducing atheroma in the aorta. Taken together, these experiments clearly demonstrated a role for LOX-1 in atherosclerosis, although the contribution of endothelial vs. smooth muscle cell or macrophage expression has yet to be determined.

The dried extract was dissolved in respective solvents prior to a

The dried extract was dissolved in respective solvents prior to assay. The total phenolic content (mg of catechin/1 mg) was determined

using Folin–Ciocalteu reagent5 and total flavonoid content (catechol equivalents/1 mg) was determined by aluminium chloride method.6 The reductive ability of the extracts was determined by potassium ferricyanide reduction method.7 The hydrogen or electron donation ability of the plant extracts was measured from bleaching of the purple colour of DPPH.8 Scavenging activity of extracts on superoxide anion radicals was determined based on the reduction of nitroblue tetrazolium (NBT).9 Hydroxyl radical scavenging and the ferrous ion-chelating potential of the extracts were measured following deoxyribose assay10 and ferrozine assay11 respectively. Thiobarbituric acid reactive substance assay 3-Methyladenine supplier was employed FRAX597 clinical trial to determine anti-lipid peroxidation assay using goat liver homogenate.12 All analyses were carried

out in triplicates. Data were presented as mean ± SD. Radical scavenging activity of extracts was expressed in terms of percentage of inhibition. DPPH, superoxide radical scavenging, hydroxyl radical scavenging and metal ion-chelating assay were calculated using the following equation: % Inhibition = (Absorbance of control − Absorbance of sample)/Absorbance of control × 100, and the anti-lipid peroxidation percentage was calculated using the formula: % ALP = (Absorbance of Fe2+ induced peroxidation-Absorbance of sample)/Absorbance of Fe2+ induced peroxidation-Absorbance of control × 100. The IC50 value was determined using Easy Plot software. The total phenolic contents of aqueous and methanolic extracts of A. solanacea leaves were 0.030 ± 0.01 and 0.040 ± 0.02 mg of catechin equivalents/1 mg dried extract respectively and the corresponding flavonoid contents were 0.257 ± 0.02 and 0.404 ± 0.03 mg of catechol equivalents/1 mg dried aqueous and methanolic extracts. Both the extracts showed powerful reducing power that increased linearly with concentration. The methanolic extract demonstrated powerful reduction

potential as compared to aqueous extract (Fig. 1). The IC50 values of methanolic and aqueous extracts for DPPH radical scavenging activity were 198.43 ± 1.30 over and 378.67 ± 2.5 μg/ml (Fig. 2) respectively which showed a marked difference with ascorbic acid standard (IC50 = 7.6 ± 0.20 μg/ml). The methanolic extract exhibited superoxide radical scavenging activity (Fig. 3) with an IC50 value of 1634. 97 ± 4.08 μg/ml and showed a significant difference when compared with butylated hydroxy anisole (IC50 value of 23.6 ± 0.86 μg/ml). The percentage inhibition of hydroxyl radical scavenging activity of the aqueous and methanolic extracts was found to be 62.81% and 92.89% respectively at 2000 μg/ml. Compared to all the other assays, at the lowest concentration (25 μg/ml) tested, the methanolic extract of A. solanacea was the one that showed higher (86.71%) free radical scavenging ability.

2% homology with canine VEGF) mixed with a liposome–DNA complex

2% homology with canine VEGF) mixed with a liposome–DNA complex. Immunization produced a 30% anti-tumor response rate, but without an increase in anti-canine VEGF antibody titers. No important side effects regarding blood biochemistry or impairment in wound healing were reported. We have now tested the effects of CIGB-247 vaccination in rats, rabbits and non-human primates to determine whether: (a) immunization produced an anti-VEGF IgG response, (b) immunity is tightly regulated and B-cell memory could be induced, and (c) vaccination produced detectable clinical, biochemical and histological side effects, NVP-BGJ398 including the ability

to recover from skin deep wounds. Our results showed that CIGB-247 was able to Epigenetic high throughput screening induce an IgG immune response specific for VEGF in the three studied species with discrete IgG antibody titers, similarly to our mouse experiments [11]. The latter could be explained by the close homology of the antigen and the self-growth factor (88.7% for rats, 94% for rabbits, and 99% for monkeys), the nature of the adjuvant, or a combination of these and other factors. In rats, as in mice, the IgG response against mouse VEGF (99% homology to

the rat molecule) suggests a breakage of B cell tolerance to the self-growth factor. The addition of montanide to CIGB-247 led to the highest titers in rats and rabbits. Sera from both species impaired the binding of KDR-Fc to human VEGF. Weekly vaccination schemes were better (rats) or similar (rabbits) inhibiting

the binding in the test, a clear indication that higher titers do not necessarily correlate with the biological effect of vaccination. Our experiments in non-human primates showed that vaccination breaks B-cell tolerance to the self-growth factor and elicits a specific and dose dependent anti-VEGF IgG response. The weekly scheme in monkeys showed a trend to higher titer values and an increased ability of the sera to block the interaction of soluble KDR-Fc with human VEGF. Purification STK38 of the IgG from monkey serum increased the resulting specific blocking activity, indicating that antibodies are responsible of the observed effect. The antibody titer kinetics in monkeys was demonstrative of a well-regulated humoral response. In the weekly scheme, the significant increase in antibody titers after the boosters is a clear evidence of B-cell memory, and provides an early indication that maintenance vaccinations after an induction phase should be foreseen for the clinical testing of CIGB-247, as has been shown by others [31]. Specific cytolysis of autologous “VEGF-charged” PBMC cells was shown in non-human primates, with the highest values for two animals belonging to the weekly vaccination group. The individual variation found – including negative individuals – could be indicative of the differences that are probably to be found in open populations submitted to this type of vaccination, or may reflect technical limitations of the used assay.

Sera from individual fish were analyzed for IPNV neutralizing ant

Sera from individual fish were analyzed for IPNV neutralizing antibody PR-171 research buy titers (NAb) using a neutralization assay as previously described [17]. This assay involved incubation of 2-fold dilutions of sera with a known amount of the reference IPNV serotype Sp, and titers were reported as the reciprocal of the highest serum dilution that resulted in a 50% reduction in the viral infectivity (TCID50 ml−1) compared with negative controls. Thirty days after vaccination with 50 μl of PBS alone or containing 1 μg of the pIPNV-PP vaccine or its respective empty plasmid, trout specimens were infected with IPNV Sp (intraperitoneal injection

of 100 μl of 1 × 107 TCID50 ml−1 per fish). At 7 days check details post-infection, 5 trout from each group were sacrificed and head kidney stored in TRIzol Reagent in order to evaluate the effect of the vaccine on virus clearance or load [23]. RNA from individual samples was isolated and 1 μg of RNA retrotranscribed to cDNA as above. Detection of IPNV VP1 gene expression was also evaluated by real time PCR, using published primers [25]. Samples were incubated for 10 min at 95 °C, followed by 50 amplification cycles (30 s at 95 °C and 1 min at 56 °C) and a dissociation cycle (30 s at 95 °C, 1 min 55 °C and 30 s at 95 °C). VP1 gene expression was normalized

and expressed as indicated before. Data are expressed as mean ± SE. Analysis of variance (ANOVA) or Student-t tests were performed to determine differences between the vaccine and control groups. Significant differences were established when P < 0.05. First, after the construction of the pIPNV-PP vaccine plasmid, we verified the correct translation of the IPNV found polyprotein in a cell-free based expression

system (Fig. 1A). A band corresponding to the polyprotein (about 106 kDa) size was not seen. However, other 4 clear bands appeared after plasmid translation, which corresponded to the expected size of unprocessed VP2 (pVP2), cleaved and mature VP2 products as well as the VP3. VP4 protein was not detected. These data confirm that the vaccine is translated to a functional VP2–VP4–VP3 polyprotein and VP4-proteolytic products are detected, as previously described for IPNV [26] and the Japanese marine Aquabirnavirus closely related to IPNV [27]. Transfection of EPC cell line with the pIPNV-PP plasmid resulted in the correct transcription of the vaccine. First, we found that the EPC-transfected cultures expressed the vaccine after 72 h as evidenced by the detection of VP2 transcripts through semi-quantitative PCR (Fig. 1B). Moreover, as a consequence of IPNV polyprotein synthesis, EPC cells showed a significant up-regulation of Mx gene expression when compared to EPC cultures transfected with the empty plasmid (Fig. 1B).

Previous history of back pain has been highlighted

as a p

Previous history of back pain has been highlighted

as a prognostic Cell Cycle inhibitor indicator in other studies (Mallen et al., 2007), but this was not supported here, probably due to the very high proportion of the sample with prior back pain (87%). Although a wide range of prognostic indicators were included here, other factors have been identified elsewhere (e.g. Mallen et al., 2007 and Foster et al., 2010) and it would be useful to examine these. Replication in other samples, perhaps with recent onset back pain, would be useful, as the current sample was mixed, and contained many people with long duration of pain. A strength of this study is presentation of the contribution of prognostic factors to poor outcome through the use of adjusted PAF calculations. Whilst adjustment for confounding is considered essential for models of outcome prediction, adjustment of PAFs is rare. Table 3 demonstrates that proportions can change substantially following adjustment, and presentation of unadjusted proportions would considerably overestimate the contribution of several factors. Although there was loss to follow-up in the study, the

sample is representative of baseline responders. Attrition biases are unlikely to substantially influence the RRs reported here, as comparisons are Bleomycin molecular weight within the sample. However, as the proportions corresponding to each factor are based on associations and risk factor prevalence, these may be affected. In this analysis, 47% of the sample had high pain intensity at baseline, compared to 46% in the total baseline sample; loss to follow-up is therefore unlikely of to have affected the proportions reported. However, initial response to the survey was 65%, and it is likely that non-responders were different to baseline responders. The impact of this is difficult to assess due to lack of information, but it is likely

that the prevalence of prognostic indicators would be lower among non-responders. However, even a 10% change in the prevalence of the prognostic indicator would only make a difference in the proportion of poor outcome associated with pain intensity of around 4% (e.g. reducing high pain intensity prevalence from 47% to 37% would lead to an unadjusted proportion of 77% compared with 81% in Table 3), indicating that our results are likely to be broadly generalisable. Comparisons are also difficult to make with other samples due to the different measures used, lack of information about prevalence of prognostic indicators, and the inability to produce adjusted figures without the original data. As proportions differ according to the prevalence of exposure and strength of association, estimates of the potential contributions of prognostic indicators should be made for individual settings.