The results of this study suggest that the Canadian C-spine rule

The results of this study suggest that the Canadian C-spine rule has the potential to affect healthcare costs considerably. The Ottawa group have previously examined the acceptability of the Canadian C-spine rule to clinicians (Brehaut et al 2009). To do this, the rule

was rated using the Ottawa Acceptability CRM1 inhibitor of Decision Rules Instrument (OADRI), which ranges from 0 (least acceptable) to 6 (most acceptable). Emergency physicians in Australia, Canada, USA, and UK rated the Canadian C-spine rule between 4 and 5 on the OADRI, suggesting good acceptability. Vaillancourt et al (2009) found 100% sensitivity and 38% specificity of the Canadian C-spine rule when used by paramedics. It would be worthwhile repeating these studies with Emergency Department physiotherapists to add to the growing body of evidence to guide this arm of the profession (Jibuike et al 2003, McClellan et al 2006, Webb 2008). The participating centres were 6 teaching and 6 community hospitals. Surprisingly, the effect of implementation of the Canadian C-spine rule was less in academic centres than in community

hospitals. Several of the academic centres had participated in an earlier validation study of the rule, which may have increased their baseline use of the rule. The procedures to introduce the rule to the active hospitals in this trial were extensive. Given this and the relatively low cost of diagnostic radiography the study could have benefited from a cost effectiveness analysis. Nevertheless, this excellent study shows the efficacy and importance of clinical decision making rules. The authors are to be congratulated on the study. “
“Summary of: Thomas M, McKinley GS-1101 mw RK, Mellor S, Watkin G, Holloway E, Scullion J, et al (2009) Breathing exercises for asthma: a randomised controlled trial. Thorax 64:

55– 61. [Prepared by Mark Elkins, CAP Co-ordinator.] Question: Does breathing training improve respiratory symptoms, Oxygenase quality of life and objective markers of disease severity in adults with asthma? Design: Randomised controlled trial. Setting: Ten general practitioner (GP) practices in Leicester, UK. Participants: Adults treated for asthma in a GP practice with moderate impairment of asthma-related health status, defined as a score less than 5.5 on the Asthma Quality of Life Questionnaire (AQLQ). Smokers were excluded. Randomisation of 183 participants allotted 94 to breathing training and 89 to a control group. Interventions: Usual physicians for both groups were requested to continue baseline therapy if possible. All participants were invited to 3 sessions within one month: an initial 60-min session with 2–4 participants, followed by two individual sessions of 30–45 minutes. At these sessions, the intervention group were educated about abnormal breathing patterns and taught appropriate regular diaphragmatic and nasal breathing techniques and encouraged to practise these exercises for at least 10 min each day.

Therefore, it was suggested that the extent and duration

Therefore, it was suggested that the extent and duration

of mechanical stretch may determine the cellular fate, such as death or proliferation. Our experimental findings show that acute mechanical stretch for 4 h causes continuous RASMC death. These findings may imply that an acute rise in blood pressure leads to the death of SMCs, a main component of the aortic medial layer. However, further studies Idelalisib using in vivo experimental conditions are required to elucidate whether an acute rise in blood pressure directly causes SMC death. Next, stretch-induced changes in the intracellular signaling of RASMCs were examined. It was reported that a high level of phosphorylated JNK was observed in AAD tissues, and that degeneration and tear of the aortic media CB-839 price had occurred in the AAD lesion. (2) and (13). In addition, it was reported that inhibition of the phosphorylation of JNK lead to regression of AAD (23). In the present study, we found that acute mechanical stretch causes rapid phosphorylation of JNK and p38 (Fig. 3A and B), which may lead to SMC death. In fact, we also observed that SP600125, a JNK inhibitor, and SB203580, a p38 inhibitor, both recovered stretch-induced RASMC death evaluated based on the MTT reduction and LDH release from the cells (Fig. 5A and

B). Although we also found that ERK1/2 are phosphorylated by mechanical stretch, ERK inhibitors failed to inhibit stretch-induced Adenosine RASMC death (data not shown). Taking these observations together, mechanical stretch causes phosphorylation of JNK and p38, which may result in SMC death that

may ultimately lead to the onset of AAD. On the other hand, a previous study showed that angiotensin II acted as an agonist for a potent inducer of AAD (1). In contrast to these findings, mechanical stretch itself, which is independent of angiotensin II stimulation, phosphorylated JNK and p38, and induced SMC death in our experiments. Although we did not measure the amount of angiotensin II in the medium, angiotensin II itself is not likely involved in JNK and p38 phosphorylation because stretch-induced AT1 receptor activation was also observed in mesenteric and renal arteries from angiotensinogen-knockout mice (24). Therefore, it is conceivable that not only agonist stimulation, but also mechanical stretch could have an important role in triggering the occurrence of AAD. ARBs are used all over the world for the treatment of patients with hypertension (25). Olmesartan, one of the ARBs, is known as an inverse agonist, which inhibits basic and stretch-induced activation of the AT1 receptor (17) and (26). In our present study, we found that olmesartan inhibited phosphorylation of JNK and p38 (Fig. 4A and B), and SMC cell death (Fig. 2) induced by acute mechanical stretch. These results suggest that olmesartan inhibits stretch-induced SMC death by suppression of phosphorylation of JNK and p38.

Apart from scientific study, general morphological description li

Apart from scientific study, general morphological description like size, colour, taste,

fracture and texture facilitates in identifying plant raw drugs. Consequently macroscopic descriptions of roots were studied according to T.E. Wallis.12 The etymological derivations were compiled from ‘Namarupajnanam’. The term ‘Namarupajnanam’ that represents nama (names) and rupa (characters) developed recently as a part of ‘Dravyagunavijnana’ in which identification of plants is studied in ancient and medieval approach to describe the plants by names and synonyms.13 Physicochemical parameters were done to analyse moisture content, total ash, acid insoluble ash, alcohol solubility and water solubility as per quality standards of API.9 Phytochemical screening was performed by using standard www.selleckchem.com/p38-MAPK.html procedures14 in order to establish chemical profile. Dried, powdered (mesh size 85) root samples of the species under study were successively extracted with solvents of increasing polarity, hexane, ethyl acetate, chloroform, methanol and water at 60–70 °C for 8 complete cycles. GPCR Compound Library clinical trial All root extracts were concentrated at 40–45 °C by using a rotary evaporator (Rotavapor R-3, Buchi, Switzerland) to 50 mL and tested for the presence of chemical constituents. One gram of each powdered

root sample of Patala namely, S. chelonoides, S. tetragonum and R. xylocarpa sieved (Mesh No. 85) was refluxed in water bath with methanol (50 mL) and filtered through Whatman No. 1 filter paper. These samples were subjected to extraction until it becomes colourless with same residue. Filtered extracts were evaporated by using rotary evaporator, followed by dissolving the residue with methanol (10 mL) and aliquots were taken for HPTLC analysis. The standard p-coumaric acid (purity ≥98%) HPLC purchased

from Sigma–Aldrich was dissolved in methanol to prepare working solution of 0.1 mg/mL concentration. The qualitative HPTLC analysis was Megestrol Acetate performed with 10 μL of methanolic extracts and standard solution of different concentrations (2–10 μL containing 20–100 μg/mL) using a solvent system, Toluene: Ethyl Acetate: Acetic Acid: Formic Acid (10:10:0.2:0.2 V/V). After development, the plate was dried in an oven at 110 °C for 10 min. The Rf values of marker and the compound of interest were measured and subjected to densitometric scan at λ = 310 nm in order to check the identity of the bands corresponding to the standard marker compound. The roots of S. chelonoides, S. tetragonum, and R. xylocarpa are similar in colour, texture and taste. The comparative analyses of macroscopic character are given in Table 2. The Ayurvedic literature describes Patala as: it is a tree having black peduncles. The leaflets become very rough on maturity. The flowers are fragrant, copper coloured and look like a pitcher shape. The seeds resemble like that of a human eye ball.

, 2000 and Craig et al , 2008) This advocated approach to comple

, 2000 and Craig et al., 2008). This advocated approach to complex health interventions, including childhood obesity prevention programmes, necessitates

a deep understanding of the determinants of the problem in the target communities. The importance of the relationship between context (e.g. socio-cultural structures and practices) and 3-deazaneplanocin A in vivo health, and in particular the relationship between context and individual health-related behaviours has been highlighted in recent years (Frohlich et al., 2001). The work of Bronfenbrenner represents a major contribution to the theoretical understanding of the relationship between a child and the context within which they function. Bronfenbrenner proposed the Ecological Systems (ES) model, which depicts layers of contextual structures that influence a child, and in turn, these are influenced by the child’s actions (Bronfenbrenner, 1977). These structures are termed the microsystems (the relationships between the child and their immediate environments, e.g. home, school), mesosystems LY2109761 supplier (the interrelationships between these settings), exosystems (settings that have an indirect effect, e.g. neighbourhood), and macrosystems

(cultural and societal values that are manifested in the micro-, meso- and exosystems). The ES model articulates the complexity and interactions of the contextual structures that a child is embedded in, and acknowledges the reciprocal nature of the relationships. The model is the basis for ecological health promotion models that attempt to move the focus away from individual behaviour change (McLeroy et al., 1988). Bronfenbrenner’s model has given rise to several conceptual models of childhood obesity.

Davison MYO10 and Birch’s model depicts child weight status at the centre, surrounded by three concentric circles; child characteristics; parenting styles and family characteristics; and community, demographic and societal characteristics (Davison and Birch, 2001). A further example is the ‘Causal Web’ model for the development of obesity, proposed by the International Obesity Taskforce (IOTF), which schematically represents contextual influences on individual lifestyle ‘choices’ (Kumanyika et al., 2002). This model encompasses national and international factors (media and advertising, urbanisation etc.), akin to Bronfenbrenner’s macrosystems, but does not acknowledge the reciprocity of relationships. In this study, we report the findings from focus groups run with members of UK South Asian communities. South Asians are a particular target group for obesity prevention, as they have higher body fat than other ethnic groups, and are more vulnerable to the health consequences of obesity (Bhopal et al., 1999, Whincup et al., 2002 and WHO expert consultation, 2004). The aim of the focus groups was to access key contextual data to inform the development of an obesity prevention programme targeting South Asian children.

This, in turn, could bias the estimate of the effect of treatment

This, in turn, could bias the estimate of the effect of treatment produced by the trial. Although investigators may not intend to modify their behaviour in these ways,

such effects could even happen subconsciously. However, if the upcoming allocation is concealed from the enrolling investigator, these effects cannot occur. After a patient has been approached and has expressed some interest in participating in the trial, an investigator Dasatinib purchase must determine whether the patient meets the eligibility criteria. Some eligibility criteria (eg, age, gender, the presence of a prosthetic joint) may be clear cut with little opportunity for interpretation. However, other eligibility criteria may be more subjective. For example, in a trial of home-based exercise training for people with chronic heart failure by Chien et al (2011), one exclusion criterion was a primary musculoskeletal disease [affecting] the assessment of exercise capacity. All NVP-BEZ235 musculoskeletal diseases will fall somewhere on a spectrum from substantially impairing the assessment of exercise capacity to having no effect. In assessing each potential participant against this criterion, the enrolling investigator

may be forced to decide subjectively whether borderline impairment is negligible or not. Knowledge of the upcoming allocation could affect (consciously or subconsciously) the decision about the patient’s eligibility. Similar motivations to those discussed above could again systematically influence which patients are allocated to each group. For example, patients with a poor prognosis may be deemed ineligible when the upcoming

allocation is to the treatment group but deemed eligible otherwise. Concealment of the allocation list prevents this potential source of bias between the groups. Patients who are deemed eligible for a trial must make a fully informed decision about their willingness to participate (World Medical Association 2008). While a comprehensive description of all the salient points must be given to each interested patient, a standard text is not usually used to guide the description. Because the description can vary between patients, there is again opportunity for knowledge of the upcoming randomisation to affect how the enrolling investigator Levetiracetam describes trial participation to the patient. For example, the negative aspects of trial participation may be emphasised if the investigator wants to divert the patient away from the upcoming allocation. Such negative aspects may include the number of visits required for outcome assessment, the possibility of randomisation to the control group, and the time, effort and expense of undertaking the intervention. Conversely, positive aspects – such as the opportunity to receive the results of health-related tests that would be undertaken as part of outcome assessment – could be emphasised.

There has been little empirical investigation of the effects of a

There has been little empirical investigation of the effects of adherence on the efficacy of falls prevention interventions. Previous literature has focussed primarily on patientlevel factors that affect adherence to interventions for

the prevention of falls. The patient’s perspective of barriers and facilitators to exercise adherence has previously been reported. For example, transport to and from the venue, cost, loss of interest, and injury all influence adherence to a schedule http://www.selleckchem.com/products/Adrucil(Fluorouracil).html of exercise classes (Bunn et al 2008, de Groot and Fagerstrom 2011, Forkan et al 2006, Lee et al 2010). However, the influence of intervention-level factors extrinsic to the patient, such as exercise mode, duration, and frequency, remain widely unanalysed. Merom and colleagues (2012) conducted an observational study examining participation in different forms of exercise for the prevention of falls. However, it only identified whether participants were participating in exercise, and did not provide a numerical measure

of adherence which would be more sensitive to change. Exploration of the association between programrelated factors and adherence is paramount, as it is these factors that can be modified by program providers to enhance adherence to interventions. A recent systematic review sought to identify the likely overall participation rate in community-based interventions for the prevention http://www.selleckchem.com/products/BKM-120.html of falls, including group exercise interventions (Nyman and Victor 2012). However, this research did not specify whether the adherence rates they used were inclusive of drop-out participants,

and the pooled adherence rates calculated were not weighted for study size. Further, no analyses were undertaken to examine the factors that are associated with adherence, nor the association between adherence and the efficacy of the intervention. As this review aspires to guide future practice in developing population-wide, community-based interventions for the prevention of falls, trials conducted in high-care living facilities or hospitals were not MTMR9 examined in this review. Therefore the research questions for this study were, in community-dwelling older adults: 1. What are the program-related factors that are associated with adherence to group exercise interventions for the prevention of falls? Papers that examined the effect of group exercise interventions for the prevention of falls were sought. The search terms were developed using a modified PICO model, ie, patient, intervention, comparator and outcome. Search terms for the comparator were omitted as there was no requirement for a specific comparison group when answering the first two study questions. The ‘falls’ terms stated served as a ‘context’ rather than an ‘outcome’ group of terms, as falls prevention could be described as a component of the study or an outcome.

8% for AT and accuracy of 92 9% and precision less than 5 4% for

8% for AT and accuracy of 92.9% and precision less than 5.4% for EZ. The stability of the two drugs under various conditions is shown in Table 4. Under all conditions tested, the two drugs proved to be stable. All results were within the acceptance criteria of ±15% deviation from the nominal concentration. The mean plasma level of AT and EZ in both products A and B are shown in Fig. 4a and b. Table 5 shows the parameters for the non-compartmental pharmacokinetic

analysis. According to ANOVA results there is no significant sequence effect for both cmax and AUC0–72 h indicating that the crossover design was properly performed. The parametric point estimates and the 90% confidence intervals for ln-transformed AUC0–t, AUC0–∞, and cmax, ( Table 6) were within commonly accepted bioequivalence range of 80–125% range, thus the results reveal Compound C price this website that the bioequivalence between products A and B could be concluded. A rapid, sensitive,

and simple method for determining AT and EZ levels in human plasma was developed and validated. The UPLC–MS/MS method described herein reveals significant advantages over other techniques, including LC–MS/MS, due to the inherently increased column efficiency of UPLC, which resulted in complete analysis within 1.2 min with significantly lower limits of quantitation (0.1 ng mL−1). To the best of our knowledge, this is the first UPLC–MS/MS method for the simultaneous determination of AT and EZ in human plasma. This fully validated method was an ideal tool for high-throughput Tryptophan synthase analysis of plasma samples used in pharmacokinetic and bioequivalence study of AT and EZ between two market products. All authors have none to declare. Special thanks to Prof. Dr. Meselhy Ragab Meselhy for allowing the performance of this research in the “Center of Applied Research and Advanced Studies” (CARAS), Faculty of Pharmacy, Cairo University. “
“Treatment of tuberculosis is now very complex because of the emergence of multi drug resistant bacteria, which are resistant to first-line anti-tuberculosis drugs, pyrazinamide, isoniazid and rifampin.1 Pyrazinamide (Fig. 1) is used extensively

in the treatment of tuberculosis together with rifampicin, isoniazid and ethambutol.2 The structure of pyrazinamide is given by Fig. 1 and the structure of metronidazole is given by Fig. 2. It has a plasma half-life of 3–4 h, and is quickly absorbed from the gastrointestinal tract with peak serum concentrations of 6–8 μg/ml occurring 1.5–2.0 h after administration.3 The determination of PZA levels in biological fluids was carried out earlier by spectroscopic methods,4, 5 and 6 colorimetric methods7 and gas chromatographic–mass spectrometric technique.8 A survey of literature revealed that HPLC technique has been used for the determination of pyrazinamide in pharmaceuticals.9 A HPLC technique reported earlier had a step of very tedious extraction.

Ciprofloxacin (Micro labs, India) and Amphotericin-B (Micro labs,

Ciprofloxacin (Micro labs, India) and Amphotericin-B (Micro labs, India) were used as reference antibiotics against bacteria and fungi, correspondingly. Antimicrobial activities of the crude extracts were first screened for their zone of inhibition by the agar well-diffusion method. Briefly, crude extracts were prepared concentration of 100 mg/ml with dimethyl sulphoxide (DMSO, SD Fine, Mumbai) as a solvent. The Mueller Hinton Agar (MHA) medium (Hi Media) was prepared and sterilized at 121 °C 15 lp/sq for 20 min the autoclave. Twenty millilitres of this sterilized agar medium (MHA)

were poured into each 9 cm sterile petridishes under aseptic conditions and allowed to settle. For the preparation of the inocula 24 h culture was emulsified in 3 ml sterile saline following the McFarland turbidity to obtain a concentration of 108 cells/ml. The suspension was standardized by adjusting the optical density to 0.1 at 600 nm (ELICO MAPK inhibitor SL-244 spectrophotometer). One hundred microlitres (100 μl) of cell suspension with approximately 106–108 bacteria per millilitre was placed in petridishes and dispersed over

agar.7 In the following, a well was prepared in the plates with the help of a sterile stainless steel-borer (6 mm diameter) two holes per plates were made into the set agar containing the bacterial culture. Each well 100 μl of the plant added at the concentration of 100 mg/ml. For each bacterial strain controls were maintained where pure solvents, instead of extract as a negative control. Plant extracts

and reference drug (Ciprofloxacin 1000 μg/ml) were allowed to diffuse Cytidine deaminase for 1 h into the plates and then incubated at 37 °C for 18 h Epigenetic Reader Domain inhibitor in inverted position. The results were recorded by measuring the zone of growth inhibition (mm) surrounding the wells. Each assay was performed in triplicates and repeated twice. Diameters of inhibition zone less than 7 mm were recorded as non-active (−), and as active (+), when the mean of inhibition zone was between 7 and 10 mm. (++) Described an inhibition diameter of more than 10 mm and less than 15 mm, (+++) an inhibition diameter between 15 and 20 mm and (++++) a diameter of more than 20 mm of growth inhibition.8 All the fungal species was cultured in Sabouraud Dextrose Broth (Hi Media) for 48 h at 27 °C and Sabouraud Dextrose Agar (SDA) was employed for the agar well diffusion experiments. Fungal suspensions were adjusted to 107 cells/ml as explained above. The zone of Inhibition was determined after incubation for 48 h at 27 °C. All tests were performed in triplicates and repeated twice.9 The minimum inhibitory concentration (MIC), which is considered as the lowest concentration of the sample which inhibits the visible growth of a microbe was determined by the microbroth dilution method. The MIC method was performed as described below on extracts that showed their high efficacy against microorganisms by the well diffusion method (zone of inhibition higher than 11 mm).

1 M Na2CO3/NaHCO3, pH 9 2, 50 μL/well) at 4 °C overnight Plates

1 M Na2CO3/NaHCO3, pH 9.2, 50 μL/well) at 4 °C overnight. Plates were blocked with PBST and 3% (w/v) non-fat dry milk for

2 h at room temperature. Plates were incubated with 3-fold dilutions to endpoint titre of pooled serum samples (100 μL per well in PBST with 1% non-fat dry milk (starting concentration: 1:50 dilution) for 2 h at room temperature. Following three washes with 100 μL PBST, plates were incubated with horseradish-peroxidase-conjugated anti-mouse IgG (Santa Cruz Biotechnology, Dallas, TX) at a dilution of 1:3000 in PBST and non-fat dry milk (1%, v/v) for 1 h at room temperature. Unbound antibody was removed by three washes with 100 μL PBST and plates were developed using SigmaFAST OPD substrate (Sigma, St. Louis, MO) (100 μL/well) and stopped with 3 M HCl (50 μL/well). The colorimetric change was measured as the optical density (OD 490 nm) on a Synergy 4 (BioTek, Winooski, http://www.selleckchem.com/products/Adrucil(Fluorouracil).html VT) microplate reader. The endpoint titre was defined as the reciprocal of the highest dilution that yields an OD-value above the mean plus three standard deviations of blank wells. The hemagglutination inhibition (HI) assay was used to assess functional antibodies to the HA able to inhibit agglutination of turkey red blood cells (tRBCs). Serum samples were treated with 4 volumes of a receptor-destroying enzyme of Vibrio cholera filtrate

(Sigma, St. Louis, MO) for 18 h at 37 °C. After addition of 3 volumes of 2.5% GABA assay (v/v) sodium citrate, the serum samples were incubated at 56 °C for 30 min and diluted with PBS why to yield a 1:10 dilution of the original serum sample. Serum samples were 2-fold serially diluted in PBS (25 μL sample volume) in Nunc® 96-well polystyrene V-bottom microwell plates (Thermo Fisher Scientific, Waltham, MA) and then incubated with recombinant reassortant virus (PR8:AH1, PR8:SH1, PR8:malNL00, PR8:malAlb01 or PR8:chickJal12) at 4 HAU/25 μL in PBS for 30 min at room

temperature. Then, 50 μL 0.5% tRBCs (Lampire Biological Laboratory, Pipersville, PA) were added and the mixture was incubated for 45 min at 4 °C. Sera from all groups were assayed individually for the challenge strain PR8:SH1 for HI activity. Divergent H7 strains were assayed with pooled sera. The HI titre was calculated from the reciprocal of the highest dilution that completely inhibited hemagglutination of red blood cells and the geometric mean titre (GMT) of two independent assays was reported as the final titre. Two negative HI readings were assigned <10, single negative results were scored a value of 5 for the calculation of geometric means. In preparedness for a potential H7N9 pandemic, it is highly desirable, not only for vaccine manufacturers, but also for health care providers, to develop an influenza vaccine that at low vaccine dose, most preferably with a single administration, stimulates good immune responses.

No spots were observed in control wells containing splenocytes bu

No spots were observed in control wells containing splenocytes but no coating antigen. The percentage of peripheral blood and splenic CD8+ T cells expressing IFNγ, TNFα and IL-2 in response to 5 h stimulation with 5 μg/ml peptides 90 and 91 was assessed by intracellular cytokine staining as previously described [5]. Duvelisib purchase Surface staining was with anti-CD8α PerCP-Cy5.5 and anti-CD4 Pacific Blue while intracellular staining was with anti-IFNγ APC,

anti-TNFα FITC and anti-IL-2 PE (all supplied by eBioscience, UK). Cytokine production frequency in peptide-unstimulated control wells (which was typically <0.1%) was subtracted from the result in peptide-stimulated wells prior to further analysis. The gating strategy is illustrated in supplementary Figure 1. Total IgG and isotype ELISA were carried out as previously described using bacterially expressed GST-tagged PfMSP119 (Wellcome/FVO allele) as the coating antigen [5]. Antibody avidity was assessed by sodium thiocyanate (NaSCN)-displacement ELISA [43]. Using previously measured total IgG ELISA titers, sera were individually diluted to a level calculated to give a titer of 1:300 and plated at 50 μl/well in 16 wells of a 96 well plate. Following incubation and washing, an ascending concentration of the chaotropic agent NaSCN was added down the plate (0–7 M NaSCN). Plates were incubated for 15 min

at room temperature before washing and development as for total IgG. The intercept of the OD405 curve for each this website sample with the line of 50% reduction of the OD405 in the NaSCN-free well for each sample (i.e. the concentration of NaSCN required to reduce the OD405 to 50% of that without NaSCN) was used as a measure of avidity. Statistical analysis was carried out using Prism 5 software (GraphPad, La Jolla, CA, USA). All ELISA titers were log10 Cytidine deaminase transformed prior to analysis. Graphs indicate sample arithmetic means; error bars where present indicate 95% confidence intervals for the population arithmetic

mean. One-way ANOVA was used for comparing normally distributed data with Bonferroni’s multiple comparison post-test for comparison of specific groups; Kruskal–Wallis tests were used for comparison of non-normally distributed data with Dunn’s multiple comparison post-test for comparison of specific groups. Two-way ANOVA was used for comparison of groups differing in two factors. Two-way repeat measures ANOVA was used for comparison of responses measured for different groups at different time points, after the exclusion of the small number of mice for which replicate data were not available at all time points. P < 0.05 was taken to be statistically significant throughout. The experimental design provided replicate groups receiving AdCh63–MVA (A–M) and AdCh63–protein (A–P) sequential regimes at 57 day and 97 day intervals. Antibody and IFNγ+ CD8+ T cell responses induced by these regimes are illustrated in Fig. 1.