A red color with sodium amalgam and HCl acid. The flavone glycoside RS-2 was found to be soluble in water, ethanol and acetone and crystallized from methanol. RS-2 analyzed for molecular formula C29H34O13, m.p. 285–286° and M+ 590 (CIMS). The wavelengths of maximum absorption as observed with various shift reagents were at; λmax (MeOH) 270, 347 nm, λmax (NaOMe) 287, 395 nm, λmax (AlCl3) 278, 389, 405 nm, λmax (AlCl3 + HCl) 277, 389, 405 nm, and λmax (NaOMe) 272, 348 nm as depicted in Graph 2. The characteristic band observed in the IR spectrum of RS-2

and the structural assignments made with the help of available literature1, 2, 3 and 4 are described below: 3396.3 cm−1 (Hydrogen bonding intermolecular stretching), 2864.5 cm−1 (CH3 stretching of CH3), 1637.9 cm−1 (α,β-unsaturated C O), 1461.5 cm−1 (Aromatic ring system), 1219.0 cm−1 (C–O–C– stretching http://www.selleckchem.com/products/PD-0332991.html vibration), and 771 cm−1 (C–H out of plane bending) as portrayed in Graph 1. Significant band at Vmax (KBr) 3396.3 cm−1 as mentioned in Graph 1 in the IR spectrum of the glycoside (RS-2) indicated the presence of hydroxyl group(s) in it. The glycoside (RS-2) was acetylated with Ac2O/Pyridine to give an acetylated product having molecular formula, C41H46O19, m.p. 204–205° and M+ 842 (CIMS). The estimation of percentage of the

acetyl group (31.04%) in the acetylated derivative was given by Weisenberger method5 GSK J4 price as described by Belcher and Godbert6 which showed that there were six acetylable hydroxyl groups in the glycoside (RS-2). The appearance of band in IR spectrum of the acetyl derivative at Vmax (KBr) 1725.4 cm−1 with disappearance of band at Vmax (KBr) 3396.3 cm−1 confirmed that the acetylation of all the hydroxyl groups present

in the glycoside RS-2 was complete. 7 and 8 The IR absorption spectrum of the flavone glycoside (RS-2) displayed important band at Vmax (KBr) 2925.9 cm−1 indicating the presence of methoxyl group(s) in it. The methoxyl group estimation (16.05%) was done by Zeisel’s method 9 which confirmed the presence of three methoxyl groups in RS-2. The 1H NMR spectrum most of the flavonoidal glycoside (RS-2) showed three singlets at δ 4.0, δ 3.97 and δ 3.80 as depicted in Graph 3 each of these integrating for three protons, thereby suggesting the presence of three methoxyl groups in RS-2. Characteristic band at Vmax (KBr) 1461.5 cm−1 in the IR spectrum of glycoside RS-2 showed the presence of C C ring stretching. The structure of the glycoside (RS-2) was elucidated by its acid hydrolysis and identifying the components of hydrolyzate and the aglycone respectively. The glycoside (RS-2) on its acid hydrolysis with 7% alcoholic H2SO4 yielded an aglycone RS-2(A) as a solid residue and sugar moiety(ies) in the filtrate. They were separated by filtration and studied separately. The aglycone RS-2(A) was found to be homogenous on TLC examination (EtOAc–MeOH–H2O, 3:2:1). It crystallized from MeOH.

Enrichment of serum A on HPV31 or HPV58 VLP yielded antibodies capable of recognizing HPV16 and only the type used for enrichment. For example, the pre-treatment titers against HPV31 and HPV58 were 211 and 2696, respectively. Enrichment on HPV58 VLP increased the titer against HPV58 to 6188 but no HPV31 antibody reactivity was buy GDC-0068 detectable. Serum B which demonstrated post-enrichment neutralization activity against HPV31, HPV33, HPV35 and HPV58

appeared to comprise multiple antibody specificities that recognized HPV16 and only the indicated non-vaccine type. Enrichment of sera C and D on HPV35 VLP yielded antibodies capable of recognising HPV16 and HPV35, but not HPV31. Antibodies enriched from serum E and F exhibited cross-recognition of more than one non-vaccine type. The enrichment of serum E on HPV31 or HPV33 VLP yielded antibodies capable of recognizing HPV16, HPV31 and HPV33 pseudoviruses. Serum F when enriched on HPV31, HPV33 and HPV58 demonstrated neutralization of HPV31 pseudovirus to a comparable level, and serum F antibodies enriched on HPV31 or selleckchem HPV33 VLP had similar titers against HPV33. The HPV16 titer dropped by a median 1.8 Log10 (IQR 1.7–2.8; n = 13) fold following enrichment on non-vaccine VLP. Enriched antibody titers against HPV16 were similar to the titers observed against the type used for enrichment, for example

antibodies in serum A when enriched on HPV31 VLP neutralized HPV16 and HPV31 at titers of 861 and 795, respectively. Antibodies enriched from Thymidine kinase serum samples A–F, were also tested against L1 VLP representing the same HPV types (Supplementary material S1). Antibody binding titers further confirmed the observations that non-vaccine type antibodies are a minority species which display similar reactivity against HPV16 and non-vaccine types and again highlighted discrepancies between binding and neutralizing antibody specificity. We undertook a proof of concept study to investigate the cross-neutralizing antibody specificities generate in response to HPV vaccination. Cross-neutralizing

antibodies are elicited in response to both licensed vaccines, Cervarix® and Gardasil®[4], [11], [12] and [13] and this is coincident with differential degrees of vaccine-induced cross-protection [1] and [2], although a direct link between the two observations has not been established. The characterisation of the cross-neutralizing response beyond antibody titer has been limited to studies of avidity [23] and the vaccine-type specificity of cross-neutralizing antibodies [24]. Sera from Cervarix® vaccinees were chosen since it is this vaccine that appears to elicit the broadest cross-neutralization of non-vaccine types [4]. In the present study, sera from Cervarix® vaccinees were shown to have high antibody titers with broad reactivity against L1 VLP with homologous L1 sequences to those of the pseudoviruses.

Encapsulation efficiency of all batches was in between 90% and 100% w/w. One of the objectives of non-aqueous emulsion technique was to entrap maximum amount of metformin HCl. As discussed earlier the major drawback of other techniques (aqueous phase) was drug leakage occurred during solidification of nanoparticles. But in oil in oil method there was not a phase where metformin can leak out. Due to polymer saturated solvent and methanol immiscible with oil, polymeric matrix was immediately precipitate

out as solvent start to evaporate and gives maximum encapsulation efficiency.14 Secondly the high concentration of polymer increases viscosity of the solution and hindrance the drug diffusion within the polymer droplets. Drug-polymer ratio do not significantly CP-690550 order increased the encapsulation efficiency of metformin HCl in all three ethylcellulose polymers (p < 0.05). The encapsulated drug in all nanoparticles was already high. In EC100 and EC300 at 1:3 and 1:6 ratios encapsulation was increased slightly by 3–4% than EC45 but at 1:9 there was no significant difference in encapsulation all three polymers because nominal effect of viscosity on entrapment was concentrated at this ratio. There were also slight differences in drug content and percentage yield within same ratios of different ethylcellulose polymers. As percentage of polymers increased the drug content was also decreased.

Fig. 1 illustrates the morphology of nanoparticles of EC45, Dorsomorphin molecular weight EC100 and EC300. All particles were spherical in nature, uniform size and have tough surface texture. EC300 nanoparticles were less porous than other two polymeric nanoparticles. Smoothness of surface was due to polymer saturated internal organic phase. Fast diffusion of organic phase in

continuous phase before stable nanoparticles development can cause aggregation. 8 But in this preparation method methanol is not diffused in oil phase therefore aggregation of particles was not observed. After confirmed the physical characteristics of nanoparticles whether drug and polymer interact chemically much at processing conditions was tested by infrared spectroscopy. Actually negated drug-polymer interaction was studied before development of nanoparticles but processing conditions of nanoparticles development may affect on its chemical stability. The IR spectra of metformin HCl, ethylcellulose and drug loaded nanoparticles shown in Fig. 2. Pure metformin HCl illustrates two typical bands at 3371 cm−1 and 3296 cm−1 due to N–H primary stretching vibration and a band at 3170 cm−1 due to N–H secondary stretching. Characteristic bands at 1626 cm−1, 1567 cm−1 allocate to C N stretching. FTIR of EC showed principal peaks between 1900 cm−1 to 3500 cm−1. Of these 2980.12 cm−1 and 2880 cm−1 peaks were due to C–H stretching and a broad band at 3487.42 cm−1 was due to O–H stretching.

We establish that clearance of these bacilli requires sustained antibiotic treatment, and abrogates the cytokine producing vaccine-specific CD4 T cells derived from the spleen and the lungs. Strikingly, although substantially decreased, significant pulmonary and systemic protection was still present following clearance of bacilli. Together these data suggest BCG may induce two mechanisms of immunity: (i) dependant on the presence of viable bacilli and associated TEM; and (ii) a further mechanism, independent

of persisting bacilli and TEM. The exact details of Duvelisib the latter mechanism are yet to be elucidated, and are the subject of current investigation. The question of BCG persistence has been noted in previous studies in mice [24], [25], [27], [32], Apoptosis inhibitor [33], [34] and [35], other animal models [23] and [26] and humans [36] and [37]. In a similar study using C57BL/6 mice and M. tb challenge [27], spleen protection was reduced by 75%, but in contrast lung immunity was unaffected. This disparity with

our study could be due to: mouse strain, challenge organism, incomplete BCG bacilli clearance, or the shorter duration between chemotherapy and challenge. To date, however, no relationship between BCG persistence and the predominance of CD4 TEM responses has been reported [9], [16], [18] and [38]. Our data indicate a clear link between BCG antigen load and T cell responses, which as demonstrated here and previously, are multifunctional (IFN-γ+/IL-2+/TNF-α+, IFN-γ+/TNF-α+ and IL-2+/TNF-α+) CD62Ll°CD4 T cells which we consider TEM[9]. We also demonstrate that antigen-specific IFN-γ could used as a direct surrogate of viable bacilli (with the caveat of appropriate antigen stimulation). We cannot rule out that our antibiotic regimen did not completely eliminate the persistent BCG without performing subsequent immunosuppression

[39], which was beyond the scope of our study. However, our data clearly demonstrate reproducible elimination to a point that no BCG baciili and antigen-specific cells could be detected after 3 months of ‘rest’. isothipendyl Therefore, we consider this sufficient BCG clearance for the objectives of this study. We define these IFN-γ+/IL-2+/TNF-α+ triple- or bi-functional cells as CD4 TEM based on CD62Llo CCR7− expression [9]. As CD62L can be cleaved by metalloproteases, we previously conducted studies using the inhibitor TAPI-2 [40] to demonstrate that identification of stimulated-responder cells as CD62Llo was not due to non-specific mechanisms of CD62L down-regulation (data not shown). We have also confirmed this by sorting CD62Llo/hi cells prior to functional assay (Kaveh & Hogarth, unpublished data).

This may demonstrate that

peer-assisted learning activities can be utilised in paired student placements without reducing access to other learning activities. It may have indicated that students in peer-assisted learning were able to use their ‘downtime’ (ie, time when, in the traditional approach, they may have been waiting for their clinical educator to direct their learning) to complete the designated peer-assisted learning tasks. The rigid structure of the formal peer-assisted learning activities may have contributed to the dissatisfaction with the model, a notion that is supported by the clinical educators citing a preference for a ‘flexible peer-assisted learning’ model in the future. To ensure Galunisertib consistency in the research protocol, the formal elements of the peer-assisted learning MLN0128 molecular weight model were prescribed and did not vary throughout the placement. Principles of learning dictate that an effective teaching strategy involves a progression of increasingly complex tasks as knowledge and skill increase.29 Although it was theoretically possible to increase complexity of the task within the prescribed activities, this may have been difficult for clinical educators and students to execute, given that it was their first experience with the

tools. If paired student placement models are utilised in clinical education, it may be important to consider incorporating flexibility in the type and number of peer-assisted learning activities facilitated each week, although the results of the trial may have been different if this approach had been tested. The time allocated to familiarise students with the tools and expectations of the peer-assisted learning model in this study

may have been insufficient, which may have contributed to students’ relative dissatisfaction with the formal tools and the model Cell press itself. Students’ willingness to engage in a different learning culture to traditional, teacher-led practices can affect their engagement with peer-assisted learning19 and has been recognised as being important to clinical educators.30 To help address this, it may be of benefit to introduce the various tools in the pre-clinical period, and to invest time in orientating learners about the evidence of both the short-term and long-term benefits of working with and learning with peers.9, 10, 11, 12, 13, 14, 16, 17, 19 and 31 It is also possible that some elements of the peer-assisted learning model may have greater acceptability to students than others, and this will be the focus of ongoing investigations. The project was conducted in one health service with one group of clinical educators, which limits generalisability. Clinical educator participants were volunteers and therefore a self-selecting group. Issues may have been missed that related specifically to clinical educators who did not volunteer.

Addition of organic phase in to aqueous phase under the influence of sonication results in rapid miscibility of ethanol with water, which increases the polarity of the ethanol and decreases the solubility of curcumin leading to initiation of crystal nucleation. Concurrently, sonication process produce bubbles, whose size is near the resonant size for the applied frequency

and begins to oscillate nonlinearly and finally collapse resulting in production of extremely high temperature, high pressure, and shock wave, which inhibits the crystal growth of curcumin. However, developed curcumin nanocrystals form complex with β-cyclodextrin, which increases the stability and solubility of curcumin in the aqueous phase. Subsequently, sodium lauryl sulfate get adsorbed on the curcumin and offer negative charge to the surface. Negatively charged particles repel each other Luminespib order and develop

an electrostatic force, which maintains the nanoparticles in Brownian motion and overcomes the Van der Waals force of attraction and gravitational force resulting in the prevention of nanoparticle aggregation and sedimentation. Prepared SLS/βCD-curcumin nanosuspension was characterized for mean particle size, surface area, span (distribution width), and uniformity as these parameters determines the solubility, stability, cellular uptake and consistency of performance.8 5-FU price In the Phosphoprotein phosphatase present study, we have prepared nine formulations to optimize various concentrations of SLS and βCD. Prepared SLS/βCD-curcumin nanosuspension was characterized for mean particle size, surface area, distribution width (span), and uniformity and the results are summarized in Table 1. Increase in concentration of SLS and βCD from 25 mg to 50 mg have shown increase in mean particle size. However, equal amount of SLS and βCD at low concentration (i.e. 25 mg) has produced mean particle size of 270 nm with

the surface area of 47 m2/g, span of 4.574 and uniformity of 1.250. Similarly, equal amount of SLS and βCD at high concentration (i.e. 50 mg) has produced mean particle size of 206 nm with surface area of 53.4 m2/g, span of 4.365 and uniformity of 1.020. Out of nine formulations, FC1 has produced a mean particle size of 176 nm with surface area of 56.8 m2/g, span of 1.456 and uniformity of 0.779. In spite of least mean particle size, span, uniformity and higher surface area, FC1 does not contain β-cyclodextrin, which may leads to curcumin instability in aqueous nanosuspension. Hence, we have preferred formulation FC3 with mean particle size of 206 nm, surface area of 53.4 m2/g, span of 4.365 and uniformity of 1.020 (Fig. 1).

We would like to acknowledge the investigators, nurses, field workers and other personnel who contributed to the conduct of this trial; Mary Rusizoka, Beatrice Kamala, Wilbroad Shangwe, Francesca Lemme, Serafina Soteli, Clemens Masesa, and the HPV-021 trial team in Mwanza; Pius Magulyati, and the laboratory staff of the National Institute for Medical Research (NIMR) Mwanza Research Centre laboratory; the administrative staff of the Mwanza Intervention Trials Unit (MITU), NIMR Mwanza Research Centre, and Sekou Toure Hospital; Lucy Bradshaw, Gillian Devereux, Jayne Gould and Sue Napierala Mavedzenge and the research support staff at the London School of Hygiene and Tropical Medicine

(LSHTM). We thank Peter Hughes and the Clinical Diagnostic Laboratory of the MRC/UVRI Uganda Research Unit in Entebbe, Selleck GSK1349572 and David Warhurst and the Department of Pathogen Molecular Biology at LSHTM for their contributions to this work. We are grateful to the Ministry of Health and Social Welfare for granting permission to conduct this study. Conflict of interest statement Dr. Watson-Jones and Dr. Mayaud have received grant support through their institutions from GlaxoSmithKline Biologicals SA. During the trial, partial salary support for Drs. Watson-Jones,

Andreasen, Brown and Kavishe came from GSK Biologicals. There are no other conflicts of interest. Dr. Brown is supported by NIH-NIHM 1K01MH100994-01 and NIH-NCATS 8KL2TR000143-08. Richard Hayes, Saidi Kapiga, Selinexor and Kathy Baisley receive support from the MRC and DFID (G0901756, MR/K012126/1). “
“Human papillomavirus (HPV) vaccines induce type-specific neutralizing antibodies which correlate with immunity to the corresponding HPV types [1], and World Health Organization guidelines recommend that assays which assess neutralization be used as the reference standard for measuring HPV vaccine responses [2]. Quadrivalent HPV (Q-HPV) also vaccine (Gardasil®, Merck Laboratories) consists of HPV 6, 11, 16 and 18 virus-like particles (VLP) and is licensed for a 3-dose

regimen. Post-Gardasil® antibody responses are typically measured by a proprietary multiplex competitive Luminex immunoassay (cLIA) [3], which is based on competitive binding of type-specific HPV antibodies in human sera with labelled monoclonal antibodies directed against neutralizing epitopes of the respective VLP types (HPV 6, 11, 16 and 18). It has been reported that HPV antibodies measured by the cLIA may decline to become undetectable over time, especially for HPV 18, despite continued vaccine efficacy in preventing infections [4] and [5]. The significance of the loss of detectable antibodies is unknown as protective levels of HPV antibodies remain undefined [1], [6] and [7] and vaccine efficacy remains near 100%. Recently, Merck Laboratories developed a total IgG Luminex immunoassay (TIgG) which measures antibodies against the entire VLP, i.e.

4) on a magnetic stirrer at 37 ± 0.5° at 100 rpm. 5 ml

quantity of sample was withdrawn at different time periods and same volume of dissolution medium was replaced in the flask to maintain Ulixertinib datasheet sink condition. The withdrawn samples were filtered and then the filtrate was diluted with phosphate buffer (pH 7.4). The samples were analyzed for drug release by measuring the absorbance at 249 nm using UV–visible spectrophotometer. The in vitro drug release studies were carried out in triplicate for each formulation. The in vitro release data of all the formulation were fitted with various kinetics models such as zero order, first order, Higuchi model and Korsmeyer–Peppas, 9 in order to predict kinetics and mechanism of drug release. The release constant was calculated from the slope of plots and regression

coefficient (r2), diffusion exponent (n) was determined. The stability study of freeze dried nanoparticles was carried out for D1 (1:2) to assess the stability of drug in nanoparticles. For this purpose the samples were taken in borosilicate vials and sealed and the vials were stored in room temperature (25°–30 °C) and refrigerator (3°–5 °C) over a period of 3 months. After specified period 0, 1, 2 and 3 months, the samples were checked for their physical appearance and drug content by UV spectrophotometer, as well as chemical stability by Fourier transform infrared (FTIR) studies. The biodistribution studies8 of ddi loaded albumin PD0332991 mw nanoparticles were carried out on healthy adult Wistar rats weighing 200–250 g and after obtaining approval from the local animal ethics committee and CPCSEA (DSCP/PH.D PHARM/IAEC/49/2010-2011). All animals were provided with proper care, food, water ad libitum

and were maintained under well ventilated in large spacious cages throughout the study. The rats were divided randomly into three groups with three animals per group and they were fasted at least 12 h before experimentation. Group 1 was injected with ddi (which was dispersed in water for injection) into the tail vein of rats, Group 2 was received ddi loaded albumin nanoparticles and Group 3 was administered polysorbate 80 coated albumin nanoparticles. All the formulations were given in a dose level equivalent to 20 mg/kg body weight. 7 One hour after injection, the rats were sacrificed by euthanized and organs such as liver, lung, kidney, through lymph nodes, spleen, brain and blood were isolated. The organs were washed with clean buffer saline and absorbed dry with filter paper and then weighed. Prior to the analysis organs homogenates were prepared and was digested with 10% v/v trichloroacetic acid and was treated with 10 ml of acetonitrile to extract didanosine. Didanosine content in the various organs was estimated by reverse-phase HPLC method. BSA nanoparticles were prepared and loaded with didanosine by desolvation techniques with ethanol as it does not require an increase in temperature.

The main supporting themes describing the lack of knowledge are presented Pomalidomide ic50 here. Both girls and their parents had limited understanding about HPV and cervical cancer. Their knowledge was described in three main areas related to HPV: what HPV is, how HPV is transmitted, and the HPV and cervical cancer connection. Many of the girls and parents answered with uncertainty when asked about what they thought HPV was. Their answers both implied

confusion and explicitly expressed this confusion and lack of knowledge about HPV. Many girls simply replied “no” when asked if they knew what HPV was. A girl in one focus group responded, “I know the V stands for vaccination…” (H, FG1). Many other girls mentioned herpes when Rigosertib asked about HPV. Herpes was not the only sexually transmitted infection confused with HPV, though.

When asked what the girls knew about HPV, one girl answered “I think of AIDS” (F, FG2). Strikingly absent in their discussions of HPV was genital warts. Many parents could articulate the phrase “human papillomavirus,” but not much more. Some parents, though not as often as girls, also simply responded “no” to regarding whether they had heard of HPV. Knowledge surrounding HPV transmission was varied. While approximately half of the parents and girls mentioned “sex,” it was often followed by qualifiers such as “I think.” The uncertainty about HPV transmission was also discussed. Some girls mentioned that HPV could be transmitted genetically, through blood (via shared needles) or saliva. Only one parent mentioned skin contact as a route of transmission. Responses from girls about their knowledge of HPV transmission included: “I reckon it’s like hereditary” (E, FG1). There was some discussion about

sex, but confusion was still present. “…I think if you’re sexually active, then that’s when, it like makes your body trigger that you can have you can contract the virus. But if you’re still like a virgin, then you can’t get it…” (D, FG2). Even though there was some Unoprostone knowledge of HPV being related to sex, the role males played in transmission was unclear to the girls. When a girls’ focus group was asked if boys could catch HPV, all of the girls answered “no” and then explained “They can get AIDS” and “They can get diseases.” The moderator prompted “So HPV is sexually transmitted, but you can’t get it from boys?” The girls then said “That doesn’t make sense” and “I think it’s if you sleep with too many boys” and “If guys don’t get it, how do we get it then?” (G, FG3). Many parents had knowledge that sexual behaviours were related to HPV, but were unsure about the relationship. Some parents attributed HPV to a high number of sexual partners. “I don’t know how it’s transmitted.

The device used, the ventilation mode while training, training pressure, duration, frequency, and progression of training were recorded for the experimental group and for the control group if it received sham training. The method of inspiratory muscle training (isocapnic/normocapnic hyperpnoea, inspiratory resistive training, threshold pressure loading, or adjustment of ventilator pressure trigger sensitivity) was also recorded. Outcome measures: Primary outcome measures were measures of inspiratory muscle strength at a controlled lung volume (eg,

maximal inspiratory pressure at residual volume), inspiratory Temsirolimus muscle endurance, the duration of unassisted breathing periods, weaning success (ie, proportion of patients successfully weaned, defined as spontaneous breathing without mechanical support for at least 48 hours), weaning duration (ie, from the identification of readiness to wean, as determined by the authors and/or commencement of inspiratory muscle training, to the discontinuation of mechanical ventilation) and reintubation (ie, proportion of extubated patients who were reintubated within the follow-up period of the study). Secondary outcomes were tracheostomy (ie, proportion GS-7340 cell line of extubated patients tracheostomised after the commencement

of training), survival, adverse effects, and length of stay in hospital or the intensive care unit. The relevant data including study characteristics and outcome data were extracted from the eligible studies by two reviewers (LM and JR) using a standard form and the third author (ME) arbitrated in cases of disagreement. The reviewers extracted information about the method (design, participants,

and intervention) and outcome data for the experimental and control groups. Authors were contacted where there was difficulty in interpreting or extracting data. The data analysis was performed using Revman 5.1 (Revman 2011). A fixed-effect model was used unless there was substantial heterogeneity (I2 > 50%), when a random-effects model was used. Continuous outcomes were reported as weighted mean differences with next 95% CIs, while dichotomous outcomes were reported as risk ratios with 95% CIs. The search retrieved 816 studies. After screening titles and abstracts, 797 were excluded and 19 full text articles were identified. After evaluation of the full text, nine studies were excluded on the basis of participants not meeting the inclusion criteria. A further three were excluded on the basis of the intervention not meeting the inclusion criteria. Therefore seven papers (Cader et al 2010, Caruso et al 2005, Martin et al 2006a, Martin et al 2006b, Martin et al 2007, Martin et al 2009, Martin 2011) met the inclusion criteria for the review. One trial was reported across five publications (Martin et al 2006a, Martin et al 2006b, Martin et al 2007, Martin et al 2009, Martin et al 2011), so the seven included papers provided data on three trials.